ࡱ> @B=>?[ N9bjbj zΐΐc-| | 8 È[z  """$)iW,"""W,W, }>>>W,N   >W,>>jqhv L 9Xs2ڇ0Ès<dhvhvJw("%>' )7"""i>."""ÈW,W,W,W,"""""""""| :  DEVELOPMENT OF NEW ANALYTICAL METHODS FOR QUANTITATIVE ESTIMATION OF A NEW DRUG M. Pharm. Dissertation Protocol Submitted to the Rajiv Gandhi University of Health Sciences, Karnataka. Bangalore.  by NAMRATA BASNET B. Pharm. Under the guidance of Dr. SYEDA HUMAIRA M Pharm. Ph.D P.G. Dept. of Quality Assurance.  DEPARTMENT OF QUALITY ASSURANCE LUQMAN COLLEGE OF PHARMACY, GULBARGA 2012-2013 RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA, BANGALORE ANNEXURE II PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION  Name of the candidate And address NAMRATA BASNET Luqman college of pharmacy, old jewargi road, gulbarga-585102  Name of the institution Luqman college of pharmacy, old jewargi road, gulbarga-585102  Course of study and subject m.PHARM (QUALITY ASSURANCE)  Date of Admission of course 9-05-2012   Title of the Topic:  DEVELOPMENT OF NEW ANALYTICAL METHODS FOR QUANTITATIVE ESTIMATION OF A NEW DRUG6 Brief Resume of the intended work: 6.1 Need for study: Chemically Terbinafine Hydrochloride is (E)-N-(6,6-dimethyl-2-hepten-4-ynyl)-N-methyl-1- naphthalenemethanamine hydrochloride. [1] [2] [3]. Terbinafine hydrochloride is a new potent orally and topically active drug of  HYPERLINK "http://en.wikipedia.org/wiki/Allylamine" \o "Allylamine" allylamine derivative with HYPERLINK "http://en.wikipedia.org/wiki/Antifungal_medication" \o "Antifungal medication"antifungal [4] activity. It is found to be potent inhibitor of squalene epoxidase, an important enzyme required in ergosterol biosysnhesis. The drug is indicated for both oral and topical treatment of mycoses [5][6][7]. The raw material of Terbinafine Hydrochloride is official in British Pharmacopoeia.[3]. Considering basic drugs capacity to generate ion-pairs with azoic dyes, there have been investigated the experimental conditions in order to develop new Spectrophotometric method for the assay of Terbinafine Hydrochloride in bulk and in dosage forms. Simple, rapid Spectrophotometric method and HPLC method may be useful for day to day routine analysis of the product. They are recommended for quality control and routine analysis where time, cost effectiveness and high specificity of analytical techniques are of great importance. Investigations of some new instrumental methods are in need for the quantitative estimation of Terbinafine Hydrochloride in bulk drug and its pharmaceutical dosage forms with high sensitivity, accuracy, precision and economic too. Few simple new validated analytical methods are required for the quantitative estimation of Terbinafine Hydrochloride in various pharmaceutical dosage forms. Review of Literature: Literature survey revealed that, chemically Terbinafine Hydrochloride is (E)-N-(6,6-dimethyl-2-hepten-4-ynyl)-N-methyl-1-naphthalenemethanamine hydrochloride.[1] [2] [3] Molecular formula- C21H26CIN HCl Molecular weight - 327.90 Characteristic -A white to off-white fine crystalline powder. Solubility -Freely soluble in methanol and dichloromethane, freely soluble in anhydrous ethanol, soluble in ethanol, slightly and very slightly soluble in water, and slightly soluble in acetone[2],[3],[4] Melting Point- 204C to 208C. [2]  INCLUDEPICTURE "http://www.drugfuture.com/Pharmacopoeia/USP32/pub/data/images/v32270/cas-78628-80-5.gif" \* MERGEFORMATINET  Chemical structure of Terbinafine Hydrochloride .[1] Literature Survey reveals that previously, the drug has been determined in various dosage forms using Spectrophotometric methods[8],[9],[10],11],[12],[13], Microbiological assay [14] in tablets and creams and by HPLC[15],[16],[17],[18]. HPTLC methods, RP-HPLC methods [19],[20][21] have also been developed. The drug has also been determined in biological fluids (plasma, urine), tissues, nails and hair by HPLC mathod [22],[23],[24]. 