桦褐孔菌次生代谢产物清除自由基活性成分的分离纯化及初步鉴定

Separation,surification and preliminary identification for eliminating free radical activity components from metabolites of inonotus obliouus

首先采用二苯基苦基苯肼自由基薄层实验法对桦褐孔菌次生代谢产物进行清除自由基活性的研究,发现桦褐孔菌菌株的次生代谢产物具有较强清除自由基的活性。并采用中压液相色谱(MPLC)、凝胶柱层析(SephadexLH-20)、硅胶柱层析和重结晶多种方法对其活性成分进行分离纯化,得到具有清除自由基活性的纯化合物LWZc和LWZf。利用薄层层析(TLC)、高效液相色谱(HPLC)、核磁共振(1H-NMR)和液质联用(HPLC-MS)等方法对LWZc和LWZf进行纯度鉴定和结构解析,结果表明:LWZf为一种已知化合物对羟基苯甲酸(C7H6O3),具有较强的清除自由基活性,从桦褐孔菌中分离得到该化合物是首次报道;初步推断LWZc化合物的可能结构为3-乙酰基-4-氨基-苯基-核糖甙(C13H17NO6),是一种新化合物,该化合物的纯品清除自由基的能力相对较弱。

The strong scavenging activity of metabolites radical scavenging assay in this study. At the same n inonotus obliquus were investigated through the free time,the activing ingredients LWZc and LWZf were separated and purified with MPLC,SephadexLH-20,silica gel column chromatography,secondary crystal and so on. Two pure compounds Lwzc and Lwzf were obtained. Meanwhile,their purity was tested with TLC, 1H-NMR and HPLC methods. These pure compounds were submitted to the structural analysis with thick solution HPLC-TOFMS and 1H-NMR. The structure identification result revealed that the molecular formula of LWZf and LWZc were O7H6O3 and C13H17N1O6 respectively. The structure of LWZf was identified as 4-hydroxy Benzoic acid. Though 4-hydroxy Benzoic acid was a known compound of strong anti-oxidant activity,it was the first time to isolated the compound from. inonotus obfiquus. The probably new structure of LWZc was deduced as 3-acetyl-4-amido-phenyl-ribotide,which activity was not high.