Development and GC-MS Validation of a Highly Sensitive Recombinant G6PDH-Based Homogeneous Immunoassay for the Detection of Buprenorphine and Norbuprenorphine in Urine

G. Wang, M. Vincent, W. Rodrigues, A. Agrawal, C. Moore, R. Barhate, E. Abolencia, C. Coulter, J. Soares, Y.-F. Zheng, C. Taylor, N. Morjana
2007 Journal of Analytical Toxicology  
L Abstract J Buprenorphine is now increasingly prescribed as an alternative to methadone for the treatment of heroin addiction. Because of its potency (dosage usages from 0.2 mg to 8 rag), the drug concentrations in body fluids are normally very low. Here, we report the first recombinant glucose-6-phosphate dehydrogenase (G6PDH)-based homogeneous immunoassay (EMIT-type assay) for free buprenorphine and free norbuprenorphine in urine. The antibody used in this assay cross-reacts nearly
more » ... ts nearly identically with buprenorphine and norbuprenorphine and, at the same time, has less than 1% cross-reactivity with a wide range of commonly prescribed opiates, particularly those structurally related compounds such as morphine, codeine, and dihydrocodeine. More importantly, this assay has a low detection limit of I ng/mL for buprenorphine or norbuprenorphine. Further evaluation of this technique using gas chromatography-mass spectrometry (GC-MS) of authentic urine samples demonstrated that the accuracy of the assay is greater than 95 %. Because this assay is designed to measure the free drugs in urine, it resulted in simplification for GC-MS or liquid chromatography-MS confirmation methods that did not require urine hydrolysis before solid-phase or liquid-liquid extraction. * This recombinant G6PDH-based immunoassay is under development and not available for sale. Reproduction (photocopying) of editorial content of this journal is prohibited without publisher's permission.
doi:10.1093/jat/31.7.377 pmid:17725885 fatcat:ykbes7o2gjbwjjybdd4yfozm5m