重楼属植物甾体皂甙的高效液相色谱分析

应用高效液相层析技术,对重楼属十八个种植物的甾体皂甙进行了定性、定量分析。植物用甲醇提取,抽出物经DIAION柱,以90％甲醇洗出总甙。在HPLC上,用ODS柱先将总甙以用醇:水(9:1)洗脱分为三馏段,每馏再在ODS柱或Rp-8柱上以甲醇:水(8:2;7:3.5)洗脱,使各个皂甙成分完全分离。被分离的每个色谱峰与已知重楼皂甙的保留时间进行比较并配合HPLC的加入法,TLC分析及用HPLC制备少量样品做MS测定来加以定性鉴定。定量采用内标及校正曲线法。

THE DETERMINATION OF THE STEROIDAL SAPONIN FROM PARIS PLANTS BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

The steroidal saponin of 18 species of Paris were qualitatively and quantitatively analysed by means of HPLC. Methanolic extracts obtained from the dried roots of Paris plants were combined and concentrated in vacuo. The residue was dissolved in H2O and then the solution chromatographed on a DIAION HP-20 column eluting with H2O and 90% MeOH successively. The 90% MeOH fraction was concentrated in vacuo to give a residue crude saponins. This residue was chromatographed on a ODS column with MeOH-H2O (9:1) to divide into three fractions. The each fraction was further separated and purified by ODS or R - 8 column chlumatography using MeOH-H2O with increasing H2O content as cluent to give complefely separated each chemical constituents. The identification of the each chromatographic peak separation on HPLC was compared with retention time and TLC as well as MS detection of pure compound separated from HPLC. The internal standard and calibration curve method were used for the quantitative analysis.