四极杆飞行时间串联质谱辅助薄层色谱鉴别重楼中多种重楼皂苷

Identification of saponins in Paridis rhizoma by thin layer chromatography with quadrupole time of flight mass spectrometry

目的:建立重楼中皂苷类成分的薄层色谱-质谱联用分析法。方法:样品经乙醇回流提取,点样于硅胶G预制板上,以三氯甲烷-乙酸乙酯-甲醇-水(17∶40∶21∶10)10℃以下放置的下层溶液为展开剂展开,喷以10%硫酸乙醇溶液,于105℃加热至条斑清晰,置紫外光灯(365 nm)下检视并成像。使用重楼皂苷Ⅰ、重楼皂苷Ⅱ、重楼皂苷Ⅵ和重楼皂苷Ⅶ对照品以及四极杆飞行时间串联质谱(QTof MS)进行成分识别。结果:紫外光灯(365 nm)下,重楼色谱可见23个荧光条斑,其中以薯蓣皂苷元为母核的皂苷显棕色,以偏诺皂苷元为母核的皂苷显淡蓝色,使用对照品和质谱仪共鉴别8个条斑,对应成分为重楼皂苷Ⅰ、重楼皂苷Ⅱ、重楼皂苷Ⅲ、重楼皂苷Ⅴ、重楼皂苷Ⅵ、重楼皂苷Ⅶ、重楼皂苷D和纤细薯蓣皂苷。结论:该法简便、专属,适用于重楼的快速鉴别并可为重楼药材整体质量评价提供参考。

Objective： To establish a method for identification of saponins in Paridis Rhizoma by thin layer chromatography -mass spectrometry. Methods： The samples were extracted by reflux with ethanol, and the chromatography was carried out on silica gel G pre - coated plates, using the lower layer of a mixture of chloroform - ethyl ace- tate -methanol -water（ 17：40： 21： 10） as mobile phase; the temperature was controlled below 10 ℃, and the samples were examined in ultraviolet light at 365 nm after derivatization with 10% sulfuric acid alcoholic solution. The bands for compounds were identified by reference substances of Chonglou saponin I , Chonglou saponin Ⅱ , Chonglou saponin VI and Chonglou saponin Ⅷ and quadrupole time of flight mass spectrometry（QTofMS）. Results： In the chromatogram of Paridis Rhizoma, twenty three bands were visualized by UV light at 365 nm, eight bands were identified, including Chonglou saponin I , Chonglou saponin Ⅱ , Chonglou saponin m, Chonglou saponin V, saponin Ⅵ, Chonglou saponin Ⅶ, Chonglou saponin D amd gracillin. The bands of diosgenin were brown, and the bands of pennogenin were light blue. Conclusion： The established method is simple and specific, which can be used for rapid detection and quality evaluation of Paridis Rhizoma.