The increasing awareness and public concern with hazard exposure to endocrine-disrupting chemicals calls for methods capable to handle numerous samples in short analysis time. In this present study, a novel method combining air-assisted liquid-liquid microextraction and liquid chromatography coupled to mass spectrometry was developed and validated for the extraction, preconcentration, and determination of 7 bisphenols (bisphenol A, bisphenol S, bisphenol AP, bisphenol P, bisphenol F, bisphenol AF, bisphenol Z), 7 parabens (methyl-, ethyl-, propyl-, butyl-, benzyl-paraben, methyl-protocatechuic acid, and ethyl-protocatechuic acid), 5 benzophenones (benzophenone-1, benzophenone-2, benzophenone-3, benzophenone-8, and 4-hydroxybenzophenone), and two antimicrobials (triclosan and triclocarban) in human urine samples. Type and volume of solvent, extraction time (cycles), pH sample, ionic strength, agitation, and needle dimensions were evaluated. The matrix-matched calibration curves of all analytes were linear with correlation coefficients higher than 0.99 in the range level of 1.0–20.0 ng mL−1. The relative standard deviation, precision, at three concentrations (1.0, 10.0 and 20.0 ng mL−1) was lower than 15% with accuracy ranging from 90% to 114%. The biomonitoring capability of the new proposed method was confirmed with the analysis of 50 human urine samples randomly collected from Brazilian children. High urinary concentrations of several EDCs associated with usage of personal care products were found.