HPLC-MS/MS法同时测定人血浆中3种抗结核药物的浓度

目的 建立快速、灵敏的高效液相色谱-质谱/质谱联用（HPLC-MS/MS）法测定人血浆中异烟肼、利福平和左氧氟沙星的浓度.方法 血浆样品以加替沙星为内标采用甲醇沉淀蛋白法萃取,采用C4色谱柱（250 mm×4.6 mm,5.0 μm,Welch materials,USA）,流动相为0.05%甲酸溶液-甲醇（v/v）梯度洗脱,流速1.0 mL·min-1,柱温25℃,进样量20μL,采用ESI＋多反应选择离子检测（MRM）.结果 异烟肼在0.18～9.0μg·mL-1与峰面积线性关系良好（r=0.999 8）,利福平在0.8～40μg·ml-1与峰面积线性关系良好（r=0.999 6）,左氧氟沙星在0,09～4,5μg·mL-1与峰面积线性关系良好（r=0.999 0）,萃取回收率均〉70%,方法回收率在85%～115%,日内、日间精密度RSD均〈9.3%（n=5）.结论 该方法专属性强、灵敏准确,适用于同时测定人血浆样品中异烟肼、利福平和左氧氟沙星的浓度.

Simultaneous determination of 3 anti-tuberculosis drugs in human plasma by HPLC-MS/MS

Objective To develop an HPLC-MS/MS method for the simultaneous determination of isoniazid,rifampicin and levofloxacin in human plasma.Methods Gatifloxacin was used as the internal standard.The compounds and internal standard were extracted from the plasma by a protein precipitation procedure with methanol.The LC separation of the analytes was performed on a Welch material RP C4 column（250 mm×4.6 mm,5.0 μm,USA） at 25 ℃,using a gradient elution program with solvents consisting of 0.05% formic acid and methanol（v/v） at 1.0 mL·min-1 and an injection volume of 20 μL.The compounds were ionized in the electrospray ionization（ESI） ion source of the mass spectrometer and were detected in multiple reaction monitoring mode.Results Calibration curves in spiked plasma were linear at 0.18-9.0,0.8-40,and 0.09-4.5 μg·mL-1 for isoniazid,rifampicin,and levofloxacin respectively.The extraction recoveries were〉70%,the methodology recoveries were 85%-115%,and the intra-and inter-day RSDs were 〈9.3%.Conclusion The method is specific,sensitive and accurate,and it can be used for the simultaneous determination of isoniazid,rifampicin,and levofloxacin in human plasma.