To preserve human health, persistent organic pollutants (POPs) and current-use pesticides (CUPs) should be monitored in fatty tissues, including breast milk. Therefore, this study aimed to optimize sample preparation conditions using a 32 factorial design for the determination of POPs and CUPs by gas chromatography coupled to mass spectrometry with negative chemical ionization (GC-NCI-MS). The method was validated for 57 POP and CUP compounds using hexane:acetone for extraction and clean-up by dispersive solid phase extraction (d-SPE) with Florisil®. The matrix effect was compensated by extracted analytical calibration. Method validation showed satisfactory results with limits of detection of 3 to 13 ng g-1 of fat. The method presented adequate accuracy (recoveries from 72 to 117%) and precision (relative standard deviation (RSD) ≤ 18%) and was applied to breast milk samples from Rio Grande do Sul State, southern Brazil, wherein all samples contained at least one compound. With principal component analysis, it was possible to associate the pesticides detected with the city of origin of the samples and the number of pregnancies of nursing mothers. Additionally, the analytical method was effective for the determination of trace levels of POPs and CUPs in breast milk and can be applied in biomonitoring studies.
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