ࡱ> +.0 !"#$%&'()*+,-v{e u  udfhMo7 bbjbjUU P 7|7|65 l^^^t,x--t|=ȱȱȱL4 |==ڀfELEE@EGJ|@ $ % x,RTGGf rF**EEFFFz*<Ex,EZFTFfF)BrL+|x,nE 61l|=Ltȱ`nn$A 0=pEFEnF.6h$X** Monroe L. WeberShirk Leonard W. Lion James J. Bisogni, Jr. Cornell University School of Civil and Environmental Engineering Ithaca, NY 14853 Laboratory Research in Environmental Engineering Laboratory Manual Laboratory Research in Environmental Engineering Laboratory Manual Monroe L. WeberShirk Instructor  HYPERLINK "mailto:mw24@cornell.edu" mw24@cornell.edu Leonard W. Lion Professor  HYPERLINK "mailto:lwl3@cornell.edu" lwl3@cornell.edu James J. Bisogni, Jr. Associate Professor  HYPERLINK "mailto:jjb2@cornell.edu" jjb2@cornell.edu School of Civil and Environmental Engineering Cornell University Ithaca, NY 14853 Fifth Edition Cornell University 2001 Educational institutions may use this text freely if the title/author page is included. We request that instructors who use this text notify one of the authors so that the dissemination of the manual can be documented and to ensure receipt of future editions of this manual. Table of Contents  TOC \o "1-2" \h \z  HYPERLINK \l "_Toc502112484" Table of Contents  PAGEREF _Toc502112484 \h 5  HYPERLINK \l "_Toc502112485" Preface  PAGEREF _Toc502112485 \h 9  HYPERLINK \l "_Toc502112486" Laboratory Safety  PAGEREF _Toc502112486 \h 10  HYPERLINK \l "_Toc502112487" Introduction  PAGEREF _Toc502112487 \h 10  HYPERLINK \l "_Toc502112488" Personal Protection  PAGEREF _Toc502112488 \h 10  HYPERLINK \l "_Toc502112489" Laboratory Protocol  PAGEREF _Toc502112489 \h 12  HYPERLINK \l "_Toc502112490" Use of Chemicals  PAGEREF _Toc502112490 \h 13  HYPERLINK \l "_Toc502112491" References  PAGEREF _Toc502112491 \h 19  HYPERLINK \l "_Toc502112492" Questions  PAGEREF _Toc502112492 \h 19  HYPERLINK \l "_Toc502112493" Laboratory Measurements and Procedures  PAGEREF _Toc502112493 \h 20  HYPERLINK \l "_Toc502112494" Introduction  PAGEREF _Toc502112494 \h 20  HYPERLINK \l "_Toc502112495" Theory  PAGEREF _Toc502112495 \h 20  HYPERLINK \l "_Toc502112496" Experimental Objectives  PAGEREF _Toc502112496 \h 22  HYPERLINK \l "_Toc502112497" Experimental Methods  PAGEREF _Toc502112497 \h 22  HYPERLINK \l "_Toc502112498" Prelab Questions  PAGEREF _Toc502112498 \h 24  HYPERLINK \l "_Toc502112499" Questions  PAGEREF _Toc502112499 \h 25  HYPERLINK \l "_Toc502112500" Data Sheet  PAGEREF _Toc502112500 \h 27  HYPERLINK \l "_Toc502112501" Lab Prep Notes  PAGEREF _Toc502112501 \h 29  HYPERLINK \l "_Toc502112502" Reactor Characteristics  PAGEREF _Toc502112502 \h 30  HYPERLINK \l "_Toc502112503" Introduction  PAGEREF _Toc502112503 \h 30  HYPERLINK \l "_Toc502112504" Reactor Classifications  PAGEREF _Toc502112504 \h 30  HYPERLINK \l "_Toc502112505" Reactor Modeling  PAGEREF _Toc502112505 \h 30  HYPERLINK \l "_Toc502112506" Mass Conservation  PAGEREF _Toc502112506 \h 34  HYPERLINK \l "_Toc502112507" Conductivity Measurements  PAGEREF _Toc502112507 \h 35  HYPERLINK \l "_Toc502112508" Procedures  PAGEREF _Toc502112508 \h 36  HYPERLINK \l "_Toc502112509" Prelab Questions  PAGEREF _Toc502112509 \h 39  HYPERLINK \l "_Toc502112510" Data Analysis  PAGEREF _Toc502112510 \h 39  HYPERLINK \l "_Toc502112511" Lab Prep Notes  PAGEREF _Toc502112511 \h 41  HYPERLINK \l "_Toc502112512" Acid Precipitation and Remediation of Acid Lakes  PAGEREF _Toc502112512 \h 43  HYPERLINK \l "_Toc502112513" Introduction  PAGEREF _Toc502112513 \h 43  HYPERLINK \l "_Toc502112514" Experimental Objectives  PAGEREF _Toc502112514 \h 47  HYPERLINK \l "_Toc502112515" Experimental Apparatus  PAGEREF _Toc502112515 \h 48  HYPERLINK \l "_Toc502112516" Experimental Procedures  PAGEREF _Toc502112516 \h 48  HYPERLINK \l "_Toc502112517" Prelab Questions  PAGEREF _Toc502112517 \h 53  HYPERLINK \l "_Toc502112518" Data Analysis  PAGEREF _Toc502112518 \h 53  HYPERLINK \l "_Toc502112519" Questions  PAGEREF _Toc502112519 \h 54  HYPERLINK \l "_Toc502112520" References  PAGEREF _Toc502112520 \h 54  HYPERLINK \l "_Toc502112521" Lab Prep Notes  PAGEREF _Toc502112521 \h 55  HYPERLINK \l "_Toc502112522" Measurement of Acid Neutralizing Capacity  PAGEREF _Toc502112522 \h 57  HYPERLINK \l "_Toc502112523" Introduction  PAGEREF _Toc502112523 \h 57  HYPERLINK \l "_Toc502112524" Theory  PAGEREF _Toc502112524 \h 57  HYPERLINK \l "_Toc502112525" Procedure  PAGEREF _Toc502112525 \h 60  HYPERLINK \l "_Toc502112526" Prelab Questions  PAGEREF _Toc502112526 \h 61  HYPERLINK \l "_Toc502112527" Questions  PAGEREF _Toc502112527 \h 61  HYPERLINK \l "_Toc502112528" References  PAGEREF _Toc502112528 \h 62  HYPERLINK \l "_Toc502112529" Lab Prep Notes  PAGEREF _Toc502112529 \h 63  HYPERLINK \l "_Toc502112530" Phosphorus Determination using the Colorimetric Ascorbic Acid Technique  PAGEREF _Toc502112530 \h 64  HYPERLINK \l "_Toc502112531" Introduction  PAGEREF _Toc502112531 \h 64  HYPERLINK \l "_Toc502112532" Experimental Objectives  PAGEREF _Toc502112532 \h 66  HYPERLINK \l "_Toc502112533" Experimental Procedures  PAGEREF _Toc502112533 \h 66  HYPERLINK \l "_Toc502112534" Prelab Questions  PAGEREF _Toc502112534 \h 67  HYPERLINK \l "_Toc502112535" Data Analysis  PAGEREF _Toc502112535 \h 67  HYPERLINK \l "_Toc502112536" Questions  PAGEREF _Toc502112536 \h 68  HYPERLINK \l "_Toc502112537" References  PAGEREF _Toc502112537 \h 68  HYPERLINK \l "_Toc502112538" Lab Prep Notes  PAGEREF _Toc502112538 \h 69  HYPERLINK \l "_Toc502112539" Soil Washing to Remove Mixed Wastes  PAGEREF _Toc502112539 \h 70  HYPERLINK \l "_Toc502112540" Objective  PAGEREF _Toc502112540 \h 70  HYPERLINK \l "_Toc502112541" Introduction  PAGEREF _Toc502112541 \h 70  HYPERLINK \l "_Toc502112542" Theory  PAGEREF _Toc502112542 \h 71  HYPERLINK \l "_Toc502112543" Apparatus  PAGEREF _Toc502112543 \h 77  HYPERLINK \l "_Toc502112544" Experimental Procedures  PAGEREF _Toc502112544 \h 78  HYPERLINK \l "_Toc502112545" Prelab Questions  PAGEREF _Toc502112545 \h 82  HYPERLINK \l "_Toc502112546" Data Analysis  PAGEREF _Toc502112546 \h 82  HYPERLINK \l "_Toc502112547" References  PAGEREF _Toc502112547 \h 82  HYPERLINK \l "_Toc502112548" Lab Prep Notes  PAGEREF _Toc502112548 \h 85  HYPERLINK \l "_Toc502112549" Oxygen Demand Concepts and Dissolved Oxygen Sag in Streams  PAGEREF _Toc502112549 \h 87  HYPERLINK \l "_Toc502112550" Introduction  PAGEREF _Toc502112550 \h 87  HYPERLINK \l "_Toc502112551" Theory  PAGEREF _Toc502112551 \h 87  HYPERLINK \l "_Toc502112552" Streeter Phelps Equation Development  PAGEREF _Toc502112552 \h 88  HYPERLINK \l "_Toc502112553" Zero Order Kinetics  PAGEREF _Toc502112553 \h 92  HYPERLINK \l "_Toc502112554" Experimental Objectives  PAGEREF _Toc502112554 \h 93  HYPERLINK \l "_Toc502112555" Experimental Methods  PAGEREF _Toc502112555 \h 94  HYPERLINK \l "_Toc502112556" Prelab Questions  PAGEREF _Toc502112556 \h 95  HYPERLINK \l "_Toc502112557" Data Analysis  PAGEREF _Toc502112557 \h 96  HYPERLINK \l "_Toc502112558" References  PAGEREF _Toc502112558 \h 97  HYPERLINK \l "_Toc502112559" Lab Prep Notes  PAGEREF _Toc502112559 \h 98  HYPERLINK \l "_Toc502112560" Methane Production from Municipal Solid Waste  PAGEREF _Toc502112560 \h 100  HYPERLINK \l "_Toc502112561" Introduction  PAGEREF _Toc502112561 \h 100  HYPERLINK \l "_Toc502112562" Theory  PAGEREF _Toc502112562 \h 100  HYPERLINK \l "_Toc502112563" Experiment description  PAGEREF _Toc502112563 \h 110  HYPERLINK \l "_Toc502112564" Experimental methods  PAGEREF _Toc502112564 \h 112  HYPERLINK \l "_Toc502112565" Prelab questions  PAGEREF _Toc502112565 \h 114  HYPERLINK \l "_Toc502112566" Data analysis  PAGEREF _Toc502112566 \h 114  HYPERLINK \l "_Toc502112567" References  PAGEREF _Toc502112567 \h 115  HYPERLINK \l "_Toc502112568" Lab Prep Notes  PAGEREF _Toc502112568 \h 117  HYPERLINK \l "_Toc502112569" Volatile Organic Carbon Contaminated Site Assessment  PAGEREF _Toc502112569 \h 119  HYPERLINK \l "_Toc502112570" Introduction  PAGEREF _Toc502112570 \h 119  HYPERLINK \l "_Toc502112571" Experiment Description  PAGEREF _Toc502112571 \h 119  HYPERLINK \l "_Toc502112572" Experimental Procedures  PAGEREF _Toc502112572 \h 120  HYPERLINK \l "_Toc502112573" Procedure (short version)  PAGEREF _Toc502112573 \h 122  HYPERLINK \l "_Toc502112574" Prelab Questions  PAGEREF _Toc502112574 \h 123  HYPERLINK \l "_Toc502112575" Data Analysis  PAGEREF _Toc502112575 \h 123  HYPERLINK \l "_Toc502112576" References  PAGEREF _Toc502112576 \h 123  HYPERLINK \l "_Toc502112577" Lab Prep Notes  PAGEREF _Toc502112577 \h 124  HYPERLINK \l "_Toc502112578" Volatile Organic Carbon Sorption to Soil  PAGEREF _Toc502112578 \h 126  HYPERLINK \l "_Toc502112579" Introduction  PAGEREF _Toc502112579 \h 126  HYPERLINK \l "_Toc502112580" Theory  PAGEREF _Toc502112580 \h 126  HYPERLINK \l "_Toc502112581" Analysis of the Unsaturated Distribution Coefficient ( EMBED Equation.DSMT4 )  PAGEREF _Toc502112581 \h 131  HYPERLINK \l "_Toc502112582" Analysis of the Saturated Distribution Coefficient ( EMBED Equation.DSMT4 )  PAGEREF _Toc502112582 \h 133  HYPERLINK \l "_Toc502112583" Experimental procedures  PAGEREF _Toc502112583 \h 135  HYPERLINK \l "_Toc502112584" Procedure (short version)  PAGEREF _Toc502112584 \h 136  HYPERLINK \l "_Toc502112585" Prelab Questions  PAGEREF _Toc502112585 \h 137  HYPERLINK \l "_Toc502112586" Data Analysis  PAGEREF _Toc502112586 \h 137  HYPERLINK \l "_Toc502112587" References  PAGEREF _Toc502112587 \h 138  HYPERLINK \l "_Toc502112588" Additional References Relevant to Data Reduction  PAGEREF _Toc502112588 \h 139  HYPERLINK \l "_Toc502112589" Symbol List  PAGEREF _Toc502112589 \h 140  HYPERLINK \l "_Toc502112590" Lab Prep Notes  PAGEREF _Toc502112590 \h 141  HYPERLINK \l "_Toc502112591" Enhanced Filtration  PAGEREF _Toc502112591 \h 142  HYPERLINK \l "_Toc502112592" Introduction  PAGEREF _Toc502112592 \h 142  HYPERLINK \l "_Toc502112593" Theory  PAGEREF _Toc502112593 \h 142  HYPERLINK \l "_Toc502112594" Previous Research Results  PAGEREF _Toc502112594 \h 144  HYPERLINK \l "_Toc502112595" Filter Performance Evaluation  PAGEREF _Toc502112595 \h 145  HYPERLINK \l "_Toc502112596" Experimental Objectives  PAGEREF _Toc502112596 \h 145  HYPERLINK \l "_Toc502112597" Experimental Methods  PAGEREF _Toc502112597 \h 145  HYPERLINK \l "_Toc502112598" Prelab Questions  PAGEREF _Toc502112598 \h 147  HYPERLINK \l "_Toc502112599" Data Analysis  PAGEREF _Toc502112599 \h 147  HYPERLINK \l "_Toc502112600" Questions for Discussion  PAGEREF _Toc502112600 \h 148  HYPERLINK \l "_Toc502112601" References  PAGEREF _Toc502112601 \h 148  HYPERLINK \l "_Toc502112602" Lab Prep Notes  PAGEREF _Toc502112602 \h 149  HYPERLINK \l "_Toc502112603" Gas Transfer  PAGEREF _Toc502112603 \h 150  HYPERLINK \l "_Toc502112604" Introduction  PAGEREF _Toc502112604 \h 150  HYPERLINK \l "_Toc502112605" Theory  PAGEREF _Toc502112605 \h 150  HYPERLINK \l "_Toc502112606" Experimental Objectives  PAGEREF _Toc502112606 \h 152  HYPERLINK \l "_Toc502112607" Experimental Methods  PAGEREF _Toc502112607 \h 153  HYPERLINK \l "_Toc502112608" Prelab Questions  PAGEREF _Toc502112608 \h 154  HYPERLINK \l "_Toc502112609" Data Analysis  PAGEREF _Toc502112609 \h 154  HYPERLINK \l "_Toc502112610" References  PAGEREF _Toc502112610 \h 155  HYPERLINK \l "_Toc502112611" Lab Prep Notes  PAGEREF _Toc502112611 \h 156  HYPERLINK \l "_Toc502112612" Instrument Instructions  PAGEREF _Toc502112612 \h 157  HYPERLINK \l "_Toc502112613" Compumet software  PAGEREF _Toc502112613 \h 157  HYPERLINK \l "_Toc502112614" pH Probe Calibration  PAGEREF _Toc502112614 \h 157  HYPERLINK \l "_Toc502112615" pH Probe Storage  PAGEREF _Toc502112615 \h 158  HYPERLINK \l "_Toc502112616" Procedure for Cleaning pH Gel-Filled Polymer Electrode  PAGEREF _Toc502112616 \h 158  HYPERLINK \l "_Toc502112617" Dissolved Oxygen Probe  PAGEREF _Toc502112617 \h 158  HYPERLINK \l "_Toc502112618" Gas Chromatograph  PAGEREF _Toc502112618 \h 160  HYPERLINK \l "_Toc502112619" UVVis Spectrophotometer  PAGEREF _Toc502112619 \h 160  HYPERLINK \l "_Toc502112620" Index  PAGEREF _Toc502112620 \h 161 Preface Continued leadership in environmental protection requires efficient transfer of innovative environmental technologies to the next generation of engineers. Responding to this challenge, the Cornell Environmental Engineering faculty redesigned the undergraduate environmental engineering curriculum and created a new seniorlevel laboratory course. This laboratory manual is one of the products of the course development. Our goal is to disseminate this information to help expose undergraduates at Cornell and at other institutions to current environmental engineering problems and innovative solutions. A major goal of the undergraduate laboratory course is to develop an atmosphere where student understanding will emerge for the physical, chemical, and biological processes that control material fate and transport in environmental and engineered systems. Student interest is piqued by laboratory exercises that present modern environmental problems to investigate and solve. The experiments were designed to encourage the process of learning around the edges. The manifest purpose of an experiment may be a current environmental problem, but it is expected that students will become familiar with analytical methods in the course of the laboratory experiment (without transforming the laboratory into an exercise in analytical techniques). It is our goal that students employ the theoretical principles that underpin the environmental field in analysis of their observations without transforming the laboratories into exercises in process theory. As a result, students can experience the excitement of addressing a current problem while coincidentally becoming cognizant of relevant physical, chemical, and biological principles. At Cornell, student teams of two or three carry out the exercises, maximizing the opportunity for a handson experience. Each team is equipped with modern instrumentation as well as experimental reactorxe "reactor" apparatus designed to facilitate the study of each topic. Computerized data acquisition and instrument control are used extensively to make it easier for students to learn how to use new instruments and to eliminate the drudgery of manual data acquisition. Software was developed at Cornell to use computers as virtual instruments that interface with a pH meter/ion (Accumet 50), gas chromatographxe "gas chromatograph" (HP 5890A), UVVis Spectrophotometer (HP 8452) This code is available at the course web site. The development of this manual and the accompanying course would not have been possible without funds from the National Science Foundation, the DeFrees Family Foundation, the Procter and Gamble Fund, the School of Civil and Environmental Engineering and the College of Engineering at Cornell University. Monroe L. WeberShirk Leonard W. Lion James J. Bisogni, Jr. Ithaca, NY  TIME \@ "MMMM d, yyyy" December 22, 2000 Laboratory Safety Introduction  MACROBUTTON MTEditEquationSection Equation Section 1 SEQ MTEqn \r \h \* MERGEFORMAT  SEQ MTSec \r 1 \h \* MERGEFORMAT Safety is a collective responsibility that requires the full cooperation of everyone in the laboratory. However, the ultimate responsibility for safety rests with the person actually carrying out a given procedure. In the case of an academic laboratory, that person is usually the student. Accidents often result from an indifferent attitude, failure to use common sense, or failure to follow instructions. Each student should be aware of what the other students are doing because all can be victims of one individual's mistake. Do not hesitate to point out to other students that they are engaging in unsafe practices or operations. If necessary, report it to the instructor. In the final assessment, students have the greatest responsibility to ensure their own personal safety. This guide provides a list of do's and don'ts to minimize safety and health problems associated with experimental laboratory work. It also provides, where possible, the ideas and concepts that underlie the practical suggestions. However, the reader is expected to become involved and to contribute to the overall solutions. The following are general guidelines for all laboratory workers: Follow all safety instructions carefully. Become thoroughly acquainted with the location and use of safety facilities such as safety showers, exits and eyewash fountains. Become familiar with the hazards of the chemicals being used, and know the safety precautions and emergency procedures before undertaking any work. Become familiar with the chemical operations and the hazards involved before beginning an operation. Personal Protection Eye Protection All people in the laboratory including visitors must wear eye protection at all times, even when not performing a chemical operation. Wearing of contact lenses in the laboratory is normally forbidden because contact lenses can hold foreign materials against the cornea. Furthermore, they may be difficult to remove in the case of a splash. Soft contact lenses present a particular hazard because they can absorb and retain chemical vapors. If the use of contact lenses is required for therapeutic reasons fitted goggles must also be worn. In addition, approved standing shields and face shields that protect the neck and ears as well as the face should be used when appropriate for work at reduced pressure or where there is a potential for explosions, implosions or splashing. Normal prescription eyeglasses, though meeting the Food and Drug Administration's standards for shatter resistance, do not provide appropriate laboratory eye protection. Clothing Clothing worn in the laboratory should offer protection from splashes and spills, should be easily removable in case of accident, and should be at least fire resistant. Nonflammable, nonporous aprons offer the most satisfactory and the least expensive protection. Lab jackets or coats should have snap fasteners rather than buttons so that they can be readily removed. Highheeled or opentoed shoes, sandals, or shoes made of woven material should not be worn in the laboratory. Shorts, cutoffs and miniskirts are also inappropriate. Long hair and loose clothing should be constrained. Jewelry such as rings, bracelets, and watches should not be worn in order to prevent chemical seepage under the jewelry, contact with electrical sources, catching on equipment, and damage to the jewelry. Gloves Gloves can serve as an important part of personal protection when they are used correctly. Check to ensure the absence of cracks or small holes in the gloves before each use. In order to prevent the unintentional spread of chemicals, gloves should be removed before leaving the work area and before handling such things as telephones, doorknobs, writing instruments, computers, and laboratory notebooks. Gloves may be reused, cleaned, or discarded, consistent with their use and contamination. A wide variety of gloves is available to protect against chemical exposure. Because the permeability of gloves of the same or similar material varies from manufacturer to manufacturer, no specific recommendations are given here. Be aware that if a chemical diffuses through a glove, that chemical is held against the worker's hand and the individual may then be more exposed to the chemical than if the glove had not been worn. Personal Hygiene Everyone working in a chemistry laboratory should be aware of the dangers of ingesting chemicals. These common sense precautions will minimize the possibility of such exposure: Do not prepare, store (even temporarily), or consume food or beverages in any chemical laboratory. Do not smoke in any chemical laboratory. Additionally, be aware that tobacco products in opened packages can absorb chemical vapors. Do not apply cosmetics in a laboratory. Wash hands and arms thoroughly before leaving the laboratory, even if gloves have been worn. Wash separately from personal laundry, lab coats or jackets on which chemicals have been spilled. Never wear or bring lab coats or jackets into areas where food is consumed. Never pipettexe "pipette" by mouth. Always use a pipette aid or suction bulb. Laboratory Protocol The chemistry laboratory is a place for serious learning and working. Horseplay cannot be tolerated. Variations in procedures including changes in quantities or reagents may be dangerous. Such alterations may only be made with the knowledge and approval of the instructor. Housekeeping In the laboratory and elsewhere, keeping things clean and neat generally leads to a safer environment. Avoid unnecessary hazards by keeping drawers and cabinets closed while working. Never store materials, especially chemicals, on the floor, even temporarily. Work spaces and storage areas should be kept clear of broken glassware, leftover chemicals and scraps of paper. Keep aisles free of obstructions such as chairs, boxes and waste receptacles. Avoid slipping hazards by keeping the floor clear of ice, stoppers, glass beads or rods, other small items, and spilled liquids. Use the required procedure for the proper disposal of chemical wastes and solventsxe "solvents". Cleaning Glassware Clean glassware at the laboratory sink or in laboratory dishwashers. Use hot water, if available, and soap or other detergent. If necessary, use a mild scouring powder. Wear appropriate gloves that have been checked to ensure that no holes are present. Use brushes of suitable stiffness and size. Avoid accumulating too many articles in the cleanup area. Usually work space around a sink is limited and piling up dirty or cleaned glassware leads to breakage. Remember that the turbid water in a sink may hide a jagged edge on a piece of broken glassware that was intact when put into the water. A pair of heavy gloves may be useful for removing broken glass, but care must be exercised to prevent glove contamination. To minimize breakage of glassware, sink bottoms should have rubber or plastic mats that do not block the drains. Avoid the use of strong cleaning agents such as nitric acid, chromic acid, sulfuric acid, strong oxidizers, or any chemical with a "per" in its name (such as perchloric acid, ammonium persulfate, etc.) unless specifically instructed to use them, and then only when wearing proper protective equipment. A number of explosions involving strong oxidizing cleaning solutions, such as chromic sulfuric acid mixtures, have been reported. The use of flammable solventsxe "solvents" should be minimized and, when they are used, appropriate precautions must be observed. Unattended Operation of Equipment Reactions that are left to run unattended overnight or at other times are prime sources for fires, floods and explosions. Do not let equipment such as power stirrers, hot plates, heating mantles, and water condensers run overnight without failsafe provisions and the instructor's consent. Check unattended reactions periodically. Always leave a note plainly posted with a phone number where you and the instructor can be reached in case of emergency. Remember that in the middle of the night, emergency personnel are entirely dependent on accurate instructions and information. Fume Hoods and Ventilation A large number of common substances present acute respiratory hazards and should not be used in a confined area in large amounts. They should be dispensed and handled only where there is adequate ventilation, such as in a hood. Adequate ventilation is defined as ventilation that is sufficient to keep the concentration of a chemical below the threshold limit value or permissible exposure limit. If you smell a chemical, it is obvious that you are inhaling it. However, odor does not necessarily indicate that a dangerous concentration has been reached. By contrast, many chemicals can be present at hazardous concentrations without any noticeable odor. Refrigerators Chemicals stored in refrigerators should be sealed, double packaged if possible, and labeled with the name of the material, the date placed in the refrigerator, and the name of the person who stored the material A current inventory should be maintained. Old chemicals should be disposed of after a specified storage period. Household refrigerators should not be used for chemical storage. If used for storage of radioactive materials, a refrigerator should be plainly marked with the standard radioactivity symbol and lettering, and routine surveys should be made to ensure that the radioactive material has not contaminated the refrigerator. Food should never be stored in a refrigerator used for chemical storage. These refrigerators should be clearly labeled "No Food". Conversely food refrigerators, which must be always outside of, and away from, the chemical work area, should be labeled "Food OnlyNo Chemicals". Radioactive Materials Radioactive materials are used in the Environmental Engineering laboratories. Doors of rooms containing radioactive materials are clearly labeled. Areas where radioactive materials are used are clearly delineated with labeling tape and signs. All equipment within areas labeled radioactive are potentially contaminated and should not be touched or removed. Do not place anything into or take anything from an area labeled radioactive. Working Alone Avoid working alone in a building or in a laboratory. Use of Chemicals Before using any chemical you need to know how to safely handle it. The safety precautions taken are dependent on the exposure routes and the potential harmful effects. Routes of Exposure ingestion inhalation absorbed through skin eye contact Each potential exposure route requires different precautions. Chemical exposure may have acute (immediate, short term) or chronic (long term potentially cumulative) affects. Information on health hazards can be found on chemical labels and in Material Safety Data Sheets. Material Safety Data Sheets MSDS sheets for most chemicals used in the laboratory are located on the bookshelf in the entrance hallway of the Environmental Laboratory. Electronic versions (potentially more current) can be found using the world wide web at:  HYPERLINK "http://www.cee.cornell.edu/safety/" http://www.cee.cornell.edu/safety/ MSDS provide extensive information on safe handling, first aid, toxicity, etc. Following is a list of terms used in MSDS: TLVThreshold Limit Valueare values for airborne toxic materials that are to be used as guides in control of health hazards. They represent concentrations to which nearly all workers (workers without special sensitivities) can be exposed to for long periods of time without harmful effect. TLV's are usually expressed as parts per million (ppm). TLV's are also expressed as mg of dust or vapor/m3 of air. TDLoToxic Dose Lowthe lowest dose of a substance introduced by any route, other than inhalation, over any given period of time and reported to produce any toxic effect in humans or to produce carcinogenic, neoplastigenic, or teratogenic effects in animals or humans. TCLoToxic Concentration Lowthe lowest concentration of a substance in air to which humans or animals have been exposed for any given period of time and reported to produce any toxic effect in humans or to produce carcinogenic, neoplastigenic, or teratogenic effects in animals or humans. TDLoLethal Dose Lowthe lowest dose (other than LD50) of a substance introduced by any route, other than inhalation, over any given period of time in one or more divided portions and reported to have caused death in humans or animals. LD50Lethal Dose Fiftya calculated dose of a substance that is expected to cause the death of 50% of an entire defined experimental animal population. It is determined from the exposure to the substance by any route other than inhalation of a significant number from that population. Table  SEQ table \r11. NFPA hazard code ratings.CodeHealthFire Reactivity 4Very short exposure can cause death or major residual injuryWill rapidly or completely vaporize at normal pressure and temperatureCapable of detonation or explosive reaction at normal temperatures and pressures 3Short exposure can cause serious temporary or residual injuryCan be ignited under almost all ambient temperaturesCapable of detonation or explosive reaction buy requires a strong initiating source or must be heated under confinement before initiation 2Intense or continued exposure can cause temporary incapacitation or possible residual injuryMust be moderately heated or exposed to high temperature before ignitionUndergoes violent chemical change at elevated temperatures and pressures or reacts violently with water. 1Can cause irritation but only minor residual injuryMust be preheated before ignitionNormally stable but can become unstable at elevated temperatures and pressures. 0During a fire offers no hazard beyond combustionWill not burnStable even under fire conditions.LCLoLethal Concentration Lowthe lowest concentration of a substance in air, other than LC50, that has been reported to have caused death in humans or animals. The reported concentrations may be entered for periods of exposure that are less than 24 hours (acute) or greater than 24 hours (subacute and chronic). LC50Lethal Concentration Fiftya calculated concentration of a substance in air, exposure to which for a specified length of time is expected to cause the death of 50% of an entire defined experimental animal population. It is determined from the exposure to the substance of a significant number from that population. Chemical Labels All chemicals must be labeled. Unlabeled containers of mystery chemicals or chemical solutions are a nightmare for disposal as well as a potential safety hazard. The OSHA Hazard Communication Standard and the OSHA Lab Standard have specific requirements for the labeling of chemicals. In a laboratory covered under the Lab Standard, if a chemical is designated as a hazardous material, that is having the characteristics of corrosivity, ignitability, toxicity (generally meaning a highly toxic material with an LD50 of 50 mg/kg or less), reactivity, etc., and if it is made into a solution or repackaged as a solid or liquid in a concentration greater than 1% (0.1% for a carcinogen) it needs to have a so called RightToKnow (RTK) label that duplicates the hazard warnings, precautions and first aid steps found on the original label. All other chemicals must have at minimum a label with chemical name, concentration, and date prepared. "Right to Know Labels" will be made available for your use when necessary. National Fire Protection Association (NFPA) ratings are included to indicate the types and severity of the hazards. The NFPA ratings are on a scale of 04 with 0 being nonhazardous and 4 being most hazardous. The ratings are described in Table 1. Chemical Storage There has been much concern, and some confusion, about the proper storage of laboratory chemicals. Here proper means the storage of chemicals in such a manner as to prevent incompatible materials from being accidentally mixed together in the event of the breakage of one or more containers in the storage area or to prevent the formation of reactive vapors that may require vented chemical storage areas. Below is a concise guide to the storage of common laboratory chemicals. Perchloric acid is separated from all other materials. Hydrofluoric acid is separated from all other materials. Concentrated nitric acid is separated from all other materials. Highly toxic materials (LD50 of 50 mg/kg or less) are stored separately. Carcinogenic chemicals are stored separately. Inorganic acids (except for 1, 2, 3 above) are stored separately. Bases are stored separately. Strong oxidizing agents are stored separately. Strong reducing agents are stored separately. Water reactive, pyrophoric and explosive materials are stored separately. Flammable organic materials (solventsxe "solvents", organic acids, organic reagents) are stored separately. Guidelines for separating incompatible chemicals: Place the chemicals to be stored separately in a heavy gauge Nalgene (or similar plastic) tub. Plastic secondary containers must be compatible with the material being stored. Strong acids, especially perchloric, nitric and hydrofluoric are best stored in plastic containers designed to store strong mineral acids. These are available from lab equipment supply houses. Bottleinacan type of containers are also acceptable as secondary containment. Small containers of compatible chemicals may be stored in a dessicator or other secure container. Secondary containment is especially useful for highly toxic materials and carcinogens. Dry chemicals stored in approved cabinets with doors may be grouped together by compatibility type on separate shelves or areas of shelves separated by taping off sections of shelving to designate where chemicals of one type are stored. Physically separated cabinets may be used to provide a barrier between groups of stored incompatible chemicals. Strong mineral acids may be stored in one cabinet and strong bases stored in a second cabinet, for example. Flammable solventsxe "solvents" should be stored in a rated flammable storage cabinet if available. If you are uncertain of the hazardous characteristics of a particular chemical refer to the MSDS for that material. A good MSDS will not only describe the hazardous characteristics of the chemical, it will also list incompatible materials. Transporting Chemicals Transport all chemicals using the containerwithinacontainer concept to shield chemicals from shock during any sudden change of movement. Large containers of corrosives should be transported from central storage in a chemically resistant bucket or other container designed for this purpose. Stairs must be negotiated carefully. Elevators, unless specifically indicated and so designated, should not be used for carrying chemicals. Smoking is never allowed around chemicals and apparatus in transit or in the work area itself. When moving in the laboratory, anticipate sudden backing up or changes in direction from others. If you stumble or fall while carrying glassware or chemicals, try to project them away from yourself and others. When a flammable liquid is withdrawn from a drum, or when a drum is filled, both the drum and the other equipment must be electrically wired to each other and to the ground in order to avoid the possible buildup of a static charge. Only small quantities should be transferred to glass containers. If transferring from a metal container to glass, the metal container should be grounded. Chemical Disposal The Environmental Protection Agency (EPA) classifies wastes by their reaction characteristics. A summary of the major classifications and some general treatment guidelines are listed below. Specific information may be found in the book, Prudent Practices for Disposal of Chemicals from Laboratories, as well as other reference materials. Ignitability: These substances generally include flammable solventsxe "solvents" and certain solids. Flammable solvents must never be poured down the drain. They should be collected for disposal in approved flammable solvent containers. In some cases it may be feasible to recover and reuse solvents by distillation. Such solvent recovery must include appropriate safety precautions and attention to potentially dangerous contamination such as that due to peroxide formation. Corrosivity: This classification includes common acids and bases. They must be collected in waste containers that will not ultimately corrode and leak, such as plastic containers. It often may be appropriate to neutralize waste acids with waste bases and where allowed by local regulations, dispose of the neutral materials via the sanitary sewer system. Again, the nature of the neutralized material must be considered to ensure that it does not involve an environmental hazard such as chromium salts from chromic acid neutralization. Reactivity: These substances include reactive metals such as sodium and various water reactive reagents. Compounds such as cyanides or sulfides are included in this class if they can readily evolve toxic gases such as hydrogen cyanide. Their collection for disposal must be carried out with particular care. When present in small quantities, it is advisable to deactivate reactive metals by careful reaction with appropriate alcohols and to deactivate certain oxygen or sulfur containing compounds through oxidation. Specific procedures should be consulted. Toxicity: Although the EPA has specific procedures for determining toxicity, all chemicals may be toxic in certain concentrations. Appropriate procedures should be established in each laboratory for collection and disposal of these materials. The handling of reaction byproducts, surplus and waste chemicals, and contaminated materials is an important part of laboratory safety procedures. Each laboratory worker is responsible for ensuing that wastes are handled in a manner that minimizes personal hazard and recognizes the potential for environmental contamination. Most instructional laboratories will have clear procedures for students to follow in order to minimize the generation of waste materials. Typically reaction byproducts and surplus chemicals will be neutralized or deactivated as part of the experimental procedure. Waste materials must be handled in specific ways as designated by federal and local regulations. University guidelines for waste disposal can be found in chapter 7 of the Chemical Hygiene Plan (available at  HYPERLINK "http://www.cee.cornell.edu/safety/" http://www.cee.cornell.edu/safety/ ) Some general guidelines are: Dispose of waste materials promptly. When disposing of chemicals one basic principle applies: Keep each different class of chemical in a separate clearly labeled disposal container. Never put chemicals into a sink or down the drain unless they are deactivated or neutralized and they are allowed by local regulation in the sanitary sewer system. [See Chemical Hygiene Plan for list of chemicals that can be safely disposed of in the sanitary sewer.] Put ordinary waste paper in a wastepaper basket separate from the chemical wastes. If a piece of paper is contaminated, such as paper toweling used to clean up a spill, put the contaminated paper in the special container that is marked for this use. It must be treated as a chemical waste. Broken glass belongs in its own marked waste container Broken thermometers may contain mercury in the fragments and these belong in their own special sealed "broken thermometer" container. Peroxides, because of their reactivity, and the unpredictable nature of their formation in laboratory chemicals, have attracted considerable attention. The disposal of large quantities (25 g or more) of peroxides requires expert assistance. Consider each case individually for handling and disposal. A complete list of compounds considered safe for drain disposal can be found in Chapter 7 of the Chemical Hygiene Plan ( HYPERLINK "http://www.cee.cornell.edu/safety/" http://www.cee.cornell.edu/safety/). Disposal techniques for chemicals not found in this list must be disposed of using techniques approved of by Cornell Environmental Health and Safety. When possible, hazardous chemicals can be neutralized and then disposed. When chemicals are produced that cannot be disposed of using the sanitary sewer, techniques to decrease the volume of the waste should be considered. References Safety in Academic Chemistry Laboratories. A publication of the American Chemical Society Committee on Chemical Safety. Fifth edition. 1990 Cornell University Chemical Hygiene Plan: Guide to Chemical Safety for Laboratory Workers. A publication of the Office of Environmental Health, 2000. ( HYPERLINK "http://www.ehs.cornell.edu/lrs/CHP/chp.htm" http://www.ehs.cornell.edu/lrs/CHP/chp.htm) OSHA Laboratory Standard One of the best books to get started with regulatory compliance is a publication from the American Chemical Society entitled, "Laboratory Waste Management. A Guidebook." Questions Why are contact lenses hazardous in the laboratory? What is the minimum information needed on the label for each chemical? When are right to know labels required? Why is it important to label a bottle even if it only contains distilled water? Find an MSDS for sodium nitrate. a) Who created the MSDS? b) What is the solubility of sodium nitrate in water? c) Is sodium nitrate carcinogenic? d) What is the LD50 oral rat? e) How much sodium nitrate would you have to ingest to give a 50% chance of death (estimate from available LD50 data). f) How much of a 1 M solution would you have to ingest to give a 50% chance of death? g) Are there any chronic effects of exposure to sodium nitrate? You are in the laboratory preparing chemical solutions for an experiment and it is lunchtime. You decide to go to the student lounge to eat. What must you do before leaving the laboratory? Where are the eyewash station, the shower, and the fire extinguishers located in the laboratory? Laboratory Measurements and Procedures Introduction  MACROBUTTON MTEditEquationSection Equation Section (Next) SEQ MTEqn \r \h \* MERGEFORMAT  SEQ MTSec \h \* MERGEFORMAT Measurements of masses, volumes, and preparation of chemical solutions of known composition are essential laboratory skills. The goal of this exercise is to gain familiarity with these laboratory procedures. You will use these skills repeatedly throughout the semester. Theory Many laboratory procedures require preparation of chemical solutions. Most chemical solutions are prepared on the basis of mass of solute per volume of solution (grams per liter or Moles per liter). Preparation of these chemical solutions requires the ability to accurately measure both mass and volume. Preparation of dilutions is also frequently required. Many analytical techniques require the preparation of known standards. Standards are generally prepared with concentrations similar to that of the samples being analyzed. In environmental work many of the analyses are for hazardous substances at very low concentrations (mg/L or g/L levels). It is difficult to weigh accurately a few milligrams of a chemical with an analytical balance. Often dry chemicals are in crystalline or granular form with each crystal weighing several milligrams making it difficult to get close to the desired weight. Thus it is often easier to prepare a low concentration standard by diluting a higher concentration stock solution. For example, 100 mL of a 10 mg/L solution of NaCl could be obtained by first preparing a 1 g/L NaCl solution (100 mg in 100 mL). One mL of the 1 g/L stock solution would then be diluted to 100 mL to obtain a 10 mg/L solution. Absorption spectroscopy is one analytical technique that can be used to measure the concentration of a compound. Solutions that are colored absorb light in the visible range. The resulting color of the solution is from the light that is transmitted. According to Beer's law the attenuation of light in a chemical solution is related to the concentration and the length of the path that the light passes through.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 2. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 1 where c is the concentration of the chemical species, b is the distance the light travels through the solution, e is a constant Po is the intensity of the incident light, and P is the intensity of the transmitted light. Absorptionxe "absorption", A, is defined as:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 2. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 2 In practice Po is the intensity of light through a reference sample (such as deionized water) and thus accounts for any losses in the walls of the sample chamber. From equation  GOTOBUTTON ZEqnNum460856 \* MERGEFORMAT  REF ZEqnNum460856 \! \* MERGEFORMAT 2.1 and  GOTOBUTTON ZEqnNum428291 \* MERGEFORMAT  REF ZEqnNum428291 \! \* MERGEFORMAT 2.2 it may be seen that absorptionxe "absorption" is directly proportional to the concentration of the chemical species.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 2. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 3 Table  SEQ \r 1 table1. Wavelengths of lightcolorwavelength (nm)ultra violet190380violet380450blue450490green490560yellow560590orange590630red630760The instrument you will use to measure absorbance is a Hewlett Packard (HP) model 8452A diode array spectrophotometer. The diode arrayxe "diode array" spectrophotometerxe "spectrophotometer" uses a broad-spectrum XE "spectrum"  source of incident light from a deuterium lamp. The light passes through the sample and is split by a grating into a spectrum of light that is measured by an array of diodes. Each diode measures a bandwidth of 2 nm with 316 diodes covering the range from 190 nm to 820 nm. The wavelengths of light and their colors are given in Table 1. The light path for the diode array spectrophotometer is shown in  REF _Ref365247940 \* MERGEFORMAT Figure 1.  