UPLC法同时测定人血浆中异烟肼、吡嗪酰胺、利福平的浓度

Simultaneous Determination of Isoniazid, Pyrazinamide and Rifampicin in Human Plasma by UPLC

目的:建立同时测定人血浆中异烟肼、吡嗪酰胺、利福平浓度的方法。方法:色谱柱:ACQUITY UPLC®BEH C18 (2.1 ×50 mm, 1.7 μm),柱温:40℃,流动相分别为乙腈及磷酸二氢钾溶液(0.025 mol∙L−1),血浆采用蛋白沉淀法前处理,流速0.3 ml/min,进样量1 μL。进样含有异烟肼、吡嗪酰胺、利福平对照品的血浆,检测方法的专属性、线性关系、回收率、精密度及稳定性,并通过该方法测定联用异烟肼、吡嗪酰胺、利福平后的结核患者血浆中的药物浓度。结果:血浆样本中异烟肼、吡嗪酰胺、利福平均独立出峰,互不干扰;线性关系良好(均r2 > 0.999);回收率均在80%~100%;精密度RSD值均

Objective: To establish the method in order to simultaneous determine concentration of isoniazid, pyrazinamide and rifampicin in human plasma. Methods: The chromatographic column was ACQUITY UPLC®BEH C18 (2.1 ×50 mm, 1.7 μm), the column temperature was 40˚C, the mobile phase was acetonitrile and potassium dihydrogen phosphate solution (0.025 mol∙L−1), the plasma was pretreated by protein precipitation method, the flow rate was 0.3 mL/min, and the inject volume was 1 μL. The plasmacontaining isoniazid, pyrazinamide and rifampicin was injected. Then we tested the specificity, linear relationship, recovery, precision and stability. The drug concentration in the plasma of tuberculosis patients who took isoniazid, pyrazinamide and rifampicin was determined by UPLC-UV method. Results: The peaks of isoniazid, pyrazinamide and rifampicin were independent and did not interference of each other, and the linear relationship was good (r2 > 0.999). The recoveries was between 80%~100%, the intra- and inter-day precision was less than 10%, and the stability was was less than 11%. Among the 17 patients with pulmonary tuberculosis, except 1 case of isoniazid below the linear range and 1 case of pyrazinamide was higher than the linear range, the rest were in the detection range. Conclusion: This method is convenient, rapid, accurate and stable, and can be used for the determination of isoniazid, pyrazinamide and rifampicin in plasma of patients with clinical tuberculosis.