HPLC法测定结核病患者血浆中异烟肼、吡嗪酰胺、利福平和左氧氟沙星的质量浓度

目的建立同时测定人血浆中异烟肼、吡嗪酰胺、利福平和左氧氟沙星的质量浓度的高效液相色谱法。方法血浆样品采用甲醇沉淀蛋白法萃取,采用C18色谱柱(150mm×4.6mm,5.0μm),用甲醇,乙腈,0.05mol·L-1 KH2PO3水溶液(三乙胺调pH6.0)进行梯度洗脱,柱温40℃,进样量20μL,采用PDA紫外检测器。结果以峰面积外标法定量,异烟肼在0.20²5.6μg·mL-1范围内与峰面积线性关系良好(r=0.999 7),吡嗪酰胺在0.2025.6μg·mL-1范围内与峰面积线性关系良好(r=0.999 7),吡嗪酰胺在0.20²5.6μg·mL-1范围内与峰面积线性关系良好(r=0.999 9),利福平在0.4025.6μg·mL-1范围内与峰面积线性关系良好(r=0.999 9),利福平在0.40²5.6μg·mL-1范围内与峰面积线性关系良好(r=0.999 3),左氧氟沙星在0.2025.6μg·mL-1范围内与峰面积线性关系良好(r=0.999 3),左氧氟沙星在0.20²5.6μg·mL-1范围内与峰面积线性关系良好(r=0.999 4),回收率为85%25.6μg·mL-1范围内与峰面积线性关系良好(r=0.999 4),回收率为85%¹15%,日内、日间精密度RSD均<9.6%(n=5)。结论该方法专属性强、灵敏度高,准确性好,适用于同时测定人血浆样品中异烟肼、吡嗪酰胺、利福平和左氧氟沙星的质量浓度。

Simultaneous determination of isoniazid,pyrazinamide,rifampicin and levofloxacin in human plasma by HPLC

Objective To develop an HPLC method for the simultaneous determination of isoniazid ,pyrazinamide ,rifampicin and levofloxacin in human plasma .Methods The compounds were extracted from the plasma by a protein precipitation procedure with meth-anol .The LC separation of the analytes was performed on a C18 column （150 mm × 4 .6 mm ,5 .0 μm） at 40 ℃ ,using a gradient elution program with solvents consisting of methanol ,acetonitrile and 0 .05 mol ？ L - 1 KH2 PO3-H2 O .The injection volume was 20 μL .The compounds were detected by PDA .Results Calibration curves in spiked plasma were linear in the range of 0 .20-25 .6 ,0 .20-25 .6 , 0 .40-25 .6 and 0 .20-25 .6 μg ？ mL - 1 for isoniazid ,pyrazinamide ,rifampicin and levofloxacin ,respectively .The recoveries were between 85% -115% ,and the intra- and inter-day RSDs were 〈 9 .6% .Conclusion The method is specific ,sensitive and accurate ,and can be used for the simultaneous determination of isoniazid ,pyrazinamide ,rifampicin and levofloxacin in human plasma .