高效液相色谱-串联质谱法测定人血清中万古霉素的浓度

Determination of concentration of vancomycin in human serum by high performance liquid chromatographytandem mass spectrum

目的建立高效液相色谱-串联质谱法(HPLC-MS/MS)测定人血清万古霉素浓度的方法。方法人血清样品用乙腈蛋白沉淀后,通过色谱柱Acquity UPLC?BEH C18(1.7μm IVD 2.1 mm×50.0 mm),以流动相(A:含0.1%甲酸的水溶液,B:含0.1%甲酸的甲醇溶液)梯度洗脱,用电喷雾离子源,正离子方式,扫描方式为多反应监测(MRM),用于监测的离子反应为m/z 725.6→144.1(万古霉素)和m/z 1145.0→100.2(万古霉素杂质C)。考察该方法的专属性、稳定性、精密度与回收率、标准曲线与定量下限、携带污染。结果万古霉素在1.5~60.0 mg/L内线性关系良好(r2=0.9992),标准曲线方程为Y=2.12×10-1X+2.29×10-3,日内精密度和日间精密度均小于10%,高、中、低3个浓度质控的回收率分别为100.30%、100.28%、95.00%,稳定性均良好。结论该方法专属性好,灵敏度高,操作便捷,可用于人血清中万古霉素血药浓度的测定。

Objective To establish an high performance liquid chromatography-tandem mass spectrum(HPLC-MS/MS)method for the determination of concentration of vancomycin in human serum.Methods Human serum samples were precipitated with acetonitrile protein and passed through a chromatographic column of Acquity UPLC?BEH C18(1.7μm IVD 2.1 mm×50.0 mm)in mobile phase(A:aqueous solution containing 0.1%formic acid,B:methanol solution containing 0.1%formic acid)for gradient elution,using electrospray ionization,positive ion mode,with scanning mode of multiple-reaction monitoring(MRM).The ion reactions used for monitoring were m/z 725.6→144.1(vancomycin)and m/z 1145.0→100.2(vancomycin impurities C).The specificity,standard curve and lower limitation of quantitation,precision and recovery rate,carrying contamination and stability of the method were investigated.Results Vancomycin has a good linear relationship in the range of 1.5 to 60.0 mg/L(r2=0.9992),and the standard curve equation was Y=2.12×10-1X+2.29×10-3.The inter-and intra-day precisions(relative standard deviation,RSD)were both less than 10%.The average absolute recoveries were 100.30%,100.28%and 95.00%for high,middle and low concentration samples,with good stability.Conclusion?This method has good specificity,high sensitivity and easy operation,which is suitable for the determination of vancomycin in human serum.