中生菌素原药有效成分高效液相色谱-串联质谱分析方法的建立

建立了采用高效液相色谱-串联质谱(HPLC-ESI-MS/MS)分析中生菌素(zhongshengmycins) 原药中各有效成分(链丝菌素A~F)的方法。原药用去离子水超声辅助提取后经离心过滤,反相离子对高效液相色谱分离,二级质谱检测,标准样品定量离子外标法定量。结果表明:在15.63~500 μg/mL质量浓度范围内,中生菌素原药各组分的仪器响应值与进样质量浓度之间呈良好的线性关系,相关系数 R2>0.990 5;方法具有较好的精密度和准确度,6次重复进样,其相对标准偏差(RSD)在0.33% ~1.96%之间;在0.1、0.5和1 μg/mg 3个添加水平下,各组分的回收率在98.2% ~101.1%之间。样品实测结果表明:供试原药中链丝菌素D的含量最高,为297.65 μg/g;其次是链丝菌素B、C和F,含量分别为247.77、285.64和115.92 μg/g;链丝菌素A和E的含量较低,分别为15.63和19.60 μg/g。该方法能满足中生菌素原药中各有效成分定性及定量分析的要求。

High-performance liquid chromatography-electrospray ionization-tandem mass spectrometry method for determination of active ingredients in technical zhongshengmycin

A reliable method for determination of active ingredients in technical zhongshengmycin material was established using high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-ESI-MS/MS).The active ingredients in technique material were extracted by the ultrasound-assisted method,and then the aqueous extract was cleaned up by centrifugation and filtration.After separated by reverse-phase ion-pair chromatography,the characteristic MS/MS ions were detected by mass spectrometer and quantified with external standard method.A good linear relationship between response and concentration was obtained in the range of 15.63-500 μg/mL,and its correlation coefficient was above 0.990 5.The relative standard derivations for different ingredients were in the range of 0.33% -1.96%.The recovery rates for different ingredients were 98.2% -101.1% in the spiking levels of 0.1,0.5 and 1 μg/mg.The contents of streptothricins D,B,C,F,A and E in the tested technical material were 297.65,247.77,285.64,115.92,15.63 and 19.60 μg/g,respectively.The method could be used to qualitative and quantitative analysis of active ingredients in technical zhongshengmycin.