Figure  SEQ \r 1 Figure1. Diagram of light path in diode arrayxe "diode array" spectrophotometerxe "spectrophotometer". The HP 8452A spectrophotometerxe "spectrophotometer" has a photometric range of 0.002  3.3 absorbance units. In practice absorbance measurements greater than 2.5 are not very meaningful as they indicate that 99.7% of the incident light at that wavelength was absorbed. Conversely, an absorbance of 0.002 means that 0.5% of the incident light at that wavelength was absorbed. When measuring samples of known concentration the Spectrophotometer software (page  PAGEREF _Ref406553258 \h 160) calculates the relationship between absorbance and concentration at a selected wavelength. The slope (m), intercept (b) and correlation coefficientxe "correlation coefficient" (r) are calculated using equation  GOTOBUTTON ZEqnNum136506 \* MERGEFORMAT  REF ZEqnNum136506 \! \* MERGEFORMAT 2.4 through  GOTOBUTTON ZEqnNum563537 \* MERGEFORMAT  REF ZEqnNum563537 \! \* MERGEFORMAT 2.6. The slope of the best fit line is  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 2. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 4 The intercept of the line is  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 2. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 5 The correlation coefficientxe "correlation coefficient" is defined as  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 2. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 6 where x is the concentration of the solute (methylene blue in this exercise)xe "methylene blue", y is the absorbance, and n is the number of samples. Experimental Objectives To gain proficiency in: Calibrating and using electronic balances Digital pipetting Preparing a solution of known concentration Preparing dilutions Measuring concentrations using a UVVis spectrophotometerxe "spectrophotometer" Experimental Methods Mass Measurements Mass can be accurately measured with an electronic analytical balance. Perhaps because balances are so easy to use it is easy to forget that they should be calibrated on a regular basis. It is recommended that balances be calibrated once a week, after the balance has been moved, or if excessive temperature variations have occurred. In order for balances to operate correctly they also need to be level. Most balances come with a bubble level and adjustable feet. Before calibrating a balance verify that the balance is level. The environmental laboratory is equipped with balances manufactured by Denver Instruments. To calibrate the Denver Instrument balances: Zero the balance by pressing the tare button. Press the MENU key until "MENU #1" is displayed. Press the 1 key to select Calibrate. Note the preset calibration masses that can be used for calibration on the bottom of the display. Place a calibration mass on the pan (handle the calibration mass using a cotton glove or tissue paper). The balance will automatically calibrate. A short beep will occur and the display will read CALIBRATED for three seconds, and then return to the measurement screen. Dry chemicals can be weighed in disposable plastic "weighing boats" or other suitable containers. It is often desirable to subtract the weight of the container in which the chemical is being weighed. The weight of the chemical can be obtained either by weighing the container first and then subtracting, or by "zeroing" the balance with the container on the balance. Temperature Measurement Use the Accumet"! pH/ion meter to measure the temperature of distilled water. The temperature probe is the 4mm diameter metallic probe. Place the probe in a 100mL plastic beaker full of distilled water. Wait at least 15 seconds to allow the probe to equilibrate with the solution. Pipette Technique Use Figure 2 to estimate the mass of 990 L of distilled water (at the measured temperature). Use a 1001000 L digital pipettexe "pipette" to transfer 990 L of distilled water to a tared weighing boat on the 100 g scale. Record the mass of the water and compare with the expected value ( REF _Ref365261331 \* MERGEFORMAT Figure 2). Repeat this step if necessary until your pipetting error is less than 2%, then measure the mass of 5 replicate 990 L pipette samples. Calculate the mean ( EMBED Equation.DSMT4  defined in equation  GOTOBUTTON ZEqnNum643596 \* MERGEFORMAT  REF ZEqnNum643596 \! \* MERGEFORMAT 2.7), standard deviation (s defined in equation  GOTOBUTTON ZEqnNum602904 \* MERGEFORMAT  REF ZEqnNum602904 \! \* MERGEFORMAT 2.8), and coefficient of variationxe "coefficient of variation", s/ EMBED Equation.DSMT4 , for your measurements. The coefficient of variation (c.v.) is a good measure of the precision of your technique. For this test a c.v. < 1% should be achievable.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 2. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 7  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 2. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 8  Figure  SEQ Figure2. Density of water vs. temperature. Measure Density Weigh a 100 mL volumetric flask with its cap (use the 400 g or 800 g balance). Prepare 100 mL of a 1 M solution of sodium chloride in the weighed flask. Make sure to mix the solution and then verify that you have exactly 100 mL of solution. Note that the volume decreases as the salt dissolves. Weigh the flask (with its cap) plus the sodium chloride solution and calculate the density of the 1 M NaCl solution. Prepare methylene bluexe "methylene blue" standards of several concentrations A methylene bluexe "methylene blue" stock solution of 1 g/L has been prepared. Use it to prepare 100 mL of each of the following concentrations: 1 mg/L, 2 mg/L, 3 mg/L, 4 mg/L, and 5 mg/L. Note any errors in transfer of mass as you prepare these dilutions (the color will make it easy to see). Prepare a standard curve and measure an unknown See page  PAGEREF _Ref406553312 \h 160 for instructions on using the UVVis Spectrophotometer software. Measure the absorbance of the methylene bluexe "methylene blue" solutions using a UVVis spectrophotometerxe "spectrophotometer". Analyze the 5 methylene blue samples plus a distilled water sample (0 mg/L methylene blue) as standards. Select Measure Standards from the computer control palette. Fill in the information for the six samples (starting with distilled water and ending with the highest concentration of methylene blue) and follow instructions as you are prompted. Save the data as \\enviro\enviro\Courses\453\fundamentals\netid_blue. Record the absorbance at 660 nm for each of the solutions. You can drag the blue cursor on the standard graph to the wavelength of choice and read the exact absorbance (and wavelength) in the digital display to the right of the graph. Note that you can do this after you have analyzed all of the standards. Record the correlation coefficientxe "correlation coefficient" (equation  GOTOBUTTON ZEqnNum137225 \* MERGEFORMAT  REF ZEqnNum137225 \! \* MERGEFORMAT 2.6), slope (equation  GOTOBUTTON ZEqnNum252391 \* MERGEFORMAT  REF ZEqnNum252391 \! \* MERGEFORMAT 2.4), and intercept (equation  GOTOBUTTON ZEqnNum304763 \* MERGEFORMAT  REF ZEqnNum304763 \! \* MERGEFORMAT 2.5) for the absorbance at 660 nm vs. methylene bluexe "methylene blue" concentration. These values are shown next to the calibration graph and correspond to the wavelength selected using the blue cursor on the standard graph. Measure the absorbance of a methylene bluexe "methylene blue" solution of unknown concentration. Select Measure Samples from the control palette. Save the data as \\enviro\enviro\Courses\453\fundamentals\netid_unknown. Record its absorbance at 660 nm and the calculated concentration. These values are given in the digital displays in the bottom left of the window. Print the results by selecting Print from the control palette. Export your standards spectra to the \\enviro\enviro\Courses\453\fundamentals folder. Turn on the pump and place the sipper tube in distilled water to clean out the sample cell by selecting Run Pump from the control palette. Prelab Questions You need 100 mL of a 1 M solution of zinc that you will use as a standard to calibrate an atomic adsorptionxe "adsorption" spectrophotometerxe "spectrophotometer". Find a source of zinc ions combined either with chloride or nitrate (you can use the world wide web or any other source of information). What is the molecular formula of the compound that you found? Zinc disposal down the sanitary sewer is restricted at Cornell. How does the disposal restriction for zinc influence how you prepare the zinc standard? How would you prepare this standard using techniques readily available in the environmental laboratory? Note that we have pipettes that can dispense volumes between 10 mL and 1 mL and that we have 100 mL and 1 L volumetric flasks. Include enough information so that you could prepare the standard without doing any additional calculations. Consider your ability to accurately weigh small masses. Explain your procedure for any dilutions. The density of sodium chloride solution XE "density of sodium chloride solution" s as a function of concentration is approximately 0.6985C + 998.29 (kg/m3) (C is kg of salt/m3). What is the density of a 1 M solution of sodium chloride? Questions Create a graph of absorbance at 660 nm vs concentration of methylene blue in Excel using the exported data file. Does absorbance at 660 nm increase linearly with concentration of methylene bluexe "methylene blue"? Plot e as a function of wavelength for each of the standards on a single graph. Make sure you include units and axis labels on your graph. If Beer s law is obeyed what should the graph look like? Did you use interpolation or extrapolation to get the concentration of the unknown? What colors of light are most strongly absorbed by methylene bluexe "methylene blue"? What measurement controls the accuracy of the density measurement? What should the accuracy be? What was your percent error in measuring the density of the 1 M NaCl solution? Data Sheet Balance Calibration Balance ID Mass of calibration mass 2nd mass used to verify calibration Measured mass of 2nd mass Temperature Measurement Distilled water temperature Pipette Technique (use DI100 balance) Density of waterxe "density of water" at that temperature Actual mass of 990 L of pure water Mass of 990 L of water (rep 1) Mass of 990 L of water (rep 2) Mass of 990 L of water (rep 3) Mass of 990 L of water (rep 4) Mass of 990 L of water (rep 5) Average of the 5 measurements Standard deviation of the 5 measurements Precision Percent coefficient of variationxe "coefficient of variation" of the 5 measurements Accuracy average percent error for pipetting  EMBED Equation.DSMT4  Where actual mass is calculated from the density of waterxe "density of water" and the setting of the pipettexe "pipette" and measured mass is measured with the balance. Measure Density (use DI800 balance) Molecular weight of NaCl Mass of NaCl in 100 mL of a 1M solution Measured mass of NaCl used Measured mass of empty 100 mL flask Measured mass of flask + 1M solution Mass of 100 mL of 1 M NaCl solution Density of 1 M NaCl solution Prepare methylene bluexe "methylene blue" standards of several concentrations Volume of 1 g/L MB diluted to 100 mL to obtain: 1 mg/L MB 2 mg/L MB 3 mg/L MB 4 mg/L MB 5 mg/L MB Measure absorbance at 660 nm using a spectrophotometerxe "spectrophotometer". Spectrophotometerxe "spectrophotometer" (computer name)? Absorbance of distilled water Absorbance of 1 mg/L methylene bluexe "methylene blue" Absorbance of 2 mg/L methylene bluexe "methylene blue" Absorbance of 3 mg/L methylene bluexe "methylene blue" Absorbance of 4 mg/L methylene bluexe "methylene blue" Absorbance of 5 mg/L methylene bluexe "methylene blue" Slope at 660 nm (m) Intercept at 660 nm (b) Correlation coefficientxe "correlation coefficient" at 660 nm (r) Absorbance of unknown at 660 nm Calculated concentration of unknown Lab Prep Notes Table  SEQ table2. Reagent list. Description SupplierCatalog numberNaClFisher Scientific BP3581 Methylene blueFisher ScientificM29125Table  SEQ table3. Equipment list Description SupplierCatalog numberCalibra 1001095 LFisher Scientific137075Calibra 10109.5 LFisher Scientific137073DI 100 analytical toploaderFisher Scientific019131ADI800 ToploaderFisher Scientific019131C100 mL volumetricFisher Scientific1019850BUVVis spectrophotometerxe "spectrophotometer"HewlettPackard Company8452ATable  SEQ table4. Methylene Blue Stock Solution DescriptionMW (g/M)conc. (g/L)100 mLC16H18N3SCl319.871100.0 mg Setup Prepare stock methylene bluexe "methylene blue" solution and distribute to student workstations in 15 mL vials. Prepare 100 mL of unknown in concentration range of standards. Divide into two bottles (one for each spectrophotometer XE "spectrophotometer" ). Verify that spectrophotometers are working (prepare a calibration curve as a test). Verify that balances calibrate easily. Disassemble, clean and lubricate all pipettes. Reactor Characteristics Introduction  MACROBUTTON MTEditEquationSection Equation Section (Next) SEQ MTEqn \r \h \* MERGEFORMAT  SEQ MTSec \h \* MERGEFORMAT Chemical, biological and physical processes in nature and in engineered systems usually take place in what we call "reactors." Reactors are defined by a real or imaginary boundary that physically confines the processes. Lakes, segments of a river, and settling tanks in treatment plants are examples of reactors. Most, but not all, reactors experience continuous flow (in and out). Some reactors, experience flow (input and output) only once. These are called "batch" reactors. It is important to know the mixing level and residence time in reactors, since they both affect the degree of process reaction that occurs while the fluid (usually water) and its components (often pollutants) pass through the reactorxe "reactor". Tracer studies can be used to determine the hydraulic characteristics of a reactorxe "reactor" such as the disinfection contact tanks at water treatment plants. The results from tracerxe "tracer" studies are used to obtain accurate estimates of the effective contact time. Reactor Classifications Mixing levels give rise to three categories of reactors; completely mixed flowxe "completely mixed flow" (CMF), plug flowxe "plug flow" (PF) and flow with dispersion XE "flow with dispersion"  (FD). The plug flow reactorxe "reactor" is an idealized extreme not attainable in practice. All real reactors fall under the category of FD or CMF. Reactor Modeling Both the CMF and the PF reactors are limiting cases of the FD reactorxe "reactor". Therefore the FD reactor model will be developed first. Equation  GOTOBUTTON ZEqnNum846802 \* MERGEFORMAT  REF ZEqnNum846802 \! \* MERGEFORMAT 3.1 is the governing differential equation for a conservative (i.e., nonreactive) substance in a reactor that has advective transport (i.e., flow) and some mixing in the direction of flow (x  dimension).  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 3. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 1 C = concentration of a conservative substance U = average fluid velocity in the x direction Dd = longitudinal dispersion coefficientxe "dispersion coefficient" t = time The dispersion coefficientxe "dispersion coefficient" is a measure of the mixing in a system. Flow with Dispersion One of the easiest methods to determine the mixing (dispersion) characteristics of a reactorxe "reactor" is to add a spike input of a conservative material and then monitor the concentration of the material in the reactor effluent. Assuming complete mixing in yz plane then transport occurs only in the x direction and the concentration of tracerxe "tracer" for any x and t (after t=0) the solution to equation  GOTOBUTTON ZEqnNum586560 \* MERGEFORMAT  REF ZEqnNum586560 \! \* MERGEFORMAT 3.1 gives:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 3. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 2 where M = mass of conservative material in the spike, Dd = axial dispersion coefficientxe "dispersion coefficient" [L2/T], x' = x  Ut, U = longitudinal advective velocity in the reactorxe "reactor", and A is the crosssectional area of the reactor. A measure of dispersion can be obtained directly from equation  GOTOBUTTON ZEqnNum332361 \* MERGEFORMAT  REF ZEqnNum332361 \! \* MERGEFORMAT 3.2. From this equation we expect a maximum value of C at t = x/U. At this time EMBED Equation.DSMT4 . If the mass of the tracerxe "tracer" input (M) and reactor crosssectional area (A) are known, then Dd can be estimated. The form of equation  GOTOBUTTON ZEqnNum524826 \* MERGEFORMAT  REF ZEqnNum524826 \! \* MERGEFORMAT 3.3 is exactly like the normal distribution curve:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 3. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 3 where  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 3. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 4 The variance in concentration over space ( EMBED Equation.DSMT4 ) is the variance in concentrations taken from many different positions in the reactorxe "reactor" at some single moment in time, t. The variance in x ( EMBED Equation.DSMT4 ) has dimensions of length squared. The variance of tracerxe "tracer" concentration versus time ( EMBED Equation.DSMT4 , with dimensions of time squared) can be measured by sampling at a single point in the reactorxe "reactor" at many different times and can be computed using the following equations.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 3. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 5 where  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 3. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 6 For discrete data points:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 3. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 7 and  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 3. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 8 Inlet and outlet boundary conditions affect the response obtained from a reactorxe "reactor". Closed reactors have little dispersion across their inlet and outlet boundaries whereas "open" reactors can have significant dispersion across their inlet and outlet boundaries. Typically open systems have no physical boundaries in the direction of flow. An example of an open system would be a river segment. Closed systems have small inlets and outlets that minimize dispersion across the inlet and outlet regions. An example of a closed system is a tank (or a lake) with a small inlet and outlet. The reactors used in the lab are closed. The  EMBED Equation.DSMT4  in equation  GOTOBUTTON ZEqnNum533013 \* MERGEFORMAT  REF ZEqnNum533013 \! \* MERGEFORMAT 3.8 is the measured average residence time for the tracerxe "tracer" in the reactor. For ideal closed reactors the measured residence time,  EMBED Equation.DSMT4 , is equal to the theoretical hydraulic residence timexe "hydraulic residence time" (q = reactor volume/flow rate). The above equations suggest that from the reactorxe "reactor" response to a spike input we can compute the dispersion coefficientxe "dispersion coefficient" for the reactor. We have two options for measuring reactor response: synoptic measurements: at a fixed time sampling many points along the axis of the reactorxe "reactor" will yield a Gaussian curve of concentration vs. distance. In practice synoptic measurements are difficult because it requires sampling devices that are timecoordinated. By combining equations  GOTOBUTTON ZEqnNum810521 \* MERGEFORMAT  REF ZEqnNum810521 \! \* MERGEFORMAT 3.4,  GOTOBUTTON ZEqnNum278707 \* MERGEFORMAT  REF ZEqnNum278707 \! \* MERGEFORMAT 3.7, and  GOTOBUTTON ZEqnNum854946 \* MERGEFORMAT  REF ZEqnNum854946 \! \* MERGEFORMAT 3.8 it is possible to estimate the dispersion coefficientxe "dispersion coefficient" from synoptic measurements. single point sampling: measure the concentration at a fixed position along the x axis of the reactorxe "reactor" for many times. If the reactor length is fixed at L and measurements are made at the effluent of the reactor (observe the concentration of a tracerxe "tracer" at x = L as a function of time) then x is no longer a variable and C(x,t) becomes C(t) only. The response curve obtained through single point sampling is skewed. The curve spread changes during the sampling period and the response curve is skewed. Peclet Numberxe "Peclet number" The dimensionless parameter Pe (Peclet numberxe "Peclet number") is used to characterize the level of dispersion in a reactorxe "reactor". The Peclet number is the ratio of advective to dispersive transport.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 3. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 9 In the limiting cases when Pe = 0 (very high dispersion) we have a complete mix regime (CMFR) and when PE = " (Dd = 0, no dispersion) we have a plug flowxe "plug flow" reactorxe "reactor" (PF). For single point sampling of the effluent response curve, skew increases as the dispersion level in the reactorxe "reactor" increases. The degree of skew depends on the dispersion coefficientxe "dispersion coefficient", the velocity in the xdirection, and the length of the reactor. Peclet values in the range 100<Pe<" result in a symmetric response curve. Response curve skew makes the assumption of a symmetrical normal distribution curve inappropriate and a new relationship between the variance and the dispersion coefficientxe "dispersion coefficient" (or Pe) has to be determined. Boundary conditions affect the determination of the dispersion coefficient. The relationship between the Peclet numberxe "Peclet number" and variance for open systems is given by:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 3. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 10 For closed systems the relationship is:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 3. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 11 The term  EMBED Equation.DSMT4 in equations  GOTOBUTTON ZEqnNum752411 \* MERGEFORMAT  REF ZEqnNum752411 \! \* MERGEFORMAT 3.10 and  GOTOBUTTON ZEqnNum326350 \* MERGEFORMAT  REF ZEqnNum326350 \! \* MERGEFORMAT 3.11 is dominant for Peclet numberxe "Peclet number"s much greater than 10 as is shown in  REF _Ref365247940 \* MERGEFORMAT Figure 1. The additional terms in equations  GOTOBUTTON ZEqnNum752411 \* MERGEFORMAT  REF ZEqnNum752411 \! \* MERGEFORMAT 3.10 and  GOTOBUTTON ZEqnNum326350 \* MERGEFORMAT  REF ZEqnNum326350 \! \* MERGEFORMAT 3.11 are corrections for skewedness in the response curve. These skewedness corrections are not very significant for Peclet numbers greater than 10. Thus for Peclet numbers greater than 10 the Peclet number can be determined using equation  GOTOBUTTON ZEqnNum266069 \* MERGEFORMAT  REF ZEqnNum266069 \! \* MERGEFORMAT 3.12 for both open and closed systems.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 3. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 12 EMBED Word.Picture.8 Figure  SEQ \r 1 Figure1. Relationship between equations  GOTOBUTTON ZEqnNum304324 \* MERGEFORMAT  REF ZEqnNum304324 \! \* MERGEFORMAT 3.10 through  GOTOBUTTON ZEqnNum337228 \* MERGEFORMAT  REF ZEqnNum337228 \! \* MERGEFORMAT 3.12. Flow through Porous Media Flow through porous mediaxe "porous media" (such as groundwaterxe "groundwater" through soil) is a type of flow with dispersion XE "flow with dispersion" . The above equations can be applied by recognizing that the relevant water velocity is the pore water velocity. The pore water velocity is U =  EMBED Equation.DSMT4 where A is the cross sectional area of the porous media and e (volume of voids/total volume) is the porosity of the porous media. Completely Mixed Flow Reactor In the case of CMF reactors, there is not an analytical solution to the advective dispersionxe "advective dispersion" equation so we revert to a simple mass balancexe "mass balance". For a completely mixed reactorxe "reactor" a mass balance on a conservative tracerxe "tracer" yields the following differential equation:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 3. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 13 where Q is the volumetric flow rate and V is the volume of the reactorxe "reactor". Equation  GOTOBUTTON ZEqnNum739367 \* MERGEFORMAT  REF ZEqnNum739367 \! \* MERGEFORMAT 3.13 can be used to predict a variety of effluent responses to tracerxe "tracer" inputs such as the pulse input used in this experiment. If a mass of tracer is discharged directly into a reactorxe "reactor" so that the initial concentration of tracer in the reactor is C0 =  EMBED Equation.DSMT4 and the input concentration is zero (Cin = 0) the solution to the differential equation is:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 3. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 14 If a reactorxe "reactor" has a complete mix flow regime its response (C/C0 vs. time) to a pulse input should plot as a straight line on a semilogarithmic plot. The slope of this plot is the negative inverse of the average hydraulic residence timexe "hydraulic residence time",  EMBED Equation.DSMT4 , of the reactor. Complete mix flow regimes can be approximated quite closely in practice. Plug Flow Reactor Plug flowxe "Plug flow" regimes are impossible to attain because mass transport must be by advection alone. There can be no differential displacement of tracerxe "tracer" relative to the average advective velocity. In practice some mixing will occur due to molecular diffusionxe "diffusion", turbulent dispersion, and/or fluid shear. For the case of the plug flowxe "plug flow" reactorxe "reactor" the advective dispersionxe "advective dispersion" equation  GOTOBUTTON ZEqnNum636911 \* MERGEFORMAT  REF ZEqnNum636911 \! \* MERGEFORMAT 3.1 reduces to:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 3. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 15  Figure  SEQ Figure2. Pulse and step input in a plug flowxe "plug flow" reactorxe "reactor". The velocity U serves to transform the directional concentration gradient into a temporal concentration gradient. In other words, a conservative substance moves with the advective flow of the fluid. The solutions to this differential equation for a pulse input and for a step input are shown graphically in  REF _Ref365260004 \* MERGEFORMAT Figure 2. Mass Conservation When a pulse of conservative tracerxe "tracer" is added to a continuous flow reactorxe "reactor", all of the tracer is expected to leave the reactor eventually. The mass of a substance that has left the reactor is given in equation  GOTOBUTTON ZEqnNum840307 \* MERGEFORMAT  REF ZEqnNum840307 \! \* MERGEFORMAT 3.16.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 3. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 16 where Q is the flow rate and M is the mass of any substance whose concentration is given by C. If Q and "t are constant, then equation  GOTOBUTTON ZEqnNum716292 \* MERGEFORMAT  REF ZEqnNum716292 \! \* MERGEFORMAT 3.16 can be rewritten as  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 3. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 17 Equation  GOTOBUTTON ZEqnNum937963 \* MERGEFORMAT  REF ZEqnNum937963 \! \* MERGEFORMAT 3.17 can be used to determine if all of the tracerxe "tracer" was measured in the reactorxe "reactor" effluent. Conductivity Measurements We will use a tracerxe "tracer" containing salt (NaCl) and red dye # 40 (for visualization). The concentration of NaCl will be monitored using a conductivityxe "conductivity" probe. Conductivity is the measure of a material's ability to conduct electric current. Conductivity is measured by passing an electrical current between two electrodes and then measuring the voltage. The electrodes can be made of platinum, titanium, goldplated nickel, or graphite. Conductivity is defined as:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 3. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 18 where G is conductivityxe "conductivity", I is the current, and E is the measured voltage. If the current is held constant, as the conductivityxe "conductivity" of the solution increases the voltage between the electrodes will decrease. For a given current, the measured voltage will increase as the size of the electrodes decreases and as the distance between the electrodes increases. We are interested, however, in measuring properties of the solution, not properties of the conductivity probe! Specific conductivity, C, is a property of the solution.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 3. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 19 where L is the distance between the electrodes and A is the area of the electrodes. The term  EMBED Equation.DSMT4 is a property of the conductivityxe "conductivity" cell and is called the cell constant. In practice, the cell constant is determined during calibration by measuring the conductivity (G) of a solution with known specific conductivity (C). The units of specific conductivity are Siemens/cm where Siemens are the inverse of Ohms. For a solution to be conductive, it must have ions that can transport the charge between the electrodes. In pure water, the only ions available are H+ and OH. Adding species that disassociate into charged ions increases both the concentration of ions and the specific conductivityxe "conductivity". At low concentrations, specific conductivity increases linearly with the concentration of ions. At very high concentrations ionion interactions become significant and the relationship is no longer linear. The specific conductivity of several common solutions is given in Table 1. Table  SEQ \r 1 table1. Conductivity of some common solutions.SolutionSpecific Conductivitypure water0.055 S/cmdistilled water0.5 S/cmdeionized water0.110 S/cmtypical drinking water0.51.0 mS/cmwastewater0.99.0 mS/cmmaximum drinking water1.5 mS/cmocean water53 mS/cm10% NaOH355 mS/cm495 mg/L NaCl1 mS/cm Conductivity measurements are temperature dependent. The conductivityxe "conductivity" of a solution will increase as the temperature increases. The Accumet"! meter that you will use in this laboratory compensates for this effect by also measuring the temperature and reporting the solution specific conductivity at 25C. In this lab sodium chloride will increase the specific conductivityxe "conductivity" of the water in the reactors. The concentration of sodium chloride will be low enough so that specific conductivity will be linearly related to the concentration of sodium chloride. Procedures A conservative tracerxe "tracer" will be used to characterize each of the reactors. A conservative tracer with 20 g NaCl/L and 4 g red dye # 40/L will be used. The salt will increase the conductivityxe "conductivity" of the water and conductivity will be measured to monitor the salt concentration. The red dye was added to the tracer to make it possible to see the tracer. A common problem when using tracers is that the tracerxe "tracer" may have a different density than the fluid that is in the reactors. In this case the salt and dye add significantly to the density of the tracer. The tracer would tend to sink to the bottom of the reactors. To compensate for this problem the density of the water being pumped into some reactors will be increased by using a glucose solution (37 g glucose/L). Glucose is nonionic and thus will not increase the conductivityxe "conductivity" of the solution. Calibrate Conductivityxe "conductivity" probe Calibrate the conductivityxe "conductivity" probe by placing it in a 495 mg NaCl/L standard. Press the conductivity button on the Accumet"! meter if it is not already in the conductivity mode. Press standardize and enter 1000 S/cm. Press enter and the meter will calibrate and return to the normal display mode. Measure Conductivityxe "conductivity" of tracerxe "tracer" Prepare a calibration curve for conductivityxe "conductivity" vs. concentration of the tracerxe "tracer" (expressed as mg/L of NaCl). The tracer has 20 g/L of NaCl. Measure the conductivity of tracer diluted with distilled water so that the final concentrations of NaCl are 500, 200, and 100 mg/L. As a zero point measure the conductivity of distilled water. Measure Reactor Response to Pulse Input For each reactor XE "reactor"  add a pulse input of sodium, measure conductivity XE "conductivity"  vs. time in the reactor effluent and use the Compumet XE "Compumet" "! software to monitor the conductivity vs. time (see discussion below). Save the collected data for later analysis using a spreadsheet program. The experimental setup is shown in  REF _Ref365259988 \* MERGEFORMAT Figure 3. Specific instructions for each type of reactor are detailed below.  Figure  SEQ Figure3. Reactor schematic. Only one reactorxe "reactor" at a time will be connected to the peristaltic pump. Porous Media The porous media XE "porous media"  column is 2.5 cm in diameter, 60 cm long and contains 60 cm of glass beads. The overall porosity including headspace and underdrains is approximately 0.4. Use this information to estimate the volume of water in the reactor XE "reactor" . The conductivityxe "conductivity" probe should be plumbed into the effluent line. Verify that the flow rate is set to 10 mL/min. Inject 10 mg NaCl (0.5 mL of tracerxe "tracer") into the influent line. Select Set Method from the Compumet XE "Compumet" "! control palette. Use automatic data transmission with a timed interval of 1 second. Set channel A to Conductivity and channel B to Off. Select Monitor Sample from the control palette. Start the pump and press the enter key on the keyboard to begin data acquisition. Measure the actual flow rate by collecting a timed sample from the effluent. To get an accurate flow rate you should collect a sample for several minutes. Estimate the width of the tracer XE "tracer"  pulse when the pulse nears the top of the reactorxe "reactor" and record the corresponding time. This information will be used to obtain an estimate of the dispersion coefficientxe "dispersion coefficient". Turn off the pump when the conductivity XE "conductivity"  returns to the baseline conductivity. Stop data acquisition by clicking on the Stop Sampling button. Save the data to \\Enviro\enviro\Courses\453\reactors\netid_porousmedia by selecting Save data from the control palette. The data will be saved in a file (tab delimited format) that can be opened by any spreadsheet program. Completely Mix Flow Reactor (CMFR) Verify that the flow rate is set to 300 mL/min. Fill the CMFR with distilled water to within about 2 mm of the overflow drain . Measure the conductivityxe "conductivity" of the distilled water. Set the stirrer to the highest setting that doesn't cause splashing (setting 8) and place the conductivity XE "conductivity"  probe near the stir bar. Add 800 mg NaCl (40 mL tracerxe "tracer") directly to the CMFR. Select Set Method from the Compumet XE "Compumet" "! control palette. Use automatic data transmission with a timed interval of 10 second. Set channel A to Conductivity and channel B to Off. Select Monitor Sample from the control palette. Start the pump and press the enter key on the keyboard to begin data acquisition. Record the time when water begins flowing out the overflow (this is your actual time zero!) Measure the flow rate by collecting a timed sample from the effluent. To get an accurate flow rate you should collect a sample for several minutes. Turn off the pump after 2 residence times. Stop data acquisition by clicking on the Stop Sampling button. Save the data to \\Enviro\enviro\Courses\453\reactors\netid_CMFR by selecting Save data from the control palette. The data will be saved in a file (tab delimited format) that can be opened by any spreadsheet program. Determine the volume of water in the CMFR. Baffled Tank Reactor The baffled tank reactorxe "reactor" is a simple attempt to reduce mixing and short-circuiting. The channels are approximately 4.5 cm wide, 4.8 cm deep and have a total length of 80 cm. Verify that the flow rate is set to 300 mL/min. Determine the volume of water in the baffled tank. Fill the baffled tank with glucose water. Measure the conductivityxe "conductivity" of the glucose water. Select Set Method from the Compumet XE "Compumet" "! control palette. Use automatic data transmission with a timed interval of 10 second. Set channel A to Conductivity and channel B to Off. Select Monitor Sample from the control palette. Inject 200 mg NaCl (10 mL of tracerxe "tracer") into the influent line with a syringe. Start the pump and press the enter key on the keyboard to begin data acquisition. During data acquisition, it is important to gently move the conductivityxe "conductivity" probe up and down to continually bring the probe into contact with the changing solution. While moving the probe up and down, do not lift the probe so high that the platinum contacts leave the solution Measure the actual flow rate by collecting a timed sample from the effluent. To get an accurate flow rate you should collect a sample for several minutes. Turn off the pump when the conductivity XE "conductivity"  returns to the baseline conductivity. Stop data acquisition by clicking on the Stop Sampling button. Measure the average conductivity XE "conductivity"  of the remaining solution in the baffled tank. Save the data to \\Enviro\enviro\Courses\453\reactors\netid_baffled by selecting Save data from the control palette. The data will be saved in a file (tab delimited format) that can be opened by any spreadsheet program. Pipe Flow The pipe flow setup consists of 15.24 m of 6 mm ID tubing. Verify that the flow rate is set to 50 mL/min. Fill the pipe with glucose water. Select Set Method from the Compumet XE "Compumet" "! control palette. Use automatic data transmission with a timed interval of 1 second. Set channel A to Conductivity and channel B to Off. Select Monitor Sample from the control palette. Inject 10 mg NaCl (0.5 mL of tracerxe "tracer") into the influent line with a syringe. Start the pump and press the enter key on the computer keyboard to begin data acquisition. Measure the actual flow rate by collecting a timed sample from the effluent. To get an accurate flow rate you should collect a sample for several minutes. Turn off the pump when the conductivity XE "conductivity"  returns to the baseline conductivity. Stop data acquisition by clicking on the Stop Sampling button. Save the data to \\Enviro\enviro\Courses\453\reactors\netid_pipe by selecting Save data from the control palette. The data will be saved in a file (tab delimited format) that can be opened by any spreadsheet program. Prelab Questions Why is a 37 g/L glucose solution used for the plug flow XE "plug flow"  reactor XE "reactor" ? Why is the glucose solution not needed for the completely mixed flow XE "completely mixed flow"  reactor? Why is the conductivityxe "conductivity" of pure water not zero? Data Analysis Use a consistent set of units throughout your data analysis and include the units in your spreadsheet and report! Derive an equation relating concentration of NaCl in the tracerxe "tracer" to conductivityxe "conductivity" based on the 4 point calibration curve. Use the slope from the equation and the baseline conductivity of each of the reactors to convert the conductivity data to concentration of NaCl for each reactorxe "reactor". Perform a mass balancexe "mass balance" on the salt. When applicable include the measurements of residual salt left in the reactors at the end of your experiments. Use equation  GOTOBUTTON ZEqnNum937963 \* MERGEFORMAT  REF ZEqnNum937963 \! \* MERGEFORMAT 3.17 to calculate the mass of NaCl measured in the effluent from each reactorxe "reactor" and compare with the mass of NaCl added. Calculate the volume of the pipe flow and porous media XE "porous media"  reactors based on their dimensions and porosity. From the data determine  EMBED Equation.DSMT4  for each reactor XE "reactor"  (use equation  GOTOBUTTON ZEqnNum653284 \* MERGEFORMAT  REF ZEqnNum653284 \! \* MERGEFORMAT 3.8 for the baffled tank, pipe flow, and porous media XE "porous media"  column and use equation  GOTOBUTTON ZEqnNum193836 \* MERGEFORMAT  REF ZEqnNum193836 \! \* MERGEFORMAT 3.14 for the CMFR) and compare with the hydraulic residence timexe "hydraulic residence time" (q = V/Q). Discuss any discrepancies. The width of the plume XE "plume"  as measured by eye for the porous media XE "porous media"  column is approximately 4 standard deviations (2 s on each side of the peak). Use your measurement of the width of the plume and equation  GOTOBUTTON ZEqnNum593778 \* MERGEFORMAT  REF ZEqnNum593778 \! \* MERGEFORMAT 3.4 to estimate the dispersion coefficient XE "dispersion coefficient"  for the porous media reactorxe "reactor". Estimate the Peclet numberxe "Peclet number"s (equation  GOTOBUTTON ZEqnNum795450 \* MERGEFORMAT  REF ZEqnNum795450 \! \* MERGEFORMAT 3.12) and the dispersion coefficientxe "dispersion coefficient"s (equation  GOTOBUTTON ZEqnNum631166 \* MERGEFORMAT  REF ZEqnNum631166 \! \* MERGEFORMAT 3.9) for the baffled tank, pipe flow, and porous media XE "porous media"  column. Compare the dispersion coefficient for the porous media reactorxe "reactor" with the estimate obtained in the previous step. Plot actual and theoretical effluent tracerxe "tracer" concentration versus time for the three reactors. Use the calculated dispersion coefficientxe "dispersion coefficient" (equation  GOTOBUTTON ZEqnNum631166 \* MERGEFORMAT  REF ZEqnNum631166 \! \* MERGEFORMAT 3.9) substituted into equation  GOTOBUTTON ZEqnNum925603 \* MERGEFORMAT  REF ZEqnNum925603 \! \* MERGEFORMAT 3.2 to model the baffled tank, pipe flow, and porous media XE "porous media"  columnxe "reactor". On the graphs also show q, and  EMBED Equation.DSMT4 (these can be added to your graph in Excel as additional plots). Use the CMFR volume to obtain the theoretical value of the initial concentration. Compare your results with theory. Give possible reasons for deviations from theoretical expectations. Discuss what you learned. Lab Prep Notes Table  SEQ table2. Equipment list DescriptionSupplierCatalog numberAccumet"! 50 meterFisher Scientific1363550floating stir barFisher Scientific1451199Amagnetic stirrerFisher Scientific115007S6 L containerFisher Scientific0348422Conductivity Cell 1/cmFisher Scientific13620160columnFisher ScientificK4208306010glass shot 297420 mSunbelt IndustriesMil 5 6 L container with bafflesCEE shop3port leur manifoldCole ParmerH0646482variable flow digital driveCole ParmerH0752330EasyLoad pump headCole ParmerH0751800PharMed tubing size 18Cole ParmerH064851820 liter HDPE JerricanFisher Scientific0296150CTable  SEQ table3. Reagent list DescriptionSupplier/SourceCatalog numberNaClFisher Scientific BP3581  red dye #40MG Newell077041glucoseAldrich15,89681000 S solution495 mg NaCl/LTracer 20 g NaCl/L 4 g red dye #40/Lglucose feed solution37 g glucose/LPrepare glucose solution for the baffled tank and pipe flow reactorsxe "reactor" in Jerricans. Prepare 1 L of tracer.xe "tracer" Prepare 1 L of 1000 S/cm solution (conductivityxe "conductivity" standard). Distribute tracerxe "tracer" and conductivityxe "conductivity" standard for each setup. Use # 18 tubing for CMFR and baffled tank. Use #16 tubing for pipe flow and porous media XE "porous media"  reactors. Place all conductivity XE "conductivity"  probes in distilled water so they are conditioned. Otherwise the readings will drift. 0.5 mL tracerxe "tracer"/100 mL = 100 mg/L. EMBED Excel.Sheet.8 Figure  SEQ Figure4. Density of glucose solution as a function of glucose concentration. EMBED Excel.Sheet.8 Figure  SEQ Figure5. Density of sodium chloride solution as a function of concentration. The concentration of glucose required to achieve the same density as a sodium chloride solution is 1.848 times as great. Acid Precipitation and Remediationxe "Remediation" of Acid Lakes Introduction  MACROBUTTON MTEditEquationSection Equation Section (Next) SEQ MTEqn \r \h \* MERGEFORMAT  SEQ MTSec \h \* MERGEFORMAT Acid precipitation has been a serious environmental problem in many areas of the world for the last few decades. Acid precipitation results from the combustion of fossil fuels, that produces oxides of sulfur and nitrogen that react in the earth's atmosphere to form sulfuric and nitric acid. One of the most significant impacts of acid rain is the acidification of lakes and streams. In some watersheds the soil doesnt provides ample acid neutralizing capacityxe "acid neutralizing capacity" to mitigate the effect of incident acid precipitationxe "acid precipitation". These susceptible regions are usually high elevation lakes, with small watersheds and shallow noncalcareous soils. The underlying bedrock of acidsensitive lakes tends to be granite or quartz. These minerals are slow to weather and therefore have little capacity to neutralize acids. The relatively short contact time between the acid precipitation and the watershed soil system exacerbates the problem. Lakes most susceptible to acidification: 1) are located downwind, sometimes hundreds of miles downwind, from major pollution sourceselectricity generation, metal refining operations, heavy industry, large population centers; 2) are surrounded by hard, insoluble bedrock with thin, sandy, infertile soil; 3) have a high runoff to infiltration ratio; 4) have a low watershedxe "watershed" to lake surface area. Isopleths of precipitation pH are depicted in  REF _Ref365260235 \* MERGEFORMAT Figure 1.  