基于LC-MS拟靶向代谢组学的山西北柴胡质量分析

Quality evaluation of Bupleuri Radix in Shanxi Province by LC-MS based pseudotargeted metabonomics

柴胡是中医临床常用中药材,皂苷是其重要的活性成分。本研究首先利用超高效液相色谱-三重四极杆复合线性离子阱质谱(UHPLC-QTrap-MS)在智能化分时间段-多反应选择离子监测(scheduled multiple reaction monitoring,sMRM)模式下建立了25种柴胡皂苷的拟靶向相对定量方法,所建立的方法具有良好的精密度、线性、重复性和稳定性。将该方法应用于37批不同产地北柴胡样本的比较,结果显示山西不同产地北柴胡皂苷差异不大,表明北柴胡生长适应力强,受产地环境因素影响小,适合广泛种植。夏季采收的柴胡与传统的春秋采收期采收的柴胡的皂苷类成分存在一定差异,说明传统采收经验的合理性。相关性分析表明,柴胡皂苷a和d与部分皂苷类成分存在显著的正相关,通过变异系数分析确定了clinoposaponin XII等4种波动较大的皂苷类成分。本研究建立的LC-MS拟靶向代谢组学方法,可以应用于柴胡皂苷类成分的全面检测,为柴胡的质量评价研究提供新的方法。

Bupleuri Radix is commonly used in the traditional Chinese medicine,and saikosaponins are the important active ingredients.In this study,we first established a relative quantitative method for 25 saikosaponins using ultra high performance liquid chromatography-triple quadrupole mass spectrometry(UHPLC-QTrap-MS)in the scheduled multiple reaction monitoring(sMRM)mode.The established method showed good intra-day and intra-day precision,linearity,repeatability and stability.Then the method was applied to compare 37 batches of Bupleuri Radix from different planting areas.The results showed that there was no significant difference in the saikosaponins composition of Bupleuri Radix from different planting areas in Shanxi Province,which indicating that Bupleuri Radix is well adapted to the environment,so it is suitable for widely planting.However,Bupleuri Radix harvested at spring and autumn were differed from those harvested at summer,which indicated that the traditional harvesting experience was reasonable.Correlation analysis showed that saikosaponins a and d were positively correlated with some saponins,and 4 saponins(such as clinoposaponin XII)showed bigger content variation were identified by coefficient of variation analysis.The LC-MS based pseudotargeted metabonomic method established in this study can be applied to the comprehensive detection of saikosaponins,which providing new method for the quality evaluation of Bupleuri Radix.