HPLC指纹图谱技术在灵芝组织分离试验中的应用

利用HPLC指纹图谱技术研究从同一灵芝子实体不同部位组织分离获得的菌株三萜化合物的差异。用常规组织分离方法获得不同菌株,在A、B、C、D四种培养基上进行出菇试验,运用HPLC指纹图谱技术获得各子实体三萜提取物HPLC指纹图谱,计算图谱间的相似度,分析三萜指纹的差异。结果表明,从子实体不同部位分离获得的四个菌株在同一培养基相同条件下培养获得的灵芝子实体的三萜指纹图谱相似度均大于0.99;同一部位分离获得的菌株在不同培养基相同培养条件下获得的子实体,其粗三萜HPLC图谱相似度均大于0.96;不同的组织分离部位和不同的培养基对灵芝菌株三萜指纹图谱的影响均不显著。18号菌株在C培养基上培养获得的子实体三萜含量最高,上层菌肉为最优组织分离部位,C培养基为最优培养基。灵芝的三萜组成不受生长环境的影响,而生长环境会对其三萜化合物含量产生一定的影响。本文首次应用HPLC指纹图谱方法对灵芝组织分离获得的菌株的差异性进行了研究。

Application of high-performance liquid chromatographic fingerprinting in the study of tissue isolation of Ganoderma lucidum

Using high-performance liquid chromatographic fingerprinting, the differences of the strains isolated from different parts of the same fruiting body of Ganoderma lucidum were investigated. Four parts of fruiting bodies were used for tissue isolation and the isolated strains were cultured on four media. The similarities between chromatograms of these samples were calculated. The results are as follows: under the same culture condition, the similarities of HPLC fingerprints of the strains isolated from different parts were all higher than 0.99. Meanwhile, the similarities of HPLC fingerprints of the strains isolated from the same part were all higher than 0.96 in different media. The effects of different isolation tissues and media to the HPLC chromatograms of the fruiting bodies were not distinctive. The triterpenoid content of strain 18 cultured on medium C is the highest among the tested fruiting bodies. The best isolated part is the upper layer of pileus and medium C is the best medium. The triterpenoid composition in the fruiting bodies of Ganoderma lucidum is mainly decided by their genetic factors; the growth condition will affect the content of triterpenoid compound and this effect can be showed by different peak-area ratios of the chromatogram. This is the first report of HPLC fingerprinting method applied to the analysis of tissue isolation of Ganoderma lucidum.