HPLC-ELSD法联合UPLC-MS/MS法检测鱼腥草注射液中非法添加的庆大霉素

摘要

A primary screening method of gentamicin in houttuynia injection was established by high performance liquid chromatography with evaporative light scattering detector(HPLC-ELSD), using 0.2 mol/L trifluoroacetic acid and methanol(92 : 8)as the mobile phase. A qualitative and quantitative method of gentamicin in houttuynia injection was established by ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS),using water(0.1% formic acetic)and acetonitrile(0.1% formic acetic)(90 : 10)as the mobile phase at a flow rate of 0.3 mL/min. At positive electrospray ionization mode,selected reaction monitoring of the precursor-product ion transitions m/z of 478.6→157.1(C1)for gentamicin. The linear calibration curve of gentamicin was y=5519.3x+879.24(R=0.9965),obtained in a concentration range of 0.15~1.35 μg/mL with a limit of detection of 0.15 g/L in houttuynia injection.%为检测鱼腥草注射液中非法添加的庆大霉素,建立了以HPLC-ELSD进行初筛,UPLC-MS/MS进行确证并定量的系统方法.十八烷基键合硅胶为填充剂,0.2 mol/L 三氟醋酸 -甲醇(92 : 8)流动相作为HPLC-ELSD初筛条件.以0.1%甲酸乙腈溶液-0.1%甲酸水溶液(10 : 90)为流动相,流速0.3 mL/min,在电喷雾离子化电离源上以多反应监测方式进行正离子检测,建立UPLC-MS/MS检测方法,用于定量分析的离子对为m/z 478.6→157.1(庆大霉素C1).结果表明,庆大霉素C1在0.15~1.35 μg/mL 浓度范围内,峰面积与浓度呈良好的线性关系,线性方程为y=5519.3x+879.24,R=0.9965.庆大霉素C1在鱼腥草注射液中检测限为0.15 g/L.