崂山蘑菇抗氧化成分提取及其活性研究

Evaluation of Antioxidant Property of Extracts Obtained from Laoshan Mushroom

用1:1的氯仿:甲醇超声提取崂山蘑菇成分,并用石油醚、乙酸乙酯和正丁醇依次进行萃取,利用DPPH法、α-脱氧核糖法和铁氰化钾还原法进行抗氧化实验。同时与BHT、没食子酸丙酯(PG)、没食子酸(GA)和抗坏血酸(AscA)4种合成抗氧化剂进行对照。结果显示乙酸乙酯提取物(EAF)具有较高的抗氧化活性。对其进行硅胶柱层析分离,得到6种纯化的组分(EAF1-EAF6),其中EAF4具有最高的抗氧化活性,在50μg/ml浓度时,其清除DPPH自由基能力仅比AscA稍弱,比BHT、PG和GA强;清除羟自由基能力在所有测定浓度下(0.4、2、10、50μg/ml)均比4种合成抗氧化剂强;还原能力在所有测定浓度下均比BHT强;在10μg/ml时,比PG强;50μg/ml时,与PG相当。测定了崂山蘑菇各提取物总酚含量,结果发现总酚含量与抗氧化活性之间具有很好的相关性。

Laoshan mushroom was extracted with methanol：chloroform（1：1）, and the extraction was then partitioned with petroleum ether, CHCl3, EtOAc and BuOH successively. Antioxidant activity （AA） of the extractions was evaluated in three different assays, namely, the 1,1-dipbenyl-2-picrylhydrazyl （DPPH） radical scavenging assay, the deoxyribose assay and the potassium ferricyanide reduction methods, and compared with that of butylated hydroxytoluene （BHT）, gallic acid （GA）, propyl gallate（PG） and ascorbic acid （AscA）. The results showed that the ethyl acetate-soluble fraction exhibited the highest AA at all of four concentration levels tested （from 0.4 to 50μg/ml）, and was further fractionated into six subfractions EAF1 - EAF6 by silica gel vacuum liquid chromatography. EAF4 was found to be the most effective subfraction in three assays, which was greater AA than BHT,PG and GA and similar to AscA in the DPPH assay at the concentration of 50μg/ml, while, in the deoxyribose assay, the EAF4 exhibited higher AA than four kinds of synthetic antioxidants at all of four concentration levels tested and in the potassium ferricyanide reduction methods exhibited higher AA than PG at the concentration of 10μg/ml and similar to PG at the concentration of 50μg/ml. The total phenolic content was determined using the Folin - Ciocalteu. Statistical analysis indicated a significant association between the antioxidant potency and total phenolic content.