LC-MS/MS-IDA-EPI法同时测定中草药及中毒样品中16种吲哚生物碱

Simultaneous determination of 16 indole alkaloids in Chinese herbal medicine and poisoning samples by LC-MS/MS-IDA-EPI method

目的建立液相色谱/三重四极杆质谱-多反应监测-信息依赖性采集-增强产物离子扫描(liquid chromatography tandem mass spectrometry-multiple reaction monitoring-information dependent acquisition-enhanced product ion,LC-MS/MS-IDA-EPI)方法,用于同时测定中草药及中毒样品中16种吲哚类生物碱,包括钩吻碱、吴茱萸生物碱、钩藤生物碱、育亨宾和士的宁等。方法用氨水溶液-甲醇(体积分数为0.01%)作为流动相,以0.4 mL/min的流速进行梯度洗脱,16种吲哚类生物碱在Phenomenex Kinetex C_(18)色谱柱上分离。采用多反应监测-信息依赖性采集-增强产物离子(multiple reaction monitoringinformation dependent acquisition-enhanced product ion,MRM-IDA-EPI)扫描模式进行检测。结果阳性样品的EPI谱图搜库匹配度均>70%且与标准品的保留时间偏差在±0.1 min内。所有化合物在相应的浓度范围内线性关系良好,相关系数均>0.998,检出限为0.01~4.00(μg/L或μg/kg),定量限为0.04~13.00(μg/L或μg/kg)。平均回收率为83.43%~109.41%,相对标准偏差(relative standard deviations,RSD)为1.70%~4.51%(n=6)。结论所建立的方法简便快捷、准确、可靠,可用于中草药和中毒样品中16种吲哚类生物碱的快速定性和定量分析。

Objective To establish a liquid chromatography tandem mass spectrometry-multiple reaction monitoring-information dependent acquisition-enhanced product ion(LC-MS/MS-IDA-EPI)method for simultaneous determination of 16 indole alkaloids in Chinese herbal medicine and poisoning samples,including gelsemium elegans alkaloids,evodia rutaecarpa alkaloids,U.rhynchophylla alkaloids,yohimbine,strychnine,etc.Methods The 16 indole alkaloids were separated on a Phenomenex Kinetex C_(18) column using mobile phases of 0.01%ammonia(v/v)aqueous solution and methanol for gradient elution at a flow rate of 0.4 mL/min.The multiple reaction monitoring-information dependent acquisition-enhanced product ion(MRM-IDA-EPI)scanning modes was used for detection.Results The matching degree of the EPI spectra for the positive samples was greater than 70%,with a deviation in retention time within±0.1 min.All compounds had good linear relationships within the corresponding concentration range,with correlation coefficients>0.998.The limits of detection and the limit of quantification were 0.01-4.00(μg/L orμg/kg)and 0.04-13.00(μg/L orμg/kg),respectively.The average recovery rate was 83.43%to 109.41%,and the relative standard deviations(RSD)were 1.70%to 4.51%(n=6).Conclusion The established method is simple,rapid,accurate,and reliable,and can be used for the rapid qualitative and quantitative analysis of 16 indole alkaloids in Chinese herbal medicine and poisoning samples.