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

Determination of Gentamicin Illegally Added in Houttuynia Injection by HPLC-ELSD and UPLC-MS/MS

关键词:  庆大霉素  鱼腥草注射液  液相色谱-蒸发光散射检测器  超高效液相色谱-串联质谱

Key words:  Gentamicin  Houttuynia Injection  HPLC-ELSD  UPLC-MS/MS

摘要:

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。以十八烷基键合硅胶为填充剂,0.2 mol/L三氟醋酸 -甲醇(92:8)为流动相作为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,流速为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 μg/ml～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。

Abstract:

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 μg/ml～1.35 μg/ml with a limit of detection of 0.15 g/L in Houttuynia Injection .