While phthalates monoesters have been recognized as the bioactive metabolites of phthalates, the knowledge on their environmental occurrence and sources is limited. In this study, monomethyl phthalate (MMP), monoethyl phthalate (MEP), mono-iso-butyl phthalate (MiBP), mono-n-butyl phthalate (MnBP), and mono-2-ethylhexyl phthalate (MEHP) were frequently detected in water samples from the Taihu Lake in China using an improved SPE-LC-MS-MS method. The mean concentrations for MMP, MEP, MiBP, MnBP, and MEHP were 51.7 ± 25.2, 6.0 ± 4.8, 19.6 ± 14.6, 42.2 ± 64.7, and 33.0 ± 37.4 ng/L, respectively, while those of their corresponding parent chemicals, DMP, DEP, DiBP, and DnBP and DEHP, were 36.54 ± 43.22, 42.64 ± 66.66, 246.8 ± 311.1, 524.7 ± 586.9, and 208.1 ± 223.5 ng/L, respectively. Three secondary monoesters of DEHP, mono (2-ethyl-5-hydroxyhexyl) phthalate (MEHHP), mono (2-ethyl-5-oxohexyl) phthalate (MEOHP), and mono (2-ethyl-5-carboxypentyl) phthalate (MECPP) were for the first time detected with mean concentration of 1.27 ± 1.33, 1.33 ± 1.54, and 0.73 ± 0.79 ng/L, respectively. The percentage of the sum concentration of MEOHP, MEHHP, and MECPP relative to total concentration of DEHP metabolites was 5.3–12.4%. DEHP was identified to be biodegraded into secondary phthalate monoesters in water from the Taihu Lake, but their contribution to the total concentration of DEHP metabolites was 1.2–3.6%, lower than those in the Taihu Lake. Primary and secondary DEHP monoesters were also detected in influents and effluents of two sewage treatment plants adjacent to the Taihu Lake, the percentages of secondary DEHP monoesters in influents were 5.8% and 11.3%, similar with those in the Taihu Lake. Taken together with their relatively high concentrations in influents, the discharging of domestic wastewater may be an important contributor to the occurrence of phthalate monoesters in the Taihu Lake.