Abstract
A liquid chromatography-tandem mass spectrometry method for the simultaneous quantification of buprenorphine (BUP), norbuprenorphine (NBUP), naloxone (NAL), and their glucuronide conjugates BUP-G, NBUP-G, and NAL-G in urine samples was developed. The method, omitting a hydrolysis step, involved non-polar solid-phase extraction, liquid chromatography on a C18 column, electrospray positive ionization, and mass analysis by multiple reaction monitoring. Quantification was based on the corresponding deuterium-labelled internal standards for each of the six analytes. The limit of quantification was 0.5μg/L for BUP and NAL, 1μg/L for NAL-G, and 3μg/L for NBUP, BUP-G, and NBUP-G. Using the developed method, 72 urine samples from buprenorphine-dependent patients were analysed to cover the concentration ranges encountered in a clinical setting. The median (maximum) concentration was 4.2μg/L (102 μg/L) for BUP, 74.7μg/L (580 μg/L) for NBUP, 0.9μg/L (85.5μg/L) for NAL, 159.5μg/L (1370 μg/L) for BUP-G, 307.5μg/L (1970 μg/L) for NBUP-G, and 79.6μg/L (2310 μg/L) for NAL-G.
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