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Published:
Journal of Analytical Toxicology,
ISSN 0146-4760,
Volume 30, Issue 4, May 2006,
pp.238-244
Quantitative Analysis of Buprenorphine and Norbuprenorphine
in Urine using Liquid Chromatography Tandem Mass Spectrometry
E.J. Fox[1], V.A. Tetlow[2], and K.R. Allen[1]
[1]Department of Clinical Biochemistry, Leeds Teaching Hospitals NHS Trust,
Britannia House, Britannia Road, Morley, Leeds, LS27 0DQ, U.K. and
[2]Department of Clinical Biochemistry, Salford Royal NHS Trust, Hope Hospital,
Stott Lane, Salford, M6 8HD, U.K.
Buprenorphine is an opioid analgesic drug that is used as an alternative
to methadone to treat heroin addiction. Established methods for the analysis
of buprenorphine and its metabolites in urine such as gas chromatography–mass
spectrometry (GC–MS) involve complicated sample extraction procedures.
The aim of the present study was to develop a sensitive yet straightforward
method for the simultaneous analysis of buprenorphine and norbuprenorphine in
urine using liquid chromatography–MS–MS. The method comprised an
enzymatic hydrolysis using Patella vulgata b-glucuronidase, followed by centrifugation
and direct analysis of the supernatant. The limits of detection and quantitation
were < 1 µg/L for buprenorphine and < 1 and 4 µg/L, respectively,
for norbuprenorphine. Assay coefficients of variation (CVs) were < 15%, with
the exception of concentrations close to the limit of quantitation, where CVs
were below 20%. In direct comparison with an established GC–MS protocol,
the method showed minimal negative bias (8.7% for buprenorphine and 1.8% for
norbuprenorphine) and was less susceptible to sample carryover. The extent of
conjugation in unhydrolyzed urine was investigated and found to be highly variable,
with proportions of unconjugated buprenorphine and norbuprenorphine of 6.4%
[range 0% to 67%; standard deviation (SD) 9.7%] and 34% (range 0% to 100%; SD
23.8%), respectively.
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