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Published:
Journal of Analytical Toxicology,
ISSN 0146-4760,
Volume 27, Number 7, October 2003,
pp. 464-470
Analysis of Buprenorphine, Norbuprenorphine, and Their
Glucuronides in Urine by Liquid Chromatography–Mass Spectrometry
Robert Kronstrand, Tor G. Seldén, and Martin
Josefsson
National Board of Forensic Medicine, Department of Forensic Chemistry, University
Hospital, SE-581 85 Linköping, Sweden
Buprenorphine is used for the treatment of chronic pain and
also in treatment of heroin addiction as an alternative to methadone. As the
availability of buprenorphine increases, so does the risk for abuse and the
pressure on forensic and clinical laboratories to analyze for it. Buprenorphine
and its dealkylated metabolite are excreted in urine, almost exclusively as
glucuronides. The aim of the present study was to evaluate electrospray liquid
chromatography tandem mass spectrometry (LC–MS–MS) for the rapid
screening and quantitation of buprenorphine and its metabolites in urine. Three
approaches were evaluated: (1) direct injection of diluted urine for measurement
of glucuronides, (2) direct injection of diluted urine after enzymatic hydrolysis
for the quantitation of buprenorphine and norbuprenorphine, and (3) quantitation
of buprenorphine and norbuprenorphine after enzymatic hydrolysis and solid-phase
extraction (SPE). One hundred six samples were subjected to procedure 1 and,
when positive, further quantitated using procedure 2. Only samples with low
analyte concentrations (< 20 µg/L) were subject to SPE. Concentrations
of buprenorphine and norbuprenorphine in patients (N = 16) ranged between 31
and 1080 µg/L and 48–2050 µg/L, respectively. In suspected
abusers (N = 33), the ranges were 2.3–796 µg/L and 5.0–2580 µg/L.
In four of the authentic samples, both the buprenorphine and norbuprenorphine
concentrations were below the 20-µg/L cutoff. We concluded that LC–MS–MS
analysis of the glucuronides provided an adequate screening method, but that
the direct method for quantitation sometimes had to be complemented with a
concentration by SPE, providing increased sensitivity, thus lowering the cutoff
from 20 to 1 µg/L urine. Reproduction
of editorial content of this journal is prohibited without publisher’s
permission.
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