title: Development and practical application of accelerated solvent extraction for the isolation of cocaine/crack biomarkers in meconium samples.
abstract: 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 17ng/g for all analytes. The limits of quantification (LOQ) were 30ng/g for anhydroecgonine methyl ester, and 20ng/g for cocaine, benzoylecgonine, ecgonine methyl ester and cocaethylene. Linearity ranged from the LOQ to 1500ng/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 samples 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.
authors.affiliation: Faculty of Pharmaceutical Sciences-University of São Paulo, Av. Prof. Lineu Prestes, 580-Bloco 13B, Cidade Universitária, São Paulo CEP 05508-000, Brazil. Electronic address: cinthia.mantovani@usp.br.
authors.forename: Cínthia de Carvalho
authors.initials: Cde C
authors.lastname: Mantovani
authors.affiliation: Faculty of Pharmaceutical Sciences-University of São Paulo, Av. Prof. Lineu Prestes, 580-Bloco 13B, Cidade Universitária, São Paulo CEP 05508-000, Brazil.
authors.forename: Marcela Bittar
authors.initials: MB
authors.lastname: Lima
authors.affiliation: Faculty of Pharmaceutical Sciences-University of São Paulo, Av. Prof. Lineu Prestes, 580-Bloco 13B, Cidade Universitária, São Paulo CEP 05508-000, Brazil.
authors.forename: Carolina Dizioli Rodrigues de
authors.initials: CD
authors.lastname: Oliveira
authors.affiliation: Faculty of Pharmaceutical Sciences-University of São Paulo, Av. Prof. Lineu Prestes, 580-Bloco 13B, Cidade Universitária, São Paulo CEP 05508-000, Brazil.
authors.forename: Rafael de Almeida
authors.initials: Rde A
authors.lastname: Menck
authors.affiliation: University Hospital-University of São Paulo, Av. Prof. Lineu Prestes, 2565, Cidade Universitária, São Paulo CEP 05508-000, Brazil.
authors.forename: Edna Maria de Albuquerque
authors.initials: EM
authors.lastname: Diniz
authors.affiliation: Faculty of Pharmaceutical Sciences-University of São Paulo, Av. Prof. Lineu Prestes, 580-Bloco 13B, Cidade Universitária, São Paulo CEP 05508-000, Brazil.
authors.forename: Mauricio
authors.initials: M
authors.lastname: Yonamine
mesh_terms: D015415:Biomarkers / Q000032:analysis / Q000737:chemistry; D005591:Chemical Fractionation / Q000379:methods*; D016578:Crack Cocaine / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D007231:Infant, Newborn; D057230:Limit of Detection; D016014:Linear Models; D008470:Meconium / Q000737:chemistry*; D015997:Neonatal Screening / Q000379:methods*
chemical_list: D015415:Biomarkers; D016578:Crack Cocaine
keywords: Accelerated solvent extraction; Cocaine; Crack; Fetal exposure; Gas chromatography–mass spectrometry; Meconium