1. HYPERLINK "http://www.phmethods.org/searchresult.asp?search=&author=Pritam+S+Jain&journal=Y&but_search=Search&entries=10&pg=1&s=0" \t "_blank"Pritam S Jain, HYPERLINK "http://www.phmethods.org/searchresult.asp?search=&author=Amar+J+Chaudhari&journal=Y&but_search=Search&entries=10&pg=1&s=0" \t "_blank" Amar J Chaudhari, HYPERLINK "http://www.phmethods.org/searchresult.asp?search=&author=Stuti+A+Patel&journal=Y&but_search=Search&entries=10&pg=1&s=0" \t "_blank" Stuti A Patel, HYPERLINK "http://www.phmethods.org/searchresult.asp?search=&author=Zarana+N+Patel&journal=Y&but_search=Search&entries=10&pg=1&s=0" \t "_blank" Zarana N Patel, HYPERLINK "http://www.phmethods.org/searchresult.asp?search=&author=Dhwani+T+Patel&journal=Y&but_search=Search&entries=10&pg=1&s=0" \t "_blank" Dhwani T Patel (2011)[8] had developed and validated UV-spectrophotometric method for determination of Terbinafine Hydrochloride in bulk and in formulation using water as the solvent. The spectrum was recorded at 283 nm. 2. Marinela Florea, Crina-Maria Monciu,(2008)[9] had developed Spectrophotometric determination of Terbinafine through ion-pair complex formation with Methyl Orange . 3.  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Abdel-Moety%20EM%5BAuthor%5D&cauthor=true&cauthor_uid=12426813" Abdel-Moety EM,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Kelani%20KO%5BAuthor%5D&cauthor=true&cauthor_uid=12426813" Kelani KO,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Abou%20al-Alamein%20AM%5BAuthor%5D&cauthor=true&cauthor_uid=12426813" Abou al-Alamein AM (2003)[10] carried out Spectrophotometric methods for the quantitative determination of Terbinafine Hydrochloride in presence of its photodegradation products. The methods make use of the first spectral derivation (1D) and the first derivative of the ratio spectra (1DD) of the named drug and its photodegradants at different justified wavelengths. The proposed methods showed good linearity, precision and reproducibility. 4. Karuna T., Neelima K., Venkateshwarlu G., Swamy P.Y.(2006)[11] had developed Spectrophotometric determination of drugs with iodine. 5. Cardoso SG, Schapoval EE (1999)[12] had worked on Ultraviolet Spectrophotometric and a non aqueous volumetric method of analysis for determining Terbinafine Hydrochloride in pharmaceutical formulations.( raw materials, tablets, and creams) The method was tested for linearity (0.8-2.8 micrograms/mL, r = 0.9997), recovery (102.00% for creams and 99.90% for tablets) and precision (101.3%, CV = 0.96%, n = 9, for creams; 100.25%, CV = 1.08%, n = 9, for tablets). The volumetric method involves titration of Terbinafine with 0.05M Perchloric Acid with Crystal Violet as indicator. This method was used for quantitative determination of Terbinafine in raw materials and tablets. Mean recovery and precision were, respectively, 100.41 and 101.18% (CV = 1.64%, n = 9) for Terbinafine tablets. 6.  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=El-Saharty%20YS%5BAuthor%5D&cauthor=true&cauthor_uid=12008136" EL-Saharty YS,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Hassan%20NY%5BAuthor%5D&cauthor=true&cauthor_uid=12008136" Hassan NY,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Metwally%20FH%5BAuthor%5D&cauthor=true&cauthor_uid=12008136" Metwally FH:(2002)[13] had developed Simultaneous determination of Terbinafine Hydrochloride and Triamcinolone Acetonide by UV Derivative Spectrophotometry and Spectrodensitometry . A binary mixture of Terbinafine Hydrochloride and Triamcinolone Acetonide was determined by three different methods. The first one concerned with determination of both drugs using first derivative (D(1)) Spectrophotometric technique at 297 and 274 nm over concentration ranges of 5-30 and 4-24g/ml, with mean percentage accuracies 99.900.67 and 100.250.49, respectively. The second method depends on ratio-spectra 1st derivative (RSD(1)) Spectrophotometry at 298 and 248 nm over the same concentration ranges with mean percentage accuracies 100.220.51 and 99.930.56, respectively. 