Evaluation of the Microgenics buprenorphine urine assay ...
EVALUATION OF THE MICROGENICS BUPRENORPHINE URINE ASSAY VERSUS LC-ES-MS
V. Cirimele, P. Kintz, S. Lohner, B. Ludes
Institut de Médecine Légale, Laboratoire de toxicologie, 11, rue Humann, 67085 Strasbourg Cedex, France
Buprenorphine is an analgesic with a long duration of action, 25-40 times more potent than morphine. In France, buprenorphine is proposed as a substitute treatment for heroin addicts under the name Subutex®. Since 1996, numerous deaths have been observed with buprenorphine, always in association with psychotropic drugs.
Even though confirmation methods such as LC-MS are needed for identification, a rapid and sensitive test like an immunoassay is essential for screening. Recently, Microgenics proposed the Diagnostix SinglestepTM ELISA test for the qualitative and semi-quantitative determination of buprenorphine in urine samples. Twenty µl of urine are incubated with an enzyme conjugate. After the washing steps, substrate is added for the final color development process.
In order to test the capacity of commercial immunoassay to detect buprenorphine, 76 real urine samples were simultaneously screened by the Diagnostix SinglestepTM ELISA test and by our validated procedure for the determination of buprenorphine and norbuprenorphine in urine by HPLC-ES-MS (1).
From the 76 urine samples tested, 33 were found positive (43.4%) after ELISA screening (cut-off at 2 ng/ml) and confirmed by LC/MS (buprenorphine 1.2 to 1052 ng/ml) and 37 were found negative (48.7%) by both procedures. For 4 urine specimens, the positive ELISA test was not confirmed by LC/MS (LOQ of 1 ng/ml). Only one urine specimen gave a false negative ELISA test.
Spiked urines with norbuprenorphine up to 500 ng/ml lead to the absence of response with the ELISA test (0.3% cross reactivity at 1 mg/L).
An enzymatic hydrolysis of the specimens with beta-glucuronidase does not modify the results, indicating that the binding site of the antibody is not linked with the glucuronide.
These results support that the Diagnostix SinglestepTM ELISA test of Microgenics can be considered as a screening procedure to detect buprenorphine in urine samples. In the case of a positive ELISA test, a confirmatory analysis is needed.
References
|1. |A. Tracqui, P. Kintz, P. Mangin, HPLC/MS determination of buprenorphine and norbuprenorphine in biological fluids and |
| |hair samples, J. Forensic Sci. 1997, 42, 111-4. |
Keywords: buprenorphine, immunoassay, LC/MS
(Vincent.Cirimele@medecine.u-strasbg.fr)
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