Figure  SEQ \r 1 Figure1. The pH of precipitation in 1999. In acidsensitive lakes the major parameter of concern is pH (pH = log{H+}, where {H+} is the hydrogen ion activity, and activity is approximately equal to concentration in moles/L). In a healthy lake, ecosystem pH should be in the range of 6.5 to 8.5. In most natural freshwater systems, the dominant pH buffering (controlling) system is the carbonate systemxe "carbonate system". The carbonate buffering system is composed of four components: dissolved carbon dioxide ( EMBED Equation.DSMT4 ), carbonic acid ( EMBED Equation.DSMT4 ), bicarbonate ( EMBED Equation.DSMT4 ), and carbonate ( EMBED Equation.DSMT4 ). Carbonic acid exists only at very low levels in aqueous systems and for purposes of acid neutralization is indistinguishable from dissolved carbon dioxide. Thus to simplify things we define  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 1 The  EMBED Equation.DSMT4  >>  EMBED Equation.DSMT4  and thus  EMBED Equation.DSMT4  (all terms enclosed in [] are in units of moles/L). The sum of all the molar concentration of the components of the carbonate systemxe "carbonate system" is designated as CT as shown in equation  GOTOBUTTON ZEqnNum795403 \* MERGEFORMAT  REF ZEqnNum795403 \! \* MERGEFORMAT 4.2.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 2 The carbonate systemxe "carbonate system" can be considered to be a "volatile" system or a "nonvolatile" system depending on whether or not aqueous carbon dioxide is allowed to exchange and equilibrate with atmospheric carbon dioxide. Mixing conditions and hydraulic residence timexe "hydraulic residence time" determine whether an aquatic system is volatile or nonvolatile relative to atmospheric carbon dioxide equilibrium. First, consider the "nonvolatile" system. Nonvolatile System For a fixed CT, the molar concentration of each species of the carbonate systemxe "carbonate system" is determined by pH. Equations  GOTOBUTTON ZEqnNum867607 \* MERGEFORMAT  REF ZEqnNum867607 \! \* MERGEFORMAT 4.3 GOTOBUTTON ZEqnNum296591 \* MERGEFORMAT  REF ZEqnNum296591 \! \* MERGEFORMAT 4.8 show these functional relationships.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 3 where  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 4  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 5 where  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 6  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 7 where  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 8 K1 and K2 are the first and second dissociation constants for carbonic acid and a0, a1, and a2 are the fraction of CT in the form  EMBED Equation.DSMT4 ,  EMBED Equation.DSMT4 , and  EMBED Equation.DSMT4  respectively. Because K1 and K2 are constants (K1 = 106.3 and K2 = 1010.3), a0, a1, and a2 are only functions of pH. A measure of the susceptibility of lakes to acidification is the acid neutralizing capacityxe "acid neutralizing capacity" (ANCxe "ANC") of the lake water. In the case of the carbonate system,xe "carbonate system" the ANC is exhausted when enough acid has been added to convert the carbonate species  EMBED Equation.DSMT4 , and  EMBED Equation.DSMT4  to  EMBED Equation.DSMT4 . A formal definition of total acid neutralizing capacity is given by equation  GOTOBUTTON ZEqnNum987939 \* MERGEFORMAT  REF ZEqnNum987939 \! \* MERGEFORMAT 4.9.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 9 ANCxe "ANC" has units of equivalents per liter. The hydroxide ion concentration can be obtained from the hydrogen ion concentration and the dissociation constant for water Kw.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 10 Substituting equations  GOTOBUTTON ZEqnNum963298 \* MERGEFORMAT  REF ZEqnNum963298 \! \* MERGEFORMAT 4.5,  GOTOBUTTON ZEqnNum513434 \* MERGEFORMAT  REF ZEqnNum513434 \! \* MERGEFORMAT 4.7, and  GOTOBUTTON ZEqnNum419395 \* MERGEFORMAT  REF ZEqnNum419395 \! \* MERGEFORMAT 4.10 into equation  GOTOBUTTON ZEqnNum237669 \* MERGEFORMAT  REF ZEqnNum237669 \! \* MERGEFORMAT 4.9, we obtain  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 11 For the carbonate system,xe "carbonate system" ANCxe "ANC" is usually referred to as alkalinityxe "alkalinity". Volatile Systems: Now consider the case where aqueous  EMBED Equation.DSMT4  is volatile and in equilibrium with atmospheric carbon dioxide. Henry's Lawxe "Henry's Law" can be used to describe the equilibrium relationship between atmospheric and dissolved carbon dioxide.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 12 where KH is Henry's constant for CO2 in moles/Latm and PCO2 is partial pressure of CO2 in the atmosphere (KH = 101.5 and PCO2 = 103.5). Because  EMBED Equation.DSMT4  is approximately equal to  EMBED Equation.DSMT4  and from equations  GOTOBUTTON ZEqnNum541463 \* MERGEFORMAT  REF ZEqnNum541463 \! \* MERGEFORMAT 4.1 and  GOTOBUTTON ZEqnNum712728 \* MERGEFORMAT  REF ZEqnNum712728 \! \* MERGEFORMAT 4.3  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 13  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 14 Equation  GOTOBUTTON ZEqnNum974308 \* MERGEFORMAT  REF ZEqnNum974308 \! \* MERGEFORMAT 4.14 gives the equilibrium concentration of carbonate species as a function of pH and the partial pressure of carbon dioxide. The acid neutralizing capacityxe "acid neutralizing capacity" expression for a volatile system can be obtained by combining equations  GOTOBUTTON ZEqnNum155289 \* MERGEFORMAT  REF ZEqnNum155289 \! \* MERGEFORMAT 4.14 and  GOTOBUTTON ZEqnNum229180 \* MERGEFORMAT  REF ZEqnNum229180 \! \* MERGEFORMAT 4.11.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 15 In both nonvolatile and volatile systems, equilibrium pH is controlled by system ANCxe "ANC". Addition or depletion of any ANC component in equation  GOTOBUTTON ZEqnNum293220 \* MERGEFORMAT  REF ZEqnNum293220 \! \* MERGEFORMAT 4.11 or  GOTOBUTTON ZEqnNum640102 \* MERGEFORMAT  REF ZEqnNum640102 \! \* MERGEFORMAT 4.15 will result in a pH change. Natural bodies of water are most likely to approach equilibrium with the atmosphere (volatile system) if the hydraulic residence timexe "hydraulic residence time" is long and the body of water is shallow. Lake ANCxe "ANC" is a direct reflection of the mineral composition of the watershedxe "watershed". Lake watersheds with hard, insoluble minerals yield lakes with low ANC. Typically watersheds with soluble, calcareous minerals yield lakes with high ANC. ANC of freshwater lakes is generally composed of bicarbonate, carbonate, and sometimes organic matter ( EMBED Equation.DSMT4 ). Organic matter derives from decaying plant matter in the watershed. When organic matter is significant, the ANC becomes (from equations  GOTOBUTTON ZEqnNum293220 \* MERGEFORMAT  REF ZEqnNum293220 \! \* MERGEFORMAT 4.11 and  GOTOBUTTON ZEqnNum640102 \* MERGEFORMAT  REF ZEqnNum640102 \! \* MERGEFORMAT 4.15):  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 16  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 17 where equation  GOTOBUTTON ZEqnNum350399 \* MERGEFORMAT  REF ZEqnNum350399 \! \* MERGEFORMAT 4.16 is for a nonvolatile system and equation  GOTOBUTTON ZEqnNum762339 \* MERGEFORMAT  REF ZEqnNum762339 \! \* MERGEFORMAT 4.17 is for a volatile system. During chemical neutralization of acid, the components of ANCxe "ANC" associate with added acid to form protonated molecules. For example:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 18 or  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 19 In essence, the ANCxe "ANC" of a system is a result of the reaction of acid inputs to form associated acids from basic anions that were dissolved in the lake water. The ANC (equation  GOTOBUTTON ZEqnNum237669 \* MERGEFORMAT  REF ZEqnNum237669 \! \* MERGEFORMAT 4.9) is consumed as the basic anions are converted to associated acids. This conversion is near completion at low pH (approximately pH 4.5 for the bicarbonate and carbonate components of ANC). Neutralizing capacity to another (probably higher) pH may be more useful for natural aquatic systems. Determination of ANC to a particular pH is fundamentally easy simply add and measure the amount of acid required to lower the sample pH from its initial value to the pH of interest. Techniques to measure ANC are described under the procedures section of this lab. Neutralization of acid precipitationxe "acid precipitation" can occur in the watershedxe "watershed" or directly in the lake. How much neutralization occurs in the watershed versus the lake is a function of the watershed to lake surface area. Generally, watershed neutralization is dominant. Recently engineered remediation XE "remediation"  of acid lakes has been accomplished by adding bases such as limestone, lime, or sodium bicarbonate to the watershed or directly to the lakes. Experimental Objectives Phase I: Acid Neutralization by Soil Column In this experiment, we will study the effect of watershedxe "watershed" soil on neutralization of acid precipitationxe "acid precipitation". Simulated acid precipitation (dilute sulfuric acid) will be applied to a column of watershed soil that will discharge to a volume of water that represents a lake. The column will contain an artificial soil amended with CaCO3 representing a highly calcareous soil. Influent and effluent lake pH and ANCxe "ANC" will be measured over time to document the depletion of the ANC of the soil column and the subsequent acidification of the lake. Organic matter as an ANC component will not be incorporated into these experiments. Phase II. Acid Lake Remediationxe "Remediation" Remediationxe "Remediation" of acid lakes involves addition of ANCxe "ANC" so that the pH is raised to an acceptable level and maintained at or above this level for some design period. In this experiment sodium bicarbonate (NaHCO3) will be used as the ANC supplement. Since ANC addition usually occurs as a batch addition, the design pH is initially exceeded. ANC dosage is selected so that at the end of the design period pH is at the acceptable level. Care must be taken to avoid excessive initial pH high pH can be as deleterious as low pH. The most common remediation XE "remediation"  procedure is to apply the neutralizing agent directly to the lake surface, instead of on the watershedxe "watershed". We will follow that practice in this lab experiment. Sodium bicarbonate will be added directly to the surface of the lake that has an initial ANCxe "ANC" of 50 eq/L and is receiving acid rain with a pH of 3. After the sodium bicarbonate is applied, the lake pH and ANC will be monitored for approximately one hour. Experimental Apparatus The experimental apparatus consists of an acid rain storage reservoir, peristaltic pump, soil column, and lake ( REF _Ref365260269 \* MERGEFORMAT Figure 2). The pH of the lake influent and of the lake will be monitored using pH probexe "pH probe"s connected to a pH meter. The soil column will be removed for phase II of the experiment.  Figure  SEQ Figure2. Schematic drawing of the experimental setup. The soil column is filled with glass beads. The ANCxe "ANC" of the soil column is the result of adding CaCO3 slurry to the top of the soil column. Undissolved CaCO3 quickly settles to the glass beads and thus CaCO3 is only removed from the column as it dissolves. For this experiment, 250 mg of CaCO3 has been added to the soil column. Detail of the soil column is shown in  REF _Ref365260299 \* MERGEFORMAT Figure 3.  Figure  SEQ Figure3. Soil column used for neutralizing acid rain. Experimental Procedures Calibration of pH Meter Calibrate both pH probexe "pH probe"s attached to the pH meter. To calibrate the probe attached to channel A toggle the display by pressing Channel until only the pH from channel A is in the display. (The display toggles between channel A, channel B, and both channels.) Then follow the calibration procedure outlined in the Appendix of this manual (page  PAGEREF _Ref409401688 \h 157). Repeat the procedure for the channel B probe (if you are doing the soil column experiment). Acid Neutralization by Soil Column Experiment In this experiment, the "acid rain" will pass through the soil column before running into the lake. Flow should be controlled to give a 15 minutes hydraulic retention time in the lake. With a lake volume of 5 liters, the required flow rate is 334 mL/min. Since surface waters are seldom at equilibrium or steady state, this experiment will be run under dynamic conditions. The lake will start out with no ANC XE "ANC" . Data will be taken from the onset of flow. Measure the volume of the lake (The lake weighs too much for the 5 kg balance but it can be weighed in two parts. Weigh the empty lake container, fill the lake, pour half of the lake into a weighed container, weigh both the lake container and the second container, add the weights of the containers with water and subtract the container weights.) Fill lake with distilled water. Add 1 mL of bromocresol green indicator to the lake. Preset pump to give desired flow rate of 334 mL/min (5 L/15 minutes). Measure the pH of the acid rain. Prepare to monitor the pH of two probes using the Compumet XE "Compumet" "! software. Set the method for automatic sampling of pH probexe "pH probe"s on both channel A and channel B every 10 seconds. See section on "Compumet"! software" (page  PAGEREF accumetsoftware \h 157) for information on using the Computer"! software to monitor pH. Place the probe attached to channel A to monitor influent to the lake. Place the probe attached to channel B to monitor the pH of the lake. Label sample bottles (see step 13). Set stirrer speed to setting 8. Observe the CaCO3 on top of the soil column. At time equal zero start the pump and begin monitoring pH at the lake influent and in the lake. Measure the flow rate using 100mL volumetric flask and a stopwatch. Take 50mL grab samples from the lake influent and effluent at 5, 10, 15, and 20 minutes. The sample volumes do not need to be measured. Collect the samples in 125mL bottles. Observe the depletion of the CaCO3 on top of the soil column. After the 20minute sample turn off the pump and stop sampling pH. Save the pH data to \\Enviro\enviro\Courses\453\acid\netid_column by selecting Save data from the control palette. The data will be saved in a file (tab delimited format) that can be opened by any spreadsheet program. Acid Lake Remediation Experimentxe "Remediation" In this experiment sodium bicarbonate will be added to a lake to mitigate the deleterious effect of acid rain. Usually sodium bicarbonate is added in batch doses (as opposed to metering in). The quantity of sodium bicarbonate added depends on how long a treatment is desired, the acceptable pH range and the quantity and pH of the incident rainfall. For purposes of this experiment, a 15minute design period will be used. That is, we would like to add enough sodium bicarbonate to keep the lake at or above its original pH and alkalinityxe "alkalinity" for a period of 15 minutes (i.e. for one hydraulic residence timexe "hydraulic residence time"). By dealing with ANCxe "ANC" instead of pH as a design parameter, we avoid the issue of whether the system is at equilibrium with atmospheric carbon dioxide. Keep in mind that eventually the lake will come to equilibrium with the atmosphere. In practice, neutralizing agent dosages may have to be adjusted to take into account nonequilibrium conditions. We must add enough sodium bicarbonate to equal the negative ANCxe "ANC" from the acid precipitationxe "acid precipitation" input plus the amount of ANC lost by outflow from the lake during the 15minute design period. Initially (following the dosing of sodium bicarbonate) the pH and ANC will rise, but over the course of 15 minutes, both parameters will drop. Calculation of required sodium bicarbonate dosage requires performing a mass balancexe "mass balance" on ANC around the lake. This mass balance will assume a completely mixed lake and conservation of ANC. Chemical equilibrium can also be assumed so that the sodium bicarbonate is assumed to react immediately with the incoming acid precipitation.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 20 where: ANCxe "ANC"out = ANC in lake outflow at any time t (for a completely mixed lake the effluent ANC is the same as the ANC in the lake) ANCxe "ANC"in = ANC of acid rain input V = volume of reactorxe "reactor" Q = acid rain input flow rate. If the initial ANCxe "ANC" in the lake is designated as ANC0, then the solution to the mass balancexe "mass balance" differential equation is:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 21 where: q = V/Q We want to find ANCxe "ANC"0 such that ANCout = 50 eq/L when t is equal to q. Solving for ANC0 we get  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 22 The ANCxe "ANC" of the acid rain (ANCin) can be estimated from its pH. Below pH 6.3 most of the carbonates will be in the form  EMBED Equation.DSMT4  and thus for pH below about 4.3 equation  GOTOBUTTON ZEqnNum925011 \* MERGEFORMAT  REF ZEqnNum925011 \! \* MERGEFORMAT 4.9 simplifies to  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 23 An influent pH of 3.0 implies the ANCxe "ANC"in =  EMBED Equation.DSMT4  = 0.001 Substituting into equation  GOTOBUTTON ZEqnNum703302 \* MERGEFORMAT  REF ZEqnNum703302 \! \* MERGEFORMAT 4.22:  EMBED Equation.DSMT4  = 1.854 meq/L The quantity of sodium bicarbonate required can be calculated from: [NaHCO3]0 =ANC0xe "ANC"  MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 24 where [NaHCO3]0 = moles of sodium bicarbonate required per liter of lake water  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 25 Verify that the system is plumbed so that the acid rain is pumped directly into the lake. Take a 50mL sample from the acid rain container. Collect the sample in a 125mL bottle. Preset pump to give desired flow rate of 334 mL/min (5 L/15 minutes). Fill lake with distilled water. Set stirrer speed to 8. Add 1 mL of bromocresol green indicator solution to the lake. Weigh out 779 mg (not grams!) NaHCO3. Add NaHCO3 to the lake. After the lake is well stirred take a 100 mL sample from the lake. Prepare to monitor the pH of one probe using the Compumet XE "Compumet" "! software. Set the method for automatic sampling of the pH probexe "pH probe" on channel A every 10 seconds. See section on "Compumet"! software" (page  PAGEREF Accumetsoftware 157) for information on using the computer to monitor pH. Place the probe attached to channel A to monitor the pH of the lake. Label sample bottles (see step 13). At time equal zero start the pump and begin monitoring the lake pH. Take 100mL grab samples from the lake effluent at 5, 10, 15, and 20 minutes. The sample volumes do not need to be measured. Collect the samples in 125mL bottles. Measure the flow rate. After the 20minute sample turn off the pump and stop sampling pH. Save the pH data to \\Enviro\enviro\Courses\453\acid\netid_remediate by selecting Save data from the control palette. The data will be saved in a file (tab delimited format) that can be opened by any spreadsheet program. Measure the lake volume. Analytical Procedures pH. pH (log{H+}) is usually measured electrometrically with a pH meter. The pH meter is a nullpoint potentiometer that measures the potential difference between an indicator electrode and a reference electrode. The two electrodes commonly used for pH measurement are the glass electrode and a reference electrode. The glass electrode is an indicator electrode that develops a potential across a glass membrane as a function of the activity ,~ molarity, of H+. Combination pH electrodes, in which the H+sensitive and reference electrodes are combined within a single electrode body will be used in this lab. The reference electrode portion of a combination pH electrode is a [Ag/AgCl/4M KCl] reference. The response (output voltage) of the electrode follows a "Nernstian" behavior with respect to H+ ion activity.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 26 where R is the universal gas constant, T is temperature in Kelvin, n is the charge of the hydrogen ion, and F is the Faraday constant. E0 is the calibration potential (Volts), and E is the potential (Volts) measured by the pH meter between glass and reference electrode. The slope of the response curve is dependent on the temperature of the sample and this effect is normally accounted for with simultaneous temperature measurements. The electrical potential that is developed between the glass electrode and the reference electrode needs to be correlated with the actual pH of the sample. The pH meter is calibrated with a series of buffer solutions whose pH values encompass the range of intended use. The pH meter is used to adjust the response of the electrode system to ensure a Nernstian response is achieved over the range of the calibration standards. Refer to the appendix (page  PAGEREF _Ref409401925 \h 157) for specific procedures. To measure pH the electrode(s) are submersed in at least 50 mL of a sample. Samples are generally stirred during pH reading to establish homogeneity, to prevent local accumulation of reference electrode filling solution at the interface near the electrode, and to ensure the diffusive boundary layer thickness at the electrode surface is uniform and small. ANCxe "ANC". The most common method to determine ANC for aqueous samples is titrationxe "titration" with a strong acid to an endpoint pH. A pH meter is usually used to determine the endpoint or "equivalence point" of an ANC titration. Determination of the endpoint pH is difficult because it is dependent on the magnitude the sample ANC. Theoretically this endpoint pH should be the pH where all of the ANC of the system is consumed, but since the ANC is not known apriori, a true endpoint cannot be predetermined. However, if most of the ANC is composed of carbonate and bicarbonate this endpoint is approximately pH = 4.5 for a wide range of ANC values. A 100mL sample is usually titrated under slowly stirred conditions. Stirring is accomplished by a magnetic stirrer. pH electrodes are ordinarily used to record pH as a function of the volume of strong acid titrant added. The volume of strong acid required to reach the ANCxe "ANC" endpoint (pH 4.5) is called the "equivalent volume" and is used in the following equation to compute ANC.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 4. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 27 A more accurate technique to measure ANCxe "ANC" is the Gran plotxe "Gran Plot" XE "Gran plot"  analysis. This is the subject of a subsequent experiment. All ANC samples should be labeled and stored for subsequent Gran analysis. Prelab Questions How many grams of NaHCO3 would be required to keep the ANCxe "ANC" levels in a lake above 50 eq/L for 3 hydraulic residence timexe "hydraulic residence time"s given an influent pH of 3.0 a lake volume of 5 L, if the current lake ANC is 50 eq/L? Data Analysis K1 = 106.3, K2 = 1010.3, KH = 101.5mol/atmL, PCO2=103.5 atm, and Kw = 1014. Soil Column Experiment Plot pH of both lake and soil column effluent versus time. How long did it take for the ANCxe "ANC" of the soil column to be depleted? (At a pH of 4.5 the ANC is approximately zero.) How long did it take for the ANCxe "ANC" of the lake to be depleted? Acid Lake Remediation XE "Remediation"  Experiment Plot measured pH of the lake versus time. Given that ANCxe "ANC" is a conservative parameter and that the lake is essentially in a completely mixed flowxe "completely mixed flow" regime equation  GOTOBUTTON ZEqnNum970368 \* MERGEFORMAT  REF ZEqnNum970368 \! \* MERGEFORMAT 4.21 applies. Graph the predicted ANC based on the completely mixed flow reactorxe "reactor" equation with the plot labeled (in the legend) as conservative ANC. Derive an equation for CT (the concentration of carbonate species) as a function of time based on the input of NaHCO3 and its dilution in the completely mixed lake assuming a nonvolatile system (the equation will be the same form as equation  GOTOBUTTON ZEqnNum970368 \* MERGEFORMAT  REF ZEqnNum970368 \! \* MERGEFORMAT 4.21). Combine your equation for CT with equation  GOTOBUTTON ZEqnNum934642 \* MERGEFORMAT  REF ZEqnNum934642 \! \* MERGEFORMAT 4.11 and plot the predicted ANCxe "ANC" of the lake vs. time for a nonvolatile system based on the measured pH (plot on the same graph as # REF ANCconservativequestion \n 2) with plot labeled as nonvolatile model). Plot the predicted ANCxe "ANC" of the lake vs. time for a volatile system using equation  GOTOBUTTON ZEqnNum534116 \* MERGEFORMAT  REF ZEqnNum534116 \! \* MERGEFORMAT 4.15 based on the measured pH (plot on the same graph as # REF ANCconservativequestion \n 2) with plot labeled as volatile model). Compare the plots and determine whether the lake is best modeled as a volatile or nonvolatile system. What changes could be made to the lake to bring the lake into equilibrium with atmospheric CO2? Questions What do you think would happen if enough NaHCO3 were added to the lake to maintain an ANCxe "ANC" greater than 50 eq/L for 3 residence times with the stirrer turned off? What are some of the complicating factors you might find in attempting to remediate a lake using CaCO3? Below is a list of issues to consider. extent of mixing solubility of CaCO3 (find the solubility and compare with NaHCO3) density of CaCO3 slurry (find the density of CaCO3) References Driscoll, C.T., Jr. and Bisogni, J.J., Jr., "Weak Acid/Base Systems in Dilute Acidified Lakes and Streams of the Adirondack Region of New York State," in Modeling of Total Acid Precipitation Impacts J.L. Schnoor (ed.), Butterworth, Stoneham, MA., 5372 (1983). Driscoll, C.T., Baker, J.P., Bisogni, J.J., And Schofield, C.L., "Aluminum Speciation and Equilibria in Dilute Surface Waters of the Adirondack Region of New York State," in Geological Aspects of Acid Deposition O.P. Bricker (ed.), Butterworth, Stoneham, MA., 5575 (1984). Barnard. T.E., And Bisogni, J.J., Jr., "Errors in Gran Function Analysis of Titration Data for Dilute Acidified Water," Water Research, 19, No. 3 393399 (1985). Bisogni, J.J., Jr. and Barnard, T.E., "Numerical Technique to Correct for Weak Acid Errors in Gran Function Analysis of Titration Data," Water Research, 21, No. 10, 12071216 (1987). Bisogni, J.J., Jr., "Fate of Added Alkalinity During Neutralization of an Acid Lake," Journal Environmental Engineering, ASCE, 114, No. 5, 12191224 (1988). Bisogni, J.J., Jr., and Kishbaugh, S.A., "Alkalinity Destruction by Sediment Organic Matter Dissolution During Neutralization of Acidified Lakes," Water, Air and Soil Pollution, 39, 8595 (1988). Bisogni, J.J., Jr. and Arroyo, S.L., "The Effect of Carbon Dioxide Equilibrium on pH in Dilute Lakes," Water Research, 25, No. 2, 185190 (1991). Olem, H. Liming Acidic Surface Waters. Lewis Publishers, Chelsea, MI. (1991). Stumm, W. and Morgan, J.J., Aquatic Chemistry, John Wiley & Sons, Inc. NY, NY 1981. Lab Prep Notes Table  SEQ \r 1 table1. Reagents DescriptionSupplierCatalog numberHCL 5.0 NFisher ScientificLC153602H2SO4 5NFisher ScientificLC258402CaCO3Fisher ScientificC633Na2CO3Fisher ScientificS263500BufferPacFisher ScientificSB105NaHCO3Fisher ScientificS233500Bromocresol GreenFisher ScientificB3835ethanolFisher ScientificA962P4 Table  SEQ table2. Equipment list DescriptionSupplierCatalog numbermagnetic stirrerFisher Scientific115007Sfloating stir barFisher Scientific1451199AAccumet"! 50 pH meterFisher Scientific13635501001095 L pipettexe "pipette"Fisher Scientific13707510109.5 L pipettexe "pipette"Fisher Scientific137073pH electrodeFisher Scientific136201086 L container (lake)Fisher Scientific0348422Easy load pump headCole ParmerH0751800digital pump driveCole ParmerH0752330_PharMed tubing size 18Cole ParmerH0648517soil columnsee design schematic,  REF _Ref365260299 \* MERGEFORMAT Figure 320 liter HDPE JerricanFisher Scientific0296150CBromocresol Green Indicating Solution Prepare solution of 400 mg Bromocresol green/100 mL ethanol. Add 0.2 mL of indicator solution per liter of acid rain or lake. Acid rain Acid rain is at pH 3.0. Prepare from distilled water. Add 1 meq H2SO4/L ([H+] at pH 3.0) to obtain a pH of 3.0. To acidify 20 liters of distilled water using 5 N H2SO4:  EMBED Equation.DSMT4  Add 4 mL of bromocresol green indicating solution to 20 L of acid rain solution. Flow Rate The residence time of the lake should be 15 minutes. The lake volume is 5 L. thus the flow rate is 334 mL/min. Use # 18 PharMed tubing. Neutralizing Column Neutralization of the acid rain will be accomplished in a porous mediaxe "porous media" column amended with CaCO3. The porous media will be glass beads or sand (approximately 200m diameter) with no ANCxe "ANC". If new glass beads are used, they should be washed with distilled water to remove ANC. To neutralize the acid rain 1 meq/L of CaCO3 will be added to the column. This will be enough ANC to raise the pH to near neutral. To neutralize 5 liters (15 minutes):  EMBED Equation.DSMT4  The porous mediaxe "porous media" column amended with CaCO3 will be prepared as follows. Add 6 cm of glass beads (approximate) to 10 cm diameter column. Fill from bottom of column with distilled water to 1 cm above glass bead surface. Attach cover and fill column to top with distilled water. Prepare CaCO3 slurry (approximately 25 mL distilled water + 250 mg CaCO3). Pour slurry into 50 mL syringe. Connect syringe to manifold at influent of column. Inject slurry into column 15 minutes before use. If prepared too long in advance the CaCO3 washes out of the column. Allow CaCO3 to settle for 10 minutes before beginning to use. Setup Prepare 20L acid rain for each group. Prepare neutralizing columns. Prepare bromocresol green solution if necessary. Attach one EasyLoad pump head to the pump drives and plumb with #18 tubing. Plumb Jerrican to pump to soil column to lake using quick connectors (see  REF _Ref365260269 \* MERGEFORMAT Figure 2). Verify that pH probexe "pH probe"s are operational, stable, and can be calibrated. If you are doing the soil column experiment you need 2 probes per group. Verify that buffers (pH = 4, 7, 10) are distributed to each student group. Provide a mount for the pH probexe "pH probe" in the lake. Set up some lakes with aeration! Measurement of Acid Neutralizing Capacity Introduction  MACROBUTTON MTEditEquationSection Equation Section (Next) SEQ MTEqn \r \h \* MERGEFORMAT  SEQ MTSec \h \* MERGEFORMAT Acid neutralizing capacity (ANCxe "ANC") is a measure of the ability of water to neutralize acid inputs. Lakes with high ANC (such as Cayuga Lake) can maintain a neutral pH even with some acid rain input whereas lakes with an ANC less than the acid input will not maintain a neutral pH. In the Adirondack region of New York State, lakes typically receive large inputs of acids during the spring thaw when the accumulated winter snow melts and runs off into the lakes. The ANC of Adirondack lakes is not always sufficient to neutralize these inputs. Theory The ANCxe "ANC" for a typical carbonatecontaining sample is defined as:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 5. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 1 This equation can be derived from a charge balance if ANCxe "ANC" is considered to be the cation contributed by a strong base titrant and if other ions present do not contribute significantly. Determination of ANCxe "ANC" or Alkalinity involves determination of an equivalence point. The equivalence point is defined as the point in the titrationxe "titration" where titrant volume that has been added equals the "equivalent" volume (Ve). The equivalent volume is defined as:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 5. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 2 where: Ns = normality (in this case Alkalinity or ANCxe "ANC") of sample, equivalents/L Vs = volume of sample, liters Nt = normality of titrant, equivalents/L. The titrationxe "titration" procedure involves incrementally adding known volumes of standardized normality strong acid (or base) to a known volume of unknown normality base (or acid). When enough acid (or base) has been added to equal the amount of base (or acid) in the unknown solution we are at the "equivalence" point. (Note: the point at which we add exactly an equivalent or stoichiometric amount of titrant is the equivalence point. Experimentally, the point at which we estimate to be the equivalence point is called the titration endpoint). There are several methods for determining Ve (or the equivalence point pH) from titrationxe "titration" data (titrant volume versus pH). The shape of the titrationxe "titration" curve (Vt versus pH) can reveal Ve. It can be shown that one inflection point occurs at EMBED Equation.DSMT4 . In the case of monoprotic acids, there is only one inflection in the pH range of interest. Therefore, an effective method to find the equivalence volume is to plot the titration curve and find the inflection point. Alternately, plot the first derivative of the titration plot and look for a maximum. Gran Plotxe "Gran Plot" Another method to find the ANCxe "ANC" of an unknown solution is the Gran plotxe "Gran plot" technique. When an ANC determination is being made, titrationxe "titration" with a strong acid is used to "cancel" the initial ANC so that at the equivalence point the sample ANC is zero. The Gran plot technique is based on the fact that further titration will result in an increase in the number of moles of H+ equal to the number of moles of H+ added. Thus after the equivalence point has been reach the number of moles of H+ added equals the number of moles of H+ in solution.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 5. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 3 Solving for the hydrogen ion concentration:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 5. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 4 Equation  GOTOBUTTON ZEqnNum148748 \* MERGEFORMAT  REF ZEqnNum148748 \! \* MERGEFORMAT 5.4 can be solved directly for the equivalent volume.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 5. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 5 Equation  GOTOBUTTON ZEqnNum723466 \* MERGEFORMAT  REF ZEqnNum723466 \! \* MERGEFORMAT 5.5 is valid if enough titrant has been added to neutralize the ANCxe "ANC". A better measure of the equivalent volume can be obtained by rearranging equation  GOTOBUTTON ZEqnNum100677 \* MERGEFORMAT  REF ZEqnNum100677 \! \* MERGEFORMAT 5.4 so that linear regression on multiple titrant volume  pH data pairs can be used.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 5. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 6 We define F1 (First Gran function) as:  Figure  SEQ Figure \r11. Gran plotxe "Gran plot" from titrationxe "titration" of a weak base with 0.05 N acid. Ct = 0.001 moles of carbonate and sample volume is 48 mL. The equivalent volume is 4.8 mL. From equation  GOTOBUTTON ZEqnNum911611 \* MERGEFORMAT  REF ZEqnNum911611 \! \* MERGEFORMAT 5.9 the ANCxe "ANC" is 5 meq/L.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 5. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 7 If F1 is plotted as a function of Vt the result is a straight line with slope =  EMBED Equation.DSMT4  and abscissa intercept of Ve ( REF _Ref365261982 \* MERGEFORMAT Figure 1). The ANCxe "ANC" is readily obtained given the equivalent volume. At the equivalence pt:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 5. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 8 Equation  GOTOBUTTON ZEqnNum740028 \* MERGEFORMAT  REF ZEqnNum740028 \! \* MERGEFORMAT 5.8 can be rearranged to obtain ANCxe "ANC" as a function of the equivalent volume.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 5. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 9 pH Measurements The pH can be measured either as activity ({H+} as measured approximately by pH meter) or molar concentration ([H+]). The choice only affects the slope of F1 since [H+] = {H+}/g.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 5. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 10 where g is the activity correction factor and the slope is Nt/V0. If H+ concentration is used then  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 5. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 11 where the slope is  EMBED Equation.DSMT4 . (This analysis assumes that the activity correction factor doesn't change appreciably during the titrationxe "titration"). There are many other Gran functions that can be derived. For example, one can be derived for Acidity or the concentration of a single weak or strong acid or base. To facilitate data generation and subsequent Gran plotxe "Gran plot" construction and analysis pH versus titrant volume can be read directly into a computer, that can be programmed to analyze the data using the Gran, plot theory. The program generates the Gran function for all data and then systematically eliminates data until the Gran function (plot) is as linear as possible. The line is then extrapolated to the abscissa to find the equivalent volume. ANCxe "ANC" Determination for Samples with pH < 4 After the equivalence point has been reached (adding more acid than ANCxe "ANC" = 0) the only significant terms in equation  GOTOBUTTON ZEqnNum478566 \* MERGEFORMAT  REF ZEqnNum478566 \! \* MERGEFORMAT 5.1 are  EMBED Equation.DSMT4  and ANC.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 5. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 12 When the pH is 2 pH units or more below the pKs of the bases in the system the only species contributing significantly to ANCxe "ANC" is the hydrogen ion (equation  GOTOBUTTON ZEqnNum603114 \* MERGEFORMAT  REF ZEqnNum603114 \! \* MERGEFORMAT 5.12) and thus the ANC is simply  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 5. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 13 For a sample containing only carbonates, if the pH is below 4 the ANCxe "ANC" is approximately equal to [H+] and no titrationxe "titration" is necessary. Titration Techniques Operationally, the first few titrant volumes can be relatively large increments since the important data lies at pH values less than that of the equivalence point (approximately pH = 4.5 for an Alkalinity titrationxe "titration"). As the pH is lowered by addition of acid the ionic strength of the solution increases and the activity of the hydrogen ion deviates from the hydrogen ion concentration This effect is significant below pH 3 and thus the effective linear range is generally between pH 4.5 and pH 3.0. The maximum incremental titrant volume ("Va) that will yield n points in this linear region is obtained as follows. If Vs Vt then equation  GOTOBUTTON ZEqnNum460197 \* MERGEFORMAT  REF ZEqnNum460197 \! \* MERGEFORMAT 5.3 reduces to  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 5. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 14 Let [H+]e be the concentration of hydrogen ions at the equivalence point and [H+]f be the final concentration of hydrogen ions at the end of the titrationxe "titration".  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 5. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 15 Thus the volume of acid added to go from [H+]e to [H+]f is  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 5. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 16 To obtain n data points between [H+]e  [H+]f requires the incremental titrant volume ("Vt) be 1/n times the volume of acid added between the equivalence point and the final titrant volume. Thus by substituting n"Vt, and typical hydrogen ion concentrations of [H+]e = 104.5 and [H+]f = 103.0 into equation  GOTOBUTTON ZEqnNum801057 \* MERGEFORMAT  REF ZEqnNum801057 \! \* MERGEFORMAT 5.16 the maximum incremental titrant volume is obtained.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 5. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 17 Procedure Calibrate the pH Meter Calibrate the pH meter using a pH probexe "pH probe" connected to channel A. Use 3 standards (pH = 4, 7, and 10). Determine ANCxe "ANC" of a Known Standard Weigh a 100 mL plastic beaker. Add approximately 50 mL of a 2.5 mM solution of Na2CO3 to the beaker. Weigh the beaker again to determine the exact volume of Na2CO3 solution. Place the beaker on the magnetic stirrer, add a stir bar and stir slowly. Place both the pH electrode and the temperature probe in the Na2CO3 solution using the probe holding arm attached to the Accumet"! meter. Analyze the sample using Gran plotxe "Gran plot" analysis as detailed at  HYPERLINK "http://www.cee.cornell.edu/mws/Software/Compumet.htm" http://www.cee.cornell.edu/mws/Software/Compumet.htm) Add 0.05 N HCl (the titrant) using a digital pipette in increments of 0.25 mLxe "pipette". Save the pH data to \\Enviro\enviro\Courses\453\acid\netid_gran by selecting Save data from the control palette. The data will be saved in a file (tab delimited format) that can be opened by any spreadsheet program. You will use this data to plot a titrationxe "titration" curve and to verify that the Gran technique accurately measures the ANCxe "ANC" of a sample. Record the ANCxe "ANC" and the equivalent volume. Determine ANCxe "ANC" of Acid Rain Samples Determine ANCxe "ANC" for all samples collected from the previous week's lab. Use the same technique as outlined above (Determine ANC of known standard) except substitute the samples collected last week and use titrant increment of 0.1 mL in the linear region. For samples that have a high ANC you can reduce the analysis time by adding titrant in larger volumes initially until the pH approaches 5. If the initial pH is less than 4.5 no titrationxe "titration" is necessary and equation  GOTOBUTTON ZEqnNum854850 \* MERGEFORMAT  REF ZEqnNum854850 \! \* MERGEFORMAT 5.13 can be used to calculate the ANCxe "alkalinity". Record the initial pH (prior to adding any titrant) and initial sample volume. After the Gran plotxe "Gran plot" analysis record the alkalinity (ANCxe "ANC")xe "alkalinity" and equivalent volume for each sample. There is no need to save the data to disk. Prelab Questions Compare the ability of Cayuga lake and Wolf pond (an Adirondack lake) to withstand an acid rain runoff event (from snow melt) that results in 20% of the original lake water being replaced by acid rain. The acid rain has a pH of 3.5 and is in equilibrium with the atmosphere. The ANCxe "ANC" of Cayuga lake is 1.6 meq/L and the ANC of Wolf Pond is 70 eq/L. Assume that carbonate species are the primary component of ANC in both lakes, and that they are in equilibrium with the atmosphere. What is the pH of both bodies of water after the acid rain input? Remember that ANC is the conservative parameter (not pH!). What is the ANCxe "ANC" of a water sample containing only carbonates and a strong acid that is at pH 3.2? Why is [H+] not a conserved species? Questions Plot the titrationxe "titration" curve of 2.5 mM Na2CO3 with 0.05 N HCl (plot pH as a function of titrant volume). Label the equivalent volume of titrant. Label the 2 regions of the graph where pH changes slowly with the dominant reaction that is occurring. Note that in a third region of slow pH change no significant reactions are occurring (added hydrogen ions contribute directly to change in pH). Prepare a Gran plotxe "Gran plot" using the data from the titrationxe "titration" curve of the 2.5 mM Na2CO3. Use linear regression on the linear region or simply draw a straight line through the linear region of the curve to identify the equivalent volume. Compare your calculation of Ve with that calculated by the Compumet XE "Compumet" "! computer program. Compare the measured ANCxe "ANC" with the theoretical value for the 2.5 mM Na2CO3 solution. Note that ANC can be defined as the excess of positive charges over the anions of strong acids. Plot the ANCxe "ANC" of the influent and the lake from phase I of the previous lab. Plot the ANCxe "ANC" of the lake from phase II of the previous lab on the same graph as was used to plot the conservative ANC model (see questions  REF ANCconservativequestion \n 2) to  REF _Ref405018997 \r 5) on page  PAGEREF ANCconservativequestion 53). Did the measured ANC values agree with the conservative ANC model? References Sawyer, C.N., P.L. McCarty and G.F. Parkin Chemistry for Environmental Engineering, 4th ed., McGrawHill (1994). Pankow, J.F. Aquatic Chemistry Concepts, Lewis Publishers (1991). Morel, F.M.M. and J.G. Hering Principles and Applications of Aquatic Chemistry WileyInterscience (1993). Stumm, W. and J.J. Morgan Aquatic Chemistry 2nd ed. Wiley Interscience (1981). Lab Prep Notes Table  SEQ \r 1 table1. Reagent list. DescriptionSupplierCatalog numberHCl 5.0 NFisher ScientificLC153602BufferPacFisher ScientificSB105Na2CO3Fisher Scientific BP3571Table  SEQ table2. Equipment list DescriptionSupplierCatalog numberAccumet"! 50 pH meterFisher Scientific1363550 pH electrodeFisher Scientific136201087x7 stirrerFisher Scientific115007Sstirbar 1/2" longFisher Scientific1451162100 mL Fisher beakerFisher Scientific0259350BSetup Prepare 1 L of the known standard (2.5 mM solution of Na2CO3). The MW is 105.99 g/mole.  