Journal of Chromatography B, 957 (2014) 14–23 Contents lists available at ScienceDirect Journal of Chromatography B jou rn al hom epage: www.elsevier.com/locate/chromb Development and practical application of accelerated solvent extraction for the isolation of cocaine/crack biomarkers in meconium samples Cínthia de Carvalho Mantovani a,∗ , Marcela Bittar Lima a , Carolina Dizioli Rodrigues de Oliveira a , Rafael de Almeida Menck a , Edna Maria de Albuquerque Diniz b , Mauricio Yonamine a a Faculty of Pharmaceutical Sciences—University of São Paulo, Av. Prof. Lineu Prestes, 580—Bloco 13B, Cidade Universitária, São Paulo CEP 05508-000, Brazil b University Hospital—University of São Paulo, Av. Prof. Lineu Prestes, 2565, Cidade Universitária, São Paulo CEP 05508-000, Brazil a r t i c l e i n f o Article history: Received 19 December 2013 Accepted 22 February 2014 Available online 5 March 2014 Keywords: Accelerated solvent extraction Fetal exposure Cocaine Crack Meconium Gas chromatography–mass spectrometry a b s t r a c t A method using accelerated solvent extraction (ASE) for the isolation of cocaine/crack biomarkers in meconium samples, followed by solid phase extraction (SPE) and the simultaneous quantification by gas chromatography–mass spectrometry (GC–MS) was developed and validated. Initially, meconium samples were submitted to an ASE procedure, which was followed by SPE with Bond Elut Certify I cartridges. The analytes were derivatizated with PFP/PFPA and analyzed by GC–MS. The limits of detection (LOD) were between 11 and 17 ng/g for all analytes. The limits of quantification (LOQ) were 30 ng/g for anhydroec- gonine methyl ester, and 20 ng/g for cocaine, benzoylecgonine, ecgonine methyl ester and cocaethylene. Linearity ranged from the LOQ to 1500 ng/g for all analytes, with a coefficients of determination greater than 0.991, except for m-hydroxybenzoylecgonine, which was only qualitatively detected. Precision and accuracy were evaluated at three concentration levels. For all analytes, inter-assay precision ranged from 3.2 to 18.1%, and intra-assay precision did not exceed 12.7%. The accuracy results were between 84.5 and 114.2% and the average recovery ranged from 17 to 84%. The method was applied to 342 meconium sam- ples randomly collected in the University Hospital—University of São Paulo (HU—USP), Brazil. Cocaine biomarkers were detected in 19 samples, which represent 5.6% of exposure prevalence. Significantly lower birth weight, length and head circumference were found for the exposed newborns compared with the non-exposed group. This is the first report in which ASE was used as a sample preparation technique to extract cocaine biomarkers from a complex biological matrix such as meconium samples. The advantages of the developed method are the smaller demand for organic solvents and the minor sample handling, which allows a faster and accurate procedure, appropriate to confirm fetal exposure to cocaine/crack. © 2014 Elsevier B.V. All rights reserved. 1. Introduction The abuse of illicit drugs during pregnancy is a relevant pub- lic health problem that affects not only the mother but also the infant. In the United States, according to the 2010 National Survey on Drug Use and Health performed by the Substance Abuse and Mental Health Services Administration (SAMSHA), 4.4% of pregnant women aged 15 to 44 years old reported they were currently illicit drug users [1]. In Brazil, in spite of epidemiological data about this ∗ Corresponding author. Tel.: +55 11 3091 2194. E-mail addresses: cinthia.mantovani@usp.br, cinthia mantovani@hotmail.com (C.d.C. Mantovani). phenomenon are still scarce, it seems that the problem is not differ- ent. In a study performed with 1000 Brazilian adolescent pregnant women (aged 11 to 19 years old), hair toxicological analysis con- firmed that 6.0% of them had used cocaine and/or marijuana during the last trimester of pregnancy. None of the mothers had reported the use of these substances in their interviews with healthcare professionals [2]. The negative impact of cocaine/crack abuse during pregnancy is widely described in the scientific literature. Low birth weight, intra-uterine growth restriction, preterm birth, neurobehavioral and developmental effects are associated with cocaine use dur- ing gestation [3–5]. Therefore, the assessment of in utero exposure would be very important for proper medical and psychological http://dx.doi.org/10.1016/j.jchromb.2014.02.039 1570-0232/© 2014 Elsevier B.V. All rights reserved.