白术多糖指纹图谱和抗氧化活性的谱效关系分析

Analysis of Spectrum-Efficacy Relationship Between Fingerprint and Antioxidant Activity of Polysaccharide in Atractylodes Macrocephala

将高效液相色谱(HPLC)指纹图谱、抗氧化活性研究与多元统计分析相结合,对不同产地的38批白术多糖进行质量评价.以三氟乙酸水解白术多糖,用1-苯基-3-甲基-5-吡唑啉酮进行衍生化,采用高效液相色谱法测定其中单糖的衍生物,得到指纹图谱.以1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除能力、羟自由基清除能力和Fe^(3+)总还原能力为指标测定抗氧化活性,并对结果进行相似度分析、层次聚类分析和偏最小二乘法-判别分析等多元统计分析.结果表明,白术多糖指纹图谱鉴别了5个共有峰,分别为鼠李糖、半乳糖醛酸、葡萄糖、半乳糖和阿拉伯糖.基于多元统计分析,浙江、安徽、河北和四川的白术样品分离显著.此外,选择半乳糖醛酸和半乳糖作为潜在的化学标记物,DPPH自由基清除能力对应的抗氧化活性可作为辅助鉴别的候选检测指标.

The quality of 38 batches of Atractylodes macrocephala polysaccharides from different producing regions was evaluated by the method of combining high performance liquid chromatography(HPLC)fingerprint,antioxidant activity and multivariate statistical analysis.Atractylodes macrocephala polysaccharides were hydrolyzed with trifluoroacetic acid and then derivatized with 1-phenyl-3-methyl-5-pyrazolinone.The derivatives of monosaccharides were determined by HPLC to obtain fingerprint.1,1-Diphenyl-2-trinitrophenylhydrazine(DPPH)radical scavenging capacity,hydroxyl radical scavenging capacity and total Fe^(3+) reduction capacity were used as indicators to determine the antioxidant activity.The results were also subjected to multivariate statistical analysis including similarity analysis,hierarchical clustering analysis and partial least square-discriminant analysis.As shown by the results,five common peaks(rhamnose,galacturonic acid,glucose,galactose and arabinose)were identified in the fingerprint of Atractylodes macrocephala.Based on the multivariate statistical analysis,the samples of Atractylodes macrocephala from Zhejiang,Anhui,Hebei and Sichuan were significantly separated.In addition,galacturonic acid and galactose were selected as potential chemical markers,and the antioxidant activity from DPPH radical scavenging capacity could be used as a candidate detection index to assist identification.