Simultaneous quantification of buprenorphine, norbuprenorphine, buprenorphine-glucuronide and norbuprenorphine-glucuronide in human umbilical cord by liquid chromatography tandem mass spectrometry.
A LCMS method was developed and validated for the simultaneous determination of buprenorphine (BUP), norbuprenorphine (NBUP), buprenorphine-glucuronide (BUP-Gluc) and norbuprenorphine-glucuronide (NBUP-Gluc) in human umbilical cord. Quantification was achieved by selected ion monitoring of precursor ions m/z 468.4 for BUP; 414.3 for NBUP; 644.4 for BUP-Gluc and 590 for NBUP-Gluc. BUP and NBUP were identified by MS(2), with m/z 396, 414 and 426 for BUP, and m/z 340, 364 and 382 for NBUP. Glucuronide conjugates were identified by MS(3) with m/z 396 and 414 for BUP-Gluc and m/z 340 and 382 for NBUP-Gluc. The assay was linear 1-50 ng/g. Intra-day, inter-day and total assay imprecision (%RSD) were <14.5%, and analytical recovery ranged from 94.1% to 112.3% for all analytes. Extraction efficiencies were >66.3%, and process efficiency >73.4%. Matrix effect ranged, in absolute value, from 3.7% to 7.4% (CV<21.8%, n=8). The method was selective with no endogenous or exogenous interferences from 41 compounds evaluated. Sensitivity was high with limits of detection of 0.8 ng/g. In order to prove method applicability, an authentic umbilical cord obtained from an opioid-dependent pregnant woman receiving BUP pharmacotherapy was analyzed. Interestingly, BUP was not detected but concentrations of the other metabolites were NBUP-Gluc 13.4 ng/g, BUP-Gluc 3.5 ng/g and NBUP 1.2 ng/g.
Concheiro
Marta
M
Chemistry and Drug Metabolism, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, Baltimore, MD 21224, USA.
Shakleya
Diaa M
DM
Huestis
Marilyn A
MA
D002047:Buprenorphine / Q000031:analogs & derivatives* / Q000032:analysis* / Q000627:therapeutic use; D002853:Chromatography, Liquid; D005260:Female; D053593:Forensic Toxicology; D020719:Glucuronides / Q000032:analysis*; D006801:Humans; D009292:Narcotic Antagonists / Q000032:analysis* / Q000627:therapeutic use; D009293:Opioid-Related Disorders / Q000534:rehabilitation; D011247:Pregnancy; D053719:Tandem Mass Spectrometry; D014470:Umbilical Cord / Q000737:chemistry*
D020719:Glucuronides; D009292:Narcotic Antagonists; D002047:Buprenorphine; C043585:norbuprenorphine