UPLC-MS法同时测定人参不同部位中16种皂苷含量及差异性分析研究

Simultaneous Analysis and Determination of 16 Ginsenosides in Different Parts of Ginseng by UPLC-MS

本研究旨在建立并优化超高效液相色谱-质谱法(UPLC-MS)分析方法,同时测定人参花、人参果、人参茎叶和人参根不同部位中16种人参皂苷含量并分析其差异性,为人参各部位质量控制与评价提供科学依据。建立UPLC-MS分析方法,采用Thermo C-18色谱柱(100 mm×2.1 mm,1.7μm),柱温30℃,采用二元线性梯度洗脱,流动相为0.1%甲酸水-乙腈溶液,质谱条件采用电喷雾离子源(ESI),负离子模式(ESI-),以及母离子监测(Precursor ion)方式进行扫描,测定人参不同部位中人参皂苷的含量,并结合统计学分析方法主成分分析(PCA)人参不同部位中的特征成分对药材进行区分。利用UPLC-MS方法定性定量分析了16种人参皂苷成分,在0.1~20μg/mL范围内线性关系良好,相关系数R^(2)>0.99,该方法检出限为0.03~0.17μg/mL,定量限为0.1~0.5μg/mL。高、中、低3个水平的平均加样回收率为98.95%~100.41%,精密度RSD为0.96%~4.88%,灵敏度、准确度及精密度均符合要求。通过方法学考察UPLC-MS定量分析16种皂苷在相应的范围内线性关系良好,测定方法快速准确。本研究可同时准确测定人参不同部位中的皂苷含量并比较其含量差异及成分差异,为人参皂苷多组分同时分析和质量评价提供理论依据。

The study was aimed to establish and optimize the analysis method of Ultra-high performance liquid chromatography-mass spectrometry(UPLC-MS),and determine the content of 16 ginsenosides in different parts of ginseng flower,ginseng fruit,ginseng stem and leaves and analyze their differences,so as to provide scientific basis for the quality control and evaluation of all parts of ginseng.Establishing the UPLC-MS analysis method,Using a Thermo C-18 column(100mm×2.1mm,1.7 μm),Column temperature:30℃,Using a binary linear gradient for elution,mobile phase is 0.1% formic acid water-acetonitrile solution,MS conditions using an electrospray ion source(ESI),negative ion mode(ESI-),and parent ion monitoring(Precursor ion) mode for scanning,to determine the content of ginsenosides in different parts of ginseng,combined with the statistical analysis method of principal component analysis(PCA),the characteristic components in different parts of ginseng were distinguished.The sixteen ginsenoside components were analyzed qualitatively and quantitatively using the UPLC-MS method,with good linearly relationship from 0.1 to 20 μg/mL,correlation coefficient R^(2)>0.99,detection limit of 0.03 to 0.17μg/mL and limit of quantification from 0.1 to 0.5 μg/mL.The average recovery rate of high,medium and low was 98.95%~100.41 %,and the precision RSD was 0.96%~4.88%,and the sensitivity,accuracy and precision met the requirements.UPLC-MS quantified 16 saponins in the corresponding range,and the determination method was fast and accurate.This study can simultaneously accurately determine the content of saponins in the different parts of ginseng and compare the differences in content and composition,which provides a theoretical basis for the simultaneous analysis and quality evaluation of multicomponents of ginsenosides.