超声波辅助提取-高效液相色谱法分析干酪中9种生物胺  

Ultrasonic assisted extraction coupled high-performance liquid chromatography for analysis of 9 kinds of biogenic amines in cheese

作　　者：张慧 王瑾 唐纱丽 肖功年[1] 曹慧 ZHANG Hui;WANG Jin;TANG Shali;XIAO Gongnian;CAO Hui(School of Biological and Chemical Engineering,ZheJiang University of Science and Technology,Hangzhou 310023,Zhejiang,China;Zhejiang Institute of Product Quality and Safety Science,Hangzhou 310018,Zhejiang,China)

机构地区：[1]浙江科技学院生物与化学工程学院,杭州310023 [2]浙江省产品质量安全科学研究院,杭州310018

出　　处：《浙江科技学院学报》2023年第6期534-541,共8页Journal of Zhejiang University of Science and Technology

基　　金：浙江省基础公益研究计划项目(LGN20C200011);杭州市农业与社会发展重点研发项目(亚运)(20230701A04)。

摘　　要：【目的】建立超声波辅助提取高效液相色谱法检测干酪中9种生物胺的测定方法。【方法】首先样品经超声波辅助溶剂提取,滤液经柱前衍生,高效液相色谱分离和测定,外标法定量;然后选择提取溶剂的种类,研究不同超声时间和温度对回收率的影响,并进行方法学验证。【结果】以质量分数为5%的三氯乙酸溶液30℃超声波提取15 min,连续提取2次,滤液合并后经丹磺酰氯衍生,液相色谱测定;9种生物胺衍生物在各自的线性范围内线性关系良好,相关系数均大于0.99,回收率为76.3%~90.1%,相对标准偏差为1.09%~7.45%,检出限为0.38 mg/kg,定量限为1.25 mg/kg。将本研究提出的方法用于天然干酪样品的检测,不同样品间生物胺种类和含量差异较大,总含量为2.44~78.1 mg/kg。【结论】本研究提出的方法适用于干酪中9种生物胺的定量检测,可为生物胺形成机制和食品安全评价研究提供技术支持。[Objective]An analytical method was established for determination of 9 kinds of biogenic amines in cheeses with ultrasonic-assisted extraction coupled high-performance liquid chromatography.[Method]First,biogenic amines in cheese samples were extracted by ultrasonic-assisted solvent,and the filtrate was derived,separated and determined with high-performance liquid chromatography(HPLC),and quantified by external standard method;then,the kinds of extraction solvent were selected to probe into the influences of different ultrasonic time and temperature on the recovery rate,and verify the methodology.[Result]9 kinds of biogenic amines are extracted twice by ultrasound at 30℃for 15 min with 5%trichloroacetic acid solution,and the filtrate is derived by dansulfonyl chloride,determined by HPLC;the linear relationship turns out to be sound in their respective linear ranges,with the correlation coefficients greater than 0.99,the recovery rates 76.3%~90.1%and relative standard deviations at three spiked levels 1.09%~7.45%,the limits of detection 0.38 mg/kg and the limits of quantification 1.25 mg/kg.The method is proposed for the determination of cheese samples,whose types and contents of biogenic amines vary greatly,with the total biogenic amine content ranging from 2.44 to 78.1 mg/kg.[Conclusion]The method is applicable to the quantitative detection of 9 kinds of biogenic amines in cheese,which can provide technical support for the study of biogenic amine formation mechanism and food safety evaluation.

关 键 词：高效液相色谱 超声提取 干酪 生物胺 测定 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]