A high performance liquid chromatographic(HPLC) method was established for the simultaneous determination of 3 drugs for diabetic mellitus (DM),including metformin hydrochloride,glipizide and telmisartan,in urine.The macromolecules in urine samples of DM patients were firstly removed by acetone precipitation method,and the supernatants were dried and redissolved in methanol.Then the extracts were analyzed by HPLC coupled with a diode array detector.The separation of analytes was performed on an Inertsil ODS column by gradient elution using acetonitrile-2.5 mmol/L SDS-20 mmol/L NaH2PO4 solution (pH=4.6) as mobile phase.The recoveries of 3 diabetic mellitus drugs were in the range of 82%-113% with relative standard deviations of 1.5%-3.1%.To obtain highest sensitivity,different wavelengths were chose for different drugs.The detection limits were 13.6 μg/L for metformin hydrochloride at 232 nm,31.5 μg/L for glipizide at 276 nm and 57.0 μg/L for telmisartan at 297 nm,respectively.The linear ranges of metformin hydrochloride,glipizide and telmisartan were in the range of 1.0-1000,1.0-1000,5.0-250.0 mg/L,respectively.The method was used to detect the contents of 3 drugs in real urine samples of DM patients.The result showed the method was applicable for the study of DM pharmaco metabonomics. |