Human milk is an important source of beneficial nutrients and antibodies for newborns and infants and, under certain circumstances, its analysis may provide information on mothers' and infants' exposure to various contaminants. In the presented study, we have introduced the new analytical approach for analysis of 24 highly occurring polycyclic aromatic hydrocarbons (PAHs) in this indicator matrix. The sample preparation procedure is based on an ethyl acetate extraction of milk; the transfer of analytes into an organic layer is enhanced by addition of inorganic salts, i.e. sodium chloride and magnesium sulphate. Following the clean-up of a crude extract on silica SPE columns, gas chromatography coupled to triple quadrupole mass spectrometry is used for PAH identification and quantitation. The average recoveries of targeted PAHs from spiked samples were in the range of 68-110% with repeatabilities below 30% and method quantitation limits ranging from 0.03 to 0.3ng/g lipid weight. This newly validated method was successfully applied for analyses of 324 human milk samples collected from nonsmoking women during two sampling periods (summer and winter) in two residential areas in the Czech Republic differing in atmospheric pollution by PAHs. From 24 targeted analytes 17 were detected at least in one sample. Phenantherene, fluoranthrene, pyrene and fluorene were the most abundant compounds found at average concentration of 13.81, 1.80, 0.86, and 2.01ng/g lipid weight respectively. Comparing the data from two sampling periods, in both areas higher concentrations were measured in samples collected during winter. Also in the highly industrialized locality with heavily contaminated air PAH amounts in milk were higher than in the control locality. These first data on PAH concentrations in human milk collected in the Czech Republic are comparable with measurements for nonsmoking women reported earlier in the United States but significantly lower than results from China, Turkey or Italy.

Institute of Experimental Medicine AS CR Videnska 1083 142 20 Prague 4 Czech Republic

University of Chemistry and Technology Prague Faculty of Food and Biochemical Technology Department of Food Analysis and Nutrition Technicka 3 166 28 Prague 6 Czech Republic

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Airborne Benzo[a]Pyrene may contribute to divergent Pheno-Endotypes in children