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Development and validation of the first liquid chromatography-tandem mass spectrometry assay for antiretrovirals in meconium

Wednesday, October 10, 2012 — Poster Session II

Noon – 2:00 p.m

Natcher Conference Center, Building 45




  • S.K. Himes
  • K.B. Scheidweiler
  • K. Tassiopoulos
  • D. Kacanek
  • R. Hazra
  • K. Rich
  • M.A. Huestis


Antiretroviral (ARV) administration to HIV-infected pregnant women and neonates reduces perinatal HIV transmission to less than 2% worldwide. However, concerns have been raised about potential toxicity in some neonates following gestational ARV exposure. Precise quantification of ARV exposure by history is difficult. Quantitative analysis of meconium, the first neonatal feces, may better reflect fetal exposure during the third and perhaps second trimesters than history alone. Therefore, a novel method for the simultaneous quantification of 16 ARV drugs and 4 metabolites in meconium by liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed and validated. Blank meconium (0.25g) was fortified with tenofovir, lamivudine, emtricitabine, abacavir and its carboxylate and glucuronide metabolites, nevirapine, raltegravir, saquinavir, amprenavir, darunavir, atazanavir, ritonavir, lopinavir, nelfinavir and its bioactive hydroxyl metabolite, stavudine, efavirenz, zidovudine and its glucuronide metabolite. Samples were homogenized in methanol and subjected to solid phase extraction prior to quantification by LC-MS/MS. Quantification employed several deuterated and isotopically-labeled internal standards. Linearity (1/x2) ranged from 10-75ng/g up to 2500ng/g and 100-500ng/g up to 25000ng/g; correlation coefficients were ≥0.99. Method applicability was demonstrated by analyzing meconium from HIV-uninfected infants born to HIV-infected mothers on ARV therapy during pregnancy (collected as part of the Pediatric HIV/AIDS Cohort Study).

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