7. Simone Goncalves Cardoso and Elfrides ES Schapoval (2000) [14] had developed Microbiological assay of Terbinafine Hydrochloride in tablet and cream applying Cylinder Plate method using strain of Aspergillus Flavous ATCC 15546 as the test organism.,Terbinafine Hydrochloride at concentrations ranging from 0.125 to 0.5 g/ml could be measured in tablets and creams. Monika P. Tagliari; Gislaine Kuminek; Slvia H. M. Borgmann; Charise D. Bertol; Simone G. Cardoso; Hellen K. Stulzer (2010)[15] had developed a simple Liquid Chromatographic method optimized for the quantitative determination of Terbinafine in pharmaceutical hydroalcoholic solutions and tablets, and was also employed for a tablet dissolution test. The analysis was carried out using a RP-C18 (250 mm 4.6 mm, 5 m) Vertical column, UV-Vis detection at 254 nm, and a methanol-water (95:5, v/v) mobile phase at a flow-rate of 1.2 ml/ min. Validation parameters like linearity, precision, accuracy, robustness and specificity were investigated. 9. Majumdar T. K., Bakthiar R., Tse F. L.(2000)[16] carried out a method for determination of Terbinafine (Lamisil) in human hair by microbore liquid chromatography / tandem mass spectrometry. 10. Marinela Florea, Corina-Cristina (2009) [17] had developed an Ion-Pair Reversed Phase Liquid Chromatographic method (IP-RP-HPLC) for the assay of Terbinafine. A C18 column was used as stationary phase, and the mobile phase consisted of an aqueous acid solution. 11. R Ramesh Raju, N Bujji Babu (2011) [18] had developed High Performance Reverse Phase Liquid Chromatographic method for Terbinafine and Bezafibrate using C 18 column. 12. Suma, B,V., Kannan. K, Madhavan V., Chandini R Nayar (2011) [19] had developed and validated HPTLC method for development of Terbinafine in bulk and tablet dosage form using pre coated silica gel 60F 254 aluminium foil TLC plates as stationary phase. 13. K.K Patel , V.V Karkhanis (2012)[20] had recently validated HPTLC method for determination of Terbinafine Hydrochloride The method had proved to be reproducible and selective for the analysis of Terbinafine Hydrochloride in bulk and in pharmaceutical dosage form. 14. Patel Krupa K (2012) [21] had developed a validated RP-HPLC method for determination of Terbinafine Hydrochloride in pharmaceutical solid dosage form. Terbinafine Hydrochloride was chromatographed on a reverse phase C18 column with a mobile phase consisting of methanol: water in the ratio of 80:20% v/v. The mobile phase was pumped at a flow rate of 1ml/min. The detection is carried out at 282 nm. The retention time of the drug was 5.84 min. The linearity curved showed a correlation coefficient (r2) of 0.9974 with concentration range of 80-160g/ml. The method was validated with respect to linearity, precision, accuracy and sensivity (LOQ and LOD). 15. Denouel J, Keller HP, Schaub P, Delaborde C,Humbert H(1995)[22] had developed the method for determination of Terbinafine and its desmethyl metabolite in human plasma by high-performance liquid chromatography. 16. Zehender H, Denouel J, Roy M, Saux L Le, Schaub P(1995)[23] developed simultaneous determination of Terbinafine (Lamisil) and five metabolites in human plasma and urine by high-performance liquid chromatography using on-line solid-phase extraction 17. Kuzne J, Kozuh Erzen N, Drobnic-Kosorok M(2001) [24] had developed the method for determination of the Terbinafine Hydrochloride Levels in Cats Plasma and Hair using a New developed Isocratic HPLC Method. 6.3 Objective of the study: The objective of the present study is to develop an accurate, precise, selective sensitive and reproducible method to quantify Terbinafine Hydrochloride in bulk and pharmaceutical dosage forms. Litrature survey reveals a few Spectrophotometric methods for estimation of Terbinafine Hydrochloride but there may be chance of excipients interference. Certain groups present in the drug molecule when treated with chromogenic agent produce a colour. Therefore visible Spectrophotometric analysis is suitable for estimation. Of those cited in literature Spectrophotometric methods have been utilized for assay of drug in pharmaceutical dosage form. Hence on the basis of literature