Validation and Application of a Method for the Determination of Buprenorphine,Norbuprenorphine,and Their Glucuronide Conjugates in Human Meconium

摘要

A novel liquid chromatography tandem mass spectrometry method for quantification of buprenorphine,norbuprenorphine,and glucuronidated conjugates was developed and validated.Analytes were extracted from meconium using buffer,concentrated by solid-phase extraction and quantified within 13.5 min.In order to determine free and total concentrations,specimens were analyzed with and without enzyme hydrolysis.Calibration was achieved by linear regression with a 1/x weighting factor and deuterated internal standards.All analytes were linear from 20 to 2000 ng/g with a correlation of determination of >0.98.Accuracy was >85.7% with intra-assay and interassay imprecision < 13.9 and 12.4%,respectively.There was no interference from 70 licit and illicit drugs and metabolites.Buffer extraction followed by SPE yielded recoveries of >85.0%.There was suppression of ionization by the polar matrix;however,this did not interfere with sensitivity or analyte quantification due to inclusion of deuterated internal standards.Analytes were stable on the autosam-pler,at room temperature,at 4 °C,and when exposed to three freeze/thaw cycles.This sensitive and specific method can be used to monitor in utero buprenorphine exposure and to evaluate correlations,if any,between buprenorphine exposure and neonatal outcomes.