灵芝孢子中抗肿瘤成分的提取与分离

目的 :检测不同破壁率的灵芝孢子的醇提效果并用色谱技术对醇提物中的抗肿瘤成分进行分析。方法 :乙醇为溶剂对灵芝孢子粉进行振荡浸提 ,醇提物依次用硅胶柱和高效液相色谱分析 ,以各部分对Hela细胞的毒性作为其抗肿瘤活性的指标。结果 :未破壁孢子 ,60%～80%及 99%破壁率孢子的醇提率分别为5% ,25% ,33%。99%破壁率灵芝孢子的醇提物经硅胶柱分离 ,氯仿洗脱部分对Hela细胞无毒害作用 ;甲醇洗脱部分的抗肿瘤活性约为乙酸乙酯洗脱部分的 34倍 ;甲醇洗脱部分经高效液相色谱分析 ,得到 2种活性较强且组成较为简单的混合物(Ⅱ1和Ⅱ3 )。结论 :灵芝孢子破壁后醇提物含量远高于未破壁孢子 ;甲醇 水洗脱系统可用于灵芝孢子醇提物中抗肿瘤活性成分的RP HPLC分析。

Extraction and Separation of Antitumor Components from Ganoderma lucidum (Leyss.ex Fr.) Karst.

OBJECTIVE: To examine the variation of alcohol extraction rates of Ganoderma lucidum spores with different rates of wall-wrack, and analyze the antitumor components of alcohol extract by chromatography. METHOD: The G. lucidum spores were soaked and extracted with absolute alcohol. The alcohol extract was chromatographed on a silica gel column and HPLC in proper order, and the antitumor activity of every eluted fraction was represented by its cytotoxicity towards Hela cells. RESULT: Extraction rates 5%, 25% and 33% corresponded to wall-wrack rates 0%, 60%-80% and 99% respectively. The alcohol extract from spores with the highest wall-wrack rate was chromatographed on a silica gel column, eluting successfully with CHCl3, EtOAc and CH3OH in order. The CHCl3 fraction had not any antitumor activity, while this activity of CH3OH fraction was 34 times greater than that of EtOAc fraction. HPLC analysis found out that two mixtures(II1 and II3) possess significant antitumor activity in vitro. CONCLUSION: The weight of alcohol extract from spores with wall-wrack was far greater than that of spores without. The antitumor components of G. lucidum spores could be analyzed with methanol-water on a reverse HPLC.