EMBED Equation.DSMT4   EMBED Equation.DSMT4 = 265 mg Na2CO3/L Prepare 1 L of the titrant (0.05 N HCl from 5.0 N HCl). Dilute 10 mL of 5.0 N HCl to 1 L. Distribute 100 mL titrant to each student group. Verify that the pH probexe "pH probe"s are operational, stable, and can be calibrated. Verify that buffers (pH = 4, 7, 10) are distributed to each student group Phosphorus Determination using the Colorimetric Ascorbic Acid Technique Introduction  MACROBUTTON MTEditEquationSection Equation Section (Next) SEQ MTEqn \r \h \* MERGEFORMAT  SEQ MTSec \h \* MERGEFORMAT Phosphorus has been identified as a prime nutrient needed for algae growth in inland environments. In 1992, the EPA reported that accelerated eutrophication was one of the leading problems facing the Nation's lakes and reservoirs. Eutrophication caused by the overabundance of nutrients in water can result in a variety of water-quality problems, including fish kills, noxious tastes and odors, clogged pipelines, and restricted recreation. In freshwater, phosphorus is often the nutrient responsible for accelerated eutrophication. Many algae blooms in rivers and lakes are attributed to elevated phosphorus concentrations resulting from human activities. Phosphorus enters surface waters from agricultural and urban runoff as well as from industrial and municipal wastewater treatment plant effluent. No national criteria have been established for concentrations of phosphorus compounds in water; however, to control eutrophication, the EPA makes the following recommendations: Total phosphates should not exceed 50 mg/L (as phosphorus) in a stream at a point where it enters a lake or reservoir. Total phosphorus should not exceed 100 mg/L in streams that do not discharge directly into lakes or reservoirs. Municipal wastewater treatment plants in many areas are required to remove phosphorous in their treatment process. While the biological treatment process removes some phosphorus, in most cases precipitation as an insoluble metal phosphate is required to meet discharge regulations. This precipitation step is normally accomplished with a metallic salt such as ferric sulfate, ferric chloride or aluminum sulfate. This precipitation step may be accomplished in the primary or secondary clarifiers. Phosphorus Quantification Techniques Quantification of phosphorous requires the conversion of the phosphorus to dissolved orthophosphate followed by colorimetric determination of dissolved orthophosphate. The analysis of different phosphorous forms (e.g. particulate or organic-P) is obtained by various pretreatment steps. Pretreatment may consist of filtering to remove suspended matter or various digestion techniques designed to oxidize organic matter. Phosphorus can be present in surface waters as organic phosphorus, orthophosphate (an inorganic form of PO4), or as condensed (solid) phosphates. The phosphorus may be in solution or as a component of suspended particulates. The wet chemical colorimetric analysis of phosphorus only works for orthophosphates and thus other forms of phosphorus must be converted to this form if they are to be analyzed. Organic phosphorus can be oxidized (digested) using perchloric acid, nitric acid-sulfuric acid, or persulfate with the persulfate technique being the safest and least time consuming. The digestion methods are detailed in APHA method 4500-P B. Three techniques for colorimetric analysis of phosphorus are available. The technique most commonly used is the ascorbic acid method, which can determine concentrations of orthophosphate in most waters and wastewater in the range from 2-200 mg P/L. Ammonium molybdate and antimony potassium tartrate react in an acid medium with dilute solutions of orthophosphate-phosphorus to form an intensely colored antimony-phospho-molybdate complex. This complex is reduced to an intensely blue-colored complex by ascorbic acid. The color is proportional to the phosphorus concentration. The complex is not stable and thus analysis must be performed within 30 minutes of adding the ammonium molybdate and antimony potassium tartrate. Barium, lead, and silver interfere by forming a precipitate. The interference from silica, which forms a pale-blue complex is small and can usually be considered negligible. Arsenate is determined similarly to phosphorus and should be considered when present in concentrations higher than phosphorus. Method Detection Limit "Method detection limit" is the smallest concentration that can be detected above the noise in a procedure and within a stated confidence level. Several types of detection limits are used including instrument detection limit (IDL), method detection limit (MDL), and practical quantitation limit (PQL). The IDL is strictly instrument noise and does not include variability due to sample preparation steps. The MDL includes both instrument noise and sample preparation variability. The MDL is obtained by making a standard that is near the MDL and dividing it into at least 7 portions. Each of the portions is then processed through all sample preparation steps and then analyzed. The MDL is calculated using the following equation.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 6. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 1 where n is the sample size and a=0.01 is generally the required confidence. The student t distribution function is available in Excel as a two sided test statistic (so use TINV(2a,n-1)) and the standard deviation, s, can be computed in Excel as STDEV(). The PQL is about five times the MDL and represents a practical and routinely achievable detection limit with reasonable assurance that any reported value greater than the PQL is reliable. According to Standard Methods the method detection limit for phosphorus when using a 1 cm light path is approximately 150 mg P/L. Spectrophotometer Limitations The diode array XE "diode array"  spectrophotometer XE "spectrophotometer"  has 316 diodes that cover the wavelength range of 190 nm to 820 nm. Each diode generates a voltage output that is proportional to the number of incident photons. The voltage is then digitized, but the manufacturer of the instrument in the Cornell Environmental Laboratory, Hewlett-Packard, doesn't report the resolution of the analog to digital converter. At very low concentrations the difference between the intensity of light transmitted through the reference and the intensity of light transmitted through the sample approaches zero. At some low concentration the difference in light intensity approaches the resolution of the analog to digital converter. Another source of instrument error is drift in lamp intensity over time. The lamp intensity is measured when a reference sample is made. The light intensity recorded by the diodes will vary proportionally to any lamp intensity drift. The IDL should decrease as the number of diodes used in the analysis increases (as in Spectral analysis) for the same reason that replicate analysis of samples decreases the standard deviation. The "Spectral analysis" feature, which can be used to measure either single or multiple components, uses as much of the spectrum XE "spectrum"  as the user desires and thus potentially decreases the IDL. Spectral analysis uses general least squares regression to add multiples of extinction coefficient arrays for each component to obtain the best curve fit for the sample. The extinction coefficient arrays are obtained from the slope of the linear regression line for absorbance as a function of concentration at each wavelength. Experimental Objectives Measure the concentration of phosphorus in several samples to test the precision of the ascorbic acid technique. Compare the results obtained using conventional analysis at a single wavelength with spectral analysis. Analyze the data using spectrophotometer XE "spectrophotometer"  software outside the lab. Analyze multiple samples so that confidence intervals can be calculated. Estimate the method detection limit (MDL). Discuss methods to improve the method detection limit. Discuss method automation. Experimental Procedures Standards Preparation Method Use 100 mg P/L stock. Use a digital pipet and prepare 1 mL of each standard. Use E-pure water to dilute the 100 mg P/L stock. Reagent Addition for Samples and Standards Pipette 1 mL sample into a disposable microcuvet using a 1 mL digital pipette XE "pipette" . Add 160 mL combined reagent and mix thoroughly by swirling. After at least 10 minutes but no more than 30 minutes, measure absorbance of each sample using a reagent blank as the reference solution. Samples and Standards to Prepare Reagent blank to be used as reference samples. Prepare 6 standards containing 0 (reagent blank), 1, 3, 10, 30, 100 mg P/L. Prepare 6 additional 10 mg P/L standards. Prepare 5 samples such as Cayuga Lake water, tap water, Chem lawn runoff, local creeks Spectrophotometer Method Use Sample Cuvettes. (Make sure to orient all cuvettes with the arrow on the left because the cuvettes are not symmetrical and have different absorbance when turned 180(.) Use the reagent blank as the reference sample for all samples. Use units of mg P/L. Label all samples with descriptions that your classmates will understand! Fill in the general description with your initials and a description of the type of samples Samples Analysis Measure the reference using a reagent blank. (The reagent blank is also the 0 mg/L standard.) Analyze the reagent blank as a sample and verify that the absorbance deviates less than 0.004 AU (absorbance units) from zero. If the absorbance deviates more than 0.004 AU reanalyze the reference sample. Analyze 6 standards as standards using the spectrophotometer XE "spectrophotometer"  and save the data as \\Enviro\enviro\Courses\453\phosphorus\netid_Pstd. Analyze 6 standards as samples using the spectrophotometer XE "spectrophotometer"  and save the data as \\Enviro\enviro\Courses\453\phosphorus\netid_Pstdsam. Analyze 7 10-mg P/L standards as samples and save the data as \\Enviro\enviro\Courses\453\phosphorus\netid_10Pstdsam. Analyze 5 samples as samples using the spectrophotometer XE "spectrophotometer"  and save the data as \\Enviro\enviro\Courses\453\phosphorus\netid_Psam. Prelab Questions You will be creating 1 mL standards by diluting a stock of 100 mg P/L. Create a table showing how you will prepare 1 mL of each of the standards. All of the samples are diluted with a small amount of combined reagent. How is this dilution accounted for when calculating the concentration of samples? Data Analysis Plot the absorbance spectra of the standards. What is happening in the UV region? Are there any absorbance peaks? Choose an appropriate wavelength (perhaps an absorbance peak) and use Excel to create a calibration curve. For the calibration curve, absorbance should be a function of phosphorus concentration. Use the 10-mg/L standards that were analyzed as samples to evaluate the Method Detection Limit using single wavelength analysis. Use your Spreadsheet to calculate the concentration of each of the replicates. Use the 10-mg/L standards that were analyzed as samples to evaluate the Method Detection Limit using spectral analysis. Report the wavelength range used for your analysis. You will need to analyze each sample and copy the results into a spreadsheet. (Note that within a data file you change which sample the software is analyzing with the sample number control.) Use the method with the lowest MDL to analyze your samples. Create a bar graph showing the concentration of each of the samples. Report the MDL in the figure caption. Spreadsheet requirements Your spreadsheet must contain all of the analysis requested above as well as the following capabilities: A well-marked cell containing the analytical wavelength for single wavelength analysis. Changes to this cell must be reflected in all calculations and graphs. The graph showing the absorbance spectra of the standards must have a vertical line indicating the analytical wavelength. All of the graphs must be on the same page as the analytical wavelength control so the effect of changing the wavelength can be easily observed. A separate sheet where you answer the 4 questions. Hints If you haven't already learned how to use Vlookup() now is the time! The row() function returns the number of the row. I found it useful for this analysis! The slope() and intercept() functions eliminate the need to type equations off of graphs! Questions Which analysis technique gave the best results? Explain why. If the analytical technique didn't significantly affect the MDL, explain why not. What types of errors dominated your ability to measure phosphorus? What method modifications do you propose to improve phosphorus measurements? Total phosphorus concentration in Cayuga Lake varies between 10 and 50 ppb. Would the techniques used in this lab be able to measure these phosphorus levels? References  HYPERLINK http://wwwrvares.er.usgs.gov/nawqa/circ-1136/h6.html#PHOS http://wwwrvares.er.usgs.gov/nawqa/circ-1136/h6.html#PHOS  HYPERLINK "http://www.epa.gov/glnpo/lmmb/methods/index.html#Volume" http://www.epa.gov/glnpo/lmmb/methods/index.html#Volume 3 Standard Methods for the Examination of Water and Wastewater. Lab Prep Notes Table  SEQ \r 1 table1. Reagents DescriptionSupplierCatalog numberconcentrated H2SO4Fisher Scientific(NH4 )6 Mo7O244H2OFisher ScientificC6H8O6Fisher ScientificK(SbO)C4H4O6H2OFisher Scientificsodium lauryl sulfateKH2PO4Fisher ScientificReagents A Sulfuric acid solution, 4.9 N: Add 136 mL concentrated H2SO4 to 800 mL E-pure water. Cool and dilute to 1 L with E-pure water. B Ammonium molybdate solution: Dissolve 40 g of (NH4 )6 Mo7O244H2O in 900 mL E-pure water and dilute to 1 L. Store at 4C. Table  SEQ table2. Equipment list DescriptionSupplierCatalog #1001095 L pipettexe "pipette"Fisher Scientific13707510109.5 L pipettexe "pipette"Fisher Scientific137073Disposable cuvetsFisher Scientific14-385-942Cuvet holderFisher Scientific14-385-939UVVis spectrophotometerxe "spectrophotometer"HewlettPackard Company8452AC Ascorbic acid: Dissolve 9 g of ascorbic acid (C6H8O6) in 400 mL E-pure water and dilute to 500 mL. Store at 4C. Keep well stoppered. Prepare fresh monthly or as needed. D Antimony potassium tartrate: Dissolve 3.0 g of K(SbO)C4H4O6H2O in 800 mL E-pure water and dilute to 1 L. Store at 4C. Combined color reagent: Combine the following solutions in order, mixing (but do not entrain air as oxygen oxides the ascorbic acid) after each addition: (Prepare fresh weekly. Store at 4C) Stock A, (4.9 N H2SO4) 50 mL Stock B, (Ammonium molybdate solution) 15 mL Stock C, (Ascorbic acid solution) 30 mL Stock D, (Antimony-tartrate solution) 5 mL Water diluent solution: Add 4.0 g sodium lauryl sulfate and 5 g NaCl per L of E-pure water. Stock phosphorus standard: Dissolve 0.4394 g of Potassium phosphate monobasic (KH2PO4) (dried at 105C for one hour) in 900 mL E-pure water. Add 2 mL of concentrated H2SO4 and dilute to 1 L. 1.0 mL = 0.100 mg P (100 mg P/L). Standard phosphorus solution: Dilute 1 mL of stock solution to 1 L. (1.0 mL = 0.1 mg P) (100 mg P/L). Soil Washing to Remove Mixed Wastes Objective  MACROBUTTON MTEditEquationSection Equation Section (Next) SEQ MTEqn \r \h \* MERGEFORMAT  SEQ MTSec \h \* MERGEFORMAT The goal of this laboratory exercise is to acquaint students with some of the chemical reactions that result in the binding of inorganic and organic pollutants in subsurface materials. Extractants used by engineers to release contaminants at hazardous waste sitexe "hazardous waste site"s (where mixtures of both types of contaminants are present) may or may not prove effective, depending upon their mechanism of action. In this laboratory exercise, students will test the efficacy of a variety of proposed extractants in the removal of a mixture of an inorganic metal cation, and an organic compound from a contaminated porous medium. Introduction Many Superfundxe "Superfund" site soils are contaminated with a mixture of contaminants including toxic metals and organic compounds. A pressing environmental problem is to devise cleanup strategies that can effectively remove mixed wastes. Many kinds of contaminants bind to soils and aquifer media (collectively referred to here as porous mediaxe "porous media"). Binding reactions limit the effectiveness of pump and treatxe "pump and treat" remediation XE "remediation"  in which a contaminated porous medium is flushed with water to remove contaminants. In such cases, it can prove useful to engineer the properties of the aqueous phase to improve the mobility of the pollutants of interest. In the case of toxic metals, release of mediumbound or adsorbed metals can be enhanced by introduction to the pore solution of a dissolved compound that will bind to the metal in the aqueous phase and form a dissolved complex. Such compounds are referred to as ligands, and ligands that bind metals very strongly are called chelating agentxe "chelating agent"s. Metal solubility and adsorptionxe "adsorption" can also be strongly influenced by the oxidation state of the metal, and use of oxidants or reductants to alter the redox conditions in a porous medium can modify metal mobility both directly and indirectly. Direct effects would be observed if oxidized and reduced metal species have different adsorption characteristics (ex. Cr2O72 vs. Cr+3). Indirect effects would be observed if a metal were bound to a solid phase that would be dissolved under different redox conditions (ex. Fe(OH)3 may dissolve under reducing conditions). Addition of acids or bases could also alter metal mobility. Adsorption of metals is very sensitive to pH shifts, with a decrease in pH favoring the release of cationic metal species (ex. Cd+2, Pb+2) and an increase in pH favoring release of anionic species (ex. Cr2O72, SeO32). Organic cations and anions will have a pH dependent adsorptionxe "adsorption" behavior similar to that described above for metal ions. However, many organic pollutants of interest are nonionic and their binding to the matrix of the porous medium is not greatly influenced by pH. Hydrophobic interactions of nonionic organic compounds with organic matter in porous mediaxe "porous media" appear to be a major driving force for their binding. The addition of surfactants to the pore solution can help to release sorbed nonionic organic pollutants. Under suitable conditions many organic pollutants can be degraded by addition of oxidants or by indigenous or added bacteria [i.e.; given that the bacteria have the necessary genetic capabilities, nutrients (N, P, etc.) and a suitable electron acceptor]. Metals, however, are elements and cannot be degraded. Porous mediaxe "porous media" is not inert. The mineral and organic constituents of the porous matrix can react with added ligands, acids, bases, oxidants, reductants, and surfactants. A consequence, in some cases, is that a desired addition may be rendered impractical. Given the variability and possible dissimilarity of conditions that influence the mobility of metal vs. organic pollutants, it is a challenging task to identify a remediation XE "remediation"  strategy that will successfully treat a given medium that is contaminated with mixed wastes. In this laboratory exercise, students will evaluate the utility of several alternative extractants for remediation of a soil that is contaminated with both a metal cation and an organic compound. Theory Binding Reactions The binding reactions of pollutants to the porous matrix may be classified, at least in part, by where and how the binding reaction takes place. The term adsorptionxe "adsorption" is used for reactions that take place at the interface between the solid and the solution. All other factors being equal, solids with a greater specific surface area (ex. units: m2/gram) will adsorb greater amounts of a dissolved solute. In adsorption reactions, the surface is referred to as an adsorbent and the solute as an adsorbate. Some adsorption reactions are driven by electrostatic attraction between the surface and the solute. Ion exchange is the term used for this type of reaction. All other factors being equal, surfaces with a greater number of charged sites per unit surface area will be able to bind greater quantities of dissolved ions. The concentration of surface exchange sites is commonly quantified as an ion exchangexe "ion exchange" capacity. Surfaces with a high density of negatively charged sites (cation exchangers) will selectively bind positively charged ions while those with a high density of positively charged sites will be selective for anions. Absorption is a process in which a solute penetrates within the solid matrix. Partitioning is a term that is synonymous with absorptionxe "absorption". As an example, we would carry out a partitioning process if we were to add a pollutant to a separatory funnel containing water and an organic liquid such as octanol and then observe the resulting distribution of the contaminant between the aqueous and octanol liquid phases. The distributed contaminant would exist as a dissolved solute in each phase. As is noted below, the phase distribution behavior of nonionic organic pollutants in soils and aquifer media displays many characteristics of absorption reactions. The absorption of nonionic organics appears to be primarily into the organic matter content of the porous medium. This reaction is driven by the water loving nature of the solute, or lack thereof (i.e., pollutant hydrophobicity). All other factors being equal, porous mediaxe "porous media" with higher organic carbon contents would have greater uptake of nonionic organic pollutants. The term sorptionxe "sorption" is somewhat loosely used when the exact mechanistic nature of the pollutants distribution between the solution and the porous medium is not understood, or when both adsorptionxe "adsorption" and absorptionxe "absorption" reactions may contribute to the contaminants phase distribution. Contaminant sorptionxe "sorption" reactions result from an reaction between a material that is dissolved in an aqueous solution with a solid phase. The physical/chemical properties of the contaminant, the solution and the sorbent all influence the resulting contaminant phase distribution. These influences are discussed below. Sorbent Surface Charge As noted above, if the sorbate is an ion, then electrostatic attraction to the surface can play an important role in contaminant adsorptionxe "adsorption". Virtually all soil surfaces are charged. Oxide Minerals Surface charge can result from the ionization of surface functional groups in response to the hydrogen ion concentration of the aqueous phase. Oxide minerals are often modeled as diprotic acids (Westall and Hohl, 1980). Accordingly the surface may donate two hydrogen ions as indicated by the following reactions:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 7. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 1  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 7. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 2 where  EMBED Equation.DSMT4  represents the oxide surface that may exchange two hydrogen ions, and K1 and K2 are equilibrium constants for the first and second acid dissociation reactions. Note, each dissociation constant can be thought of as a expression of the relationship between the concentration of protonated and deprotonated surface sites and the solution hydrogen ion concentration. Accordingly:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 7. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 3  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 7. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 4 K1 therefore represents the solution hydrogen ion concentration at which the concentration of positively charged, diprotic, surface sites  EMBED Equation.DSMT4  is equal to that of surface sites containing a single proton EMBED Equation.DSMT4 . Similarly, when  EMBED Equation.DSMT4  equals K2 then  EMBED Equation.DSMT4  = EMBED Equation.DSMT4 . Although other models of the acid base behavior of oxide surface are conceivable, the above model is helpful in that it predicts that the surfaces can have both positively and negatively charged sites. With this model, H+ release from the surface will occur in response to a decrease in the solution H+ concentration (i.e., an increase in pH, where pH is defined as log EMBED Equation.DSMT4 ). Accordingly, we would expect increasingly higher solution pH conditions to favor formation of negatively charged surfaces, and this is observed. Different surfaces would have different acidity constants (K1 and K2) and would be expected to have different surface charges at the same solution pH. Each surface, at one unique pH, would have an equal concentration of  EMBED Equation.DSMT4  and  EMBED Equation.DSMT4  sites and would have no net charge. This is also observed and is referred to as the pH point of zero charge (PZC). SiO2, a common oxide in porous mediaxe "porous media" (the main component of sand), has a low PZC (H" pH 2 to 3) while iron and aluminum oxides (that commonly occur as surface coatings) have considerably higher PZCs (H" pH 7 to 8) (Parks and DeBruyn, 1962). Soil Organic Matter Another pHdependent origin of surface charge is the ionization of the acidic functional groups in soil organic matter. The carboxyl groups of humictype organic matter typically have acidity constants d" 105 (pK d" 5) and are therefore highly ionized at circumneutral pH. Isomorphic Substitution A final source of charge in soil is isomorphic substitution in the crystalline lattice of some clay minerals. Substitution of Al+3 for Si+4 and Mg+2 for Al+3 will result in a net negative charge for the clay mineral phase. The combined effects of isomorphic substitution, ionization of organic functional groups and the low PZC of silicon oxide minerals make it likely that many porous mediaxe "porous media" will have a net negative charge. Consequently, stronger binding of cationic contaminants is generally anticipated. Sorbent Ion Exchange Reactions Ion exchange reactions involve the exchange of ions of the same charge at an oppositely charge site on the solid surface. Exchange reactions are often characterized by selectivity coefficients that may be thought of as equilibrium constants for the exchange reaction. For example, in the exchange of two monovalent cations, the exchange reaction may be depicted as:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 7. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 5 where:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 7. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 6 The magnitude of the selectivity coefficient,  EMBED Equation.DSMT4 , reflects the extent to which ion x+ vs. y+ will accumulated at the surface. Ions with high selectivity coefficients can displace more weakly held ions from an exchange site. In a negatively charged soil, anionic compounds (ex. ionized organic acids, NO3, Cr2O72, etc.) will be repelled from the surface and therefore may be highly mobile. Cationic species (ex. quaternary ammonium organic compounds, divalent transition metals, etc.) will be attracted to the surface and have restricted mobility. In principle, exchangeable pollutant cations may be mobilized by introduction of high concentrations of an innocuous cation. The practicality of such an approach would be dictated by the extent to which other exchangeable cations (that are not of environmental concern) are also exchanged. Since cations such as Na+, K+, Ca+2 Mg+2 are abundant in porous mediaxe "porous media", the amount of a cation added for exchange of a trace pollutant would have to be in great excess of the pollutant cation. As a result, release of contaminant cations by an ion exchangexe "ion exchange" mechanism does not appear to be economically feasible. Sorbent Hydrophobic Interactions The mechanisms responsible for the adsorptionxe "adsorption" of charged species differ considerably from those for nonionic compounds. Adsorption of charged ions may, in some cases, involve more than the simple electrostatic attraction of ions to a surface of opposite charge. Transition metal cations, for example, will often adsorb to oxide surfaces even under solution conditions that confer a positive charge on the surface (see additional discussion below under the topic of solution characteristics). The sorptionxe "sorption" of nonionic organic pollutants behaves as if it is a partitioning process into the organic matter that is present as part of the soil matrix. Some of the general characteristics that lead to this conclusion are the observance of linear sorption isothermsxe "isotherms" at high solution concentrations (that can approach the solubility limit of solute compounds). [Note, an isotherm is simply the relationship between the quantity of pollutant that is bound (per unit mass or unit surface area of the sorbent) and the concentration of contaminant in solution.] In contrast, adsorption reactions are limited by the availability of surface sites and adsorptionxe "adsorption" isotherms are typically nonlinear at high solute concentrations. Partition reactions are also relatively free from competition (i.e., the presence of a second solute does not effect the sorptive uptake of the first) while competition for surface sites is an expected characteristic in an adsorption process. The extent of sorption of a given nonionic organic onto a variety of sorbents is highly correlated with their organic content as expressed by the weight fraction of organic carbon, foc (Karickhoff, 1984). For the same sorbent, the sorption of different nonionic solutes is highly correlated with their octanolwater partition coefficients (Kow) (Karickhoff, 1984). Collectively, these observations lead to the conclusion that the sorption of nonionic organic pollutants is primarily driven by hydrophobic interactions between the solute and the organic matter in the sorbent. Solution pH Solution conditions can have dramatic effects on the adsorptionxe "adsorption" of cationic contaminants. For example the adsorption of cationic transition metals to oxide surfaces typically increases markedly over a narrow range of 1 to 2 pH units referred to as the adsorption edge. The pH dependence of metal ion adsorption can be explicitly accounted for by writing the adsorption reaction as:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 7. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 7 where  EMBED Equation.DSMT4  is the pHdependent metal distribution coefficient, and according to Honeyman and Santschi (1988)  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 7. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 8 A plot of  EMBED Equation.DSMT4  versus pH, is referred to as a Kurbatov plot (after Kurbatov et. al., 1951), and may be used to reveal the magnitude of the exponent, x for  EMBED Equation.DSMT4  in the distribution coefficient. The ratio EMBED Equation.DSMT4  is the quantity of adsorbed metal per unit surface. Since the above reaction and its equilibrium constant, Kd, are an over simplification of the actual adsorptionxe "adsorption" mechanism, measured values of x are rarely integers. Nevertheless, x values ranging from 1 to 2 are common for adsorption of metal cations on oxide surfaces and demonstrate the strong dependence of the adsorption processes on pH. For example, if x = 2, an increase of 1 pH unit would result in a 100 fold increase in the amount of bound metal per unit surface (at the same solution concentration of metal ion). In general, adsorbed metal cations will be released as a consequence of a decrease in solution pH. Since the surfaces in the porous medium also have acid/base properties, and because many porous mediaxe "porous media" contain acidreactive components (such as carbonate minerals) a very large acid dose may be required to effectively alter the pH of the pore water. For this reason, acid extraction of adsorbed metals may not always be feasible. MetalLigandxe "ligand" Complexes Another influence of solution conditions on metal adsorptionxe "adsorption" is through the reactions of metals with ligands to form complexes. In some cases, metalligandxe "ligand" complexes adsorb weakly or not at all (ex. Cl complexes of Cd and Hg), in other cases metalligand complexes may adsorb with a binding strength greater than that of the free metal (ex. organic complexes of Cu) (Benjamin and Leckie, 1982). Judicious selection of a ligand for introduction into a porous medium may, therefore, be used to accomplish the release of adsorbed cations. Added ligands may, in some cases, undergo exchange reactions with the porous mediaxe "porous media" or react to form complexes with cations that are not of environmental concern. For this reason the dose of a ligand needed to effectively release adsorbed metals will vary with the composition of the porous media and ligand addition may not prove feasible in some cases. Oxidants and Reductants Changing solution composition by the introduction of oxidizing or reducing agents may accomplish the release of adsorbed metals. Iron oxides are strong metal binding agents and may be solubilized by reduction from ferric (Fe III) to ferrous (Fe II) iron. Many transition metals (e.g. Cd, Co, Cu, Ni, Pb, Zn) will remain as divalent cations during such a shift in redox status, and may therefore simply reabsorb to another surface. In some cases, alteration of the media redox conditions may directly influence metal mobility. For example reducing conditions would favor the presence of a cationic form of chrome (Cr+3) over the more mobile anionic form (Cr2O72). Addition of oxidants may therefore help to mobilize chrome, however the organic matter in soils and ferrous minerals will also react with added oxidants. In a manner similar to the role of iron oxides, the organic matter in porous mediaxe "porous media" can be responsible for the binding of metal cations. The reaction of an added oxidantxe "oxidant" with humictype organic matter may therefore accomplish solubilization of some metals (Lion et al., 1982). Strong oxidants will also act to break down organic contaminants. Hydroxyl Radicals One application that has been used for remediation XE "remediation"  of organic contaminated soils is the introduction of Fentons reagent. Fentons reagent is a mixture of hydrogen peroxide and ferrous iron (Fe+2). These chemicals react to produce hydroxyl radicals (OH) according to the following reaction:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 7. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 9 The hydroxyl radicals produced by Fentons reagent are highly reactive and can effectively degrade recalcitrant aromatic compounds by ring substitution followed by ring cleavage (Sedlak and Andren, 1991). Surfactants Additions of surfactants may aid in the release of sorbed nonionic organic pollutants. In the case of sorptionxe "sorption" reactions that are driven by hydrophobic interactions, surfactant additions can have two beneficial effects: 1) a decrease in the aqueous activity coefficient for the dissolved nonionic organic compound and 2) formation of micelles in the aqueous phase. The effect of the aqueous activity coefficient can be illustrated by examination of the sorptionxe "sorption" isotherm for the organic pollutant. If the isotherm is linear, then we may write:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 7. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 10 where G is the mass of solute sorbed per mass of solid,  EMBED Equation.DSMT4 is the sorptive distribution coefficient, CL is the aqueous concentration of the sorbate, and g is the activity coefficient of the dissolved sorbate. Surfactants may act to decrease the activity coefficient, g, for a nonionic molecule increasing the concentration in the aqueous phase in equilibrium with a given adsorbed amount, G. Surfactants increase the solubility of nonionic molecules because the hydrophobicnonionic molecules adsorb to the long hydrocarbon group while the ionic sulfonic group provides high solubility ( REF _Ref405629902 Figure 1).  Figure  SEQ \r 1 Figure1. Molecular structure of detergent. The hydrocarbon chain (R group) of the detergent used in this lab is C12H25. However, since surfactants are surfaceactive, they may also sorb to the porous medium, increase its organic content, and consequently increase the sorptionxe "sorption" of a nonionic organic contaminant. High concentrations of watersoluble cosolvents such as methanol and acetone can also act to decrease the activity coefficient, g, and act to solubilize sorbed nonionic organic compounds (Schwarzenbach et al., 1993). Surfactant molecules can aggregate into micelles in which their polar functional groups are oriented towards the aqueous solvent and their nonpolar tails are oriented inward toward each other. The space within the micelles therefore provides a hydrophobic refuge for nonionic contaminants (Edwards et al., 1991). Surfactants will form micelles at aqueous concentrations greater than their critical micelle concentration (CMC). Since, as noted above, surfactants will sorb at the surface of the porous mediaxe "porous media", a high dose of surfactant may be required in order to maintain an aqueous concentration greater than the CMC. Bacterial Polymers Many of the solution modifications discussed above involve the addition of synthetic agents to contaminated soil to accomplish the release of sorbed contaminants. Natural constituents that occur in soils and aquifers may also enhance contaminant transport (McCarthy and Zachara, 1989). Bacterial polymers naturally occur in soil solution and have welldocumented metal binding properties. The presence of bacterial polymers may therefore act as a natural process by which metal mobility is enhanced (Chen et al., 1995). The extracellular polymerxe "extracellular polymer"s produced by bacteria are heteropolysaccharides and have high molecular weight. Interestingly, these large molecules have also been show to be effective at binding nonionic organic pollutants and at enhancing their transport in aquifer materials (Dohse and Lion, 1994). In principle, bacterial polymers with suitable binding properties could be produced in engineered reactorxe "reactor" systems and be applied to contaminated waste sites to enhance the mobility of metal and nonionic organic contaminant mixtures. The efficacy of this type of remediation XE "remediation"  process has yet to be determined. Apparatus Students will apply a range of extractantxe "extractant" types (or mixtures of different types) to remove contaminants (Zn and methylene bluexe "methylene blue") from a porous medium. Laboratory extractions will mimic an engineered soil washing system in which the contaminated soil is actively mixed with the extractant and then separated. A rotator will be used to provide agitation of samples of the medium with extractants, and a centrifuge will be used to provide phase separation. A UV/visible spectrophotometerxe "spectrophotometer" with a diode arrayxe "diode array" detector will be used to measure the concentration of the extracted organic pollutant. Extracted metal concentrations will be measured with an atomic absorptionxe "absorption"xe "atomic absorption" (AA) spectrophotometer. Experimental Procedures Each group will develop their own hypothesis and experimental protocol. Different concentrations of extractants, different organic contaminants, and different washing techniques could be the investigation subjects. Alternate organic contaminants should be cleared with the instructor prior to the lab period. Each group should limit the investigation to approximately 10 samples and should include appropriate controls and replicates. The following protocol assumes that a common sand is employed to represent the porous medium. It is desirable, but not essential, to characterize each medium to be used (prior to the laboratory exercise) with respect to its carbon content [the Walkley Black method is one common procedure (Allison, 1965)], cation exchange capacity, and specific surface area [by sorptionxe "sorption" of ethylene glycol monoethyl ether (EGME) (Cihacek and Bremmer, 1979)]. I. Creation of a Contaminated Porous Medium A stock solution containing the soil contaminants will be provided [50 mg/L Zn and 100 mg/L methylene bluexe "methylene blue"]. For each extractantxe "extractant" used in part II below, 2 samples of contaminated sand and one sample of clean sand will be used. The following procedure is based on the assumption that each student group will evaluate 3 extractants or 3 concentrations of an extractant. Weigh out 9 aliquots of sand, 2.50.05 g each, and pour into 10 mL plastic centrifuge tube. Record the mass of the centrifuge tube with the sand (see Table 1). Add 5 mL of the contaminant stock solution to 6 of the samples. Add 5 mL distilled water to 3 of the samples (clean controls). Place all of the samples on a rotator to mix the sand and the contaminant/clean solutions. Agitate for 15 minutes. Centrifuge the suspensions at 3000 x g for 5 minutes. Pour the supernatant from the 6 contaminated sand samples into a 125 mL bottle. Pour the supernatant from the 3 clean sand samples into a separate 125 mL bottle. Weigh the centrifuge tubes with the sand and pore water. Calculate the volume of pore water by subtracting the centrifuge tube and sand masses. II. Determination of the Amount of Contaminant Sorbed by the Sand Methylene blue  UV/Vis Spectroscopy Nitrate absorbs ultraviolet light and is present in the contaminated samples from the addition of Zn(NO3)26H2O. We could account for this either by preparing a nitrate standard and using it as a component in spectral analysis or by eliminating the ultraviolet part of the spectrum XE "spectrum"  from the analysis. We will eliminate the nitrate interference by using a wavelength of 660 nm when measuring methylene bluexe "methylene blue". See page  PAGEREF _Ref406553429 \h 160 for instructions on using the UV/Vis Spectrophotometer. Measure the absorbance of 1, 5, and 10 mg/L methylene bluexe "methylene blue" solutions as Standards. Save the file as \\Enviro\enviro\Courses\453\soilwash\netid_MBstd. Measure the absorbance of the combined supernatant from the 3 clean sand samples, the combined supernatant from the 6 contaminated sand samples, and the contaminating solution (diluted by a factor of 10) as Samples. Save the file as \\Enviro\enviro\Courses\453\soilwash\netid_contamsuper. Record the concentration of methylene bluexe "methylene blue" in the clean supernatant and contaminated supernatant (see Table 2). You can drag the blue cursor on the "standard graph" to the wavelength of choice and read the exact absorbance (and wavelength) in the digital display to the left of the graph and concentration in the digital display at the bottom of the Spectrophotometer window. If the clean supernatant has significant absorbance at 660 nm then alternate analytical techniques may need to be used. The difference between the methylene bluexe "methylene blue" concentration in the contaminant solution and the concentration in the supernatant may be used to determine the sorbed contaminant concentration as:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 7. