Published: Journal of Analytical Toxicology, ISSN 0146-4760, Volume 31, Number 4, May 2007, pp. 214-216

Rapid Quantification of Buprenorphine-Glucuronide and Norbuprenorphine-Glucuronide in Human Urine by LC–MS–MS
S. Hegstad, H.Z. Khiabani, E.L. Øiestad, T. Berg, and A.S. Christophersen
Norwegian Institute of Public Health, Division of Forensic Toxicology and Drug Abuse, P.O. Box 4404 Nydalen, NO-0403 Oslo, Norway

A liquid chromatography–tandem mass spectrometry (LC–MS–MS) method was developed and validated for the determination of buprenorphine-glucuronide (BUP-G) and norbuprenorphine-glucuronide (NBUP-G) in human urine. The method included a dilution step followed by filtration through a Mini-Uniprep Filter and direct injection onto the LC column. The analytes were quantified in multiple reactions monitoring mode using one transition ion. Norbuprenorpine-d3 (NBUP-d3) was used as the internal standard. The concentration ranges were 6–161 ng/mL for BUP-G and 12–295 ng/mL for NBUP-G. Recoveries determined after filtration for the analytes were 75%. The between-day precision of the method was in the range of 4.8–11%. The limits of quantification were found to be 4.6 ng/mL for BUP-G and 11.8 ng/mL for NBUP-G. Approximately 1000 samples from law enforcement, prison inmates, probation services, and hospitals were analyzed by the presented method. The ratios of drug glucuronides versus creatinine were calculated for a selection of samples (n = 151), where there was information on treatment with buprenorphine between 16 and 20 mg/day. The majority (86%) of the samples had a ratio of BUP-G/creatinine below 570 µg/g, and 76% of the samples had NBUP-G/creatinine lower than 1060 µg/g. The LC–MS–MS method proved to be robust and specific for the determination of BUP-G and NBUP-G in urine.

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