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Environmental hormone belongs to a class of chemicals in the environment.It can affect the endocrine system of human and animals and lead to the reproductive, developmental and behavioural abnormalities, phthalate esters(PAEs)are one of the important environmental hormones, mainly used as plasticizers.In this paper, we choosed 5 phthalate monoesters which were reported greater toxicity as research objects, including MMP, MEP, MBP, MBzP and MEHP.Five Phthalate metabolites were extracted from the urine sample using liquid-liquid extraction and the amounts of urine were 20ml.The Optimization conditions of liquid-liquid extraction were that the usage amount of n-hexane was 10 ml and three times.Then flowed the elution solution to dry by nitrogen, and diluted them to 200μl with pure water.The contents of 5 kinds of phthalates metabolites were determined using RT-HPLC with the wave-length of determine was 228nm(Chart 1).HPLC condition: Sepax Technologies GP-Phenyl(5μm,4.6×250mm,120A) cartridge; acetonitrile(0.2 % acetic acid): water(0.2 % acetic acid)=45∶55 (v/v) as mobile phase.