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 11 where Cinitial is the contaminant solution concentration and Cfinal is the concentration of the supernatant. Solution volume is the volume of contaminant added initially. Zinc  Atomic Absorption Spectroscopy Calibrate the AA using the zinc standards (1, 2, and 6 mg/L). Dilute all of the following samples by a factor of 10 to ensure sufficient sample volume for the analysis and to ensure that the results are in the calibrated range. Measure and record the zinc concentration of the combined supernatant from the 3 clean sand samples. Measure and record the zinc concentration of the combined supernatant from the 6 contaminated sand samples (see Table 4). Measure and record the zinc concentration in the contaminating solution (the zinc concentration should be close to 50 mg/L). The difference between the zinc concentration in the contaminant stock and the concentration in the supernatant may be used to determine the sorbed contaminant concentration using equation  GOTOBUTTON ZEqnNum308177 \* MERGEFORMAT  REF ZEqnNum308177 \! \* MERGEFORMAT 7.11. III. Soil Washing Solutions Students may wish to experiment with extractant mixtures. Some combinations of extractantxe "extractant" solutions may react violently! All proposed mixtures of extractants should be cleared with the course instructor prior to their use. The following combinations should be avoided: mixtures of oxidants with reductants, mixtures of acids with bases, and mixtures of oxidants with organic extractants including: surfactants, chelating agentxe "chelating agent"s or cosolvents. The following extractant solutions will be available for testing. At least one group should measure the extractant capabilities of distilled water because it is by far the cheapest! Distilled water. Acid: H" 1 M solution of HCl prepared by diluting 27.4 mL of the concentrated acid to 1 L with distilled water. Cosolvent: 1:1 (v/v) mixture of acetone and distilled water. Nonionic surfactant: 10% (v/v) solution of Triton X100 prepared by diluting 100 mL to 1 L with distilled water. Note: Triton X100 is a nonionic surfactant with a CMC of 2x105 M (Edwards et al., 1991). The chemical formula for Triton X100 is:  where: x = 9 to 10, giving the surfactant a molecular weight of H" 607g/mole. Anionic surfactant: 100 mM solution of dodecyl sulfate, sodium salt (C12H25SO4Na with MW of 288.4 so 28.84 g/L). This extractant XE "extractant"  works very well at full strength; lower concentrations could be investigated. Chelating agent: H" 0.1 M solution of ethylenediaminetetraacetate (EDTA) prepared by dissolving 37.22 g of the disodium salt in distilled water and diluting to 1 L. Base: H" 1 M NaOH solution prepared by dissolving 40 g of NaOH distilled water and diluting to 1 L. Oxidant: 1:1 (v/v) mixture of 30% H2O2 and distilled water. Reductant: H" 1 M solution of Na2S2O3.5H2O prepared by dissolving 248 g in distilled water and diluting to 1 L. IV. Soil Washing Protocol The ability of each extractantxe "extractant" to remove the zinc and methylene bluexe "methylene blue" from the contaminated soil will be measured by exposing the contaminated soil to the extractant and then measuring the concentration of the zinc and methylene blue in the extractant. Add 5 mL of each extractantxe "extractant" to be tested to 2 contaminated sand samples and 1 clean sand sample. Place the sand extractantxe "extractant" mixtures on a rotator to mix for 15 minutes. Centrifuge the suspensions at 3000 x g for 5 minutes. Decant the supernatant from each centrifuge tube into labeled 15 mL bottles. A small air line can be used to help force the supernatant from the centrifuge tubes. V. Analysis of Extracted Metal and Organic Pollutants Methylene blue  UV/Vis Spectroscopy Note that it is unnecessary to measure the methylene blue XE "methylene blue"  concentration in samples that do not appear to have any blue. Samples that are visually free of methylene blue can be recorded as 0 mg/L methylene blue. Measure the absorbance of each of the clean sand extracts as Samples. All of the clean extracts can be analyzed together if desired. Save as \\Enviro\enviro\Courses\453\soilwash\netid_cleanext. Measure concentration of methylene bluexe "methylene blue" in each of the clean extracts based on the absorbance at 660 nm (see Table 2). Measure the absorbance of each of the extracts of the contaminated sand as unknowns. There will be 2 replicates for each extract. All of the contaminated extracts can be analyzed as a group so that their results are saved in a single file. Save as \\Enviro\enviro\Courses\453\soilwash\netid_contamext. Measure concentration of methylene bluexe "methylene blue" in each of the contaminated extracts based on the absorbance at 660 nm (see Table 2). Note that it may be necessary to choose a different analytical wavelength or to dilute the sample if the absorbance exceeds H"2.5 at 660 nm. Calculate the mass extracted per mass of sand as:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 7. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 12 where solution volume is the sum of residual pore water volume after decanting the contaminant plus the extractantxe "extractant" volume. Zinc  Atomic Absorption Spectroscopy Dilute all samples by a factor of 10 prior to analysis. Measure and record the absorbance of the supernatant from the 3 clean sand samples (see Table 4). Measure and record the absorbance of the supernatant from the 6 contaminated sand samples. Dilute the supernatant with distilled water if the absorbance is not less than the absorbance of the 6-mg/L standard. Calculate the concentration of zinc in each of the sand extracts. Calculate the mass of Zn removed per mass of sand using equation  GOTOBUTTON ZEqnNum832151 \* MERGEFORMAT  REF ZEqnNum832151 \! \* MERGEFORMAT 7.12. The results for any extractantxe "extractant" must justify the cost of its use. The results obtained by extraction of contaminated soil using distilled water serve as a basis for comparison to which results obtained with extractants should be compared. Results from all extractants or extractant combinations evaluated should be compared to provide an intercomparison of their relative effects of the removal of metal and organic pollutants. Prelab Questions The point of zero charge for SiO2 is approximately at pH = 2.5. Is the charge of SiO2 positive or negative at a pH of 7? Do cations or anions generally bind most strongly to soil? Develop a hypothesis concerning soil washing, and write an experiment protocol to test your hypothesis that you can do in a lab period. Include detail of concentrations of extractants and contaminants for each vial. You may want to work with your lab partner(s) because this will be your experiment! Design your experiment to use no more than 9 vials. Data Analysis Report the contaminated sand concentration (grams of contaminant/gram of sand) for zinc and methylene bluexe "methylene blue". Calculate the fractional removal of zinc and methylene bluexe "methylene blue" for each extractant or extractant concentrationxe "extractant". The fractional removal based on the amount of contaminant initially sorbed is  EMBED Equation.DSMT4  where G is defined in equation  GOTOBUTTON ZEqnNum230062 \* MERGEFORMAT  REF ZEqnNum230062 \! \* MERGEFORMAT 7.11 and "G is defined in equation  GOTOBUTTON ZEqnNum843981 \* MERGEFORMAT  REF ZEqnNum843981 \! \* MERGEFORMAT 7.12. Present this using an appropriate graph. Discuss which extractantxe "extractant" performed best at removal of the zinc. Which was best at removing the test organic? Which extractant worked best at removing both contaminants? Discuss these results in terms of the chemical change that the extractant was designed to accomplish. (Note that these questions may need to be modified based on the samples you analyzed.) Discuss any difficulties in evaluating extractantxe "extractant" effectiveness and propose improved analytical techniques. Discuss your results and their implications for the hypothesis that you developed. Analyze your results including the reproducibility of replicate analyses in terms of possible sources of error. Suggest options for additional research. References Allison, L. E., Organic carbon, in: Soil Analysis Part 2: Chemical and Microbiological Properties, C. A. Black (ed.), Amer. Soc. Agronomy, Madison, WI, p 1367, 1965. Benjamin, M.M. and J.O. Leckie, Effects of complexation by Cl, SO4 and S2O3 on adsorptionxe "adsorption" behavior of Cd on oxide surfaces, Env. Sci. & Tech. 16(3), pp. 162170, 1982. Chen, J.H., L.W. Lion, W.C. Ghiorse, and M.L. Shuler, Trace metal mobilization in soil by bacterial extracellular polymerxe "extracellular polymer"s; Water Research, (1995, in press). Cihacek, L.J. and J.M. Bremmer, A simplified ethylene glycol monoethyl ether procedure for assessment of soil surface area, J. Soil Sci. Soc. Am. 43 pp. 821822, 1979. Dohse, D.M., L.W. Lion, The effect of microbial polymers on the partitioning and transport of phenanthrene in a lowcarbon sand; Environmental Sci. & Tech. 28(4), 541547 (1994). Edwards, D.A., R.G. Luthy, and Z. Liu, Solubilization of polycyclic aromatic hydrocarbons in micellar nonionic surfactant solutions, Env. Sci. & Tech. 25, pp. 127133, 1991. Honeyman, B.D. and P.H. Santschi, Metals in aquatic systems, Env. Sci. & Tech. 22(8), pp. 862871. Karickhoff, S.W., "Organic Pollutant Sorption in Aquatic Systems", J. Hydraulic Engrg., 110(6), p. 707, 1984. Kurbatov, M.H., G.B. Wood, and J.D. Kurbatov, Isothermal adsorptionxe "adsorption" of cobalt from dilute solutions, J. Phys. Chem. 55, pp. 11701182, 1951. Lion, L.W., R.S. Altmann, and J.O. Leckie, "Trace metal adsorptionxe "adsorption" characteristics of estuarine particulate matter: Evaluation of contributions of Fe/Mn oxide and organic surface coatings," Env. Sci. and Tech. 16(10), pp. 660666 (1982). McCarthy, J. F. and J. M. Zachara, Subsurface transport of contaminants, Env. Sci. & Tech. 23, pp. 496502, 1989. Parks, G.A. and P.L. DeBruyn, The zero point of charge of oxides, J. Phys. Chem. 66, pp. 967973, 1962. Sedlak, D.L., and A.W. Andren, Oxidationxe "Oxidation" of chlorobenzene with Fentons reagent, Env. Sci Tech. 25(4), pp. 777782, 1991. Schwarzenbach, R.P., P.M. Gschwend, and D.M. Imboden, Environmental Organic Chemistry, Wiley Interscience Publ., New York, NY; 681 pp., 1993. Standard Methods for the Examination of Water and Wastewater, L.S. Clesceri, A.E. Greenberg and R.R. Trussell (eds.) 17th edition, Am. Public Health Assoc.(Publisher), Washington, DC; 1989. Westall, J. and H. Hohl, A comparison of electrostatic models for the oxide/solution interface, Adv. Colloid Interface Sci. 12 p. 265, 1980. Table  SEQ \r 1 table1. Data table. bottle #contaminated or cleanmass sand (g)mass bottle + sand (g)mass bottle + sand + pore water (g)cleancont.cont.cleancont.cont.cleancont.cont.Table  SEQ table2. Methylene blue data table. concentrationclean sand supernatantcontaminated sand supernatantextractantxe "extractant" 1 clean sandextractantxe "extractant" 1 cont. sand rep 1extractantxe "extractant" 1 cont. sand rep 2extractantxe "extractant" 2 clean sandextractantxe "extractant" 2 cont. sand rep 1extractantxe "extractant" 2 cont. sand rep 2extractantxe "extractant" 3 clean sandextractantxe "extractant" 3 cont. sand rep 1extractantxe "extractant" 3 cont. sand rep 2Table  SEQ table3. Zinc concentration measurements data table. dilutionconcentrationclean sand supernatantcontaminated sand supernatantextractantxe "extractant" 1 clean sandextractantxe "extractant" 1 cont. sand rep 1extractantxe "extractant" 1 cont. sand rep 2extractantxe "extractant" 2 clean sandextractantxe "extractant" 2 cont. sand rep 1extractantxe "extractant" 2 cont. sand rep 2extractantxe "extractant" 3 clean sandextractantxe "extractant" 3 cont. sand rep 1extractantxe "extractant" 3 cont. sand rep 2 Lab Prep Notes Table  SEQ table4. Reagent list. DescriptionSupplierCatalog numberZn(NO3)26H2OFisher ScientificMethylene BlueFisher ScientificM29125HClFisher ScientificNaOHFisher ScientificS318500H2O2Fisher ScientificH325500Na2S2O3Fisher ScientificS445500nitrilotriacetic acidAldrichN8407Triton X100Aldrich23,4729acetoneFisher ScientificA9291FeSO47H2OAldrich31,0077alginic acid, sodium saltAldrich18,0947Fe(NO3)39H2OAldrich21,6828Humic acidAldrichH1,6752Nitric Acid 6 NFisher ScientificLC17702Dodecyl sulfate, sodium salt (C12H25SO4Na)Aldrich85,1922Zinc reference solutionFisher ScientificSZ13100Table  SEQ table5. Stock Solutions (100 mL each). DescriptionMW (g/M)conc. (g/L)100 mLC16H18N3SCl319.87101 gZn(NO3)26H2O297.4227.404822.74 g 5 g as ZnTable  SEQ table6. Contaminating Solution (1 L) DescriptionMW (g/M)conc. (mg/L)1 LC16H18N3SCl319.8710010 mL stockZn(NO3)26H2O297.4850 (as Zn)10 mL stock Table  SEQ table7. Equipment list DescriptionSupplierCatalog numberrefrigerated centrifuge MP4RFisher Scientific0500644place rotor 4BFisher Scientific050069Diode array spectrophotometerxe "spectrophotometer"HewlettPackard8452Arototorque rotatorCole ParmerH076370010 mL centrifuge vialsFisher Scientific055291Arepipet II DispensorFisher Scientific1368762BPP bottles 15 mLFisher Scientific029238GZinc Disposal Guidelines The amount of zinc that can be disposed to the sanitary sewer is limited. The wastewater treatment plant has a limit on the concentration of zinc that can be in the sludge. The Zinc stock solution should not be disposed to the sanitary sewer. Zinc that is sorbed to the sand can be dried and sent to the landfillxe "landfill" in the trash. Setup Use repipet dispensors for contaminating solution, distilled water and possibly for additives. Prepare calibration standards for the AA and for the UV-Vis spectrophotometers. Each group needs 9 centrifuge vials, 20 15-mL bottles, and 2 125-mL bottles. Connect a very fine tube to an air line at each island to be used to help empty the supernatant from the centrifuge vials. Devise technique to filter samples prior to AA analysis! Class Plan Demonstrate use of AA when samples contain particulate matter. (Keep sipper tube off of the bottom of the vial!) EDTA works well for Zn at 0.1 M Dodecyl sulfate works well for MB at 0.01 M Oxygen Demand Concepts and Dissolved Oxygen Sag in Streams Introduction  MACROBUTTON MTEditEquationSection Equation Section (Next) SEQ MTEqn \r \h \* MERGEFORMAT  SEQ MTSec \h \* MERGEFORMAT In recent years biodegradable has become a popular word. Often it is assumed that if something is biodegradable, then disposal is not a problem. We know that throwing nonbiodegradable substances into our environment leads to degradation of our planet. But disposal of biodegradable compounds also can be detrimental to the environment. The effects of improper disposal of biodegradable substances became a source of public outrage in the early 1800's. The flush toilet was becoming popular and sewage was discharged directly into the nearest waterway. The receiving waters were quickly polluted. Fish in the receiving waters died and the water had a very offensive odor. Although there are many reasons why we no longer discharge untreated sewage into the environment, (including disease transmission, sediment buildup...) one of the reasons is directly related to the fact that sewage contains much that is biodegradable. Theory Biodegradablexe "Biodegradable" means that a substance can be converted into simpler compounds by biologically mediated reactions. The second law of thermodynamics predicts that oxidation of high energy level organics (relative to low energy level CO2) is favored. Oxygen is one of the strongest oxidizing agents found in natural aquatic systems. Oxidationxe "Oxidation" reactions are thermodynamically favored, but kinetically slow unless microbially mediated. The end products of complete aerobic biodegradation are CO2 and H2O. Production of CO2 has recently come under fire as a potential cause of global warming XE "global warming" , but that is not the subject of this lab. The problem is not with the products of biodegradation, the problem is that aerobic biodegradation of a compound requires another reactant. Let's look at the biodegradation of a simple organic compound, glucose.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 1 To balance the equation oxygen is needed.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 2 The consumption of oxygen needed for biodegradation can be a problem. Oxygen is not very soluble in water. The equilibrium concentration of oxygen in water is approximately 10 mg/L (see  REF _Ref365357907 \* MERGEFORMAT Figure 1). That means that the degradation of a few mg/L of a biodegradable compound in a river could result in the depletion of dissolved oxygenxe "dissolved oxygen". Fish have a bad day when oxygen is depleted from their environment. Some species of fish such as trout begin to suffer when the dissolved oxygen concentration drops below 5 mg/L. EMBED Excel.Sheet.8 Figure  SEQ \r 1 Figure1. Equilibrium dissolved oxygenxe "dissolved oxygen" concentration as a function of water temperature. Oxygen in water is consumed during aerobic biodegradation of organic compounds and is replenished from the atmosphere. The two processes have different kinetics, but are coupled. As the oxygen is depleted by biodegradation the rate at which oxygen is transferred into the water increases because the transfer driving force increases. The rate at which oxygen is dissolved into water from the atmosphere is proportional to the deficit of oxygen in the water. The oxygen deficitxe "oxygen deficit" is simply the difference between the equilibrium oxygen concentration and the actual oxygen concentration. These two reactions (reaeration, and biochemical utilization) are modeled by the StreeterPhelps equation. In order to increase the rate at which the biodegradation occurs, the concentration of bacteria was increased for use in this laboratory experiment. Bacteria respire and thus consume oxygen even when no substrate is present. Thus an additional term for bacterial respirationxe "endogenous respiration" will be needed to model the oxygen sagxe "oxygen sag" results obtained in the laboratory. Streeter Phelpsxe "Streeter Phelps" Equation Development Well begin by developing the oxygen deficitxe "oxygen deficit" as a function of time in a completely mixed batch reactorxe "reactor" (no inflow and no outflow) with initial concentrations of Biochemical Oxygen Demandxe "Biochemical Oxygen Demand" (BODL) and dissolved oxygenxe "dissolved oxygen". We will include oxidation of BODL and reaeration from the atmosphere. These effects are coupled in equation  GOTOBUTTON ZEqnNum670626 \* MERGEFORMAT  REF ZEqnNum670626 \! \* MERGEFORMAT 8.3 where C represents oxygen concentration. The first two terms on the right are negative since oxidation of BOD and respiration consume oxygen while the third term is usually positive since reaeration increases the concentration of oxygen (except in the rare instance where the dissolved oxygen concentration is greater than the equilibrium dissolved oxygen concentration). Eventually we will make a comparison between time in a reactor and distance down a river.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 3 Oxidationxe "Oxidation" of BOD We must first develop a relationship for the change in oxygen concentration due to oxidation of organics. The rate that oxygen is used will be proportional to the rate that substrate (or biochemical oxygen demand) is oxidized. The rate of substrate utilization by bacteria is given by the Monodxe "Monod" relationship  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 4 SEQ[eq] \n \* MERGEFORMAT  where L is substrate concentration expressed as oxygen demand or BODL [mg/L], k is the maximum specific substrate utilization rate, Ks is the half velocity constant, and X is the concentration of bacteria. However, the concentration of bacteria is a function of the substrate concentration and thus application of the Monodxe "Monod" equation to a polluted river is not trivial. Often the bacterial concentration remains relatively constant. If the half velocity concentration is large relative to the concentration of substrate we obtain  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 5 where kox is a first order oxidation rate constant that includes both the approximation that the bacteria concentration is roughly constant and that the substrate concentration is smaller than the half velocity constant. Separate variables and integrate  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 6 to obtain  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 7 The rate of oxygen utilization is equal to the rate of substrate utilization (when measured as oxygen demand) and thus we have  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 8 where C is the dissolved oxygenxe "dissolved oxygen" concentration [mg/L]. Now we can substitute for L in equation  GOTOBUTTON ZEqnNum268674 \* MERGEFORMAT  REF ZEqnNum268674 \! \* MERGEFORMAT 8.8 using equation  GOTOBUTTON ZEqnNum138140 \* MERGEFORMAT  REF ZEqnNum138140 \! \* MERGEFORMAT 8.7 to obtain  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 9 Respiration Bacteria utilize oxygen for respiration and for cell synthesis. When no substrate is present the bacteria cease synthesis, but must continue respiration. This continual use of oxygen is termed "endogenous respiration." Bacteria use stored reserves for endogenous respirationxe "endogenous respiration". We can model this oxygen demand as a constant that is added to the demand for oxygen caused by substrate utilization. As a first approximation, we can assume that this oxygen demand is proportional to the concentration of bacteria. In addition, we will assume that the population of bacteria is relatively constant throughout the experiment.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 10 where b is the specific endogenous oxygen consumption rate and ke is the endogenous oxygen consumption rate. Oxygen Transfer Coefficient The rate of oxygen transfer is directly proportional to the difference between the actual dissolved oxygenxe "dissolved oxygen" concentration and the equilibrium dissolved oxygen concentration.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 11 where C* is the equilibrium oxygen concentration, C is the actual dissolved oxygenxe "dissolved oxygen" concentration, and  EMBED Equation.DSMT4  is the is the overall volumetric oxygen transfer coefficient. If reaeration is the only process affecting the oxygen concentration then equation  GOTOBUTTON ZEqnNum142088 \* MERGEFORMAT  REF ZEqnNum142088 \! \* MERGEFORMAT 8.11 can be integrated to obtain  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 12 Oxygen Deficit We now have equations for the reaction of oxygen with BODL, endogenous respirationxe "endogenous respiration", and for reaeration. Substituting into equation  GOTOBUTTON ZEqnNum423086 \* MERGEFORMAT  REF ZEqnNum423086 \! \* MERGEFORMAT 8.3 we get  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 13 We can simplify the equation by defining oxygen deficitxe "oxygen deficit" (D) as:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 14 and noting that the rate of change of the deficit must be equal and opposite to the rate of change of oxygen concentration  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 15 We must remember that the deficit can never be greater than the equilibrium concentration (D must always be less than C*)! In addition, the BOD model breaks down if the dissolved oxygenxe "dissolved oxygen" concentration is less than about 2 mg/L because the lack of oxygen will limit microbial kinetics and  EMBED Equation.DSMT4  will no longer equal  EMBED Equation.DSMT4 . If we stick to conditions under which our assumptions are valid then we can substitute equations  GOTOBUTTON ZEqnNum200777 \* MERGEFORMAT  REF ZEqnNum200777 \! \* MERGEFORMAT 8.14 and  GOTOBUTTON ZEqnNum138983 \* MERGEFORMAT  REF ZEqnNum138983 \! \* MERGEFORMAT 8.15 into equation  GOTOBUTTON ZEqnNum695483 \* MERGEFORMAT  REF ZEqnNum695483 \! \* MERGEFORMAT 8.3 to obtain  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 16 This is a first order linear differential equation. Integration with initial oxygen deficitxe "oxygen deficit" = Do @ t = 0 gives:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 17 Application to a River We are interested in the oxygen deficitxe "oxygen deficit" as a function of distance down a stream. As an approximation we can think of a cross section of a river as a completely stirred reactorxe "reactor" that is slowly moving downstream. The relation between time in a batch reactor and distance down the river is simply  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 18 where u is the stream velocity and x is distance. The StreeterPhelps model assumes a constant input of biodegradable substrate, Lo, at x = 0 and the model is valid under steadystate conditions. Of particular concern is the maximum deficit, Dc. We want to know the value of Dc and where (or when) it will occur ( EMBED Equation.DSMT4 ). This will be the "critical point." If there are going to be adverse effects (like dead fish) this will be the place. The maximum oxygen deficitxe "oxygen deficit" occurs when  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 19 We can substitute this into the first order differential equation  GOTOBUTTON ZEqnNum139336 \* MERGEFORMAT  REF ZEqnNum139336 \! \* MERGEFORMAT 8.6 to get  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 20 and solve for Dc to get  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 21 an equation with unknowns tc and Dc. The StreeterPhelps equation still holds at the critical point so we also have  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 22 also with unknowns xc and Dc. So now we have two equations in two unknowns. We can solve for tc by eliminating Dc.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 23 To find Dc given the kinetic coefficients and the initial oxygen deficitxe "oxygen deficit", first find tc using equation  GOTOBUTTON ZEqnNum383344 \* MERGEFORMAT  REF ZEqnNum383344 \! \* MERGEFORMAT 8.23. Then use equation  GOTOBUTTON ZEqnNum540802 \* MERGEFORMAT  REF ZEqnNum540802 \! \* MERGEFORMAT 8.21 to solve for Dc. Zero Order Kinetics An alternate model can be derived based on the assumptions that the concentration of bacteria is relatively constant and that the rate of substrate utilization is zero order, i.e., the concentration of the substrate is greater than the half velocity constant Ks.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 24 The change of the deficit of dissolved oxygenxe "dissolved oxygen" is equal to the change caused by microbial degradation (k0) plus change due to endogenous respirationxe "endogenous respiration" minus the reaeration ( EMBED Equation.DSMT4 ).  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 25 This equation is only valid when the substrate concentration is greater than Ks. To simplify derivation, assume that Ks is very small relative to the initial BOD added to the system and apply the zeroorder model until the substrate is completely oxidized. When the substrate concentration reaches zero a discontinuity will occur as substrate oxidation stops. Separating variables and integrating  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 26  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 27 and solving for the dissolved oxygenxe "dissolved oxygen" deficitxe "oxygen deficit"  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 28 The substrate concentration is depleted when  EMBED Equation.DSMT4 . Substituting into equation  GOTOBUTTON ZEqnNum342204 \* MERGEFORMAT  REF ZEqnNum342204 \! \* MERGEFORMAT 8.28 to get the maximum dissolved oxygenxe "dissolved oxygen" deficitxe "oxygen deficit" yields  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 29 where Dt is the dissolved oxygenxe "dissolved oxygen" deficitxe "oxygen deficit" at the transition when the substrate is all utilized. For times greater than  EMBED Equation.DSMT4  there is no longer any substrate and thus k0 = 0 and equation  GOTOBUTTON ZEqnNum929597 \* MERGEFORMAT  REF ZEqnNum929597 \! \* MERGEFORMAT 8.25 becomes  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 30 Separating variables and integrating  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 31  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 32  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 8. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 33 Equation  GOTOBUTTON ZEqnNum143112 \* MERGEFORMAT  REF ZEqnNum143112 \! \* MERGEFORMAT 8.33 is valid for all times greater than  EMBED Equation.DSMT4 . The general shapes of the two types of sag curves are shown in  REF _Ref365357945 \* MERGEFORMAT Figure 2.  Figure  SEQ Figure2. Dissolved oxygen sagxe "oxygen sag" curves obtained from zero and first order models for substrate utilization. Experimental Objectives The objectives of this lab are to: Illustrate the effects of adding biodegradable compounds to natural waters. Evaluate the StreeterPhelps dissolved oxygenxe "dissolved oxygen" sagxe "oxygen sag" model and a zero order substrate utilization model and compare with laboratory data. Explain the theory and use of dissolved oxygenxe "dissolved oxygen" probexe "oxygen probe"s. Experimental Methods EMBED Word.Picture.8 Figure  SEQ Figure3. Apparatus used to measure dissolved oxygen XE "dissolved oxygen"  consumption rates. In this lab we will examine the effects of adding a small amount of a biodegradable compound to a small batch reactorxe "reactor". We will measure the dissolved oxygenxe "dissolved oxygen" concentration over time using a dissolved oxygen probexe "oxygen probe". The apparatus is shown in  REF _Ref365357960 \* MERGEFORMAT Figure 3. The BOD measured using this technique will be lower than the BOD measured using the standard BOD test because a significant fraction of the glucose will be converted into cell material (i.e. used for synthesis instead of for respiration). This technique can be used to obtain kinetic parameters for yield, half velocity constant and maximum substrate utilization rate  ADDIN ENRef (Ellis et al., 1996). Probe Calibration Calibrate the dissolved oxygenxe "dissolved oxygen" probe (see  HYPERLINK "http://www.cee.cornell.edu/mws/Software/DOcal.htm" http://www.cee.cornell.edu/mws/Software/DOcal.htm). Oxygen Transfer Coefficient Prepare to monitor dissolved oxygenxe "dissolved oxygen". Place the dissolved oxygenxe "dissolved oxygen" probexe "oxygen probe" in the reactorxe "reactor". Pour 50 mL of deoxygenated distilled water into the batch reactorxe "reactor". Set the stirrer speed to 5. Set the airflow rate to 50 mL/min. Monitor the dissolved oxygenxe "dissolved oxygen" for 3 minutes (or longer). Save the data as \\Enviro\enviro\Courses\453\oxygen\netid_O2trans. The data will be used later to estimate the oxygen transfer coefficient. Endogenous respiration oxygen requirements Pour 50 mL of a bacterial suspension into the batch reactorxe "reactor". Place the dissolved oxygenxe "dissolved oxygen" probexe "oxygen probe" in the reactorxe "reactor". Set the stirrer speed to 5. Set the airflow rate to 250 mL/min to aerate the reactorxe "reactor" contents. Prepare to monitor dissolved oxygenxe "dissolved oxygen". After the dissolved oxygen concentration is close to saturation turn off the air and monitor the dissolved oxygenxe "dissolved oxygen" for 3 minutes (or longer). Save the data as \\Enviro\enviro\Courses\453\oxygen\netid_endog. The data will be used later to estimate the endogenous respirationxe "endogenous respiration" rate. BOD of glucose solution Set the airflow rate to 250 mL/min and aerate the bacterial suspension used previously. Prepare pipettexe "pipette" to add 75 L of glucose solution (this will provide a BOD of 15 mg/L when diluted by 50 mL bacterial suspension). Prepare to monitor dissolved oxygenxe "dissolved oxygen". Turn off the airflow. As quickly as possible, add glucose through the port in the bottle and begin monitoring the dissolved oxygenxe "dissolved oxygen" concentration. Monitor the dissolved oxygenxe "dissolved oxygen" until the dissolved oxygen concentration reaches approximately 0 mg/L. Save the data as \\Enviro\enviro\Courses\453\oxygen\netid_BOD. The data will be used later to estimate the BOD of the glucose solution. DO Sag Curves Set the airflow rate to 250 mL/min and aerate the bacterial suspension used previously. Prepare to monitor dissolved oxygenxe "dissolved oxygen". Reduce the airflow rate to 50 mL/min. Begin monitoring the dissolved oxygenxe "dissolved oxygen" in the reactorxe "reactor". Use 5 second data intervals. After H"300 seconds of monitoring add 10 mg glucose BOD/L (50 (L stock) to the reactorxe "reactor". Observe the oxygen depletion in the reactorxe "reactor". Continue monitoring until the dissolved oxygenxe "dissolved oxygen" concentration returns to within 90% of the original DO concentration. Save the data as \\Enviro\enviro\Courses\453\oxygen\netid_sag. Prelab Questions A dissolved oxygenxe "dissolved oxygen" probexe "oxygen probe" was placed in a small vial in such a way that the vial was sealed. The water in the vial was sterile. Over a period of several hours the dissolved oxygen concentration gradually decreased to zero. Why? (You need to know how dissolved oxygen probes work to answer this!) Which assumption is different between the StreeterPhelps and the zero order model? Data Analysis The rate constants can be estimated using Excel. A sample spreadsheet is available at the course web site. Oxygen Transfer Coefficient Estimate the gas transfer XE "gas transfer"  coefficient from equation  GOTOBUTTON ZEqnNum488555 \* MERGEFORMAT  REF ZEqnNum488555 \! \* MERGEFORMAT 8.12 or by using the spreadsheet model. Graph the dissolved oxygenxe "dissolved oxygen" concentration vs. time along with the theoretical curve. Endogenous Decay Estimate the endogenous oxygen consumption rate from the slope of the graph or by using the spreadsheet model. Graph the dissolved oxygenxe "dissolved oxygen" concentration for the bacteria culture in the BOD bottle without any added BOD vs. time along with the theoretical curve. BOD of Glucose Use the "DO sag" Excel spreadsheet to estimate the first or zero order oxygen utilization coefficients, and the BOD exerted by the glucose. Which model fits the data best? How long did it take for the biodegradation of the glucose to occur? What was the change in dissolved oxygenxe "dissolved oxygen" concentration during that time? How much BOD did the glucose solution exert expressed as a fraction of the BOD of the glucose added. Graph the dissolved oxygenxe "dissolved oxygen" concentration vs. time for the glucose solutions along with the theoretical curves. Identify the regions where biodegradation of the glucose was occurring. Dissolved Oxygen Sag Use the previous estimates of the oxygen transfer coefficient, endogenous respiration XE "endogenous respiration"  rate, and fraction of BOD exerted (note that a different amount of BOD was added for the sag curve than for the BOD measurement!) to plot zero and first order model predictions of the dissolved oxygen XE "dissolved oxygen"  sag XE "oxygen sag"  curve. Discuss any discrepancies. Estimate the first and zero order oxygen utilization coefficients and the BOD exerted using the spreadsheet models by minimizing the RMSE using both models with your data. Use the endogenous respiration XE "endogenous respiration"  rate and the reaeration rate estimated previously. Which model (zero or first order) fits the data best? Are the fit parameters significantly different than those obtained in the BOD of glucose analysis? Include the estimated parameters in your report. Graph the dissolved oxygenxe "dissolved oxygen" concentration vs. time for the dissolved oxygen sagxe "oxygen sag" curve along with the theoretical curves. On the graph indicate maximum dissolved oxygenxe "dissolved oxygen" sagxe "oxygen sag" and compare with the BOD added. Why is the dissolved oxygenxe "dissolved oxygen" sagxe "oxygen sag" less than the BOD added? References  ADDIN ENBib Ellis, T. G.; D. S. Barbeau; B. F. Smets and C. P. L. J. Grady. 1996. Respirometric technique for determination of extant kinetic parameters describing biodegradation Water Environment Research 68(5): 917926. Lab Prep Notes Table  SEQ \r 1 table1. Reagent list DescriptionSupplierCatalog numberpeptoneFisher ScientificBP1420100tryptoneFisher ScientificBP1421100glucoseAldrich15,8968yeast extractFisher ScientificBP1422100MgSO47H2OFisher ScientificCaCl22H2OFisher ScientificKH2PO4Fisher ScientificK2HPO4Fisher ScientificNa2HPO4 7H2OFisher ScientificNH4ClFisher ScientificFeCl3 6H2OFisher Scientific Table  SEQ table2. Equipment list DescriptionSupplierCatalog numbermagnetic stirrerFisher Scientific115007SAccumet"! 50 pH meterFisher Scientific1363550ATI Orion DO probeFisher Scientific13299856 L containerFisher Scientific0348422250 mL PP bottleFisher Scientific02925D15 mL PP bottlesFisher Scientific029238Gvariable flow digital driveCole ParmerH0752330EasyLoad pump headCole ParmerH0751800PharMed tubing size 18Cole ParmerH06485184 prong hypodermic tubing diffuserCEE shop1/4 plugCole ParmerH06372501/4 unionCole ParmerH0637250stainless steel hypodermic tubingMcMaster Carrgas diffusing stoneFisher Scientific11139BBacterial stock preparation using 20% PTYG Grow 4 liter culture of Ps. putida Heat 1 L of distilled water and dissolve media for 4 L of 20% PTYG. Dilute to 4 L in 6 L container containing aeration stone and stirrer. Thaw one cryovial containing Ps. putida and transfer into PTYG media. Stir and aerate for 24 hours. Wash/enumerate Ps. putida culture Centrifuge 4 L culture in 250 mL bottles to obtain concentrated stock (5000 rpm for 10 minutes). Resuspend total culture in 500 mL using 10x BOD dilution water (pH control is essential for bacterial growth and trace nutrients are required). Refrigerate at 4(C. Table  SEQ table3. 20% PTYG culture media. (Prepare 4 L)compoundmg/Lg/4Lpeptone10004tryptone10004yeast extract20008glucose10004MgSO44701.9CaCl22H2O700.28 Setup Prepare the Ps. putida culture starting 48 hours before lab. Table  SEQ table4. BOD dilution water stock solutions. Use 10 mL per liter of each of the 4 solutions to prepare 10x BOD dilution water. phosphate bufferM.W.g/Lmg/100 mLMKH2PO4136.098.585062.46K2HPO4174.1821.752175124.87Na2HPO4 7H2O268.0733.43340124.60NH4Cl53.491.717031.78Magnesium sulfateMgSO4120.3911110091.37Calcium chlorideCaCl2110.9927.52750247.77Ferric chlorideFeCl3 6H2O270.30.25250.925Prepare 100 mL glucose stock solution. Attach one EasyLoad pump head to the pump drives and plumb with size 18 tubing connected to the hypodermic diffuser. Verify that DO probes are operational, stable, and can be calibrated. Mount DO probes on magnetic stirrers. (Use large stirbars.) Use 100 mL plastic beakers containing 50 mL of bacteria suspension. The open tops will result in negligible oxygen transfer during the course of the experiments. Prepare 1 L of deoxygenated distilled water right before class using the techniques outlined in the gas transfer XE "gas transfer"  lab (see page  PAGEREF _Ref498828623 \h 154). Glucose Stock Solution C6H12O6 + 6O2 6CO2 + 6H2O  EMBED Equation.DSMT4  EMBED Equation.DSMT4  Glucose Dilutions  EMBED Equation.DSMT4  100 L in 100 mL will provide 10 mg/L BOD 10 L of stock solution diluted into 100 mL provides 1 mg BOD/L. Methane Production from Municipal Solid Waste Introduction  MACROBUTTON MTEditEquationSection Equation Section (Next) SEQ MTEqn \r \h \* MERGEFORMAT  SEQ MTSec \h \* MERGEFORMAT Archaeological investigations of landfills have revealed that biodegradable wastes can be found virtually intact 25 years after burial. We know that landfills contain bacteria with the metabolic capability to degrade many of the materials that are common components of municipal refuse. The persistence for decades of degradable materials in the presence of such organisms appears somewhat paradoxical. In this experiment students will explore the factors that influence biodegradation of waste materials in landfills. Although recycling has significantly reduced the amount of landfillxe "landfill" space dedicated to paper and other lignocellulosics, paper products are still a significant fraction of the solid waste stream. In this laboratory students will measure the rate and extent of anaerobic degradation of newsprint, Kraft paper, coated paper, and food scraps. Theory Table  SEQ Table \r11. Typical physical composition of residential MSW in 1990 excluding recycled materials and food wastes discharged with wastewater  ADDIN ENRf8 (Tchobanoglous, Theisen et al. 1993)ComponentRangeTypicalOrganic(% by weight)(% by weight) food wastes6189.0 paper254034.0 cardboard3106.0 plastics4107.0 textiles042.0 rubber020.5 leather020.5 yard wastes52018.5 wood142.0Organic total =SUM(ABOVE) 79.5Inorganicglass4128.0tin cans286.0aluminum010.5other metal143.0dirt, ash, etc.063.0Inorganic total =SUM(ABOVE) 20.5 Over 150 million tons of municipal solid waste (MSW) are generated every year in the United States, and more than 70% of the MSW is deposited in landfills  ADDIN ENRf8 (Gurijala and Suflita 1993). Paper constitutes the major weight fraction of MSW, and this laboratory will focus on the biodegradation of that component. Anaerobic biodegradation of paper produces methanexe "methane" and carbon dioxide. Methane is a fuel and is the major component of natural gas. Methane produced in sanitary landfills represents a usable but underutilized source of energy. Energy recovery projects are frequently rejected because the onset of methane production is unpredictable and methane yields vary from 130% of potential yields based on refuse biodegradability data  ADDIN ENRf8 (Barlaz, Ham et al. 1992). The low methane yields are the result of several factors that conspire to inhibit anaerobic biodegradation including low moisture levels, resistance to biodegradation, conditions that favor bacterial degradation pathways that do not result in methane as an end product, and poor contact between bacteria and the organic matter. Characteristics of municipal solid waste The physical composition of residential municipal solid waste (MSW) in the United States is given in  REF _Ref386866828 \* MERGEFORMAT Table 1. The fractional contribution of the listed categories has evolved over time, with a trend toward a decrease in food wastes because of increased use of kitchen food waste grinders, an increase in plastics through the growth of their use for packaging, and an increase in yard wastes as burning has ceased to be allowed by most communities  ADDIN ENRf8 (Tchobanoglous, Theisen et al. 1993). Excluding plastic, rubber, and leather, the organic components listed in  REF _Ref386866828 \* MERGEFORMAT Table 1 are, given sufficient time, biodegradable. Table  SEQ Table2. Percentage distribution by weight of paper types in MSW  ADDIN ENRf8 (Tchobanoglous, Theisen et al. 1993)Type of paperRangeTypicalnewspaper102017.7books and magazines5108.7commercial printing486.4office paper81210.1other paperboard81210.1paper packaging6107.8other nonpackaging paper81210.6tissue paper and towels485.9corrugated materials202522.7Total100.0 Although recycling efforts divert a significant fraction of paper away from landfills, paper continues to be a major component of landfilled waste. The types of paper found in MSW are listed in  REF _Ref386866926 \* MERGEFORMAT Table 2. The elemental composition of newsprint and office paper are listed in  REF _Ref386866938 \* MERGEFORMAT Table 3. Table  SEQ Table3. Elemental composition of two paper types on a dry weight basis  ADDIN ENRf8 (Tchobanoglous, Theisen et al. 1993).ConstituentNewsprintOffice PaperC49.1%43.4%H6.1%5.8%O43.0%44.3%NH4N4 ppm61 ppmNO3N4 ppm218 ppmP44 ppm295 ppmPO4P20 ppm164 ppmK0.35%0.29%SO4S159 ppm324 ppmCa0.01%0.10%Mg0.02%0.04%Na0.74%1.05%B14 ppm28 ppmZn22 ppm177 ppmMn49 ppm15 ppmFe57 ppm396 ppmCu12 ppm14 ppm The major elements in paper are carbon, hydrogen, and oxygen that together constitute 93.5% of the total solids. The approximate molecular ratios for newspaper and office paper are C6H9O4 and C6H9.5O4.5 respectively. Biodegradation of cellulose, hemicellulose, and lignin Cellulose and hemicellulose are the principal biodegradable constituents of refuse accounting for 91% of the total methanexe "methane" potential. Cellulose forms the structural fiber of many plants. Mammals, including humans, lack the enzymes to degrade cellulose. However, bacteria that can break cellulose down into its subunits are widely distributed in natural systems, and ruminants, such as cows, have these microorganisms in their digestive tract. Cellulose is a polysaccharide that is composed of glucose subunits (see  REF _Ref386867079 \* MERGEFORMAT Figure 1).  Figure  SEQ Figure \r11. Cellulose (two glucose subunits are shown). Another component of the walls of plants is hemicellulose, which sounds similar to cellulose but is unrelated other that that it is another type of polysaccharide. Hemicelluloses made up of five carbon sugars (primarily xylose) are the most abundant in nature. Lignin is an important structural component in plant materials and constitutes roughly 30% of wood. Significant components of lignin include coniferyl alcohol and syringyl alcohol subunits ( REF _Ref386867106 \* MERGEFORMAT Figure 2).  Figure  SEQ Figure2. Coniferyl (left) and syringyl (right) subunits of lignin. The exact chemical structure of lignin is not known but its reactivity, breakdown products, and the results of spectroscopic studies reveal it to be a polymeric material containing aromatic rings with methoxy groups (OCH3)  ADDIN ENRf8 (Tchobanoglous, Theisen et al. 1993). One of the many proposed structures for lignin is shown in  REF _Ref386867117 \* MERGEFORMAT Figure 3.  