HPLC-DAD法同时测定孢子菌肺炎患者血浆中的磺胺甲唑与甲氧苄啶浓度

Simultaneous determination of sulfamethoxazole and trimethoprim in human plasma in patients with pneumocystis carinii pneumonia by HPLC-DAD

目的:建立HPLC-DAD法同时测定血浆中磺胺甲唑(SMZ)与甲氧苄啶(TMP)浓度,用于临床肺孢子菌肺炎(PCP)患者治疗药物监测。方法:200μL血浆样本经60μL10%高氯酸蛋白沉淀,离心后取上清液30μL进样。色谱柱为Kromasil 100-5-C_8(4.6×150 mm,5μm),流动相为甲醇–0.1 mol·L^(-1)乙酸铵(35:65),流速为0.9 m L·min^(-1),柱温为35℃,紫外检测波长为240 nm。采集20名经口服复方新诺明(0.48 g/次,3次/d)的PCP患者不同时间点共51份血浆样本,测定浓度。结果:在所建立的色谱条件下,磺胺甲唑、甲氧苄啶与各杂质分离良好,单次分析时间为6 min/样本。两待测物线性范围均为1~300μg·m L^(-1),最低检测限为1μg·m L^(-1)。通过10名PCP患者(取血点n≥3)浓度测定结果确定1 h时间点为峰浓度取血点,统计所有患者1 h时间点血药浓度结果,磺胺甲唑峰浓度均值为(131.9±41.3)μg·m L^(-1),甲氧苄啶峰浓度均值为(6.3±2.0)μg·m L^(-1)。参照目标范围浓度,仅55.0%的患者达标,其余需剂量调整。结论:此法操作简单,准确、精密度好,适用于临床PCP患者复方磺胺甲唑的治疗药物监测。

Objective： A precise and rapid HPLC-DAD method has been developed and validated for the simultaneous determination of sulfamethoxazole（SMZ） and trimethoprim（TMP） in patients with pneumocystis carinii pneumonia（PCP）. Methods： plasma sample（200ul） was deproteinized with 60μL 10% perchloric acid, and separated on a reversed phase C_8 column with a mobile phase of methanol- 0.1 mol·L^（-1） ammonium acetate（35：65） using isocratic elution. The flow rate was 0.9 min·L^（-1） and the detection wavelength was 240 nm. 51 plasma samples drawn from 20 patients with PCP were collected and quantified. Results： The calibration curves were linear over the range of 1-300 μg·m L^（-1） for both analytes. The run time for each sample of the method was within 6 min. The peak time was at 1.0h, which is obtained from the preliminary test of 10 PCP patients samples（3-5 time-point samples from each patient）. The average plasma peak concentrations of SMZ and TMP were（131.9±41.3） μg·m L^（-1） and（6.3±2.0） μg·m L^（-1）, respectively. Comparing with the target concentration range, only 55.0% patients met the criterion and the rest of them needed dose adjustment. Conclusion： This method can be used as an effective approach in clinic field.