survey it was thought to develop a precise, accurate, simple, reliable, economical, sensitive, validated and less time consuming UV- Visible Spectrophotometric and Chromatographic methods for estimation of drugs in bulk and pharmaceutical dosage form. The objectives of the present work are: To develop validated analytical UV- Visible Spectrophotometric method for estimation of Terbinafine Hydrochloride in bulk drugs and various pharmaceutical dosage forms. To develop RP-HPLC method to estimate Terbinafine Hydrochloride in bulk drug and dosage forms. PLAN OF WORK The work has been planned to carry out on following lines: Estimation of Terbinafine Hydrochloride by UV Spectroscopy: By selecting some common solvents and studying the solubility of raw material and selecting the solvent. Study of Spectra and selection of method and wavelengths. Analysis of marketed formulation by proposed method.  7. 8 i. Terbinafine Hydrochloride is slightly soluble in water soluble in water, freely soluble in methanol, soluble in ethanol and a number of UV-Spectrophotometric methods can be developed by using buffer with acidic pH or organic solvents including chloroform for its quantitative estimation in bulk drug and pharmaceutical dosage forms.[8] Calorimetric methods for estimation of Terbinafine Hydrochloride. Selection of suitable solvent and reagents. Optimization of solvent system, reagent concentration etc. Selection of method and wavelength. Study of effect of pH, temperature, colour stability. Analysis of market formulation by proposed method. i. Since Terbinafine Hydrochloride contains tertiary amino group which is protonated in acid medium it can react with the dyes containing sulphonic groups. The dyes containing sulphonic groups dissociate in pH range 1-5. The protonated Terbinafine Hydrochloride forms ion complex with the dissociated dyes which may be quantified by extraction in chloroform by Visible Spectrophotometry. Considering basic drugs capacity to generate ion-pairs with azoic dyes, Spectrophotometric method for the assay of Terbinafine Hydrochloride in bulk and in dosage forms can be developed based on ion-pair formation..[25] ii. A number of organic acid anhydrides are capable of forming colored products with tertiary amines; malonic, citric, and aconitic acids are representative examples. A citric/acetic anhydrides can be used for the Spectrophotometric determination.[25] iii. The amino group present in the drug allows the oxidative coupling reaction with 3-methyl-2-benzothiazolinone hydrazone (MBTH) in presence of Ferric chloride / Cerric ammonium sulphate and forms colored chromogen (VI) by which drug can be estimated quantitatively by colorimetry. Estimation of Terbinafine by RP- HPLC The method involving following steps: 1.Selection of mobile phase and wavelengths 2. Selection of Chromatographic Conditions. 3. Analysis of marketed formulation by the proposed method. RP- HPLC method can be developed by using either isocratic or gradient system with stationary phase of C18 or C8 column. 7.1 Materials and Methods In the present investigation we are in need of following Materials: i. Raw Material: Working Standard of Terbinafine Hydrochloride, Raw material of Terbinafine Hydrochloride ii. Reagents: Water, Alcohol, Methanol, Dyes: Bromo Thymol blue, Bromo Cresol Purple. iii. Market samples of Terbinafine Tablets. iv. Instruments: Double Beam UV Visible Spectrophotometer, (1700) Shimadzu, Japan. High Performance Liquid Chromatography with pump. Chromatogrphic instruments and volumetric glass apparatus, Vacuum filter, Sonicator, Stirrer. Analytical Balance. All the above mentioned will be made available from Luqman College of Pharmacy, Gulbarga. 