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. 2009 Jul 1;188(1-3):144-51.
doi: 10.1016/j.forsciint.2009.04.005. Epub 2009 Apr 29.

Simultaneous quantification of buprenorphine, norbuprenorphine, buprenorphine-glucuronide and norbuprenorphine-glucuronide in human umbilical cord by liquid chromatography tandem mass spectrometry

Marta Concheiro  1 , Diaa M Shakleya, Marilyn A Huestis
Affiliations

Simultaneous quantification of buprenorphine, norbuprenorphine, buprenorphine-glucuronide and norbuprenorphine-glucuronide in human umbilical cord by liquid chromatography tandem mass spectrometry

Marta Concheiro et al. Forensic Sci Int. .

Abstract

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.

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Figures

Figure 1
Figure 1
LCMS, LCMS2 and LCMS3 chromatograms of an umbilical cord sample fortified at 3 ng/g (low quality control sample) for buprenorphine, norbuprenorphine, buprenorphine-glucuronide and norbuprenorphine-glucuronide.
Figure 2
Figure 2
LCMS, LCMS2 and LCMS3 chromatograms of a blank umbilical cord.
Figure 3
Figure 3
LCMS, LCMS2 and LCMS3 chromatograms of an authentic umbilical cord specimen positive for norbuprenorphine (1.2ng/g), buprenorphine-glucuronide (3.5ng/g) and norbuprenorphine-glucuronide (13.4ng/g). No buprenorphine was detected. The cumulative maternal buprenorphine dose over the 111 day treatment period was 1,760 mg.
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