Simultaneous Determination of Sulfamethoxazole and Trimethoprim Concen-trations in Human Serum by LC-MS/MS and Application in Therapeutic Drug Monitoring
Objective:To establish an LC-MS/MS method for the simultaneous determination of sul-famethoxazole(SMZ)and trimethoprim(TMP)concentrations in human serum and applie it to therapeutic drug monitoring.Methods:Sulfamethoxazole-13C6 and deuterated trimethoprim-2H3 were used as internal standards.Serum samples were precipitated with acetonitrile containing internal standards,and chromato-graphic separation was achieved using an HSS T3 column(2.1 mm × 50 mm,1.8 μm).Gradient elution was performed with 5 mmol·L-1 ammonium formate(containing 0.1％formic acid)and acetonitrile as mobile phase.The detection was carried out using an electrospray ionization source in multiple reaction monitoring mode.Results:SMZ and TMP showed good linearity in the ranges of 2.50-500 μg·mL-1 and 0.100-20.0 μg·mL-1,respectively.The intra-and inter-batch precision and accuracy for both SMZ and TMP were within 15.0％.The extraction recovery rates were 98.4％-100.1％for SMZ and 100.2％～107.6％for TMP,with matrix ef-fects ranging from 85％-115％.SMZ and TMP in serum samples were stable for 28 hours at room tem-perature,after three freeze-thaw cycles at-40℃ or-80℃,or after long-term freezing at-40℃ or-80℃ for 110 days.Whole blood samples were stable for 24 hours at room temperature.The test solutions were also stable for 72 hours in an autosampler(10℃)or a refrigerated storage(4℃).The monitoring results of SMZ and TMP blood concentrations in patients were all within the standard curve ranges.Conclusion:This method allows for rapid and sensitive detection of SMZ and TMP concentrations in human serum.
SulfamethoxazoleTrimethoprimTherapeutic drug monitoringLC-MS/MS
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