基于高分辨质谱和代谢组学技术对不同银杏叶制剂差异成分的快速分析

Rapid identification of constituents from different Ginkgo biloba preparations by high resolution mass spectrometry and metabolomics technology

采用高效液相色谱串联四极杆飞行时间质谱法(HPLC-Q/TOF-MS),利用代谢组学技术,比较银杏叶制剂组成成分的差异,包括银杏叶滴丸、银杏酮酯滴丸、银杏叶提取物片。采用XSELECT HSS T3色谱柱(4.6 mm×150 mm,3.5μm),以0.1%甲酸水-乙腈为流动相,柱温为40℃进行色谱分离,采用ESI-Q/TOF检测,负离子模式扫描。采用偏最小二乘判别分析方法(PLS-DA)进行数据统计,比较不同银杏叶制剂中的差异成分。结果表明,采用高分辨质谱鉴定了21种银杏叶提取物片与银杏叶滴丸的差异成分,其中滴丸中含量大于银杏叶提取物片的有7种,主要是黄烷醇类化合物,银杏叶提取物片中含量大于滴丸的有14种,多数是有机酸类化合物。鉴定了银杏叶滴丸与银杏酮酯滴丸的12种差异成分,其中银杏叶滴丸中含量大于酮酯滴丸的有7种,主要是黄烷醇类化合物,酮酯滴丸中含量大于银杏叶滴丸的有5种,主要是金松双黄酮和有机酸。结果阐明了不同厂家的银杏叶制剂之间的差异物质,为建立更完善的银杏制剂质控标准提供了科学指导。

A qualitative analysis was developed to identify different ingredients from three Ginkgo biloba preparations(including Yinxingye Diwan,extract of Ginkgo biloba leaves tablets and Yinxingtongzhi Diwan)based on high resolution mass spectrometry and metabolomics technology.An XSELECT HSS T3(4.6 mm×150 mm,3.5μm)column was used for separation,with the mobile phases consisting of acetonitrile and water containing 0.1%formic acid.The column temperature was set at 40°C.Negative ion mode was used for mass spectrometric data acquisition.Through partial least squares projection to latent structure-discriminant analysis(PLS-DA),we firstly found the different compounds among different Ginkgo biloba preparations.Subsequently,the compounds displaying different abundance levels via database searching and literature matching were identified.Finally,we identified 21 different compounds between the extract of Ginkgo biloba leaves tablets and Yinxingye Diwan,which mostly belong to the organic acids and flavonols families.Quantitative analysis showed that the ingredients which had higher abundance in extract of Ginkgo biloba leaves tablets were mainly organic acids,whereas those exhibiting higher levels in Yinxingye Diwan are flavonols.We also identified 12 different ingredients between the Yinxingye Diwan and Yinxingtongzhi Diwan,which were flavonols,sciadopitysin and organic acids.The results of this study are useful for studying different chemical constituents from distinct Ginkgo biloba preparations.