Figure  SEQ Figure3. A postulated formulation for spruce lignin (by  ADDIN ENRf8 (Brauns 1962), as cited by  ADDIN ENRf8 (Pearl 1967)). This structure is suggested by spectroscopic studies and the chemical reactions of lignin. Degradation of lignin requires the presence of moisture and oxygen and is carried out by filamentous fungi  ADDIN ENRf8 (Prescot, Harley et al. 1993). The biodegradability of lignocellulosic materials can be increased by an array of physical/chemical processes including pretreatment to increase surface area (size reduction), heat treatment, and treatment with acids or bases. Such treatments are useful when wood and plant materials are to be anaerobically degraded to produce methanexe "methane". Research on this topic has been performed by Cornell Prof. James Gossett  ADDIN ENRf8 (Gossett and McCarty 1976; Chandler, Jewell et al. 1980; Gossett, Stuckey et al. 1982; Pavlostathis and Gossett 1985a; Pavlostathis and Gossett 1985b). Three major groups of bacteria are involved in the conversion of cellulosic material to methanexe "methane"  ADDIN ENRf8 (Zehnder 1978): (1) the hydrolytic and fermentative bacteria that break down biological polymers such as cellulose and hemicellulose to sugars that are then fermented to carboxylic acids, alcohols, carbon dioxide and hydrogen gas, (2) the obligate hydrogen reducing acetogenic bacteria that convert carboxylic acids and alcohols to acetate and hydrogen, and (3) the methanogenic bacteria that convert primarily acetate and hydrogen plus carbon dioxide to methane. Sulfate reducing bacteria (SRB) may also play a role in the anaerobic mineralization of cellulosic material. In the presence of sulfate, the degradation process may be directed towards sulfate reduction by SRB with the production of hydrogen sulfide and carbon dioxide  ADDIN ENRf8 (Barlaz, Ham et al. 1992). Cellular requirements for growth The availability of oxygen is a prime determinant in the type of microbial metabolism that will occur. Microbial respiration of organic carbon is a combustion process, in which the carbon is oxidized (i.e., is the electron donor) in tandem with the reduction of an electron acceptor. The energy available to microorganisms is greatest when oxygen is used as the electron acceptor and therefore aerobic metabolic processes will dominate when oxygen is available. Some microorganisms require oxygen to obtain their energy and are termed obligate aerobes. In the absence of oxygen, other electron acceptors such as nitrate (NO3), sulfate (SO42) and carbon dioxide (CO2) can by used. Organisms that can only exist in an environment that contains no oxygen are termed obligate anaerobes. Organisms that have the ability to grow in both the presence and the absence of oxygen are said to be facultative. The availability of nutrients can limit the ability of cells to grow and consequently the extent of biodegradation. Nitrogen and/or phosphorous constitute important nutrients required for cell synthesis. Inorganic bacterial nutritional requirements also include sulfur, potassium, magnesium, calcium, iron, sodium and chloride. In addition, inorganic nutrients needed in small amounts (minor or trace nutrients) include zinc, manganese, molybdenum, selenium, cobalt, copper, nickel, vanadium and tungsten. Organic nutrients (termed growth factors) are also sometimes needed (depending on the microorganism) and include certain amino acids, and vitamins  ADDIN ENRf8 (Metcalf & Eddy 1991). Environmental conditions such as pH, temperature, moisture content, and salt concentration can have a great influence on the ability of bacteria to grow and survive. Most bacteria grow in the pH range from 4.0 to 9.5 (although some organisms can tolerate more extreme pH values), and typically grow best in the relatively narrow range from 6.5 to 7.5 (Metcalf & Eddy, 1991). Microorganisms have a temperature range over which they function best, and are loosely characterized as phychrophilic (ability to grow at 0C), mesophilic (optimal growth at 2540C) or thermophilic (optimal growth above 4550C)  ADDIN ENRf8 (Brock 1970). Many common methanogens are mesophilic. Elevated temperatures also favor faster reaction rates. While some microorganisms are very tolerant of low moisture conditions, active microbial growth and degradation of organic matter necessitates that water not be a scarce resource. Cells take water in through their semipermeable membrane surface by osmosis. This uptake mechanism requires that the solute concentration inside the cell be higher than that of the outside media. Organisms that grow in dilute solutions can not tolerate high salt concentrations because their normal osmotic gradient is reversed and they can not take in water. Some cell strains, termed halophiles are adapted for growth at very high salt concentrations. The above factors suggest that bacterial degradation of MSW to produce methanexe "methane" will occur optimally at circumneutral pH, low ionic strength, in the absence of oxygen, nitrate and sulfate, in the presence of moisture and nutrients, and under mesophilic conditions. Estimates of paper biodegradability Volatile solids (VS) content (determined by weight loss on ignition at 550C) has been used to estimate the biodegradability of MSW components, but this measure overestimates the biodegradability of paper. Paper products have a very high volatile solids content. Newsprint, office paper, and cardboard have VS of 94%, 96.4%, and 94% respectively  ADDIN ENRf8 (Tchobanoglous, Theisen et al. 1993). Paper products also can have a high content of lignocellulosic components that are only slowly degradable. Lignin constitutes approximately 21.9%, 0.4% and 12.9% respectively of the VS in newsprint, office paper, and cardboard. Lignin content and biodegradability are strongly correlated and thus lignin content can be used to estimate biodegradability and potential methanexe "methane" production. Chandler et al.  ADDIN ENRf8 (1980) found a relationship between lignin content and biodegradable volatile solids using a wide variety of waste materials. The empirical relationship suggests that not only is lignin not easily biodegraded, but that lignin also reduces the biodegradability of the nonlignin components. This reduction in biodegradability may be caused by lignin polymeric material physically preventing enzymatic access to the nonlignin components. The relationship is  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 1 Table  SEQ Table4. Biodegradability of selected components of MSW  ADDIN ENRf8 (Tchobanoglous, Theisen et al. 1993)VS/TSLignin/VSVSbiodegradable*Type of waste%%%mixed food 7150.482newsprint9421.922office paper96.40.482cardboard94.012.947* Obtained by using equation  GOTOBUTTON ZEqnNum887954 \* MERGEFORMAT  REF ZEqnNum887954 \! \* MERGEFORMAT 9.1 where VSbiodegradable is the biodegradable fraction of the volatile solids and VSlignin is the fraction of volatile solids that are lignin. From equation  GOTOBUTTON ZEqnNum188104 \* MERGEFORMAT  REF ZEqnNum188104 \! \* MERGEFORMAT 9.1 the maximum destruction of VS is limited to about 83%, a limitation due to the production of bacterial byproducts. The high concentration of lignin in newsprint makes it much less biodegradable than more highly processed office paper ( REF _Ref386853197 \* MERGEFORMAT Table 4). Energy recovery from MSW Energy could be recovered from MSW by direct combustion in an incinerator or by anaerobic biodegradation and production of methanexe "methane". Proximate analysis is used to measure moisture content, volatile matter, fixed carbon (combustible but not volatile), and ash. Proximate analysis can be used to predict ash production from incineration. The energy content is measured in a bomb calorimeter. Proximate analysis results and energy content of MSW are given in  REF _Ref386852901 \* MERGEFORMAT Table 5. Table  SEQ Table5. Proximate analysis and energy content of selected components of MSW  ADDIN ENRf8 (Tchobanoglous, Theisen et al. 1993).moisturevolatile matterfixed carbonashenergy as collectedenergy dryType of waste%%%%(MJ/kg)(MJ/kg)fats295.32.50.237.538.3mixed food 7021.43.654.213.9fruit waste78.716.640.74.018.6meat waste38.856.41.83.117.729.0cardboard5.277.5123516.417.3magazines4.166.4722.512.212.7newsprint681.111.51.418.619.7mixed paper10.275.98.45.415.817.6waxed cartons3.490.94.51.226.327.3 Gas production from anaerobic digestion is typically 30% CO2 and 70% CH4. The methanexe "methane" is a valuable fuel and has an energy content of 802.3 kJ/mol or 50 MJ/kg. The combustion of methane produces only carbon dioxide and water. Because paper products are a major fraction of MSW and paper energy content is significant, the majority of energy in MSW is contained in paper products. Incineration or methanexe "methane" production can be used to capture some of this available energy.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 2 Effect of MSW particle size The large size of pieces of MSW is suspected to decrease the ability of microbes to degrade the material. Landfill gas production has been correlated with refuse particle size  ADDIN ENRf8 (Ferguson 1993). The effect of particle size reduction was initially explained by the resultant increase in surface area available for microbial attach. Laboratory studies under saturated conditions, however, suggest that size reduction, even down to a few microns or tens of microns has little effect on the rate of degradation. According to Ferguson (1993), surface area increases only slightly with decreasing particle size for platey and fibrous particles such as paper. Thus the effect of size reduction on the methanexe "methane" production in landfills may be that relatively large pieces of plastic, paper, or other material shield the materials beneath them from infiltrating water. The shielded material may remain too dry for biodegradation. Pulverization breaks down the impermeable barriers and more of the waste is exposed to water  ADDIN ENRf8 (Ferguson 1993). Potential methanexe "methane" production from municipal solid waste Under anaerobic conditions microorganisms can produce both CO2 and CH4 (methanexe "methane") without consuming any oxygen. Other significant end products include odorous gases such as ammonia (NH3), and hydrogen sulfide (H2S) (see  REF _Ref386951277 \* MERGEFORMAT Figure 4). Because anaerobic biodegradation produces gas it is possible to monitor the extent and rate of anaerobic biodegradation by measuring gas production  ADDIN ENRf8 (Suflita and Concannon 1995).  Figure  SEQ Figure4. Reactants and products for anaerobic degradation of organic matter. Gas production Because anaerobes get relatively little energy from the organic matter their conversion of carbon to cell material (synthesis) is much lower than for aerobes. Typically 10% of the organic matter may be converted to anaerobe cell mass. Thus the majority of the biodegraded organic matter is converted to gas and the gas production can be used as a measure of biodegradation. The ideal gas law is used to determine the moles of gas produced from the pressure, volume, and temperature.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 3 The pressure in the sealed test bottles that will be used in this laboratory is initially atmospheric. Because the number of moles is a linear function of the pressure we can write  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 4 where "P is the change in pressure relative to the initial pressure in the bottle. In these experiments the bottle volume is 120 mL and the maximum recommended pressure increase is 80 kPa (12 psi). The volume of liquid in the bottles is 20 mL and the volume contributed by solids is expected to be negligible. Thus the nominal volume of gas in the bottles will be 100 mL. Solving for the number of moles of gas (CH4 and CO2) produced by anaerobic digestion  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 5 The molecular formula of cellulose is C6H10O5 and thus 27 g of cellulose has 1 mole of carbon. The relation obtained in equation  GOTOBUTTON ZEqnNum228629 \* MERGEFORMAT  REF ZEqnNum228629 \! \* MERGEFORMAT 9.5 is used to determine the maximum amount of cellulose that can be anaerobicly degraded without exceeding 80 kPa in the bottles.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 6 The mass of paper containing 84 mg of biodegradable cellulose can be obtained using  REF _Ref386853197 \* MERGEFORMAT Table 4 and the results of equation  GOTOBUTTON ZEqnNum692961 \* MERGEFORMAT  REF ZEqnNum692961 \! \* MERGEFORMAT 9.6. The mass of dry newspaper that will produce a pressure increase of 80 kPa is  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 7 Similar calculations can be performed for other types of waste. The maximum mass of glucose (CH2O has 30 g of glucose per mole of carbon) is  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 8 Although glucose is expected to be completely biodegradable, a small amount of glucose will be converted into refractory cell byproducts. The above calculations are based on the assumption that all of the gas produced is volatile and is not dissolved. Carbon dioxide is soluble and thus some of the CO2 produced will be dissolved and will not result in increased pressure. Acid neutralizing capacity requirements The high partial pressure of CO2 resulting from anaerobic biodegradation causes a high concentration of carbonic acid  EMBED Equation.DSMT4  and thus would result in a reduced pH if there were insufficient Acid Neutralizing Capacity (ANCxe "ANC"). The amount of ANC required to counteract the high partial pressure of CO2 can be obtained from the Henrys constant for dissolution of CO2, and from the dissociation constant for carbonic acid.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 9 where KH has a value of 3.12 x 104 moles/J. The first dissociation constant for carbonic acid is  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 10 where K1 has a value of 106.3. The definition of ANCxe "ANC" for a carbonate systemxe "carbonate system" in equilibrium with the gas phase is  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 11 Where a0, a1, a2 are the fractions of total carbonate present as carbonic acid  EMBED Equation.DSMT4 , bicarbonate  EMBED Equation.DSMT4 , and carbonate  EMBED Equation.DSMT4  respectively and Kw is the dissociation constant for water. At circumneutral pH the hydrogen ion, hydroxide ion, and carbonate ion concentrations are negligible and equation  GOTOBUTTON ZEqnNum323452 \* MERGEFORMAT  REF ZEqnNum323452 \! \* MERGEFORMAT 9.11 simplifies to  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 12 The ratio of bicarbonate to carbonic acid may be determined from equation  GOTOBUTTON ZEqnNum669020 \* MERGEFORMAT  REF ZEqnNum669020 \! \* MERGEFORMAT 9.10. Solving for the ratio of bicarbonate to carbonic acid:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 13 Equation  GOTOBUTTON ZEqnNum265391 \* MERGEFORMAT  REF ZEqnNum265391 \! \* MERGEFORMAT 9.13 can be substituted into equation  GOTOBUTTON ZEqnNum808652 \* MERGEFORMAT  REF ZEqnNum808652 \! \* MERGEFORMAT 9.12 to obtain  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 14 An estimate of the ANCxe "ANC" required to maintain a neutral pH under a pressure of 30 kPa of CO2 can be obtained by substituting appropriate values into equation  GOTOBUTTON ZEqnNum680961 \* MERGEFORMAT  REF ZEqnNum680961 \! \* MERGEFORMAT 9.14.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 15 The basal medium that will be used in this laboratory contains 71 meq/L ANCxe "ANC" from sodium bicarbonate. If the pressure of CO2 reaches 60 kPa (30 kPa initial pressure plus 30 kPa from the production of CO2 during an experiment) then solving equation  GOTOBUTTON ZEqnNum775902 \* MERGEFORMAT  REF ZEqnNum775902 \! \* MERGEFORMAT 9.14 for pH shows that (given the 71 meq ANC in the basal medium and a CO2 pressure of 60 kPa) the pH will drop to 6.88. Thus, the basal medium is sufficiently buffered to protect against significant pH changes. Carbon dioxide solubility At pH less than H"9 the inorganic carbon will partition into three species, gaseous  EMBED Equation.DSMT4 , aqueous EMBED Equation.DSMT4 , and aqueous EMBED Equation.DSMT4 .  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 16 The number of moles of the inorganic carbon species can be determined based on the partial pressure of  EMBED Equation.DSMT4 , the ANCxe "ANC" of the liquid and the gas and liquid volumes. The moles of gaseous  EMBED Equation.DSMT4  is obtained from the ideal gas law  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 17 The number of moles of  EMBED Equation.DSMT4  is obtained from the Henrys law constant.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 18 The concentration of bicarbonate,  EMBED Equation.DSMT4 , is equal to the ANCxe "ANC" (for pH < 9).  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 19 The total number of moles of inorganic carbon is the sum of the three species.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 20 Therefore, the number of moles of inorganic carbon in an enclosed volume is a linear function of the partial pressure of CO2 ( REF _Ref387110973 \* MERGEFORMAT Figure 5). The pH will change as the partial pressure of CO2 changes as shown in equation  GOTOBUTTON ZEqnNum775902 \* MERGEFORMAT  REF ZEqnNum775902 \! \* MERGEFORMAT 9.14. Solving for the concentration of hydrogen ions equation  GOTOBUTTON ZEqnNum775902 \* MERGEFORMAT  REF ZEqnNum775902 \! \* MERGEFORMAT 9.14 becomes  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 21 The relationship between pH and partial pressure of CO2 is shown in  REF _Ref387110973 \* MERGEFORMAT Figure 5.  Figure  SEQ Figure5. Number of moles of inorganic carbon and pH as a function of the partial pressure of carbon dioxide given gas and liquid volumes of 60 mL and ANCxe "ANC" of 71 mmoles/L. The basal medium to be used in this experiment will be purged with a 30:70 mixture of carbon dioxide and nitrogen prior to use. As shown in  REF _Ref387110973 \* MERGEFORMAT Figure 5 the pH of the basal medium is expected to rise to approximately 7.17. The headspace will also be purged with the same gas mixture and thus there will be 5.6 mmoles of inorganic carbon in the bottles initially. After the anaerobic biodegradation has gone to completion, the carbon dioxide concentration will be measured by gas chromatographxe "gas chromatograph"y and the gas pressure by pressure sensorxe "pressure sensor"s and thus the partial pressure of carbon dioxide will be known.  REF _Ref387110973 \* MERGEFORMAT Figure 5 or equation  GOTOBUTTON ZEqnNum804712 \* MERGEFORMAT  REF ZEqnNum804712 \! \* MERGEFORMAT 9.20 can be used to determine the final mass of inorganic carbon in the bottles. The difference between the initial and final inorganic carbon concentration can be used to determine the amount of organic carbon converted to carbon dioxide. Methane solubility The Henrys constant for methanexe "methane" ( EMBED Equation.DSMT4 ) at 25C is 1.48 x 105 mol/J  ADDIN ENRf8 (Mackay and Shiu 1981). Methane is significantly less soluble than carbon dioxide and does not form other soluble aqueous species. The mass of gaseous and dissolved methane is given by equations  GOTOBUTTON ZEqnNum817778 \* MERGEFORMAT  REF ZEqnNum817778 \! \* MERGEFORMAT 9.22 and  GOTOBUTTON ZEqnNum914205 \* MERGEFORMAT  REF ZEqnNum914205 \! \* MERGEFORMAT 9.23.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 22  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 23 The ratio of the mass of gaseous to dissolved methanexe "methane" gives an indication of the significance of dissolved methane.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 24 Substituting appropriate values into equation  GOTOBUTTON ZEqnNum378900 \* MERGEFORMAT  REF ZEqnNum378900 \! \* MERGEFORMAT 9.24  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 25 If the gas and liquid volumes are approximately equal there will be approximately 26 times as much methanexe "methane" in the gaseous phase as in the dissolved phase. This ratio is independent of the methane partial pressure. The total number of moles of methane can be obtained from the partial pressure of methane in the gaseous phase.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 26 The partial pressure of methanexe "methane" will be determined from the pressure in the bottle and mass of methane as measured by the gas chromatographxe "gas chromatograph". Temperature effects The temperature of the bottles directly affects the pressure of gas as well as influences the rate of gas production by the microbes. Cummings and Stewart found that methanexe "methane" production was sharply inhibited by temperatures in excess of the optimum (37C) and was undetectable at 20C  ADDIN ENRf8 (1995). However, Suflita and Concannon  ADDIN ENRf8 (1995) reported anaerobic digestion at room temperature over a period of 2 months. If desired a constant temperature water bath can be used to keep all of the digesters at a constant and optimal temperature (35C) for anaerobic degradation. Experiment description The experimental setup is a flexible system for obtaining data on the anaerobic decomposition of various organic materials by measuring the pressure of the gas produced. A schematic of the experimental setup is shown in  REF _Ref386866169 \* MERGEFORMAT Figure 6.  Figure  SEQ Figure6. Experimental setup (not to scale). Pressure generated by microbial gas production is monitored by pressure sensorxe "pressure sensor"s. Bacterial degradation of selected materials will be assayed by placing known quantities in 120 mL bottles, inoculating with an active anaerobic mixed culture, sealing the bottles with rubber septa and aluminum crimp caps, and monitoring gas pressure and composition over time. Anaerobic digester supernatant from the Ithaca Wastewater Treatment Plant will be used as a source of microbes. The bottles will be monitored for biogas production with pressure sensorxe "pressure sensor"s connected with a needle through the septa (see  REF _Ref386866169 \* MERGEFORMAT Figure 6). Gas composition will be determined by periodic analysis of methanexe "methane" (CH4) and carbon dioxide (CO2) via gas chromatographxe "gas chromatograph"y with a thermalconductivity XE "conductivity"  detector. Gas production measurements will be automated by using pressure sensorxe "pressure sensor"s in a procedure comparable to that described by Suflita and Concannon  ADDIN ENRf8 (1995). With this technique, a large number of bottles can be monitored with automated data acquisition by a single computer, allowing a wide variety of chemical and environmental parameters to be explored. The automated acquisition of gas data is necessary due to the numbers of bottles and length of incubation (ca. 4 weeks) anticipated. This experiment will be setup and left virtually unattended while other laboratory exercises continue in intervening weeks. Table  SEQ Table6. Suggested sample preparation. Each group does 2 sample types with 2 replicates. Each section does 2 water and 2 inoculum controls.Sample SizeInoculumReplicatesSample TypemgmL#environmental controlnone02bacterial controlnone52positive control (glucose)9052filter paper5052cardboard?52office paper?52newsprint?52Student selected organic materials ?5up to 4 vials Each sample type should be cut into small enough pieces to easily insert into the bottle. Students may also be interested in exploring biodegradation of other organic components of municipal solid waste (banana peels, rags, plastic bags, etc.).  REF _Ref386952779 \* MERGEFORMAT Table 6 suggests one configuration of several sample types. Each sample will receive 15 mL of basal medium. One control should be unamended (i.e., with no waste) and contain the microbial inoculum and the O2free water to monitor any gas production attributable just to the added sludge, one positive control should be the microbial inoculum, plus 90 milligrams of glucose and the O2free water (to verify that the microbial population is active in the added sludge), and one control should be plain O2free water to control for variations in temperature, and air pressure. The sample sizes of the various samples should be determined so that the bottles will not generate pressure greater than 80 kPa. Experimental methods Safety concerns Municipal wastewater sludge will be used as a source of microbes. The sludge may contain biological and/or chemical hazards and should be handled accordingly. Biological production of gas will generate pressure in a closed container. Testing has shown that this system is safe up to at least 200 kPa (30 psi). At approximately this pressure the needle is typically forced out of the septa. If the bottle is not vented the pressure can increase until the crimp cap is forced off. Bottles should not be capped for very long before the needles are inserted and pressure monitoring begins. The pressure trends should be monitored and, if excessive pressures are produced, the bottles must be vented and/or the temperature of the bath may be reduced. Sharp needles are used in the experimental setup and precautions should be taken to avoid puncturing unintended objects (including students). Analysis of moisture content and volatile solids The fraction of volatile solids in the paper samples is the maximum that could possibly be degraded. Note that paper products cannot be ashed accurately because the strong flames easily carry some of the ashes away. Steps for sample moisture content and volatile solids fraction follow: Weigh an aluminum boat Weigh the aluminum boat with an organic sample (boat + water + VS + ash) Dry in the 105C oven Cool in a desiccator Weigh (boat + VS + ash) Ash in the 550C muffle furnace Cool in a desiccator Weigh (boat + ash) Sample preparation The bottles will be purged initially with a mixture of 30% CO2 and 70% N2 to remove any O2 and to establish an initial carbon dioxide concentration so that the initial pH is not excessively high. If no carbon dioxide were present in the purging gases the carbonic acid would be stripped out of solution and the pH would rise in the basal medium. Acquisition of pressure data The biogas pressure will be measured indirectly by the pressure sensorxe "pressure sensor"s. The specified sensors work from zero to 100 kPa (6.89476 kPa/psi). They will withstand 2.5 times rated pressure (i.e., 250 kPa) but their output may be erroneous above the upper limit of the working range. The output of the pressure sensors is zero to 0.100 volts with 0.100 volts indicating approximately 100 kPa. The outputs of the sensors are fed through a 32-channel multiplexer/signal conditioner, an A/D converter board and are monitored using LabVIEW software. Gas chromatograph analysis for separation of CO2, CH4 and N2 (Optional) Permanent gases can be analyzed using a thermal conductivityxe "conductivity" detector (TCD) on a gas chromatographxe "gas chromatograph". The thermal conductivity detector measures the rate at which heat is transported from the detector. If a gas with a different thermal conductivity than the carrier gas passes through the detector a peak is detected. A flame ionization detector could be used to measure methanexe "methane", but would not be able to detect carbon dioxide or nitrogen since they do not burn. A micropacked column containing packing designed for analyses of permanent gases and light hydrocarbons (Supelco Carboxen 1004) is used to separate the gases. The TCD must be calibrated with known masses of the gases of interest. Nitrogen, carbon dioxide and methanexe "methane" are available as compressed gases and can be sampled at atmospheric pressure by opening a valve in a compressed gas line slightly and sampling the discharge with a gas tight syringe. The ideal gas law is used to calculate the moles of gas. The current atmospheric pressure in Ithaca is available through the World Wide Web at  HYPERLINK "http://cuinfo.cornell.edu/Ithaca/Weather/" http://cuinfo.cornell.edu/Ithaca/Weather/. If the atmospheric pressure is reported in inches of mercury it can be converted to Pascals by multiplying by 3386 Pascals/Inch of Hg. The temperature of the laboratory is available from the pH meters equipped with temperature probes. If a 100 L gas sample is used, the atmospheric pressure is 100 kPa, and the temperature is 22C then the number of moles of gas are calculated as:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 9. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 27 The number of moles of gas is independent of the type of gas. The relationship between peak area and moles of gas is calculated by analyzing a known number of moles of each gas. The TCD response will be different for each gas since the thermal conductivityxe "conductivity" of each gas is different. Experimental method (short version) Dry 2 2 g samples for each sample type in the 105(C oven. Take dried organic sample from the oven. Keep 1 dried sample and determine the VS of the other sample. Weigh appropriate amounts of the various dried samples for methane XE "methane"  production. Load bottles with organic samples (cut to smaller size as needed). Add 15 mL of basal medium to each of the bottles. Add 5 mL of inoculum to each of the bottles. Purge the headspace of the bottles with an oxygenfree gas stream that is 30% CO2 and 70% N2. Seal the bottles. Insert the pressure sensorxe "pressure sensor" hypodermic needle into the bottle. Sample the bottle pressures using the data acquisition software (take samples every hour and save the data as \\Enviro\enviro\Courses\453\methane XE "methane" \pressure). Gas Analysis Method Calibrate the gas chromatographxe "gas chromatograph" using methanexe "methane" and carbon dioxide and using 20 mL samples Take an initial headspace gas sample and analyze it using the gas chromatographxe "gas chromatograph". Sample gas composition after gas production has ceased using the gas chromatographxe "gas chromatograph". Prelab questions Estimate the mass of cardboard and the mass of office paper that will produce a pressure rise of 80 kPa in the sample bottles at 35C if the headspace volume is 100 mL. Use the predicted biodegradability based on the lignin content of the paper. Data analysis Perform the analysis on the data from your lab section. Calculate total gas production in moles. For each sample use the record of pressure vs. time to determine if the reaction appears to have gone to completion. For your samples, compare volatile solids (VS) and gas production by converting the mass of volatile solids to moles of carbon using an approximate molecular formula for the sample. The molecular formula for the volatile fraction of paper can be approximated by C6H10O5. Calculate and plot the fraction of VS degraded as a function of time for each sample. Compare the fuel value of the methanexe "methane" produced with the fuel value of the original sample for each of the samples. Use the estimates of the original fuel value (Table 5) and the measured methane production. The fuel value of glucose is 424.7 KJ/mole C. If you dont have the gas composition of your samples, then assume 70% of the gas produced was methane. Optional Analysis Requiring Gas Composition Calculate the moles of CO2 and CH4 produced by your samples based on the gas chromatographxe "gas chromatograph" analysis. Include the effect of carbon dioxide solubility. Use the basal medium control to subtract the initial headspace as well as any gas production by the inoculum. Calculate the final pressure based on the GC measurements and compare with the pressure transducer measurements. Remember that the pressure transducer measured gage pressure. References  ADDIN ENBib Barlaz, M. A., R. K. Ham, et al. (1992). Microbial chemical and methanexe "methane" production characteristics of anaerobically decomposed refuse with and without leachate recycling. Waste Management & Research 10(3): 257267. Brauns, F. E. (1962). Soluble native lignin, milled wood lignin, synthetic lignin and the structure of lignin. Holzforschung 16: 97102. Brock, T. D. (1970). Biology of Microorganisms. London, PrenticeHall. Chandler, J. A., W. J. Jewell, et al. (1980). Predicting Methane Fermentation Biodegradability. Biotechnology and Bioengineering Symposium No. 10, John Wiley & Sons, Inc. Cummings, S. P. and C. S. Stewart (1995). Methanogenic interactions in model landfillxe "landfill" cocultures with paper as the carbon source. Letters in Applied Microbiology 20(5): 286289. DiStefano, T. D. (1992). Biological dechlorination of tetrachloroethene under anaerobic conditions, Cornell University. Ferguson, C. C. (1993). A hydraulic model for estimating specific surface area in landfillxe "landfill". Waste Management & Research 11(3): 227248. Gossett, J. M. and P. L. McCarty (1976). Heat Treatment of Refuse for Increasing Anaerobic Biodegradability. AIChE Symposium Series 158(72): 6471. Gossett, J. M., D. C. Stuckey, et al. (1982). Heat Treatment and Anaerobic Digestion of Refuse. Journal of the Environmental Engineering Division 108: 437454. Gurijala, K. R. and J. M. Suflita (1993). Environmental factors influencing methanogenesis from refuse in landfillxe "landfill" samples. Environmental Science and Technology 27(6): 11761181. Mackay, D. and W. Y. Shiu (1981). A critical review of Henry's law constants for chemicals of environmental interest. Journal of Physical Chemical Reference Data 10(4): 11751199. Metcalf & Eddy, I. (1991). Wastewater Engineering. Treatment, Disposal, and Reuse. New York, McGrawHill, Inc. Pavlostathis, S. G. and J. M. Gossett (1985). Alkaline Treatment of Wheat Straw for Increasing Anaerobic Biodegradability. Biotechnology and Bioengineering 27: 334344. Pavlostathis, S. G. and J. M. Gossett (1985). Modeling Alkali Consumption and Digestibility Improvement From Alkaline Treatment of Wheat Straw. Biotechnology and Bioengineering 27: 345354. Pearl, I. A. (1967). The Chemistry of Lignin. New York, NY, Marcel Dekker, Inc. Prescot, L. M., J. P. Harley, et al. (1993). Microbiology. Dubuque, IA, Wm. C. Brown, Publ. Suflita, J. M. and F. Concannon (1995). Screening tests for assessing the anaerobic biodegradation of pollutant chemicals in subsurface environments. Journal of Microbiological Methods 21: 267281. Tchobanoglous, G., H. Theisen, et al. (1993). Integrated Solid Waste Management. Engineering Principles and Management Issues. New York, McGrawHill, Inc. Zehnder, A. J. B. (1978). Ecology of methanexe "methane" formation. Water Pollution Microbiology. R. Mitchell. New York, John Wiley & Sons. 2: 349376. Lab Prep Notes Table  SEQ Table7. Equipment list DescriptionVenderCatalog500 l syringe w/ valveSupelco22272side port needleSupelco22289Carboxen 1004 micropacked columnSupelco1-2846Hp 5890 Series II GCHewlettPackard5890ATCD kitHewlettPackard19232E1/8" column adapterHewlettPackardoption 095pressure regulatorsHewlettPackardL43RS232C boardHewlettPackardoption 560HeliumCornell StoresWrist action ShakerFisher Scientific14260VialsSupelco3-3111Aluminum crimp topsFisher03-375-23CButyl stopperFisher03-375-22AACrimping toolSupelco3-3280EPDM stoppers13x20 mmSigma Z166073luer lock needles 21 gaugeFisher148265BPressure transducer, 0 to 15 psigOmegaPX136-015GV12 V DC Power supplyOmegaPSS-12IncubatorFisher11690650DMultiplexerNational Instruments4 slot chassis SCXI10007765700132 channel SCXI110077657200SCXI1200 parallel port77678300Setup Use anaerobic digester supernatant as inoculum source. Place supernatant under fume hood. Use 5 mL per sample. Setup 10 port purger with CO2 and N2 gas metered through rotometers. The top ball should be at 24 mm for CO2 and at 84 mm for N2. Set the GC with 300 Kpa column pressure, 180C oven, 250C injector and detectors, and 1.2 minute run time. Use 20 mL sample. The gases should come out in the order N2, CH4, and CO2 at 0.44, 0.72, and 1 minute respectively. (Only if you are doing the optional GC analysis) 4 samples/group plus 2 inoculum blanks and 2 water blanks. Class Plan Sign up for samples Each group chooses 2 types of samples Dry samples in oven Ash 1 of the 2 samples Table  SEQ Table8. Reagents ReagentVenderCatalogbasal mediumNAglucoseAldrich15,8968paper (various types)NAWhatman Filter Paper (No. 1)Fisher Scientific098051A Table  SEQ Table9. Basal medium for anaerobic growth  ADDIN ENRf8 (DiStefano 1992). CompoundQuantity (per liter)NH4Cl200 mgK2HPO43H2O100 mgKH2PO455 mgMgCl26H2O200 mgResazurin1 mgFeCl24H2O100 mgTrace Metals Solution10 mLNa2S9H2O500 mgNaHCO36 gThe first six compounds are added to distilleddeionized water, then purged with N2 until solution turns from blue to pink. The remaining components are added, followed by a 15minute purge with the 70% N2/30% CO2 gas mixture.Table  SEQ Table10. Trace metals for anaerobic growth  ADDIN ENRf8 (DiStefano 1992).CompoundQuantity (mg/L)MnCL24H2O100CoCl26H2O170ZnCl2100CaCl22H2O251H3BO319NiCl26H2O50Na2MoO42H2O20 Table  SEQ table11. Gas chromatograph conditions gaspressureflowcarrier (He) kPa5 mL/minRef15 mL/mintemperaturesCoven (isothermal)Injector250TCD250ColumnSupplierCatalog numberCarboxen 1004 micropacked columnSupelco1-2846Volatile Organic Carbon Contaminated Site Assessment Introduction  MACROBUTTON MTEditEquationSection Equation Section (Next) SEQ MTEqn \r \h \* MERGEFORMAT  SEQ MTSec \h \* MERGEFORMAT Roughly 75 percent of the major cities in the U.S. depend, at least in part, on groundwaterxe "groundwater" for their water supply. Various estimates of the nationwide extent of groundwater contamination are stated to range from one to over two percent of the nation's usable groundwater (Council on Environmental Quality, 1981). Volatile organic compounds (VOCs) are the most frequently detected organic pollutants of groundwater in the United States. In fact, the VOCs are so ubiquitous that their analysis has been considered by the U.S. Environmental Protection Agency as a screening procedure to establish the need for more extensive characterization of groundwater samples from hazardous waste disposal sites. In the upstate region of New York (excluding Long Island), of approximately 570 groundwater contamination incidents reported by 1985, 98% involved either the volatile components of gasolinexe "gasoline" and petroleumxe "petroleum" or solventsxe "solvents" and degreasers (NY State DEC, 1985).  Figure  SEQ \r 1 Figure1. Subsurface VOC transport processes. The vadose zone is the region of the soil profile in which some pores contain gas and are therefore, unsaturated. Volatile organics may be transported in the subsurface as dissolved components in groundwaterxe "groundwater". However, by virtue of their volatility, VOCs will also exist in the gas phase of unsaturated porous mediaxe "porous media". As a result, volatile contaminants can be transported by advection and diffusionxe "diffusion" in the vapor phase. VOC transport processes are illustrated in  REF _Ref365247940 \* MERGEFORMAT Figure 1. Experiment Description Students will use soil gas samplingxe "soil gas sampling" to prospect for a VOC that has leaked from a subsurface source into an unsaturated soil system. A rectangular soil box contaminated with a combination of liquid acetone, octane and toluene will be used. A soil with high organic content (potting soil) or low organic content (sand) may be used as the box filling material (porous medium). The VOCs will be identified and measured using a gas chromatographxe "gas chromatograph" (GC). Experimental Procedures Calibration (Peak Times) Each compound will have a unique retention time in the gas chromatographxe "gas chromatograph". The time for each of the 3 VOC peaks can be obtained by injection of 100 l head space samples from crimp cap sealed vials containing a small volume liquid acetone, octane, and toluene. Use the syringe technique described below. Analyze each compound 4 times (12 samples) using a gas chromatograph (see page  PAGEREF _Ref406553492 \h 160 for information on using the gas chromatograph). These analyses will also serve to calibrate the GC by generating the peak area that corresponds to the saturated vapor concentration. Gas chromatogram peak areas may be assumed to be directly proportional to the mass of vapor injected. Syringe technique for sampling vial headspace The purpose of this syringe technique is to minimize the effects of sorptionxe "sorption" to the Teflon and glass surfaces in the syringe and to eliminate carryover of sample in the needle. Using separate needles to collect samples and to inject into the GC eliminates needle carryover of sample. Remove GC needle. Purge syringe 5 times with room air to remove any residual VOCs. Put on sample needle. Insert into sample bottle (with syringe at zero volume) Fill syringe fully with gas, wait 4 seconds, and purge syringe contents back into the source bottle (repeat 3 times). Fill syringe and adjust to 100 L. Close syringe valve and remove syringe from sample vial and remove sample needle. Put on GC needle. Instruct GC to measure sample (see page  PAGEREF _Ref406553465 \h 160 for information on using GC software). Insert needle in injection port, open syringe valve, inject sample, hit start button all as quickly as possible. Remove syringe from the GC injection port. Soil Moisture Content The dry weight of moist soil may be readily determined by placing H" 5 g moist soil into a tarred aluminum weighing pan. Weigh the pan and its contents to obtain the soil s wet weight, and place the pan into a 105oC oven for e" 1 hour. Remove the soil from the oven and place in a desiccator and allow it 5 minutes to cool. Weigh the cooled soil to obtain the dry weight. The moisture content is  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 10. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 1 Soil Density To determine the approximate density of the soil, rsoil, place a weighed quantity of dry soil (H" 30 g) into a 100 mL graduated cylinder containing 60 mL water. Record the volume occupied by the water plus the soil.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 10. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 2 where rsoil = rb/q, (b= bulk density of the soil and q = fraction of void volume. Soil Gas Sampling Table  SEQ \r 1 table1. Physical data for octane, acetone, and toluene. OctaneAcetoneTolueneAqueous solubility (g/m3)0.6very515Vapor Pressure (kPa)1.88 (1.47)243.8 (2.9)H (kPa m3/mol)3000.01590.67 EMBED Equation.DSMT4  (g/L)/(g/L)1230.00650.275Molecular FormulaCH3(CH2)6CH3CH3COCH3C6H5CH3Molecular weight114.2358.0892.14density (g/mL)0.710.78570.8669 See Table 1 for physical properties of the VOCs. See Tables 2, 3, and 4 (in the Lab Prep Notes) for necessary reagents, equipment and GC method. Prior to the laboratory the instructor will create a spill of a VOC by injecting 10 mL of liquid of two or more NAPLs into the soil box to be sampled by students. During the lab students will analyze approximately 50 soil gas samples from the soil box using the syringe technique outlined below. Results from the soil box analyses may be mapped using units of concentration (g/m3). Syringe technique for soil gas samplingxe "soil gas sampling" Remove GC needle. Purge syringe 10 times with room air to remove any residual VOCs. Put on sample needle. Insert into soil bed (with syringe at zero volume). Fill syringe and adjust to 100 l. Close syringe valve, remove syringe from soil bed and remove sample needle. Put on GC needle. Instruct GC to measure sample (using software). Insert needle in injection port, open syringe valve, inject sample, hit the enter key (or OK) all as quickly as possible. Remove syringe from the GC injection port. Analysis of Soil Gas Sampling Students will use their analysis of VOC standards to obtain the corresponding GC retention times and use this information to identify the unknown VOCs in the contaminated soil box. The vapor pressure and ideal gas law are used to estimate the mass of each compound present in the samples used to calibrate the GC.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 10. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 3 where n is the number of moles of the compound, P is the vapor pressure of the compound [kPa], V is the syringe volume [L], R is the Gas Constant (8.31  EMBED Equation.DSMT4 ), and T is the temperature of the gas in the syringe [K]. The relationship between peak area (as measured by the GC) and mass of the compound is determined from the calibration. Soil gas concentrations should be reported and plotted as contour lines on a map of the soil box (see  REF _Ref365711727 \* MERGEFORMAT Figure 2 for an illustration).  Figure  SEQ Figure2. Students will prepare a map of the surface of their soil box. The map will show isoconcentration lines for each VOC. Procedure (short version) Instructor will demonstrate syringe technique (be careful not to pull plunger out of barrel) and Gas Chromatograph technique. Place H"5 g of accurately weighed soil in oven to determine moisture content. (Weigh both the empty dish and the soil + dish.) Calibrate GC by analyzing 4 samples for each VOC. Take soil gas samples. Convert the soil gas peak areas to concentrations (g/m3). This data will be shared between groups. Finish soil moisture contentxe "soil moisture content" measurement (cool dry soil in desiccator and then weigh). Measure soil densityxe "soil density" using dry soil. Pour waste potting soil into designated waste container. Clean plasticware. Prelab Questions How are the identities of the chromatogram peaks determined when using a gas chromatographxe "gas chromatograph"? Explain why different needles are used for sampling from source vials and injecting the sample into the GC. Consider the temperature of the injection port (see Table 4) and the fact that these compounds absorb to most surfaces. Data Analysis Calculate the mass of each VOC in 100 (L of headspace. Calculate the concentration of saturated vapor for each compound in g/m3. Plot isoconcentration lines of the identified VOCs (expressed as gas concentration in g/m3) on maps of the contaminated site (see figure 2 for example). Prepare a map for each compound showing isoconcentration lines. (The Excel 3D surface plot with contour lines can be used. Note that the grid needs to have uniform distance between samples for the Excel 3D surface plot to work correctly.) Compare the saturated vapor concentration with the peak concentration observed in the sand box. Calculate the soil moisture contentxe "soil moisture content" and density. References Ashworth, R. A., G. B. Howe, and T. N. Rogers. AirWater Partitioning Coefficients of Organics in Dilute Aqueous Solutions. J. Hazard. Mater. 