HPLC法同时测定食用植物酵素中12种有机酸  

Simultaneous Determination of Twelve Organic Acids in Edible Plant Source Jiaosu by HPLC

作　　者：王珍珍 沙如意[1] 王高坚 徐成龙 戴静 葛青[1] 毛建卫[1,2] 朱姣琳 WANG Zhen-zhen;SHA Ru-yi;WANG Gao-jian;XU Cheng-long;DAI Jing;GE Qing;MAO Jian-wei;ZHU Jiao-lin(Zhejiang Provincial Key Lab for Chem&Bio Processing Technology of Farm Product,Zhejiang Provincial Collaborative Innovation Center of Agricultural Biological Resources Biochemical Manufacturing,School of Biological and Chemical Engineering,Zhejiang University of Science&Technology,Hangzhou 310023,China;Zhejiang Industry Polytechnic College,Shaoxing 312000,China;Fuyuan Biotechnology Co.,Ltd.(Ningbo),Ningbo 315194,China)

机构地区：[1]浙江省农产品化学与生物加工技术重点实验室,浙江省农业生物资源生化制造协同创新中心,浙江科技学院生物与化学工程学院,浙江杭州310023 [2]浙江工业职业技术学院,浙江绍兴312000 [3]富元生命科技(宁波)有限公司,浙江宁波315194

出　　处：《食品工业科技》2020年第19期279-285,共7页Science and Technology of Food Industry

基　　金：浙江省重点研发计划项目(2017C02009);中国博士后科学基金(2018M632475);金华市农业类重大项目(2018-2-001a);省属高校基本科研业务费专项资金项目(2019JL10)。

摘　　要：为研究食用植物酵素发酵过程中有机酸的变化规律,建立了同时检测12种有机酸含量的高效液相色谱法(HPLC),并检测了青梅酵素、沙棘酵素、火龙果酵素、博伊森莓酵素和黄金酵素5种食用植物酵素中有机酸的种类及含量,以有机酸为指标进行主成分分析和分层聚类热图分析。结果表明,优化后的色谱条件为:检测波长210 nm,流动相为甲醇:KH_2PO_4(0.01 mol/L,pH2.7)=2∶98(V/V),柱温:20℃。利用此方法,分别从上述5种酵素中检测到10、11、9、10和4种有机酸。基于R软件运用主成分分析法研究了影响5种食用植物酵素中有机酸含量的主要因子,提取了2个主成分,累计方差贡献率达77.00%。对不同原料食用植物酵素中的12种有机酸进行分层聚类热图分析,可将有机酸分成三类,一类以乳酸为主,一类以苹果酸为主,另一类主要以柠檬酸为主。本研究可以为食用植物酵素发酵过程中有机酸的检测方法提供依据。The high performance liquid chromatography(HPLC)method for simultaneous determination of 12 kinds of organic acids was established,in order to study the changes of organic acids in the fermentation process of edible plant source Jiaosu,such as Prunus mume Jiaosu,sea-buckthorn Jiaosu,pitaya Jiaosu,boysenberry Jiaosu and gold Jiaosu.The contents of organic acid in five edible plant source Jiaosu were used as indicators for principal component analysis and hierarchical cluster analysis heatmap to study the differences of organic acid.The optimal chromatographic conditions were as follows:Detection wavelength of 210 nm,the mobile phase composed of methanol and 0.01 mol/L K2HPO4 with pH adjusted to 2.7 by phosphoric acid(2∶98,V/V)at 20℃.Ten,eleven,nine,ten and four kinds of organic acids were determined in these edible plant source Jiaosu by the HPLC method,respectively.Based on R software,the main factors affecting the organic acid content were studied by principal component analysis.Two principal components were extracted,and the cumulative variance contribution rate was 77.00%.According to the hierarchical clustering and heatmap analysis of 12 kinds of organic acid metabolites in edible plant source Jiaosu from different raw materials,the organic acids could be divided into three categories,citric acid,malic acid and citric acid.This study can provide a method basis for the monitoring and detection of organic acids in fermentation of edible plant source Jiaosu.

关 键 词：酵素 有机酸 高效液相色谱(HPLC) 主成分分析 热图分析 

分 类 号：TS255[轻工技术与工程—农产品加工及贮藏工程]