灰树花水提物抑糖基化物质的分离纯化

Isolation and Purification of Compounds with Non-enzymatic Protein Glycation Inhibition Activity from Aqueous Extract of Grifola frondosa

从大孔树脂类型、洗脱剂与上样量等方面确定灰树花中抑糖基化反应化合物的初分离条件,并选取抑糖基活性最高的组分进行薄层层析分析和硅胶柱分离纯化,最后利用高效液相色谱法和傅里叶红外光谱分析化合物。结果表明:采用D101大孔树脂,25 g树脂最佳上样量为2 g,20%乙醇洗脱组分C 3的抑糖基化活性最高。用氯仿∶甲醇(v/v 1∶9)为洗脱液对C 3进行硅胶柱层析纯化,共分离出5个组分C3 5~C3 9,其中C 3 8和C3 9具有比较强的抑糖基化活性,合并C3 8和C3 9,再次用氯仿∶甲醇=(v/v 2∶8)为洗脱液进行硅胶柱层析纯化,共分离出5个组分G 1~G 5,薄层层析和HPLC分析分别显示G 3为单一斑点和单一峰型,说明G 3是一种单体化合物,根据红外光谱结果初步推断该化合物为酚类物质。

Compounds with non-enzymatic protein glycation(NEPG)inhibition activity were isolated and purified from aqueous extract of Grifola frondosa.The initial isolation conditions were determined in terms of macroporous resin type,eluent and loading amount.Then the fraction with the highest activity was further subjected to thin layer chromatography analysis(TLC)and silica gel column separation and purification.Finally,the purified compound was analyzed by HPLC and infrared absorption spectroscopy.The results showed that fraction C 3 with the highest activity were obtained under the following conditions:D101 macroporous at a loading amount of 2 g sample in 25 g resin,and eluted by 20%ethanol.Fraction C3 was further purified by silica gel column chromatography with a solution of chloroform and methanol(v/v 1∶9)as the elution agent.After this step,five components C3 5-C3 9 were obtained.Among them,C3 8 and C3 9 showed relatively stronger activity than the other components and were therefore mixed together.Then the mixture was further purified by silica gel column chromatography with a mixture solution of chloroform and methanol(v/v 2∶8)as the elution agent.As a result,five components G1-G5 in total were obtained.TLC and HPLC analyses showed that G3 was a single spot and a single peak,suggesting that G3 was a monomeric compound.Furthermore,the results of absorption spectroscopy suggested this compound a phenolic substance.