18, p. 2536, 1988. Council on Environmental Quality, "Contamination of Groundwater by Toxic Organic Chemicals", 1981. Hwang, Y., J. D. Olson, and G. E. Keller, II, Steam Stripping for Removal of Organic Pollutants from Water. 2. VaporLiquid Equilibrium Data. Ind. Eng. Chem. Res. 31, p. 17591768, 1992. Mackay, D. and W. Y. Shiu, A Critical Review of Henrys Law Constants for Chemicals of Environmental Interest, J. Phys. Chem. Ref. Data. 10, p. 11751199, 1981. New York State Department of Environmental Conservation, "Draft Upstate New York Groundwater Management Program", N.Y.S.D.E.C., Division of Water, Draft Report WM P94, January, 1985. Lab Prep Notes Table  SEQ table2. Reagents list DescriptionSourceCatalog numberOctaneFisher Scientific030081AcetoneFisher ScientificO2991tolueneFisher ScientificT324500Potting soilAgway (remove large particles by screening to 2 mm) Table  SEQ table3. Equipment list DescriptionSupplierCatalog number500 l syringe w/ valveSupelco22272side port needleSupelco222891 mL syringe w/ valveSupelco22273Hp 5890 Series II GCHewlettPackard5890ASep purgepacked/FIDHewlettPackardoption 6001/8" column adapterHewlettPackardoption 095pressure regulatorsHewlettPackardL43RS232C boardHewlettPackardoption 560Nitrogen, Air, and Hydrogen gasGeneral StoresWrist action ShakerFisher Scientific14260DesiccatorFisher Scientific0864215VialsSupelco3-3111Aluminum crimp topsSupelco3-3220SeptaSupelco3-3200Crimping toolSupelco3-3280Setup Prepare 1 soil box under fume hood. Moisten the sand but not so much that there is standing water. Pipette 10 mL of liquid acetone, octane, and toluene in sand box and record injection locations. This should be done in the morning before the lab exercise. Dry approximately 100 g of potting soil for each group that will be used for density determinations. Replace injection port septa on both GCs. Verify that GCs are working properly by injecting gas samples from each VOC source bottle. If the baseline is above 30 (as read on the computer display) then heat the oven to 200C to clean the column. Verify that sufficient gas is in the gas cylinders (hydrogen, air, nitrogen). Prepare VOC source vials that contain liquid acetone, octane, and toluene (they can be shared by two groups of students). Class Plan Setup uniform spreadsheets for data entry Make sure spreadsheet is completely filled out by end of lab Table  SEQ table4. Gas chromatograph conditions gaspressureflowcarrier (N2)320 kPa15 mL/minAir230 kPa300 mL/minHydrogen130 kPa45 mL/mintemperaturesCoven (isothermal)100Injector250FID250ColumnSupplierCatalog numberSupelcowax 10 30 metersSupelco25301Run length of 66 seconds with octane, acetone, and toluene at 0.57, 0.63, 0.96 minutes respectively. Maximum sample volume is about 100 l. Larger samples can lead to a significant broadening of the peak. Syringe clean up Disassemble and heat syringes to 45C overnight to remove residual VOCs. Place syringe barrels upside down on top of openings above fan in oven to facilitate mass transfer. Volatile Organic Carbon Sorption to Soil Introduction  MACROBUTTON MTEditEquationSection Equation Section (Next) SEQ MTEqn \r \h \* MERGEFORMAT  SEQ MTSec \h \* MERGEFORMAT Volatile organic carbon compounds (VOCs) can exist as vapors, nonaqueous phase liquids, dissolved in water, or sorbed to surfaces. Sorption is the term used to refer to the binding reactions between organic pollutants and the subsurface medium. Sorption slows the rate of transport of both dissolved and volatile organic pollutants. Laboratory experiments will be performed to evaluate the sorptionxe "sorption" of acetone, hexane, and octane vapors to a soil and to estimate the extent to which VOC transport is slowed by binding to the soil media. Theory GasLiquid Partitioning The equilibrium between gas and solution is described by the following reaction:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 1 The equilibrium constant ( EMBED Equation.DSMT4 ) for the reaction is given by:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 2 where CG is the concentration in the gas phase (Example units: g/m3) and CL is the concentration in the aqueous phase (g/m3) Alternately, the liquidvapor equilibrium is sometimes expressed as:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 3 where PG is the partial pressure of the gas (atm), and  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 4 where R is the universal gas constant  EMBED Equation.DSMT4  and T is the absolute temperature (oK). Both H and  EMBED Equation.DSMT4  are referred to as Henry's law constants and may be viewed conceptually as distribution coefficients for gases between the aqueous solution phase and the vapor phase. All other factors being equal, VOCs with higher Henry's law constants will have a greater fraction of their total mass in the gas phase of an unsaturated porous medium. LiquidSolid Partitioning The sorptionxe "sorption" of organic pollutants that are dissolved in water onto soils and aquifer materials also may be described in many cases by a distribution coefficient ( EMBED Equation.DSMT4 ):  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 5 where G is the mass of solute sorbed per mass of solid. Equation  GOTOBUTTON ZEqnNum407012 \* MERGEFORMAT  REF ZEqnNum407012 \! \* MERGEFORMAT 11.5 predicts that the amount of pollutant bound to the soil (G) will increase linearly with the concentration in the aqueous phase (CL). Any relationship between the amount bound to the soil and the concentration in solution applies at a constant temperature and is referred to as an isotherm. Equation  GOTOBUTTON ZEqnNum703957 \* MERGEFORMAT  REF ZEqnNum703957 \! \* MERGEFORMAT 11.5 is an example of a linear isotherm. The success of the linear isotherm in describing sorptionxe "sorption" of nonionic organic pollutants in saturated soils has been remarkable. Linear isothermsxe "isotherms" have been found to describe sorption of a wide array of nonionic compounds onto sediments and soils (Karickhoff et al. 1979, Chiou et al. 1979). Many investigations have demonstrated that the distribution coefficient ( EMBED Equation.DSMT4 ) for sorptionxe "sorption" of a single organic contaminant onto a variety of soil materials can be related to the organic content of the sorbent. This observation permits the definition of an organic normalized partitioning coefficient (Koc):  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 6 where foc is the weight fraction of organic carbon in the soil. Koc values for a range of different organic compounds have been shown to be related to their octanolwater partitioning coefficients (Kow) and also to their aqueous solubilities (Karickhoff, 1984). An important implication of these results is that sorptive distribution coefficients of organic pollutants in water saturated aquifers ( EMBED Equation.DSMT4 ) may be predicted given knowledge of the organic content of the soil (foc) and the octanolwater partitioning coefficient (Kow) of the contaminant or a related parameter such as its aqueous solubility. GasSolid Partitioning Sorption of gases is frequently described using the classic equation developed by Brunauer, Emmett and Teller (1938), i.e., the BETxe "BET" equation:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 7 where G is the amount of sorbed gas per unit of surface (with units such as moles/m2 or g/g if the surface area is not known), GM is the amount of sorbed gas corresponding to monolayer surface coverage, P is the partial pressure of the sorbed gas, Po is its saturated vapor pressure, and B is a parameter related to solute binding intensity; more specifically:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 8 where e is the energy of the adsorbate vaporadsorbent surface interaction (cal/mole), ev is the vaporization energy of the organic (cal/mole), k is the Boltzmann constantxe "Boltzmann constant", and T is the absolute temperature (oK). To the extent that it is valid in soil, the BETxe "BET" equation predicts vapor sorptionxe "sorption" isothermsxe "isotherms" to be nonlinear. Nonlinear isotherms have been observed for sorption of organic vapors onto dry soils at high vapor concentrations by several investigators (Chiou, 1990; Rhue, et al., 1988; and Ong and Lion, 1991c). Under conditions of low vapor pressure, P << Po, the BET equation reduces to:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 9 which is the Langmuirxe "Langmuir" adsorptionxe "adsorption" equation that applies to monolayer limited adsorption. The BETxe "BET" equation further reduces to a linear isotherm when B << (Po/P).  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 10  Figure  SEQ \r 1 Figure1. Linear, Langmuirxe "Langmuir" and BETxe "BET" sorptionxe "sorption" isothermsxe "isotherms" for the case where Gm = 10,000 and B = 1,000. A) Isotherms for P/Po < 0.3. B) Isotherms for P/Po up to 1.0.  REF _Ref365247940 \* MERGEFORMAT Figure 1 illustrates linear, Langmuirxe "Langmuir" and BETxe "BET" isothermsxe "isotherms" that share a common set of B and Gm values. Linear sorptionxe "sorption" isotherms for vapors are a reasonable expectation at low vapor concentrations (Ong and Lion, 1991a). At higher vapor concentrations, surface site limitations and the phenomenon of vapor condensation at the surface (for which the BETxe "BET" model attempts to account) will result in nonlinear VOC sorptionxe "sorption" isothermsxe "isotherms". Results obtained at Cornell (Ong et al., 1991; Ong and Lion, 1991c) have confirmed that condensation of organic vapors will occur at high vapor pressures in moist porous mediaxe "porous media". Condensed water and organic vapors compete for the available pore space. Since soils are generally wet prior to the introduction of organic contaminants, vapor condensation is expected to be limited to the pore volume not occupied by water. The extent to which organic vapor condensation is significant will, therefore, be a function of the soil moisture contentxe "soil moisture content", and the relative vapor concentration (P/P0). Since the unsaturated zone in an aquifer will typically contain condensed water, description of the sorptionxe "sorption" of organic vapors in the unsaturated zone must, at a minimum, consider a binary mixture of the organic and water vapor. The organic vapor of concern may accumulate in the unsaturated zone through at least three processes: (a) by direct sorption from the vapor phase, including vapor sorption to dry mineral surfaces (if present), vapor sorption at the gaswater interface, and vapor condensation, (b) by solubilization in condensed pore water as governed by Henry's Lawxe "Henry's Law" (equation  GOTOBUTTON ZEqnNum665083 \* MERGEFORMAT  REF ZEqnNum665083 \! \* MERGEFORMAT 11.2 or  GOTOBUTTON ZEqnNum420407 \* MERGEFORMAT  REF ZEqnNum420407 \! \* MERGEFORMAT 11.3), and (c) by sorption from condensed porewater solution onto the soil (as governed by equation  GOTOBUTTON ZEqnNum840102 \* MERGEFORMAT  REF ZEqnNum840102 \! \* MERGEFORMAT 11.5). Since, at very low vapor pressures, a linear isotherm is expected to govern vapor sorption, we may write a linear isotherm in terms of the gas concentration:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 11 where the magnitude of  EMBED Equation.DSMT4  depends on the sorbent s moisture content. The relative contributions to G of processes such as vapor dissolution into soil moisture and sorptionxe "sorption" at the liquidair interface can be assessed through the use of a mass balancexe "mass balance". The total mass of vapor sorbed onto the soil, under equilibrium conditions, can be viewed as being distributed between: (Mass sorbed at the solid-liquid interface) + (Mass dissolved in the liquid phase) + (Mass sorbed at liquid-air interface + condensation)  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 12 where Ms is the mass of the sorbent (soil or other porous mediaxe "porous media") and w is a lumped parameter that includes the effects of water surface sorptionxe "sorption" and condensation. From equation  GOTOBUTTON ZEqnNum607595 \* MERGEFORMAT  REF ZEqnNum607595 \! \* MERGEFORMAT 11.5 for liquidphase sorptionxe "sorption":  EMBED Equation.DSMT4  where G = X/Ms and Ms is the mass of the sorbent. Also, from Henry's Lawxe "Henry's Law":  EMBED Equation.DSMT4 . Substituting these two relationships and equation  GOTOBUTTON ZEqnNum462162 \* MERGEFORMAT  REF ZEqnNum462162 \! \* MERGEFORMAT 11.11 into equation  GOTOBUTTON ZEqnNum156525 \* MERGEFORMAT  REF ZEqnNum156525 \! \* MERGEFORMAT 11.12 gives:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 13 If the density of waterxe "density of water" on the soil surface is expressed as  EMBED Equation.DSMT4 , the term VL/Ms becomes  EMBED Equation.DSMT4 . Assuming water surface sorptionxe "sorption" and condensation are negligible (w H" 0), then for high moisture content equation  GOTOBUTTON ZEqnNum927428 \* MERGEFORMAT  REF ZEqnNum927428 \! \* MERGEFORMAT 11.13 will plot as a straight line, with the ordinate intercept equal to the contribution of aqueous phase sorption to vaporphase partitioning ( EMBED Equation.DSMT4 ).  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 14 Equation  GOTOBUTTON ZEqnNum474253 \* MERGEFORMAT  REF ZEqnNum474253 \! \* MERGEFORMAT 11.14 indicates that the linear vapor distribution coefficient,  EMBED Equation.DSMT4 , can be predicted from the saturated distribution coefficient and the Henrys law constant for a given VOC. Experiments by Ong and Lion (1991a) have shown that such predictions are reasonable as long as the moisture content of the soil is high enough to ensure that the VOC that is dissolved in sorbentbound water behaves as if the liquid were comparable to the water in a bulk aqueous phase. In general, a moisture content equivalent to an average surface coverage of H" 5 layers of water molecules is adequate for this assumption to be obeyed (Ong and Lion, 1991a). Many soil ambient moisture contents are in excess of this limiting value. Pollutant Transport in Porous Media The advective dispersionxe "advective dispersion" equation is used to describe pollutant movement in porous mediaxe "porous media". For onedimensional (ex., horizontal) transport of a conservative ( EMBED Equation.DSMT4 ) pollutant:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 15 where t is time, x is distance, U is the groundwaterxe "groundwater" pore velocity and E is the macroscopic dispersion coefficientxe "dispersion coefficient" (Freeze and Cherry, 1979). Since volatile organic pollutants react with the surfaces of the porous mediaxe "porous media" through which the contaminant flows, equation  GOTOBUTTON ZEqnNum328802 \* MERGEFORMAT  REF ZEqnNum328802 \! \* MERGEFORMAT 11.15 must be modified to account for the sorptionxe "sorption" reaction by addition of the term:  EMBED Equation.DSMT4 where  EMBED Equation.DSMT4  is the bulk density of the porous medium (g/cm3), and q is the volumetric moisture content (volume of liquid per unit bulk volume of the porous medium). q is equal to the porosity, f, in a saturated porous medium. From the chain rule,  EMBED Equation.DSMT4 and for a linear isotherm,  EMBED Equation.DSMT4 . Therefore, the advectiondispersion equation for a compound that experiences sorptive binding to the soil matrix becomes:   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 16   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 17 The retardation factorxe "retardation factor", R, for a pollutant in soil is defined as:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 18 Therefore the advective dispersionxe "advective dispersion" equation modified for sorptive binding becomes:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 19 From equation  GOTOBUTTON ZEqnNum112246 \* MERGEFORMAT  REF ZEqnNum112246 \! \* MERGEFORMAT 11.19 it is apparent that the velocity and dispersion of a sorbed compound will be reduced by the magnitude of retardation factorxe "retardation factor", R. So,  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 20 Note that R may be determined directly from knowledge of the medium properties ( EMBED Equation.DSMT4  and q) and from the distribution coefficient for sorptionxe "sorption" ( EMBED Equation.DSMT4  for water saturated media or  EMBED Equation.DSMT4  for a vapor). Interestingly, equation  GOTOBUTTON ZEqnNum937247 \* MERGEFORMAT  REF ZEqnNum937247 \! \* MERGEFORMAT 11.19 may also be applied to describe vapor movement in a gas chromatographxe "gas chromatograph" (GC). GC columns are selected to ensure that the components of a vapor mixture will be separated (by virtue of their different retardation factorxe "retardation factor"s) by the time they arrive at the GC detector (situated at the end of the column). In the absence of pressure gradients, transport of vapors will occur primarily through the process of diffusionxe "diffusion" and equation  GOTOBUTTON ZEqnNum564561 \* MERGEFORMAT  REF ZEqnNum564561 \! \* MERGEFORMAT 11.19 reduces to:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 21 where Es is the effective diffusionxe "diffusion" coefficient of the VOC in the porous mediaxe "porous media". Vapor diffusionxe "diffusion" coefficients in unsaturated porous mediaxe "porous media" are different from those in a bulk gas phase because the vapor must follow a tortuous path to move through the open pores. The relationship proposed by Millington and Quirk (1961) is commonly used to correct vapor diffusion coefficients for the conditions in the soil media.   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 22 where Ebulk is the vapor diffusionxe "diffusion" coefficient in air, a is the volumetric air content of the porous medium (volume of gas per unit bulk volume of medium), and f is the porosity (a + q = f). Analysis of the Unsaturated Distribution Coefficient ( EMBED Equation.DSMT4 ) A mass balancexe "mass balance" calculation will be used to determine the unsaturated vapor distribution coefficient ( EMBED Equation.DSMT4 ). After equilibration the VOC mass will be distributed between the vapor phase and the solid phase (sorbed VOC).  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 23 where  EMBED Equation.DSMT4  is the mass of sorbed VOC and equals  EMBED Equation.DSMT4  (from equation  GOTOBUTTON ZEqnNum659832 \* MERGEFORMAT  REF ZEqnNum659832 \! \* MERGEFORMAT 11.11). In control bottles there is no sorbent so the VOC mass must reside entirely in the vapor phase:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 24 where the subscript uc indicates that volumes and concentrations are those measured in the unsaturated control bottles. Setting equations  GOTOBUTTON ZEqnNum768496 \* MERGEFORMAT  REF ZEqnNum768496 \! \* MERGEFORMAT 11.23 and  GOTOBUTTON ZEqnNum211735 \* MERGEFORMAT  REF ZEqnNum211735 \! \* MERGEFORMAT 11.24 equal to one another (the mass of VOC was the same for all vials) and rearranging gives:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 25 Solving for  EMBED Equation.DSMT4   EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 26 Using the measured soil moisture contentxe "soil moisture content"s and values of  EMBED Equation.DSMT4 , students may check the validity of equation  GOTOBUTTON ZEqnNum608428 \* MERGEFORMAT  REF ZEqnNum608428 \! \* MERGEFORMAT 11.14. Unsaturated Mass Fraction Distribution The total mass of the VOC is distributed between the gas and sorbed phases (equation  GOTOBUTTON ZEqnNum546550 \* MERGEFORMAT  REF ZEqnNum546550 \! \* MERGEFORMAT 11.23).  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 27 where  EMBED Equation.DSMT4  is the fraction of the VOC mass in the gas phase and  EMBED Equation.DSMT4  is the fraction of the VOC mass sorbed to the soil. The relationship between the fraction of VOC in each phase is obtained from the definition of the unsaturated distribution coefficient (equation  GOTOBUTTON ZEqnNum568284 \* MERGEFORMAT  REF ZEqnNum568284 \! \* MERGEFORMAT 11.11).  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 28 Thus we have two equations in two unknowns. Solving we obtain  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 29  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 30 where  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 31 Determination of  EMBED Equation.DSMT4  requires a measurement of the fraction sorbed given measurements of the total mass added and the mass in the gas phase. This analysis becomes inaccurate as the magnitude of  EMBED Equation.DSMT4  decreases and approaches the coefficient of variationxe "coefficient of variation" of  EMBED Equation.DSMT4 . Analysis of the Saturated Distribution Coefficient ( EMBED Equation.DSMT4 ) A mass balancexe "mass balance" calculation will be used to determine the saturated vapor distribution coefficient ( EMBED Equation.DSMT4 ). The calculation assumes that students can reproduce the introduction of the mass of VOC (MVOC) into each sample bottle. After equilibration the VOC mass can be distributed between the vapor phase the aqueous phase and the solid phase (sorbed VOC).  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 32 where GMs is the mass of sorbed VOC and equals  EMBED Equation.DSMT4  (from equation  GOTOBUTTON ZEqnNum866166 \* MERGEFORMAT  REF ZEqnNum866166 \! \* MERGEFORMAT 11.5). In saturated control bottles there is no sorbent so the VOC mass must just be distributed between the vapor phase and the aqueous phase:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 33 where the subscript sc indicates that volumes and concentrations are those measured in the control bottles. Henrys law (equation  GOTOBUTTON ZEqnNum926326 \* MERGEFORMAT  REF ZEqnNum926326 \! \* MERGEFORMAT 11.2) can be used to eliminate  EMBED Equation.DSMT4 .  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 34 Equation  GOTOBUTTON ZEqnNum784778 \* MERGEFORMAT  REF ZEqnNum784778 \! \* MERGEFORMAT 11.34 can be used to obtain an estimate of Henrys law constant by assuming that the mass of VOC added is the same for the vials with and without water. Solving for  EMBED Equation.DSMT4  and substituting  EMBED Equation.DSMT4  for  EMBED Equation.DSMT4  we obtain:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 35 Equation  GOTOBUTTON ZEqnNum300893 \* MERGEFORMAT  REF ZEqnNum300893 \! \* MERGEFORMAT 11.32 can now be used to obtain an estimate of  EMBED Equation.DSMT4 . The mass of VOC added to the bottles was the same for all vials. Thus ( EMBED Equation.DSMT4 ) in equation  GOTOBUTTON ZEqnNum300893 \* MERGEFORMAT  REF ZEqnNum300893 \! \* MERGEFORMAT 11.32 is equal to  EMBED Equation.DSMT4 . In addition, CL and CG are interrelated through Henrys law (equation  GOTOBUTTON ZEqnNum610766 \* MERGEFORMAT  REF ZEqnNum610766 \! \* MERGEFORMAT 11.2). Substituting into equation  GOTOBUTTON ZEqnNum791109 \* MERGEFORMAT  REF ZEqnNum791109 \! \* MERGEFORMAT 11.32 gives:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 36 Solving for  EMBED Equation.DSMT4   EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 37 Measured VOC headspace concentrations are substituted into equation  GOTOBUTTON ZEqnNum412774 \* MERGEFORMAT  REF ZEqnNum412774 \! \* MERGEFORMAT 11.37. Values for Henrys law constants are given in Table 1.  EMBED Equation.DSMT4  is obtained from equation  GOTOBUTTON ZEqnNum137587 \* MERGEFORMAT  REF ZEqnNum137587 \! \* MERGEFORMAT 11.34. In vials containing soil, VG is determined by subtracting both VL (50 mL) and the volume of the sorbent from the total vial volume. The volume of the sorbent may be calculated from the sorbent weight and density as:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 38 It is instructive to calculate the phase distribution of each VOC in bottles that contain no soil. The fraction () of VOC mass in the gas phase is given by  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 39 Assuming the total volume of the bottle is 120 mL and 50 mL of liquid are added, then  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 40 values for octane, acetone, and toluene are therefore 0.994, 0.009, and 0.278 respectively indicating that only toluene has a significant mass fraction in both the gas and aqueous phases. In the absence of strong sorptionxe "sorption" by soil, octane will reside primarily in the gas phase and acetone will reside primarily in the aqueous phase. Determinations of  EMBED Equation.DSMT4  for these two compounds may therefore not be feasible using the headspace technique. Saturated Mass Fraction Distribution The total mass of the VOC is distributed between the gas, sorbed, and liquid phases (equation  GOTOBUTTON ZEqnNum216792 \* MERGEFORMAT  REF ZEqnNum216792 \! \* MERGEFORMAT 11.32).  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 41 where  EMBED Equation.DSMT4  is the fraction of the VOC mass in the liquid phase. The relationship between the fractions of VOC in the solid and liquid phases is obtained from the definition of the saturated distribution coefficient (equation  GOTOBUTTON ZEqnNum577509 \* MERGEFORMAT  REF ZEqnNum577509 \! \* MERGEFORMAT 11.5).  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 42 The relationship between the fractions of VOC in the gas and liquid phases is obtained from the definition of the Henrys law constant (equation  GOTOBUTTON ZEqnNum265829 \* MERGEFORMAT  REF ZEqnNum265829 \! \* MERGEFORMAT 11.2).  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 43 Thus we have three equations in three unknowns. Solving we obtain  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 44  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 45  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 46 where  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 11. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 47 Determination of  EMBED Equation.DSMT4  requires a measurement of the fraction sorbed given measurements of the total mass added, the mass in the gas phase, and knowledge of the Henrys law constant. This analysis becomes inaccurate when S decreases and approaches the coefficient of variationxe "coefficient of variation" of  EMBED Equation.DSMT4 . Experimental procedures VOC distribution coefficients will be determined using crimp top vials sealed with Teflon backed septa and aluminum crimp caps. VOCs will be analyzed by students using a gas chromatographxe "gas chromatograph" (GC). A data table for preparation of the necessary vials is shown in Table 4. The data table is also available in spreadsheet form as isotherm calculator. Analysis of the Unsaturated Distribution Coefficient ( EMBED Equation.DSMT4 ) The distribution coefficient for three VOCs (acetone, octane, and toluene) with the unsaturated soil medium ( EMBED Equation.DSMT4 ) will be determined using the headspace method of Peterson et al. (1988). Students will prepare 120 mL vials (actually 121.5 0.5 mL) in triplicate containing 0 and 20 g of moist soil (6 vials total). The weight of replicates need not be precisely matched, but the weight of soil should be measured accurately. Seal the vials using Teflonbacked septa (the shiny Teflon side faces the vial contents) and aluminum crimp cap. The 3 VOCs are added to the vials by introduction of 1 mL of the saturated vapor taken from the headspace of source vials (crimp capped vials containing liquid octane, toluene, and acetone). Use a dedicated syringe for each VOC and leave dedicated needles in the source bottles. The vials should be placed on a wrist action shaker to continue agitation for e" 30 minutes to permit the VOC time to sorb to the soil medium. After equilibration, vials are removed from the wrist action shaker and the head space sampled using a 500 L gas tight syringe. Use the syringe technique as described previously for sampling vial headspace. Analysis of the Saturated Distribution Coefficient ( EMBED Equation.DSMT4 ) The saturated distribution coefficient ( EMBED Equation.DSMT4 ) for three VOCs (acetone, octane, and toluene) with the soil medium will be determined using the headspace method of Garbarini and Lion (1985) as modified by Peterson et al. (1988). These vapors are selected to demonstrate the effect of VOC Henry's law constant on VOC phase distribution (see Table 1). Students will prepare 120 mL vials (actually 121.5 0.5 mL) in triplicate containing 0 and 20 g of potting soil (6 vials total). The weight of replicates need not be precisely matched, but the weight of soil should be measured accurately. Soil density should be separately measured as described below. If moist soil is used students should also determine the dry weight unless the instructor provides this information (see below). Into each vial students will add 50 mL of tap water. Seal the vials using a Teflonbacked septa (the shiny Teflon side faces the vial contents) and aluminum crimp cap. Octane and toluene are then added to the vials by introduction of 1 mL of the saturated vapor taken from the headspace of source vials (crimp capped vials containing liquid octane and toluene). Use different needles for collecting and delivering the VOC to reduce the transfer of VOC on the needles. The needle used to collect the VOC from the source bottle can be left in the source bottle and simply attached to the syringe when needed. Acetone is added by introduction of 200 l of the liquid compound using a gas tight syringe. The vials should be placed on a wrist action shaker to continue agitation for e" 30 minutes to permit the VOC time to sorb to the soil medium. After equilibration, vials are removed from the wrist action shaker and the head space sampled using a 500 L gas tight syringe. To sample the vial headspace, use a 100 mL sample and the syringe technique described previously. Procedure (short version) Weigh soil and add to isotherm vials (6 vials with 20 g). Add 50 mL tap water to 6 vials (3 each with 0 and 20 g soil). Seal 12 vials. Add octane, toluene, and acetone to all vials (1 mL gas from source vials for all VOC s except 200 L liquid acetone for vials with tap water). Place vials on shaker for 30 minutes.  Calibrate GC by analyzing 4 100-mL samples for each VOC. Take vials off of shaker. Measure VOC concentrations for each vial and record peak areas in spreadsheet. Reanalyze the VOC concentrations for any vials for which anomalous data was obtained. Remove vial caps. Pour waste potting soil into designated container. Wash vials. Prelab Questions Why does the determination of  EMBED Equation.DSMT4  become inaccurate as the magnitude of S decreases and approaches the coefficient of variationxe "coefficient of variation" of  EMBED Equation.DSMT4 ? What conditions are necessary to obtain linear isothermsxe "isotherms" for gas/solid partitioning of organic vapors? When can equation  GOTOBUTTON ZEqnNum162962 \* MERGEFORMAT  REF ZEqnNum162962 \! \* MERGEFORMAT 11.14 be used to predict the relationship between liquid/solid partitioning and gas/solid partitioning based on soil moisture contentxe "soil moisture content" and Henrys law constant? Data Analysis A Note on Units Express mass of VOC in grams (as measured by the GC). Express concentrations in g/mL. Remember to account for the fact that the syringe volume for GC analysis is 100 L Express all volumes in mL. Estimate the mass of each VOC added to the unsaturated sample vials based on  EMBED Equation.DSMT4  (from equation  GOTOBUTTON ZEqnNum968763 \* MERGEFORMAT  REF ZEqnNum968763 \! \* MERGEFORMAT 11.24). Report mean and coefficient of variationxe "coefficient of variation" (standard deviation/mean) for each VOC. Calculate the unsaturated vapor distribution coefficient ( EMBED Equation.DSMT4 ) using equation  GOTOBUTTON ZEqnNum875668 \* MERGEFORMAT  REF ZEqnNum875668 \! \* MERGEFORMAT 11.26. Report a single mean and coefficient of variationxe "coefficient of variation" for each VOC. Calculate the mass fraction associated with the soil and gas phases under unsaturated conditions for each of the VOCs. Use equations  GOTOBUTTON ZEqnNum463171 \* MERGEFORMAT  REF ZEqnNum463171 \! \* MERGEFORMAT 11.29,  GOTOBUTTON ZEqnNum378980 \* MERGEFORMAT  REF ZEqnNum378980 \! \* MERGEFORMAT 11.30, and  GOTOBUTTON ZEqnNum367607 \* MERGEFORMAT  REF ZEqnNum367607 \! \* MERGEFORMAT 11.31. Assume Ms = 20 g and bottle volume is 121.5 mL. Use the average  EMBED Equation.DSMT4  calculated in 2 above. Compare the coefficient of variationxe "coefficient of variation" of  EMBED Equation.DSMT4  to the mass fractions to evaluate which  EMBED Equation.DSMT4  determinations are potentially accurate. Create a stacked bar graph of the mass fractions for each VOC. Estimate the Henrys law constant ( EMBED Equation.DSMT4 ) for octane and toluene using equation  GOTOBUTTON ZEqnNum753920 \* MERGEFORMAT  REF ZEqnNum753920 \! \* MERGEFORMAT 11.35 (different masses of acetone were used for the saturated and unsaturated vials and thus equation  GOTOBUTTON ZEqnNum753920 \* MERGEFORMAT  REF ZEqnNum753920 \! \* MERGEFORMAT 11.35 can not be used for acetone). Report mean and coefficient of variationxe "coefficient of variation". Compare your results to the Henrys law constants reported in the table on page  PAGEREF VOCproperties 121. Estimate the mass of each VOC added to the saturated sample vials based on  EMBED Equation.DSMT4  (from equation  GOTOBUTTON ZEqnNum745661 \* MERGEFORMAT  REF ZEqnNum745661 \! \* MERGEFORMAT 11.34) using tabulated Henrys constants (see table on page  PAGEREF VOCproperties 121). Report mean and coefficient of variationxe "coefficient of variation" for each VOC. Calculate the saturated vapor distribution coefficient ( EMBED Equation.DSMT4 ) using equation  GOTOBUTTON ZEqnNum498789 \* MERGEFORMAT  REF ZEqnNum498789 \! \* MERGEFORMAT 11.37. Report a single mean and coefficient of variationxe "coefficient of variation" for each VOC. Use tabulated Henrys constants reported in the table on page  PAGEREF VOCproperties 121. Calculate the mass fraction associated with the soil, gas, and water phases under saturated conditions using equations  GOTOBUTTON ZEqnNum674845 \* MERGEFORMAT  REF ZEqnNum674845 \! \* MERGEFORMAT 11.44 -  GOTOBUTTON ZEqnNum805746 \* MERGEFORMAT  REF ZEqnNum805746 \! \* MERGEFORMAT 11.47. Assume Ms = 20 g, VL = 50 mL, and bottle volume is 121.5 mL. Use the average calculated  EMBED Equation.DSMT4  and the Henrys law constants reported in the table on page  PAGEREF VOCproperties 121. Compare the coefficient of variationxe "coefficient of variation" of  EMBED Equation.DSMT4  to the mass fractions to evaluate which  EMBED Equation.DSMT4  determinations are potentially accurate. Create a stacked bar graph of the mass fractions for each VOC. Calculate  EMBED Equation.DSMT4  for toluene using equation  GOTOBUTTON ZEqnNum520105 \* MERGEFORMAT  REF ZEqnNum520105 \! \* MERGEFORMAT 11.14 and compare with the measured value of  EMBED Equation.DSMT4 . Assuming a typical soil porosityxe "soil porosity" (q) of 0.33 and bulk density (rb) of 1.7 g/cm3, equation  GOTOBUTTON ZEqnNum886077 \* MERGEFORMAT  REF ZEqnNum886077 \! \* MERGEFORMAT 11.18 may be used to calculate the retardation of dissolved VOCs and VOC vapors. Assuming a pore water velocity of 1 m/day, how long will it take for the dissolved toluene to be transported a distance of 100 m? Include a range of the estimate based on 1 standard deviation. If toluene is removed by withdrawing vapor at a velocity of 100 m/day, how long will it take toluene vapor to travel 100 m? [Note in this case  EMBED Equation.DSMT4  replaces  EMBED Equation.DSMT4  and a replaces q in equation  GOTOBUTTON ZEqnNum886077 \* MERGEFORMAT  REF ZEqnNum886077 \! \* MERGEFORMAT 11.18. Assume the soil voids are filled with gas so a = q = 0.33.] Include a range of the estimate based on 1 standard deviation. Discuss how the results of this experiment would guide you in remediating a site contaminated with toluene, acetone, and, octane. References Brunauer, S., P.H. Emmett and E. Teller, "Adsorption of Gases in Multimolecular Layers", J. Amer. Chem. Soc., 60, p. 309, 1938. Chiou, C.T., L.J. Peters and J.H. Freed, "A Physical Concept of SoilWater Equilibria For Nonionic Organic Compounds", Science, 206, p. 831, 1979. Chiou, C.T., "Roles of Organic Matter, Minerals and Moisture in Sorption of Nonionic Compounds and Pesticides by Soils", in Humic Substances in Soil and Crop Sciences; Selected Readings, P. MacCarthy, C.E. Clapp, R.L. Malcolm and R.R. Bloom (Eds.), Am. Soc. of Agron. & Soil Sci. Soc. of Am, Madison, WI, pp. 111160, 1990. Freeze, R.A. and J.A. Cherry; Groundwater, Prentice Hall, Inc.; Englewood Cliffs, NJ, 604 pp., 1979. Karickhoff, S.W., D.S. Brown and T.A. Scott, "Sorption of Hydrophobic Pollutants on Natural Sediments", Water Res., 13, p. 241248, 1979. Karickhoff, S.W., "Organic Pollutant Sorption in Aquatic Systems", J. Hydraulic Engrg., 110(6), p. 707, 1984. Millington, R.J., J.M. Quirk, Permeability of porous solids, Trans. Faraday Soc. 57, p. 12001207, 1961. Ong, S.K., and L.W. Lion, "Sorption Equilibria and Mechanisms for Trichloroethylene onto Soil Minerals", J. Env. Qual., 20(1), p. 180188, 1991a. Ong, S.K., and L.W. Lion, "Effects of Soil Properties and Moisture on the Sorption of TCE Vapor", Water Resources Research, 25(1), p. 2936, 1991b. Ong, S.K., and L.W. Lion, "Trichloroethylene Vapor Sorption onto Soil Minerals at High Relative Vapor Pressure", Soil Sci. Soc. Am. J., 55(6), p. 15591568, 1991c. Ong, S.K., S.R. Lindner and L.W. Lion, "Applicability of Linear Partitioning Relationships for Organic Vapors onto Soil Minerals", in: Organic Substances and Sediments in Water, R.A. Baker (Ed.), Lewis Publ., Chelsea, MI, pp. 275289, 1991. Rhue, R.D., P.S.C. Rao and R.E. Smith, "Vapor Phase Adsorption of Alkylbenzenes and Water on Soils and Clays", Chemosphere, 17, p. 727741, 1988. Additional References Relevant to Data Reduction Garbarini, D.R. and L.W. Lion, Evaluation of sorptive partitioning of nonionic organic pollutants in closed systems by headspace analysis, Env. Sci. & Tech., 19(11), 11221128 (1985). Gossett, J.M. Measurement of Henry's law constants for C1 and C2 chlorinated hydrocarbons, Env. Sci. & Tech., 21(2), 202 (1987). Peterson, M.S., L.W. Lion, and C.A. Shoemaker, Influence of vapor phase sorptionxe "sorption" and diffusionxe "diffusion" on the fate of trichloroethylene in an unsaturated aquifer system. Env. Sci. & Tech. 22(5), 571578, (1988). Symbol List SymbolDefinition EMBED Equation.DSMT4 Henrys Constant EMBED Equation.DSMT4 Liquid/solid partitioning coefficient EMBED Equation.DSMT4 Gas/solid partitioning coefficientBSolute binding intensityCConcentrationEDispersion coefficientfMass fractionkBoltzmann constant XE "Boltzmann constant" KocLiquid/organic carbon partitioning coefficientKowoctanolwater partitioning coefficientsMMassPPartial pressureP0Saturated vapor pressureRUniversal gas constantRRetardation factorTAbsolute Temperature(oK).TTemperaturetTimeUGroundwater pore velocityVVolumeXMass sorbed at the solid-liquid interfacexDistanceGMass of solute sorbed per mass of solidevaporadsorbent surface interactionevvaporization energy of the organicfPorosityqVolumetric moisture contentrDensity of waterwMass sorbed at liquid-air interface + condensation SubscriptDefinitionbBulkGGas phaseLAqueous phaseMMonolayerocOrganic carbonSSoil phase or sorbentscSaturated controlucUnsaturated controlVOCVolatile organic carbon Lab Prep Notes Table  SEQ table5. Reagents list DescriptionSourceCatalog numberOctaneFisher Scientific030081AcetoneFisher ScientificO2991tolueneFisher ScientificT324500Potting soilAgway (remove large particles by screening to 2 mm) For equipment list and gas chromatographxe "gas chromatograph" method see page  PAGEREF VOCequipment 124. Setup Replace injection port septa on all GCs. Verify that GCs are working properly by injecting gas samples from each VOC source bottle. If the baseline is above 30 (as read on the computer display) then heat the oven to 200C to clean the column. Verify that sufficient gas is in the gas cylinders (hydrogen, air, nitrogen). Verify that sufficient vials/aluminum crimp tops and Teflon seals are available. Prepare VOC source vials that contain liquid acetone, octane, and toluene (they can be shared by two groups of students). Fill repipet dispensors with tap water and place on each lab bench. Table  SEQ table6. Each group of students requires the following syringes Octane (gas)1 mLToluene (gas)1 mLAcetone (gas)1 mLAcetone (liquid)0.5 mLSource vial (gas)0.5 mLIsotherm (gas)0.5 mLSyringe clean up Disassemble and heat syringes to 45C overnight to remove residual VOCs. Place syringe barrels upside down on top of openings above fan in oven to facilitate mass transfer. Enhanced Filtration Introduction  MACROBUTTON MTEditEquationSection Equation Section (Next) SEQ MTEqn \r \h \* MERGEFORMAT  SEQ MTSec \h \* MERGEFORMAT Slow sand filters have been used to remove particles from drinking water since the early 1800's. Although slow sand filtration XE "slow sand filtration"  is an old technology, the mechanisms responsible for particle removal are not well understood. Because slow sand filter performance gradually increases with time, it has often been assumed that the growth of biofilms is responsible for the gradual improvement in filter performance. Research conducted at Cornell suggests that biofilms are not responsible for significant particle removal and that most particles are removed by physicalchemical mechanisms. The particles that are captured on slow sand filters have been shown to significantly improve filter performance  ADDIN ENRef (WeberShirk and Dick, 1997). More recent research has shown that a filter aid can be extracted under acid conditions from particles harvested from Cayuga Lake or from Cayuga Lake sediment. The filter aid has been shown to greatly enhance bacteria removal. The filter aid is soluble at very low pH, and forms floc at neutral pH. This naturally occurring filter aid may be able to improve rapid sand filter performance. Theory In new slow sand filters with clean filter media, particles are initially removed by attaching to the filter media. However, as the filter media begins to be covered with removed particles, particles begin to attach to previously removed particles. If particleparticle interaction is more favorable than particlemedia interaction then particle removal efficiency increases as the media becomes covered with particles. This improvement in filter performance with time is commonly observed in slow sand filters and is referred to as filter ripening. Filter ripening often takes several weeks to several months for new slow sand filters. Slow sand filters that operate with pristine water sources may never achieve efficient particle removal because the lack of particles in the source water results in a sparse coating of the filter media. Potential mechanisms of particle removal by slow sand filters are summarized in  REF _Ref365358386 \* MERGEFORMAT Figure 1. Physicalchemical removal mechanisms are responsible for most of the particle removal that occurs in slow sand filters. The one exception is that suspension-feeding nanoflagellates attached to the filter media can capture a significant fraction of bacteria  ADDIN ENRef (WeberShirk and Dick, 1999). Thus, bacteria removal by suspension feeding predators is significant provided the influent bacteria concentration is sufficient to maintain a large predator population. Biofilms on the filter media have not been shown to significantly increase particle removal. Straining of bacteriasized particles by the filter media and attachment of bacteriasized particles to the filter media were shown to not be significant because the removal of bacteria by a clean filter column was negligible (Weber-Shirk and Dick, 1997). It is possible that straining becomes significant as filters clog and pores become smaller. Attachment of particles to previously removed particles is considered likely.  Figure  SEQ \r 1 Figure1. Particle removal mechanisms that potentially could be operative in slow sand filters. Physical-chemical filter ripening may be the result of the changes in pore geometry that enhance straining or the modification of filter media surfaces that enhance the ability of particles to attach. Decreasing the pore size to enhance straining is not a reasonable way to improve particle removal because the head loss through the filter increases rapidly as the pore size decreases. Thus, the best way to enhance physicalchemical ripening is to modify filter media surfaces. The filter aid may act by coating the surface and providing more favorable attachment sites. Filtration theory suggests that particle removal will be first order with respect to depth if the filter media is homogeneous  ADDIN ENRef (Iwasaki, 1937). In equation form the relationship between particle concentration, C, and depth is given by  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 12. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 1 where l is the filter coefficient with units of [1/L]. Setting appropriate integration limits  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 12. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 2 where L is the depth of the filter bed and Co is the influent particle concentration and integrating gives:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 12. