HPLC法测定灵芝孢子粉中三萜含量

为准确测定灵芝孢子粉中三萜的含量,运用高效液相建立适合孢子粉的分析测定方法。通过对前处理条件的优化,确定40%乙醇为孢子粉中等极性三萜酸类的最佳提取溶剂,浓缩倍数是子实体提取条件的50倍。通过色谱柱和洗脱条件的优化,建立了包括灵芝酸I、灵芝烯酸C、灵芝酸C2等13种标准品测定方法,方法学考察显示该分析方法精密度、重复性、稳定性的RSD值均小于5%,可以用于灵芝孢子粉中三萜类成分的定量检测。通过5组样品的分析发现,灵芝酸C6、灵芝酸G、灵芝酸A、灵芝酸D、灵芝酸F是灵芝孢子粉中的主要三萜类成分,其中灵芝酸A含量最高,平均占样品三萜总量的比例达19.71%;三萜类成分的溶出量与是否破壁没有相关性。三萜类成分在灵芝孢子粉和灵芝孢子油产品中的含量非常低,孢子粉的三萜含量为14.24-99.70μg/g,仅为子实体的1/100,灵芝孢子油中三萜含量也均低于50μg/g,因此三萜类成分不适合作为灵芝孢子粉及其相关产品的定量检测指标。

Determination of triterpenoids in Ganoderma lingzhi spore powder by HPLC

High-performance liquid chromatography method was established to determine the content of triterpenoids in Ganoderma lingzhi spore powder.40%ethanol was proved to be the best solvent for extracting the medium-grade triterpenic acid in spore powder,and the extract was concentrated 50 times.The determination method of 13 standard products including ganoderic acid I,ganoderenic acid C,and ganoderic acid C2 were established by optimizing the column and elution conditions.The methodological investigation showed that RSD values of method precision,repeatability and stability were less than 5%,indicating the method was appropriate for the quantitative detection of medium-polar triterpenoids in Ganoderma lingzhi spore powder.The analysis of multiple samples showed that ganoderic acid C6,ganoderic acid G,ganoderic acid A,ganoderic acid D and ganoderic acid F were the main triterpenoids in Ganoderma lingzhi spore powder,and the content of ganoderic acid A was the highest,whose average proportion in the total quantity of triterpenoids was 19.71%.There was no correlation between triterpene content and wall breaking treatment.The content of triterpenoids in spore powder was 14.24–99.70mg/g,only accounting for one percent of triterpenoid content of the fruiting body.The content of triterpenoids in the Ganoderma lingzhi spore oil was less than 50mg/g.The results suggested that the content of triterpenoids in Ganoderma lingzhi spore powder and oil was too low to be regarded as the quantitative detection index of Ganoderma lingzhi spore powder products.