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A simple,fast and sensitive LC-MS/MS method was developed and validated for the simultaneous determination of the concentrations of temsirolimus and its major metabolite,sirolimus,in human whole blood.The blood sample (100 μL) after adding temsirolimus-d7 and sirolimus-d3 intemal standards was precipitated with 0.200 mL of methanol/0.300 M zinc sulfate (70/30,v/v) and then analyzed by a Shimatzu LC system coupled to a Sciex API-5000mass spectrometer.The chromatographic separation was carried out on a BDS Hypersil C8 column (50x 3.0 mm,5 μm) at 50℃ with a mobile phase conposed of methanol/water/formic acid (72/28/0.1)(v/v/v) containing 2.50 mM ammonium acetate.