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Exposure to persistent organic pollutants begins in uterine life. The study was conducted to evaluate associations between gestational diabetes mellitus and the presence of persistent organic pollutants in placenta samples. They were derived from a birth cohort study in Ourense (NW Spain). Selective pressurized liquid extraction methodologies were used to determine targeted persistent organic pollutants in placenta samples. Cleanup of extracts was performed by solid-phase extraction using EZ-POP cartridges and detection by gas chromatography coupled to tandem mass spectrometry. Statistical calculations were performed by comparing the values obtained in the case and control groups. Statistical significance was considered as p < 0.050. Results: The concentrations of several congeners of polybrominated diphenyl ethers (PBDE) and polychlorinated biphenyls (PCB) presented lower levels in the placentas of expectant mothers with gestational diabetes mellitus when comparing them to the control cohort. This difference was statistically significant. It was revealed that this association was independent of obesity, age, parity, amenorrhoea at birth, or a family history of diabetes mellitus. To the best of our knowledge, this paper reported an inverse relationship between PBDE and PCB levels in placenta and gestational diabetes mellitus for the first time. We believe this may open a door to future studies in this field. [ABSTRACT FROM AUTHOR]