液相色谱法制备山柰酚单体

Purification of Kaempferol in Ginkgo Biloba L. by Preparative LC

银杏叶提取物用水饱和的正丁醇萃取,浓缩萃取液,水解后经过大孔树脂有机溶剂梯度分离纯化山柰酚。合并山柰酚纯度在50%以上的洗脱液,常压反相液相色谱纯化。检测色谱条件:采用YWGC18色谱柱(250mm×4.6mm,10μm);流动相为甲醇—(0.4%磷酸)(60∶40)系统;柱温为(30±1)℃;流速为1.0mL·min-1;检测波长为360nm。在此检测条件下山柰酚与其它组分的色谱峰得到基线分离。山柰酚的线性范围在0.05～0.5μg之间,相关系数r=0.9999。加样平均回收率为98.1%,相对标准偏差为2.7%(n=6)。经过两步层析,制备的山柰酚纯度为95%,得率为73%。本方法纯度和得率高,分离时间快,重复性好。

To separate and purify kaempferol in Ginkgo biloba L. by preparative HPLC. Extract of Ginkgo biloba L. was extracted by n-BuOH. The concentrated solution was hydrolyzed and separated by micro porous resins, with elution of solvent gradient. Kaempferol extract, which purity was higher than 70%, was purified by normal pressure RP liquid chromatography. Kaemperol was measured on a YWG C18 column (250mm×4.6mm, 10μm) and a mobile phase of methanol-0.4% H3PO4 (60∶40) at a flow rate of 1.0 mL·min-1 and temperature at (30±1)℃. The detection wavelength was 360 nm. In the chromatogram, kaempferol was baseline separated from other ingredient. Kaempferol had good linearity in the range of 0.05 μg to 0.5 μg with the correlation coefficients of 0.9999. The average recovery was 98.1% with RSD 2.7%(n=6). After two step chromatography, the final purity and recovery of purified kaempferol were 95% and 73%, which structure was confirm by IR, UV and MS. The method is swift and good repeat.