<p>Excessive exposure to endocrine-disrupting chemicals (EDCs) during pregnancy can have long-term health effects for the mother and the neonates. However, longitudinal data on EDC accumulation throughout gestation are limited. This prospective cohort study aimed to characterize the changes in paraben and phthalate exposure throughout gestation. We enrolled 641 healthy pregnant women and collected urine samples at four time points during routine antenatal visits, alongside clinical outcome data. Samples were analyzed using UPLC coupled with a triple quadrupole mass spectrometer. Methyl paraben was predominantly found across all time points (5.65 to 3070.87, 10.60 to 5261.46, 11.41 to 903.68 and 6.12 to 899.69 ng/ml respectively). Overall paraben concentrations peaked during the second trimester, with total parabens reaching 103.39 (IQR: 243.35–527.80). Among phthalates, monoethyl phthalate showed the highest levels throughout pregnancy (2.20 to 1541.36, 7.31 to 2530.13, 2.18 to 656.70 and 0.44 to 980.30 ng/ml respectively). In correlation analysis, certain parabens (EP, HP) and phthalate metabolites (MCPP, MMP, MiNP) showed negative correlations with birth weight, birth length, and vitamin D levels in the second trimester; however, subsequent regression analyses did not identify any statistically significant associations. This study offers a comprehensive exploration of the exposure patterns of parabens and phthalate metabolites throughout gestation emphasizing pronounced exposure during the second trimester. The elevated levels of methyl paraben observed in this Indian cohort, compared to international data, underscore the critical need for region-specific regulatory guidelines. Our findings also establish a foundational reference for EDCs in pregnant Indian women, emphasizing future public health strategies.</p> Graphical Abstract <p></p>