高速逆流色谱分离制备橘皮中多甲氧基黄酮

Preparative Isolation and Purification of Polymethoxylated Flavones from Citrus Peel by High-speed Counter-counter Chromatography

建立高速逆流色谱法分离制备橘皮中4种多甲氧基黄酮类化合物的方法。以石油醚-乙酸乙酯-甲醇-水(1∶0.8∶1∶1.2,V/V)为两相溶剂系统,上相为固定相,下相为流动相。在主机转速800 r/min、流动相流速2.0 m L/min、检测波长254 nm条件下分离,从200 mg橘皮粗提物中一步分离制备得到5,6,7,8,3’,4’-六甲氧基黄酮32.8 mg、3,5,6,7,8,3’,4’-七甲氧基黄酮10.9 mg、5,6,7,8,4’-五甲氧基黄酮43.5 mg和5-羟基-6,7,8,3’,4’-五甲氧基黄酮18.7 mg。产物纯度经HPLC检测分别为98.9%,97.8%,99.2%和99.8%。通过质谱和核磁共振氢谱、碳谱等波谱分析确定各化合物结构。

Four polymethoxylated flavones （PMFs） were prepared, isolated and purified from citrus peel by highspeed counter-counter chromatography （HSCCC）. A two-phase solvent system composed of petroleumether ： ethyl acetate： methanol：water （1：0.8：1：1.2,V/V） was used, the top phase was stationary phase and the bottom phase was mobile phase. The 32.8 mg 5,6,7,8,3＇ ,4＇-hexamethoxyflavone, 10.9 nag 3,5,6,7,8,3＇,4＇-heptamethoxyflavone, 43.5 mg 5,6,7,8,4＇ -pentamethoxyflavone and 18.7 mg 5-hydroxy- 6,7,8,3＇ ,4＇- Pentamethoxyflavone were obtained from 200 mg of the crude extract of citrus peel in one-step elution under the conditions of a flow rate of 2.0 mL/min, the rotation speed was 800 r/min and the detection wave length of 254 nm. High performance liquid chromatographic analysis of nobiletin, 3,5, 6,7,8,3＇ ,4＇ -heptamethoxyflavone, 5,6,7,8,4＇ -pentamethoxyflavone and 5 -hydro xy-6,7,8,3＇ ,4＇ -pentamethoxyflavone revealed that their purity were individually 98.9%, 97.8%, 99.2% and 99.8%. And their structures were identified by MS, 1H-NMR and 13C-NMR. The results indicate that HSCCC is a powerful technique for the purification of polymethoxylated flavones from citrus peel.