Validation and application of a method for the determination of buprenorphine, norbuprenorphine, and their glucuronide conjugates in human meconium.
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 >or=85.7% with intra-assay and interassay imprecision<or=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 >or=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 autosampler, at room temperature, at 4 degrees 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.
Chemistry and Drug Metabolism Section, Intramural Research Program, National Institute on Drug Abuse, 5500 Nathan Shock Drive, Baltimore, Maryland 21224, USA.
Sherri L
SL
Kacinko
Diaa M
DM
Shakleya
Marilyn A
MA
Huestis
D002047:Buprenorphine / Q000031:analogs & derivatives* / Q000032:analysis* / Q000627:therapeutic use; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D005260:Female; D020719:Glucuronides / Q000032:analysis* / Q000737:chemistry*; D006801:Humans; D007223:Infant; D008470:Meconium / Q000737:chemistry* / Q000378:metabolism; D009035:Mothers / Q000523:psychology; D009293:Opioid-Related Disorders / Q000188:drug therapy; D011247:Pregnancy; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
D020719:Glucuronides; D002047:Buprenorphine; C043585:norbuprenorphine