7.2. Source of Data: Data will be obtained from i Pubmed , Science Direct Medline, and other internet facilities. ii Literature search and related articles from library of Luqman College of Pharmacy Gulbarga etc. Method of Collection of Data(Including Sampling Procedures if any) The research work that will be carry out in the laboratory by various experimental technique includes: Analysis of reference raw material and their physical constant, solubility, assay analysis data will be done. Spectroscopic data will be obtained for identification of the drug molecule compound. Data of estimation of reference raw material and market samples will be obtained spectrophotometrically and through HPLC graphs. Data Collected from Journals and articles: Journal of Pharmaceutical and Biomedical Analysis. Journal of AOAC International International Journal of Pharmacy and Technology Internet Browsing: HYPERLINK "http://www.sciencedirect.com"www.sciencedirect.com HYPERLINK "http://www.google.com"www.google.com HYPERLINK "http://www.rxlist.com"www.rxlist.com HYPERLINK "http://www.pubmed.com"www.pubmed.com HYPERLINK "http://www.drugbank.com"www.drugbank.com HYPERLINK "http://www.chemicallist.com"www.chemicallist.com The Drug sample will be collected from Panas Pharmaceutical Pvt. Ltd Nepal or from other reputed company. 7.3 Does the study require any investigation or intervention to be conducted on patients or other humans ? if so please mention briefly. -NOT APPLICABLE 7.4 Has ethical clearance been obtained from your institution in case of 7.3? -NOT APPLICABLE- List of References:  HYPERLINK "http://www.chemicalland21.com/lifescience/UH/TERBINAFINE%20HYDROCHLORIDE.htm" http://www.chemicalland21.com/lifescience/UH/TERBINAFINE%20HYDROCHLORIDE.htm Terbinafine Wikipedia, The free encyclopedia; http://en.wikipedia.org/wiki/Terbinafine British Pharmacopoeia 2012, Online, 2112.  HYPERLINK "http://bp2012.infostar.com.cn/Bp2012.aspx?a=query&title=%22Terbinafine+Hydrochloride%22&tab=a-z+index&l=T&xh=1" http://bp2012.infostar.com.cn/Bp2012.aspx?a=query&title=%22Terbinafine+Hydrochloride%22&tab=a-z+index&l=T&xh=1 Budavari S. The Merck Index. 12 th ed. New Jersey, USA: Merck and Co., Inc., 1996, 1564. Clinical Pharmacology/ Biopharmaceutics Review Nov.5 1999, Centre for Drug Evaluation and Research., Application Number 21-124 Martindale. The Complete Drug Reference. 32 nd ed. Great Britain: Council of Royal Pharmaceutical Society of Great Britain, 1999, 367-87. Delgodo JN, Remers WA. Wilson and Gisvold's, Text book of Organic Medicinal and Pharmaceutical Chemistry. 9 th ed. Philadelphia: J. B. Liipincott Company; 1991.145-53. Pritam S, Jain, Amar J Chaudhari, Sruti A Patel, Zarana N Patel, Dhwani T Patel, Development and Validation of the UV- Spectrophotometric method for determination of Terbinafine Hydrochloride in bulk and in formulation. 2011, 2, ( 3),198-202.  HYPERLINK "http://www.phmethods.org/text.asp 2011/2/3/198/9364" http://www.phmethods.org/text.asp 2011/2/3/198/9364. Marinela Florea, Crina-Maria Monciu: Spectrophotometric Determination Of Terbinafine Through Ion-Pair Complex Formation With Methyl Orange, Farmacia, 2008, LVI, (4), 393.  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Abdel-Moety%20EM%5BAuthor%5D&cauthor=true&cauthor_uid=12426813" Abdel-Moety EM,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Kelani%20KO%5BAuthor%5D&cauthor=true&cauthor_uid=12426813" Kelani KO,  HYPERLINK "Abou al-Alamein AM" Abou al-Alamein AM: Spectrophotometric determination of Terbinafine in presence of its Photodegradation products. Boll Chim Farm 141 (4), 267-73(2003)PMID12426813.  HYPERLINK "http://pubget.com/profile/author/A%20M%20Abou%20al-Alamein" http://pubget.com/profile/author/A%20M%20Abou%20al-Alamein Karuna T., Neelima K., Venkateshwarlu G., Swamy P.Y., Spectrophotometric determination of drugs with iodine, Journal of Scientific & Industrial Research, 2006, 65(10), 808-811. Cardoso SG, Schapoval EE. UV Spectrophotometry and non aqueous determination of Terbinafine Hydrochloride in dosage forms. J AOAC Int 1999,82:380-3.  