Contamination of drinking water by ethylene glycol (EG) is a public health concern. EG causes adverse health effects in humans and animals, including cardiopulmonary and acute renal failure. EG and other glycols, such as propylene glycol (PG) are major components in antifreeze liquids, which may be the main source of contamination of ground water. A sensitive LC/electrospray ionization (ESI)-MS/MS method was developed to measure trace amounts of EG, diethylene glycol, and 1,2- and 1,3-PG in several water sources, including municipal tap, lake, river, and salinated water. In this method, glycols in water samples were derivatized with benzoyl chloride by the Schotten-Baumann reaction, followed by liquid-liquid extraction using pentane as the organic solvent prior to the LC/ESI-MS/MS determination. QC included analysis of a method blank and samples fortified at low and high levels. Analytical data showed excellent linear calibration for all observed glycols, with good precision and accuracy. The method detection limits for the studied glycols ranged from 1.9 to 6.1 ng/mL across the water matrixes tested. This method is suitable to help assess water quality in areas that may be prone to glycol contamination.