正相高效液相色谱法同时测定肉制品中4种羟基十八碳二烯酸异构体的含量  

Simultaneous Measurement of Four Hydroxyoctadecadienoic Acid Isomers in Meat Products by Normal-phase High Performance Liquid Chromatography

作　　者：刘婷 熊强[1] 耿志明[2] 徐为民[2,3] LIU Ting;XIONG Qiang;GENG Zhiming;XU Weimin(College of Food Science and Light Industry,Nanjing Tech University,Nanjing 211816,China;Institute of Agricultural Products Processing,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;Jiangsu Collaborative Innovation Center of Meat Production and Processing,Quality and Safety Control,Nanjing 210095,China)

机构地区：[1]南京工业大学食品与轻工学院,江苏南京211816 [2]江苏省农业科学院农产品加工研究所,江苏南京210014 [3]江苏省肉类生产与加工质量安全控制协同创新中心,江苏南京210095

出　　处：《食品科学》2021年第4期271-277,共7页Food Science

基　　金：国家自然科学基金面上项目(31671877);江苏省自然科学基金项目(BK20171324)。

摘　　要：为建立同时测定肉制品中13-羟基-9Z,11E-十八碳二烯酸(13-hydroxy-9Z,11E-octadecadienoic acid,13-Z,E-HODE)、13-羟基-9E,11E-十八碳二烯酸(13-hydroxy-9E,11E-octadecadienoic acid,13-E,E-HODE)、9-羟基-10Z,12E-十八碳二烯酸(9-hydroxy-10Z,12E-octadecadienoic acid,9-Z,E-HODE)、9-羟基-10E,12E-十八碳二烯酸(9-hydroxy-10E,12E-octadecadienoic acid,9-E,E-HODE)的高效液相色谱检测方法,肉制品中的13-Z,EHODE、13-E,E-HODE、9-Z,E-HODE和9-E,E-HODE经甲醇提取、采用Sep-Pak C18柱净化浓缩后,在硅胶柱Absolute SiO2(250 mm×4.6 mm,5μm)上以正己烷-异丙醇-乙酸(98.3∶1.6∶0.1,V/V)为流动相进行分离,选用二极管阵列检测器在234 nm进行测定。结果表明,13-Z,E-HODE、13-E,E-HODE、9-Z,E-HODE和9-E,E-HODE分别在质量浓度为0.5~20.0、0.25~10.0、0.75~12.5μg/mL和0.5~7.5μg/mL的范围内线性关系良好(R2分别为0.9994、0.9992、0.9992、0.9996),检出限分别为0.075、0.035、0.090、0.060μg/g,定量限分别为0.25、0.12、0.32、0.20μg/g,不同添加水平的平均回收率分别为89.03%、89.03%、89.33%、87.93%。对18种肉制品含量分析表明,所有样本都含有13-Z,E-HODE、13-E,E-HODE、9-Z,E-HODE和9-E,E-HODE,含量分别为1.73~9.10、0.56~5.79、2.37~11.02、0.78~5.82μg/g。综上,建立的检测方法快捷、准确、重复性好,可用于同时测定肉制品中13-Z,EHODE、13-E,E-HODE、9-Z,E-HODE和9-E,E-HODE的含量。In this study,a high performance liquid chromatography(HPLC)method was established for simultaneous determination of 13-hydroxy-9Z,11E-octadecadienoic acid(13-Z,E-HODE),13-hydroxy-9E,11E-octadecadienoic acid(13-E,E-HODE),9-hydroxy-10Z,12E-octadecadienoic acid(9-Z,E-HODE)and 9-hydroxy-10E,12E-octadecadienoic acid(9-E,E-HODE)in meat products.The analytes were extracted from samples with methanol,purified by Sep-Pak C18 column,and then separated using isocratic elution on an Absolute SiO2 column(250 mm×4.6 mm,5μm)with a mobile phase consisting of n-hexane,isopropanol and acetic acid(98.3:1.6:0.1,V/V)before being detected at 234 nm using a photo-diode array(PDA).The results indicated that the method displayed a good linearity within the concentration ranges of 0.5–20.0μg/mL(R^2=0.9994)for 13-Z,E-HODE,and 0.25–10.0μg/mL(R^2=0.9992)for 13-E,E-HODE,0.75–12.5μg/mL(R^2=0.9992)for 9-Z,E-HODE and 0.5–7.5μg/mL(R^2=0.9996)for 9-E,E-HODE,respectively.The detection limits(LODs)for 13-Z,E-HODE,13-E,E-HODE,9-Z,E-HODE and 9-E,E-HODE were 0.075,0.035,0.090 and 0.060μg/g,respectively,and the limits of quantification(LOQs)were 0.25,0.12,0.32 and 0.20μg/g,respectively.The average recoveries from spiked samples were 89.03%,89.03%,89.33%and 87.93%,respectively.When this method was used to analyze 18 meat products,all the samples contained 13-Z,E-HODE,13-E,E-HODE,9-Z,E-HODE and 9-E,E-HODE at levels of 1.73–9.10,0.56–5.79,2.37–11.02 and 0.78–5.82μg/g,respectively.In conclusion,this method was proved to be fast,accurate and reproducible,and could be used for the simultaneous quantitative analysis of 13-Z,E-HODE,13-E,E-HODE,9-Z,E-HODE and 9-E,E-HODE in meat products.

关 键 词：羟基十八碳二烯酸 脂质氧化 亚油酸 高效液相色谱 

分 类 号：TS201.6[轻工技术与工程—食品科学]