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=El-Saharty%20YS%5BAuthor%5D&cauthor=true&cauthor_uid=12008136" EL-Saharty YS,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Hassan%20NY%5BAuthor%5D&cauthor=true&cauthor_uid=12008136" Hassan NY,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Metwally%20FH%5BAuthor%5D&cauthor=true&cauthor_uid=12008136" Metwally FH: Simultaneous determination of Terbinafine Hydrochloride and Triamcinolone Acetonide by UV derivative Spectrophotometry and Spectrodensitometry , Journal of Pharmaceutical and Biomedical Analysis, 2002 28, ( 3-4), 569-580. Simone Goncalves Cardoso and Elfrides ES Schapoval (2000); Microbiological assay for Terbinafine Hydrochloride in tablet and cream, International Journal of Pharmaceutics 203(2000), 109-113 Monika P. Tagliari; Gislaine Kuminek; Slvia H. M. Borgmann; Charise D. Bertol; Simone G. Cardoso; Hellen K. Stulzer ;Terbinafine : Optimization of a LC method for quantitative analysis in Pharmaceutical formulations and its application for a tablet dissolution test. Quim, Nova, Sao Paulo, 2010, 33, (8).  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Majumdar%20TK%5BAuthor%5D&cauthor=true&cauthor_uid=10918370" Majumdar TK,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Bakhtiar%20R%5BAuthor%5D&cauthor=true&cauthor_uid=10918370" Bakhtiar R,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Melamed%20D%5BAuthor%5D&cauthor=true&cauthor_uid=10918370" Melamed D,  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed?term=Tse%20FL%5BAuthor%5D&cauthor=true&cauthor_uid=10918370" Tse FL Determination of Terbinafine (Lamisil) in human hair by Microbore Liquid Chromatography/tandem Mass Spectrometry.  HYPERLINK "http://www.ncbi.nlm.nih.gov/pubmed/10918370" \l "#" \o "Rapid communications in mass spectrometry : RCM." Rapid Commun Mass Spectrom. 2000, 14(14),1214-9. Marinela Florea, Corina-Cristina Aram, Crina-Maria Monciu: Determination of Terbinafine Hydrochloride by ion pair reverse phase liquid chromatography, Farmacia 2009, L VII,1. R. Ramesh Raju, N Bujji Babu : Simultaneous analysis of RPHPLC method development and validation of Terbinafine and Bezafibrate drugs in pharmaceutical dosage forms. Pharmacophore ( An international Research Journal) 2011, 2 (4), 232-238. Suma B.V, Kannan. K, Madhavan V., Chandini R. Nayar; HPTLC method for determination of Terbinafine in the bulk and tablet dosage form; International Journal of Chem. Tech. Research. CODEN (USA); IJCRGG ISSN: 0974-4290, 2011 3 (2), 742-748,. K.K Patel , V.V Karkhanis ; A validated HPTLC method for determination of Terbinafine Hydrochloride in pharmaceutical solid dosage form. International journal of Pharmaceutical Sciences and Research, UPSR 2012,3 (11), 4492-4495. 21. Patel Krupa K; A validated RP-HPLC method for determination of Terbinafine Hydrochloride in pharmaceutical solid dosage form. International Journal Of Pharmacy &Technology IJPT , 2012 , 4 ,(3) , 4663-4669 22. Denouel J, Keller HP, Schaub P, Delaborde C,Humbert H: Determination of Terbinafine and its desmethyl metabolite in human plasma by High-Performance Liquid Chromatography. Journal of Chromatography B: Biomedical Sciences and Applications 1995, 663(2), 353-359. 23. Zehender H, Denouel J, Roy M, Saux L Le, Schaub P: Simultaneous determination of Terbinafine (Lamisil) and five metabolites in human plasma and urine by High-Performance Liquid Chromatography using on-line solid-phase extraction. Journal of Chromatography B: Biomedical Sciences and Applications 1995,664(2),347-55. 24. Kuzne J, Kozuh Erzen N, Drobnic-Kosorok M: Determination of the Terbinafine Hydrochloride Levels in Cats Plasma and Hair Using a New Developed Isocratic HPLC Method. Biomedical Chromatography BMC 2001,15(8),497-502. 25. Website; research.cm.utexas.edu/nbauld/CHAPTER%2021.htm. Chapter 21: Amines. 9 SIGNATURE OF CANDIDATE ( NAMRATA BASNET )10REMARKS OF THE GUIDE The candidate is working under my direct supervision in laboratories of L.C.P Gulbarga-58520211NAME AND DESIGNATION OF 11.1 GUIDE  Dr. SYEDA HUMAIRA M.Pharm, Ph.D PROFESSOR11.2 SIGNATURE 11.3 CO-GUIDE (IF ANY) 11.4 SIGNATURE 11.5 HEAD OF DEPARTMENT 11.6 SIGNATUREDr. SYEDA HUMAIRA M.Pharm, Ph.D PROFESSOR DEPT. 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SYED SANAULLAH M. Pharm. 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