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Liquid Chromatography With Tandem Mass Spectrometric Detection for the Measurement of Ethyl Glucuronide and Ethyl Sulfate in Meconium: New Biomarkers of Gestational Ethanol Exposure?

Morini, Luca BSc*; Marchei, Emilia BSc; Pellegrini, Manuela BSc; Groppi, Angelo PhD*; Stramesi, Cristiana PhD*; Vagnarelli, Federica MD; Garcia-Algar, Oscar MD, PhD§; Pacifici, Roberta PhD; Pichini, Simona PhD

doi: 10.1097/FTD.0b013e31818b2fd9
Original Article

A liquid chromatography tandem mass spectrometric (LC-MS/MS) method with postcolumn addition of acetonitrile for the determination of ethyl glucuronide (EtG) and ethyl sulfate (EtS) in meconium was developed and validated using pentadeuterated EtG and pentadeuterated EtS as internal standards. The analytes were extracted from the matrix by acetonitrile, concentrated by solid phase extraction, separated using a reversed-phase chromatographic column, and quantified within 9 minutes. Lower limits of quantification were 5 and 1 ng/g meconium for EtG and EtS, respectively. Calibration curves were linear from lower limits of quantifications to 500 ng/g, with a minimum r2 > 0.999. At 3 concentrations spanning the linear dynamic range of the assay, mean recoveries ranged between 78.7% and 96.8% for EtG and between 72.1% and 95.6% for EtS. Inaccuracy was better than 8.1%, with intra-assay and interassay imprecision better than 7.2% and 10.5%, respectively. Matrix effects (ion suppression/enhancement) were found to be negligible. The analytes of interest were stable at room temperature, at 4°C, when exposed to 3 freeze-thaw cycles, and when stored at −20°C for up to 6 months. This sensitive and specific method was used to assess the presence of these alcohol biomarkers in meconium samples from 2 different city cohorts.

From the *Department of Legal Medicine and Public Health, University of Pavia, Pavia; †Department of Therapeutic Research and Medicine Evaluation, Istituto Superiore di Sanitá, Roma; ‡Arcispedale Santa Maria Nuova, Reggio Emilia, Italy; and §Unitat de Recerca Infància i Entorn, Paediatric Service, Hospital del Mar, Barcelona, Spain.

Received for publication April 30, 2008; accepted August 21, 2008.

Correspondence: Simona Pichini, Department of Therapeutic Research and Medicine Evaluation, Istituto Superiore di Sanità, V.le Regina Elena 299, 00161 Rome, Italy (e-mail:

© 2008 Lippincott Williams & Wilkins, Inc.