title: Development and validation of the first liquid chromatography-tandem mass spectrometry assay for simultaneous quantification of multiple antiretrovirals in meconium.
abstract: A novel method for the simultaneous quantification of 16 antiretroviral (ARV) drugs and 4 metabolites in meconium was developed and validated. Quantification of 6 nucleoside/nucleotide reverse transcriptase inhibitors, 2 non-nucleoside reverse transcriptase inhibitors, 7 protease inhibitors, and 1 integrase inhibitor was achieved in 0.25 g of meconium. Specimen preparation included methanol homogenization and solid-phase extraction. Separate positive and negative polarity multiple reaction monitoring mode injections were required to achieve sufficient sensitivity. Linearity ranged from 10 to 75 ng/g up to 2500 ng/g for most analytes and 100-500 ng/g up to 25,000 ng/g for some; all correlation coefficients were ≥0.99. Extraction efficiencies from meconium were 32.8-119.5% with analytical recovery of 80.3-108.3% and total imprecision of 2.2-11.0% for all quantitative analytes. Two analytes with analytical recovery (70.0-138.5%) falling outside the 80-120% criteria range were considered semiquantitative. Matrix effects were -98.3-47.0% and -98.0-67.2% for analytes and internal standards, respectively. Analytes were stable (>75%) at room temperature for 24 h, 4 °C for 3 days, -20 °C for 3 freeze-thaw cycles over 3 days, and on the autosampler. Method applicability was demonstrated by analyzing meconium from HIV-uninfected infants born to HIV-positive mothers on ARV therapy. This method can be used as a tool to investigate the potential effects of in utero ARV exposure on childhood health and neurodevelopmental outcomes.
authors.affiliation: Chemistry and Drug Metabolism Section, Intramural Research Program, National Institute on Drug Abuse, 251 Bayview Boulevard, Baltimore, Maryland 21224, United States.
authors.forename: Sarah K
authors.initials: SK
authors.lastname: Himes
authors.forename: Karl B
authors.initials: KB
authors.lastname: Scheidweiler
authors.forename: Katherine
authors.initials: K
authors.lastname: Tassiopoulos
authors.forename: Deborah
authors.initials: D
authors.lastname: Kacanek
authors.forename: Rohan
authors.initials: R
authors.lastname: Hazra
authors.forename: Kenneth
authors.initials: K
authors.lastname: Rich
authors.forename: Marilyn A
authors.initials: MA
authors.lastname: Huestis
mesh_terms: D044966:Anti-Retroviral Agents / Q000009:adverse effects / Q000032:analysis* / Q000378:metabolism; D002853:Chromatography, Liquid / Q000592:standards / Q000639:trends; D015331:Cohort Studies; D005260:Female; D015658:HIV Infections / Q000188:drug therapy / Q000378:metabolism; D006801:Humans; D008470:Meconium / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D011247:Pregnancy; D011251:Pregnancy Complications, Infectious / Q000188:drug therapy / Q000378:metabolism; D011297:Prenatal Exposure Delayed Effects / Q000139:chemically induced / Q000378:metabolism; D053719:Tandem Mass Spectrometry / Q000592:standards* / Q000639:trends
chemical_list: D044966:Anti-Retroviral Agents