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 3 Table  SEQ \r 1 table1. Typical values of filter coefficients adapted from  ADDIN ENRef (Tien and Payatakes, 1979). Filter mediumGrain size (mm)particle typeparticle size (m)approach velocity (cm/hr)l (1/cm)Calcium carbonate??Ferric floc105000.1Calcium carbonate??Ferric floc1010000.044Anthracite0.77Quartz powder2225000.064Sand0.54Chlorella55000.34Sand0.647Fuller s earth64700.363Granular carbon0.594Clay4405000.102Equation  GOTOBUTTON ZEqnNum464164 \* MERGEFORMAT  REF ZEqnNum464164 \! \* MERGEFORMAT 12.3 can be used to evaluate the filter coefficient, l. A list of previously measured filtration constants is given in Table 1. Filtration theory suggests that filter performance would be optimal if the filter aid were applied uniformly throughout the filter. Uniform application is difficult, however, because the filter aid will be captured first order with respect to depth if the filter aid is applied using normal down flow operation. It may be possible to apply the filter aid during a gentle backwash thus enabling the filter aid to distribute more uniformly. Application techniques that optimize the filter aid distribution require further study. Previous Research Results Previous research  ADDIN ENRef (WeberShirk and Dick, 1997) has shown that Cayuga Lake water particles can enhance filter performance and thus Cayuga Lake particles (CLP) from the Bolton Point Water Treatment Plant sedimentation basin were tested. Three filters were treated with 30 mL of concentrated CLP suspension from the Bolton Point Water Treatment Plant. One filter had the CLP mixed throughout the filter bed, one filter had the CLP mixed throughout the top 2 cm of the filter bed, and one filter had the CLP applied only to the top of the filter bed. The three application techniques were used because particles may improve filtration efficiency by providing surfaces to which bacteria attach more readily or because the pores within the sediment are smaller and thus more effective at straining particles. The filter with the particles distributed throughout the filter bed performed the best with approximately 99% removal of kaolin compared with 96% removal for the filter with the CLP on top of the filter bed. This result suggested that kaolin was being removed by attaching to CLP rather than by straining. CLP from the Bolton Point facility contain alum and possibly other polymers used in the water treatment process. Previous research also indicated that an acid treatment of Cayuga Lake sediment dissolves species that flocculate and attach to filter media at neutral pH. This Cayuga Lake Sediment Extract (CLSE) has been shown to rapidly ripen slow sand filters and achieve up to 6 log (99.9999%) removal of E. coli. The CLSE has also been shown to enhance E. coli removal at rapid sand filtration rates. Filter Performance Evaluation Several measurement techniques could be used to characterize filter performance. Particle concentrations could be measured using a particle counter, or measured indirectly using a turbidimeterxe "turbidimeter". If the particle suspension absorbed a significant amount of light, a spectrophotometerxe "spectrophotometer" could be used. A microscope could be used to count particles. If microorganisms are used as the source particles, they could be enumerated using standard microbiological techniques such as membrane filtration followed by growth on selective media.  Figure  SEQ Figure2. Light path in a turbidimeterxe "turbidimeter". Turbidimeters measure the amount of light scatter caused by a suspension of particles. Because absorptionxe "absorption" and scattering of light are influenced by both size and surface characteristics of the suspended material, turbidityxe "turbidity" is not a direct quantitative measurement of the concentration of suspended solids. In a turbidimeterxe "turbidimeter" the scattered light (measured at a right angle to the incident light) and the transmitted light intensities are measured ( REF _Ref365358446 \* MERGEFORMAT Figure 2). The ratio of scattered light to transmitted light is proportional to the turbidity of the sample. The constant of proportionality is determined by measuring a known standard. Experimental Objectives The purpose of this research is to evaluate the ability of the CLSE filter aid to enhance particle removal in a filter operating at rapid sand filtration rates. We will use tap water amended with kaolin, 2.5 cm diameter filter columns, and turbidimeters. Students will assembly the apparatus. Experimental Methods Setup 2.5 cm diameter filter column plumbing (Make all connections firmly and verify that the connections cant be pulled apart) including 1 L of clay suspension on a stirrer, peristaltic pump for metering in clay suspension and filter aid, flow meter, pressure reducing valve, and pressure sensor XE "pressure sensor"  for head loss. Add 8 cm of sand to the filter column (by mass). Carefully observe the sand surface as you gradually increase the flow rate from zero in backwash mode. Measure the pressure required to begin to lift the bed. Continue backwashing the filter to clean the sand until the effluent turbidity XE "turbidity"  is less than 0.5 NTU Obtain head loss (in cm) as a function of flow rate (down flow mode) over a range of 1 to 25 m/hr (8.2 to 204 mL/min) using at least 5 data points. Use the rotometer to measure the flow rate. Challenge the filter with a kaolin suspension (approximately 5 NTU) for 30 minutes to determine baseline filter performance. Backwash the filter Add the filter aid (the amount and method of application will be discussed during lab) Set the down flow rate to 5 m/hr. Measure the head loss to see if the filter aid increased the head loss  Figure  SEQ Figure3. Picture of experiment setup. Pump a clay suspension into the filter influent so that the influent concentration is 10-mg/L kaolin. Measure effluent turbidity XE "turbidity"  and head loss as a function of time for 30 minutes. Take turbidity measurements every 5 minutes and measure the head loss continuously using the Signal Monitor XE "Signal Monitor"  software. Backwash the filter. If you have time test the filter again to see if the filter aid improved filter performance even after backwashing. Table  SEQ table \s1 2. Filtration parameters.  EMBED Excel.Sheet.8  Prelab Questions How much water is required to operate one of the laboratory filters for 2 hours? Dont include the water required to fill the filter initially. Given the dimensions for the filter column, a glass density of 2.65 g/cm3, and filter porosity of 0.4, estimate the mass of glass beads in one filter column. (Show your calculations.) Draw a plumbing schematic of a filter column that allows you to do the following: Measure the pressure drop across the column using a pressure sensor XE "pressure sensor" , reverse the flow of water for backwash, and maintain a high pressure in the filter column to avoid dissolution of gasses. Explain how you will switch the filter from down flow to back wash mode. Data Analysis Compare the pressure required to begin to lift the bed with the calculated value based on fluid statics. Plot head loss vs. flow rate for a clean bed and estimate the hydraulic conductivity XE "conductivity"  of the sand. Is the flow laminar or turbulent? What technique did you use to determine the flow regime? Plot the fraction of influent particles remaining in the effluent vs. time for each run on a single graph. Plot head loss as a function of time for each run on a single graph. Calculate the filter coefficient (equation  GOTOBUTTON ZEqnNum421621 \* MERGEFORMAT  REF ZEqnNum421621 \! \* MERGEFORMAT 12.3) for the filter with and without the filter aid. Questions for Discussion Did the filter aid make a significant difference in filter performance? How was the head loss affected by the addition of the filter aid? The laboratory filter columns were 8 cm deep. Rapid sand filters have 60 cm of media. Estimate the fractional bacteria removal for a 60 cm deep filter of media. What assumptions did you make to predict the performance of a 60 cm column? What further experimentation do you recommend? References  ADDIN ENBib Iwasaki, T. 1937. Some Notes on Sand Filtration Journal American Water Works Association 29: 1591. Liljestrand, H. M.; I. M. C. Lo and Y. Shimizu. 1992. Sorption of humic materials onto inorganic surfaces for the mitigation of facilitated pollutant transport processes Proceedings Of The Sixteenth Biennial Conference Of The International Association On Water Pollution Research And Control, Washington, D.C., USA, May 26(111): 12211228. Tien, C. and A. Payatakes. 1979. Advances in Deep Bed Filtration AIChE Journal 25(5): 737. WeberShirk, M., and R. I. Dick. 1997. PhysicalChemical Mechanisms in Slow Sand Filters. Jour. AWWA. 89:87100. Weber-Shirk, M. L. and R. I. Dick (1999). Bacterivory by a Chrysophyte in Slow Sand Filters. Water Research 33(3): 631-638. Lab Prep Notes Table  SEQ table3. Equipment list DescriptionSupplierCatalog numbermagnetic stirrerFisher Scientific115007Svariable flow digital driveCole ParmerH0752330EasyLoad pump headCole ParmerH0751802Filter columns1001095 l pipettexe "pipette"Fisher Scientific13707510109.5 l pipettexe "pipette"Fisher Scientific1370732100P TurbidimeterHach Company46500002100N TurbidimeterHach Company4700000high pressure flow cellHach Company47451020 liter HDPE JerricanFisher Scientific0296150CSetup Attach two EasyLoad pump heads to the pump drives. Setup turbidimeters and verify that the vials are clean. Gas Transfer Introduction  MACROBUTTON MTEditEquationSection Equation Section (Next) SEQ MTEqn \r \h \* MERGEFORMAT  SEQ MTSec \h \* MERGEFORMAT Exchange of gases between aqueous and gaseous phases is an essential element of many environmental processes. Wastewater treatment plants require enhanced transfer of oxygen into activated sludge tanks to maintain aerobic degradation. Water treatment plants require gas transfer XE "gas transfer"  to dissolve chlorine gas or ozone. Gas transferxe "Gas transfer" can also be used to remove unwanted volatile chemicals such as carbon tetrachloride, tetrachloroethylene, trichloroethylene, chloroform, bromdichloromethane, and bromoform from water  ADDIN ENRef (Zander et al., 1989). Exchange of a dissolved compound with the atmosphere is controlled by the extent of mixing in the aqueous and gaseous phase, the surface area of the interface, the concentration of the compound in the two phases, and the equilibrium distribution of the compound. Technologies that have been developed to enhance gas transfer include: aeration diffusers, packedtower air stripping, and membrane stripping. Each of these technologies creates a high interface surface area to enhance gas transfer. Theory Oxygen transfer is important in many environmental systems. Oxygen transfer is controlled by the partial pressure of oxygen in the atmosphere (0.21 atm) and the corresponding equilibrium concentration in water (approximately 10 mg/L). According to Henrys Law, the equilibrium concentration of oxygen in water is proportional to the partial pressure of oxygen in the atmosphere. Natural bodies of water may be either supersaturated or undersaturated with oxygen depending on the relative magnitude of the sources and sinks of oxygen. Algae can be a significant source of oxygen during active photosynthesis and can produce supersaturation. Algae also deplete oxygen levels during the night. At high levels of supersaturation dissolved gas will form microbubbles that eventually coalesce, rise, and burst at the water surface. The bubbles provide a very efficient transfer of supersaturated dissolved gas to the gaseous phase, a process that can be observed when the partial pressure of carbon dioxide is decreased by opening a carbonated beverage. Bubble formation by supersaturated gasses also occurs in the environment when cold water in equilibrium with the atmosphere is warmed rapidly. The equilibrium dissolved oxygenxe "dissolved oxygen" concentration is a function of temperature and as the water is warmed the equilibrium concentration decreases ( REF _Ref365247940 \* MERGEFORMAT Figure 1).  Figure  SEQ \r 1 Figure1. Dissolved oxygenxe "dissolved oxygen" concentrations in equilibrium with the atmosphere as a function of water temperature. Supersaturation of dissolved gases can also occur when water carrying gas bubbles from a water fall or spillway plunges into a deep pool. The pressure increases with depth in the pool and gasses carried deep into the pool dissolve in the water. When the water eventually approaches the surface the pressure decreases and the dissolved gases come out of solution and form bubbles. Bubble formation by supersaturated gases can kill fish (similar to the bends in humans) as the bubbles form in the bloodstream. Gas transferxe "Gas transfer" rate can be modeled as the product of a driving force (the difference between the equilibrium concentration and the actual concentration) and an overall volumetric gas transfer XE "gas transfer"  coefficient (a function of the geometry, mixing levels of the system and the solubility of the compound). In equation form  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 13. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 1 where C is the dissolved gas concentration, C* is the equilibrium dissolved gas concentration and  EMBED Equation.DSMT4  is the overall volumetric gas transfer XE "gas transfer"  coefficient . Although  EMBED Equation.DSMT4  has dimensions of 1/T, it is a function of the interface surface area (A), the liquid volume (V), the oxygen diffusionxe "diffusion" coefficient in water (D), and the thickness of the laminar boundary layer (d) through which the gas must diffuse before the much faster turbulent mixing process can disperse the dissolved gas throughout the reactorxe "reactor".  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 13. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 2  Figure  SEQ Figure2. Single film model of interphase mass transfer of oxygen. The overall volumetric gas transfer XE "gas transfer"  coefficient is system specific and thus must be evaluated separately for each system of interest  ADDIN ENRef (Weber and Digiano, 1996). A schematic of the gas transfer XE "gas transfer"  process is shown in  REF _Ref440510781 \h Figure 2. Fickian diffusionxe "diffusion" controls the gas transfer in the laminar boundary layer. The oxygen concentration in the bulk of the fluid is assumed to be homogeneous due to turbulent mixing and the oxygen concentration above the liquid is assumed to be that of the atmosphere. The gas transfer XE "gas transfer"  coefficient will increase with the interface area and the diffusionxe "diffusion" coefficient and will decrease with the reactorxe "reactor" volume and the thickness of the boundary layer. The functional form of the relationship is given by  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 13. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 3 Equation  GOTOBUTTON ZEqnNum863953 \* MERGEFORMAT  REF ZEqnNum863953 \! \* MERGEFORMAT 13.1 can be integrated with appropriate initial conditions to obtain the concentration of oxygen as a function of time. However, care must be taken to ensure that the overall volumetric gas transfer XE "gas transfer"  coefficient is not a function of the dissolved oxygenxe "dissolved oxygen" concentration. This dependency can occur where air is pumped through diffusers on the bottom of activated sludge tanks. Rising air bubbles are significantly depleted of oxygen as they rise through the activated sludge tank and the extent of oxygen depletion is a function of the concentration of oxygen in the activated sludge. Integrating equation  GOTOBUTTON ZEqnNum934240 \* MERGEFORMAT  REF ZEqnNum934240 \! \* MERGEFORMAT 13.1 with initial conditions of C = C0 at t = t0  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 13. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 4  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 13. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 5 This equation can be linearized so that  EMBED Equation.DSMT4  is the slope of the line.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 13. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 6 The simple gas transfer XE "gas transfer"  model given in equation  GOTOBUTTON ZEqnNum181906 \* MERGEFORMAT  REF ZEqnNum181906 \! \* MERGEFORMAT 13.5 is appropriate when the gas transfer coefficient is independent of the dissolved gas concentration. This requirement can be met in systems where the gas bubbles do not change concentration significantly as they rise through the water column. This condition is met when the water column is shallow, the bubbles have large diameters, or the difference between the concentration of dissolved gas and the equilibrium concentration is small. Experimental Objectives The objectives of this lab are to: illustrate the dependence of gas transfer XE "gas transfer"  on gas flow rate. develop a functional relationship between gas flow rate and gas transfer XE "gas transfer" . Explain the theory and use of dissolved oxygenxe "dissolved oxygen" probes. See the appendix of this manual (page  PAGEREF _Ref406553633 \h 158) for information on how the dissolved oxygen probexe "oxygen probe" works and how to calibrate it. A small reactorxe "reactor" that meets the conditions of a constant gas transfer XE "gas transfer"  coefficient will be used to characterize the dependence of the gas transfer coefficient on the gas flow rate through a simple diffuser. The gas transfer coefficient is a function of the gas flow rate because the interface surface area (i.e. the surface area of the air bubbles) increases as the gas flow rate increases. In order to measure the reaeration rate it is necessary to first remove the oxygen from the reactorxe "reactor". This can be accomplished by bubbling the solution with a gas that contains no oxygen. Nitrogen gas is typically used to remove oxygen from laboratory reactors. Alternately, a reductantxe "reductant" can be used. Sulfite is a strong reductant that will reduce dissolved oxygenxe "dissolved oxygen" in the presence of a catalyst.  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 13. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 7 If complete deoxygenation is desired a 10% excess of sulfite can be added. The sulfite will continue to react with oxygen as oxygen is transferred into the solution. The oxygen concentration can be measured with a dissolved oxygenxe "dissolved oxygen" probexe "oxygen probe" or can be estimated if the temperature is known and equilibrium with the atmosphere assumed ( REF _Ref365247940 \* MERGEFORMAT Figure 1). Experimental Methods EMBED Word.Picture.8 Figure  SEQ Figure3. Apparatus used to measure reaeration rate. The reactorxe "reactor" is a 250 mL polypropylene narrow mouth bottle with an additional port drilled near the neck to allow a hypodermic tubing diffuser to extend into the bottle ( REF _Ref365247987 \* MERGEFORMAT Figure 3). The hypodermic tubing diffuser consists of 4 stainless steel hypodermic tubes that are glued to a plug that mates to 1/4 tubing connectors. The hypodermic tubes are bent so that they are spaced at 90 intervals around the perimeter of the bottle. The DO probe funnel/mixer assembly is inserted into the bottle opening. The bottle is placed on a magnetic stirrer and the DO probe is inserted into the funnel/mixer assembly. The spacing of the hypodermic tubing at the perimeter of the bottle is designed to prevent rising bubbles from touching the DO probe membrane. The hypodermic diffuser is connected to a peristaltic pump (or other source of regulated air flow). Calibrate the DO probe (See page  PAGEREF DOcalibration \h 159). Prepare to monitor the dissolved oxygenxe "dissolved oxygen" concentration using the Compumet XE "Compumet" "! software. Use 5 second data intervals and monitor pH on channel A. The Accumet"! meter and the Compumet"! software think the dissolved oxygen probexe "oxygen probe" is a pH probexe "pH probe". Although the output of the meter is pH, it should be interpreted as mg/L of dissolved oxygen. See page  PAGEREF _Ref406553594 \h 157 for instructions on using Compumet"! software. Add 50 mL of distilled water to a 100 mL beaker. Set the stirrer speed to 5. Add H"10 mg CoCl2 6H2O (note this only needs to be added once because it is the catalyst). A stock solution of CoCl2 6H2O (100 mg/mL  thus add 100 mL) has been prepared to facilitate measurement of small cobalt doses. Set the air flowrate to 50 mL/min (or to the desired flow rate). Turn the air off. Add enough sodium sulfite to deoxygenate XE "deoxygenate"  the solution. A stock solution of sodium sulfite (100 mg/mL) has been prepared to facilitate measurement of small sulfite doses. (50 mL of water at 10 mg O2/L = 0.5 mg O2, therefore add 5 mg sodium sulfate or 20 mL of stock solution.) Turn the air on and start collecting data using the Compumet XE "Compumet" "! software. Monitor the dissolved oxygenxe "dissolved oxygen" concentration until it reaches 80% of saturation value. Save the data as \\Enviro\enviro\Courses\453\gastran\netid_100 for later analysis. Repeat steps 611 using flow rates of 100, 200, 300, and 500 mL/min. Prelab Questions Calculate the mass of sodium sulfite needed to reduce all the dissolved oxygenxe "dissolved oxygen" in 50 mL of pure water in equilibrium with the atmosphere and at 30C. Sketch your expectations for dissolved oxygenxe "dissolved oxygen" concentration as a function of time for the flow rates used on a single graph. The graph can be done by hand and doesnt need to have any numbers on the time scale. Sketch your expectations for  EMBED Equation.DSMT4  as a function of gas flow rate. Do you expect a perfectly straight line or do you expect some nonlinearities? Why? What do you expect  EMBED Equation.DSMT4  to be when the gas flow rate is zero? Data Analysis Eliminate the data from each data set when the dissolved oxygenxe "dissolved oxygen" concentration was less than 0.5 mg/L. This will ensure that all of the sulfite has reacted. Set t0 to the time at the beginning of the remaining data. Subtract t0 from each of the times so the remaining data now starts at zero. Plot the 5 data sets with the corrected times on a single graph. Estimate  EMBED Equation.DSMT4  using linear regression and equation  GOTOBUTTON ZEqnNum200278 \* MERGEFORMAT  REF ZEqnNum200278 \! \* MERGEFORMAT 13.6 for each data set. Show a graph with the linearized data and the best fit lines. Graph  EMBED Equation.DSMT4  as a function of gas flow rate. Comment on results and compare with your expectations and with theory. References  ADDIN ENBib Weber, W. J. J. and F. A. Digiano. 1996. Process Dynamics in Environmental Systems. New York, John Wiley & Sons, Inc. Zander, A. K.; M. J. Semmens and R. M. Narbaitz. 1989. Removing VOCs by membrane stripping American Water Works Association Journal 81(11): 7681. Lab Prep Notes Table  SEQ \r 1 table1. Reagent listDescriptionSupplierCatalog numberNa2SO3Fisher ScientificS430500CoCl2 6H2OFisher ScientificC371100Setup Prepare the sodium sulfite immediately before class and distribute to groups in 15 mL PP bottles to minimize oxygen dissolution and reaction with the sulfite. Table  SEQ table2. Stock solutions listreagentM.W.g/100 mLmg/ mLmL/ groupsolubility g/LNa2SO3126.0410 g10010125CoCl2 6H2O237.9210 g1001770The cobalt solution can be prepared anytime and stored long term. Distribute to student stations in 15 mL PP bottles. Table  SEQ table3. Equipment listDescriptionSupplierCatalog numbermagnetic stirrerFisher Scientific115007S1001095 L pipettexe "pipette"Fisher Scientific13707510109.5 L pipettexe "pipette"Fisher Scientific13707315 mL PP bottlesFisher Scientific029238Gvariable flow digital driveCole ParmerH0752330EasyLoad pump headCole ParmerH0751800PharMed tubing # 18Cole ParmerH06485184 prong hypodermic tubing diffuserCEE shop1/4 plugCole ParmerH06372501/4 unionCole ParmerH0637250stainless steel hypodermic tubingMcMaster CarrAttach two EasyLoad pump heads to the pump drives and plumb with size 18 tubing joined and connected to the hypodermic diffuser. Verify that DO probes are operational, stable, and can be calibrated. The solution behind the membranes should be clear and free of bubbles. If necessary replace membranes. Mount DO probes on magnetic stirrers. Instrument Instructions Compumet XE "Compumet"  software Information on use of the Compumet XE "Compumet"  software is available at  HYPERLINK "http://www.cee.cornell.edu/mws/Software/Compumet.htm" http://www.cee.cornell.edu/mws/Software/Compumet.htm. pH Probe Calibration Select U.S. Standard Buffers From the main screen, press the setup key. Setup 1 menu is shown. Press the 1 key to select Buffer Recognition. Observe display of Setup Buffer Recognition. Press the 1 key again to select U.S. standard buffers. Press the enter key to implement the selection. Press the clear key to return to the main screen. Full Calibration (done once a day) This will calibrate both the slope and the null point. Use either 2 or 3 buffers. Clear Existing Buffers Press the pH key to select pH measurement. Observe display of the current pH standardization points. Press the standardization key. A menu of standardization options appears. Press the 2 key to select Clear Existing Standards. The meter returns to the main screen, but with all pH standardization points (for the current channel) cleared from memory. The electrode slope is reset to 100% or 59.16 mV/pH. Add Buffers (can use buffers at pH of 4, 7, and 10) Press the standardization key. A menu of standardization options is displayed. Press the 1 key to select Update or Add a Standard. The Prepare Buffer/Standard screen appears. Make sure the electrode is in the buffer solution. Press the enter key. The meter will wait until an electrode stability criterion is reached. Then it will automatically read the signal and calibrate. The meter returns to the main screen with the added buffer point shown. Repeat these steps for each of the buffers. Updating the Standardization (done every hour) Use a single buffer to adjust the null point of the calibration curve. The calibration slope will remain the same as from the last full calibration. Press the standardization key. A menu of standardization options is displayed. Press the 1 key to select Update or Add a Standard. The Prepare Buffer/Standard screen appears. Make sure the electrode(s) is in the buffer solution. Press enter. The meter will wait until a predetermined electrode stability is reached, then it will automatically read the signal and calibrate. pH Probe Storage Preparation for storage Rinse with distilled water Air dry Store for months as needed Preparation for use Heat in 60C water and stir for 15 minutes Place in pH buffer 4 or 7 at room temperature for 15 minutes Standardize Probes that fail to standardize may require cleaning Procedure for Cleaning pH Gel-Filled Polymer Electrode Warm distilled water to 40 - 60 (C Suspend the electrode in the warm water for about 15 min while stirring with a magnetic stirrer. This will loosen any material attached to the probe Add 1/2 tsp of detergent Terg-A-Zyme( to the water. Keep stirring for 15 min Rinse well with distilled water Store probe in pH 4 buffer for at least 3 hr (the longer the better). Although less preferable, the probe can also be stored in pH 7 buffer Standardize Probes that fail to standardize may need to be discarded Note: Terg-A-Zyme( - Fischer cat. no. 04-322--11A. Dissolved Oxygen Probe Theory The probe makes use of the fact that an applied potential of 0.8 V can reduce O2 to H2O:  EMBED Equation.DSMT4   MACROBUTTON MTPlaceRef \* MERGEFORMAT  SEQ MTEqn \h \* MERGEFORMAT  SEQ MTSec \c \* Arabic \* MERGEFORMAT 13. SEQ MTEqn \c \* Arabic \* MERGEFORMAT 8 The cell is separated from solution by a gas permeable membrane that allows O2 to pass through (Figure 1). The concentration of O2 in the cell is kept very low by reduction to H2O. The rate at which oxygen diffuses through the gas permeable membrane is proportional to the difference in oxygen concentration across the membrane. The concentration of oxygen in the cell is H"0 and thus the rate at which oxygen diffuses through the membrane is proportional to the oxygen concentration in the solution. Oxygen is reduced to water at a silver (Ag) cathode of the probe. Oxygen reduction produces a current that is measured by the meter. The probe is calibrated using a two-point calibration and can be temperature compensated if desired. Directions can be obtained at  HYPERLINK "http://www.cee.cornell.edu/mws/Software/DOcal.htm" http://www.cee.cornell.edu/mws/Software/DOcal.htm. Monitoring  Figure 1. Schematic of dissolved oxygenxe "dissolved oxygen" probexe "oxygen probe". The dissolved oxygen XE "dissolved oxygen"  probe output is in the Pico amp (10-12 A) range. This current must be converted to a voltage in order to be measured by the laboratory data acquisition system. An analog circuit is used to convert the dissolved oxygen probe XE "oxygen probe"  into a voltage. The voltage is monitored using the Signal Monitor XE "Signal Monitor"  software. Information on using the Signal Monitor software is available at  HYPERLINK "http://www.cee.cornell.edu/mws/Software/signal_monitor.htm" http://www.cee.cornell.edu/mws/Software/signal_monitor.htm. The 8-pin plug from the signal-conditioning box that is attached to the dissolved oxygen XE "dissolved oxygen"  probe XE "oxygen probe"  needs to be plugged into the middle row of ports on the lab bench. Calibration A calibration routine is available in the Signal Monitor XE "Signal Monitor"  software. Follow the instructions in the software and use the help as needed.  HYPERLINK "http://www.cee.cornell.edu/mws/Software/DOcal.htm" http://www.cee.cornell.edu/mws/Software/DOcal.htm. Dissolved Oxygen Probe Storage Remove membrane from probe. Rinse electrode with distilled water. Store electrode covered with electrode cap (dry). Gas Chromatograph Information on the spectrophotometer XE "spectrophotometer"  analysis software is available at  HYPERLINK "http://www.cee.cornell.edu/mws/Software/gas_chromatograph.htm" http://www.cee.cornell.edu/mws/Software/gas_chromatograph.htm. UVVis Spectrophotometer Information on the spectrophotometer XE "spectrophotometer"  analysis software is available at  HYPERLINK "http://www.cee.cornell.edu/mws/Software/Spectrophotometer.htm" http://www.cee.cornell.edu/mws/Software/Spectrophotometer.htm. Index  INDEX \h "A" \c "2"  A absorption, 20, 71, 72, 78, 145 acid neutralizing capacity, 43, 45, 46 acid precipitation, 43, 47, 50 adsorption, 24, 70, 71, 72, 74, 75, 82, 83, 128 advective dispersion, 33, 34, 130, 131 alkalinity, 45, 50, 61 ANC, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 57, 58, 59, 60, 61, 62, 107, 108, 109 atomic absorption, 78 B BET, 127, 128 Biochemical Oxygen Demand, 88 Biodegradable, 87 Boltzmann constant, 128, 140 C carbonate system, 44, 45, 107 chelating agent, 70, 80 coefficient of variation, 23, 133, 135, 137, 138 completely mixed flow, 30, 39, 53 Compumet, 37, 38, 39, 49, 51, 62, 153, 154, 157 conductivity, 35, 36, 37, 38, 39, 41, 111, 113, 147 correlation coefficient, 21, 24, 28 D density of sodium chloride solution, 25 density of water, 129 deoxygenate, 154 diffusion, 34, 119, 131, 139, 151, 152 diode array, 21, 65, 77 dispersion coefficient, 30, 31, 32, 33, 37, 40, 130 dissolved oxygen, 87, 88, 89, 90, 92, 93, 94, 95, 96, 97, 150, 151, 152, 153, 154, 159 E endogenous respiration, 88, 89, 90, 92, 95, 96 extracellular polymer, 77, 82 extractant, 77, 78, 79, 80, 81, 82, 84 F flow with dispersion, 30, 33 G gas chromatograph, 9, 109, 110, 111, 113, 114, 115, 120, 123, 131, 135, 141 gas transfer, 96, 99, 150, 151, 152 Gas transfer, 150, 151 gasoline, 119 global warming, 87 Gran plot, 53, 58, 59, 61, 62 Gran Plot, 53, 58 groundwater, 33, 119, 130 H hazardous waste site, 70 Henry's Law, 46, 129 hydraulic residence time, 32, 34, 40, 44, 46, 50, 53 I ion exchange, 71, 74 isotherms, 74, 127, 128, 137 L landfill, 86, 100, 115 Langmuir, 128 ligand, 75 M mass balance, 33, 40, 50, 129, 131, 133 methane, 100, 101, 102, 103, 104, 105, 109, 110, 111, 113, 114, 115, 116 methylene blue, 22, 23, 24, 25, 28, 29, 77, 78, 79, 80, 81, 82 Monod, 88, 89 O oxidant, 76 Oxidation, 83, 87, 88 oxygen deficit, 88, 90, 91, 92, 93 oxygen probe, 93, 94, 95, 152, 153, 159 oxygen sag, 88, 93, 96, 97 P Peclet number, 32, 33, 40 petroleum, 119 pH probe, 48, 49, 51, 56, 60, 63, 153 pipette, 11, 23, 55, 61, 66, 69, 95, 149, 156 plug flow, 30, 32, 34, 39 Plug flow, 34 plume, 40 porous media, 33, 37, 40, 41, 55, 56, 70, 71, 73, 74, 75, 76, 77, 119, 129, 130, 131 pressure sensor, 109, 111, 113, 114, 145, 147 pump and treat, 70 R reactor, 9, 30, 31, 32, 33, 34, 35, 37, 38, 39, 40, 41, 50, 53, 77, 88, 91, 94, 95, 151, 152, 153 reductant, 153 remediation, 47, 48, 70, 71, 76, 77 Remediation, 43, 47, 49, 53 retardation factor, 130, 131 S Signal Monitor, 146, 159 slow sand filtration, 142 soil density, 122 soil gas sampling, 119, 121 soil moisture content, 122, 123, 129, 132, 137 soil porosity, 138 solvents, 12, 16, 17, 119 sorption, 71, 72, 74, 76, 77, 78, 120, 126, 127, 128, 129, 130, 131, 134, 139 spectrophotometer, 21, 22, 24, 28, 29, 65, 66, 67, 69, 77, 86, 145, 160 spectrum, 21, 66, 78 Streeter Phelps, 88 Superfund, 70 T titration, 52, 57, 58, 59, 60, 61, 62 tracer, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42 turbidimeter, 145 turbidity, 145, 146 W watershed, 43, 46, 47, 48   Prepared by Tom Shelley (Chemical Hygiene Officer) 7/96. For additional information or questions you may have, please contact Tom Shelley at 54288.  Alkalinity can be expressed as equivalents/L or as mg/L (ppm) of CaCO3. 50,000 mg/L CaCO3 = 1 equivalent/L.  Radicals contain an odd number of electrons.  PAGE 128  PAGE 127 CEE 453: Laboratory Research in Environmental Engineering Spring 2001  PAGE iii  STYLEREF "Heading 1,1" Volatile Organic Carbon Sorption to Soil PAGE \# "'Page: '#' '"  Need a reference for these equations. PAGE \# "'Page: '#' '"  simplify measurements and data analysis PAGE \# "'Page: '#' '"  Potential design question Regulatory limit of x M Zn/g sand and y M methylene blue/g sand. Give cost of each extractant. Assume fractional removal is independent of fraction remaining (linear isotherm). identify lowest cost option for remediating site with original concentrations xi Zinc and yi methylene blue. It may be desirable to use a combination of extractants. Assume that all extractants are equally recyclable so that the cost of each extractant is proportional to the volume of extractant required times the cost of the extractant. PAGE \# "'Page: '#' '"  Add section on modeling sag curves using Labview. 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2  Oxygen (ppm)-----'--- c---'--- c-- 45--}---- --- -' c '  'SummaryInformation(5DocumentSummaryInformation87X_1034429836Fa1la1lOle A|HP X`hp x B  Sheet1Sheet2Sheet3Sheet4Sheet5Sheet6Sheet7Sheet8Sheet9Sheet10Sheet11Sheet12Sheet13Sheet14Sheet15Sheet16Chart1  WorksheetsCharts 6> _PID_GUIDAN{788F54C3-6290-11D1-B2E4-006097C28853}* DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A   "C"t==  "C oxidaObjInfoBEquation Native CF_1034429838F@n1l@n1lOle Mtion "t++  "C respiration "t++  "C reaeration "t2 DSMT4WinAllBasicCodePagesObjInfoNEquation Native ON_1034429841F{1l{1lOle UTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A   dLdt==  "-kLXK s ++L DSMT4WinAllBasicCodePagesObjInfoVEquation Native W_1034429844F1l1lOle _Times New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A   dLdt==  "-kXLK s ++LE"@  "-kXK s []LE"@"-k ox LObjInfo`Equation Native a_1034429846F`1l1lOle ht DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  q dLL L o L +" = q("-k ox )dt 0t +" DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  L==L o e  "-k ox tObjInfoiEquation Native j;_1034429849F1l@1lOle oObjInfopEquation Native q_1034429851F)1l1lOle xz DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A   "C oxidation "t=  dLdt= -k ox L DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A   "C oxidation "t= -k ox L o eObjInfoyEquation Native z_1034429854F 71l 71lOle   "-k ox tc DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A   "C respiration "ObjInfoEquation Native _1034429856F˯1lQ1lOle t=-bX =-k e DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A   "C reaeration "tObjInfoEquation Native _1034429859h/F@ز1l@ز1lOle = k  v,l C *  - C() DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  k  v,lObjInfoEquation Native _1034429862F1l1lOle  DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  ln C * "-CC * "-C ObjInfoEquation Native _1034429864F1l1lOle 0 ==k  v,l (t"-t 0 ) DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A   "C"tObjInfoEquation Native  _1034429867F'1l'1lOle = -k ox L o e  "-k ox t  k e  + k  v,l C *  - C() DSMT4WinAllBasicCodePagesObjInfoEquation Native  _1034429869F1lP41lOle Times New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  D=C * -C DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APObjInfoEquation Native 5_1034429872F1l1lOle G_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A   dCdt =  "-dDdt DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_EObjInfoEquation Native _1034429874F01l01lOle _A   dLdt DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  -k ox LObjInfoEquation Native _1034429877FN1lp1lOle  DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A   "D"t==k e ++k ox L o e  "-k oObjInfoEquation Native _1035367702_xF\1l\1lOle x t  - k  v,l D   FMathType 4.0 Equation MathType EFEquation.DSMT49q@6M6GHDSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APCompObjiObjInfoEquation Native ._1034429882F1lv1lG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  D== k e k  v,l ++D o  k e k  v,l [] e "-k  v,l t ++ k ox L o k  v,l "-k ox  e  "-k ox t  - e -k  v,l t []  DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_EOle ObjInfoEquation Native  _1034429885FЃ1lЃ1l_A  t =  xu DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  t c = Ole ObjInfoEquation Native $_1034429887F1l1lx c u DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A   "D"t= 0Ole ObjInfoEquation Native _1034429890F1lP1lOle ObjInfoEquation Native _1034429892F$1l1l DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  0==k e ++k ox L o e  "-k ox t c   !"#$%&'()*+/012347:;<=ABCDEIJKLMNOSTUVWXYZ^_`abcdeijklpqrstuvwx|}~  - k  v,l D c DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  D c == Ole ObjInfoEquation Native _1035367729F021lи1lk e ++k ox L o e  "-k ox t c k  v,l FMathType 4.0 Equation MathType EFEquation.DSMT49qOle  CompObj iObjInfoEquation Native tX@6M6GHDSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  D c = k e k  v,l ++D o "- k e k  v,l [] e  "-k  v,l t c ++ k ox L o k  v,l "-k ox  e  "-k ox t c  - e -k  v,l t c []_1034429898F1l1lOle ObjInfoEquation Native   DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  t c = 1k  v,l -k ox  ln k e -k  v,l D o ()k  v,l -k ox ()L o k ox2 ++ k  v,l k ox []f DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/AP_1034429900FL1lL1lOle ,ObjInfo-Equation Native .G_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A   dLdt= "-kLXK s ++LE"@-kX =-k 0 FMathType 4.0 Equation MathType EFEquation.DSMT49q_1035367898IFY1lY1lOle 5CompObj6iObjInfo8@6M6GHDSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  k  v,l DEquation Native 9 _1034429903Fpt1lpt1lOle >ObjInfo?Equation Native @w_1034429905) F1l1lOle FObjInfo  G[ DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A   dDdt= k 0  + k e"- k  v,l DEquation Native H_1034429908FP1l1lOle PObjInfoQ DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  q dDk 0 ++k e"- k  v,l D D o D +" = qdt 0t +"                          ! 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"2 -DB f\%(`!T -DB f 8XJ"xڍSOPyI&[InAqb0XSl5@նR{rx!<.}aAE"qkey7H>'G!Y$]SNYQ`e y nptL 5u9*@o EG3zl/% AyZa 7Qf/w}衢 >NPT H?H:Y],*/ NYNp-b8.u#~) Up_P v @J(&=#\S%;h {/0Og>z$QlDA?hV2enrf Z5Y0[Ferguson, 1993 #801]D2&?hV2enrf Z6& 0[Ferguson, 1993 #801]D]&>hV2enrf`a [Suflita, 1995 #790] Dd%J  C A"2  IWr͡*m#ܞk ά5Qs̜bhNIe T~:ѳkc\"5y^خSr7-W<ʒ`-"dʙ7<~3zVD.wZ(j}TlD%(Y28dQeEQcuc]EzcF<-1l)fŠ^3A 3.i25= b1p=?\" GVznk Kݺ-)@Qn.ԻO<$i>4vJ? 1`uYzX[OOjP iz4ѩ2͇ݴޢ0] _'AC2A[8qaŴU20*/>V3" 9|Bb:_ڧόanUl#A'Ԙ[!h6@2Vk脐N : /eiAm2ith0TQJ.D; v> ~2Rkh`fc34 ,ıV!XXlƥVrq 8lRqw-~lK+D}mk^_|5 v [^{Z`uFkZa VX 0Fyn8,ED6Ɉj?P4C?7G8^_NW ?oa=BvfBlaOl8Įn=W$7T}׊zr} ֢ȩ!;coIdVq*[IqዸA,8`Lc'hLWi5{˞u9ma?kpΫvyGjU~Z+p?_|#R&FL/a@boF,>c^,]' vLdO!lNўTv'=lVmo䲾 eNgX C\젓 C3y9ls`#ρ <| ~%,c_sy1p= O)x~J+-h)\Sϓ_oUW{>׃[L1Ft/FL@Ca6!]pT`HRn'jN6C}DelZѸ34.rh5Ю8}p Pp!E,4Q|%Ja,f"<2d99+ag,Ty.7Kz!Ua,r!&A<7OBsnΆkAs0Cw*z@*x˛>k4FՖ! hܰ:ҎYin}D<4_vkĻ?F0d9OR;rî[VQYT%z^SSY-C ׌Ole ObjInfoXZEquation Native  _1034429438]F^S1l^S1l DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  n== PVRT DSMT4WinAllBasicCodePagesOle ObjInfo\^Equation Native _1035361389RaF@kV1l@kV1lTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  Dn== DPVRT FMathType 4.0 Equation MathType EFEquation.DSMT49qOle CompObj`biObjInfocEquation Native     !"#$%&),-./0126789=>?@ABFGHIJKOPQRSTUVZ[\]abcdhijkopqrswxyz{|}~f@6M6GDSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  Dn== 80x10 3 Pa()100x10 "-6 m 3 ()8.31 Pa"m 3 mol"K()308K()==3.13 mmole C FMathType 4.0 Equation MathType EFEquation.DSMT49q@6M6GHDSMT4WinAllBasicCodePages_1035361244nfFxY1l Z1lOle  CompObjeg iObjInfohEquation Native _1035361402kF\1l` ^1lOle CompObjjliTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  3.13 mmole C" 27 mg cellulosemmole C==84 mg cellulose FMathType 4.0 Equation MathType EFEquation.DSMT49q@6M6GHDSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A   84 mg VS biodegraObjInfomEquation Native _1035361358pF_1l_1lOle 'dable 80 kPa" VS0.22 VS biodegradable " TS0.94 VS== 400 mg dry newspaper80 kPaCompObjoq(iObjInfor*Equation Native +_1034429450[wuF&d1l&d1l FMathType 4.0 Equation MathType EFEquation.DSMT49q@6M6GHDSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  3.13 mmole C" 30 mg glucosemmole C==94 mg glucose DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_EOle 3ObjInfotv4Equation Native 5,_1034429453yF 4g1lh1l_A  H 2 CO 3* []f DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  K H == Ole :ObjInfoxz;Equation Native <_1034429456s}Fk1lNm1lH 2 CO 3* []P CO 2  DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_EOle CObjInfo|~DEquation Native 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New RomanSymbolCourier NewMT Extra!/ED/APOle tObjInfouEquation Native v _1034429473F`1l21lG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A   HCO 3"- []H 2 CO 3* []== a 1 a 0 == K 1 H ++ []Ole ObjInfoEquation Native _1034508914F1l@?1ll DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  ANC== P CO 2  K H K 1 H ++ []Ole CompObjiObjInfoEquation Native r FMathType 4.0 Equation MathType EFEquation.DSMT49qzV@6M6GHDSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  ANCE"@ 3x10 4 Pa()3.12x10 "-4  molesN"m()10 "-6.3 M()10 "-7 MANCE"@47meq/L_1034429480Fň1lL1lOle ObjInfoEquation Native  DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  CO 2 DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/AP      !"#e$f&'()*+,-./0123456789:;<=>?@ABCDEFGHIJKLMNOPQRSTUVWXYZ[\]^_`abcdvghjilknmoprqstwxyz{|}~_1034429483FY1lY1lOle ObjInfoEquation 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Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  H 2 CO_1034429498FÜ1lÜ1lOle ObjInfoEquation Native  3*p DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  n H 2 CO 3*  ==P C_1034429500FPП1lPП1lOle ObjInfoEquation Native O 2  K H V l DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  HCO 3_1034429503FV1lV1lOle ObjInfoEquation Native "-/ DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  n HCO 3"-  ==ANC"V _1034429505F0d1l0d1lOle ObjInfoEquation Native Kl  DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  n CO 2  (total)  =_1034429508F1l1lOle ObjInfoEquation Native (= P CO 2  V g RT++P CO 2  K H ++ANC()V ll DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/AP_1034429510QF1l1lOle ObjInfoEquation Native G_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  H ++ []== P CO 2  K H K 1 ANC  DSMT4WinAllBasicCodePages_1034429513F~1lP1lOle ObjInfoEquation Native %   "#$%&'()*+/0123456:;<=>?@ABEHIJKLMNRSTUVW[\]abcdhijknqrstxyz~Times New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  K HCH 4 ()b DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/AP_1034429515F1l1lOle ObjInfoEquation Native ~G_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  n CH 4 (g) == P CH 4  V g RT DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/AP_1034429518F-1l-1lOle  ObjInfo Equation Native G_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A  n CH 4 (aq) ==P CH 4  K HCH 4 () V l DSMT4WinAllBasicCodePages_1034429520FG1lG1lOle ObjInfoEquation Native Times New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A   n CH 4 (g) n CH 4 (aq) == V g V l K HCH 4 () RT_1034429523Fb1l1lOle ObjInfo Equation Native !| DSMT4WinAllBasicCodePagesTimes New RomanSymbolCourier NewMT Extra!/ED/APG_APAPAE%B_AC_AE*_HA@AHA*_D_E_E_A   n CH 4 (g) n CH 4 (aq) == 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