6-O-异丙酰乙二胺荧光素衍生HPLC分离荧光检测脂肪酸

6-O-异丙酰乙二胺荧光素衍生HPLC分离荧光检测脂肪酸 Analysis of Fatty Acids as 6-Oxy-（Isopropionyl Ethylenediamine） Derivatives by High Performance Liquid Chromatography with Fluorescence Detection 蔡丽华 CAI Lihua1 1 first-author 杜小兰 DU Xiaolan2 2 张华山 ZHANG Huashan2 2 中南民族大学工商学院 中南民族大学工商学院 Engineering and Commerce College,South-Central University for Nationalities Engineering and Commerce College,South-Central University for Nationalities 武汉大学化学与分子科学学院 武汉大学化学与分子科学学院 College of Chemistry and Molecular Sciences,Wuhan University College of Chemistry and Molecular Sciences,Wuhan University 应用6-O-异丙酰乙二胺荧光素（IPEF）作为柱前衍生试剂,结合高效液相色谱（HPLC）荧光检测,成功地衍生、分离并检测了食用油中12种脂肪酸:戊酸（C5）、己酸（C6）、庚酸（C7）、辛酸（C8）、壬酸（C9）、癸酸（C10）、月桂酸（C12）、肉豆蔻酸（C14）、棕榈酸（C16）、油酸（C18∶1）、硬脂酸（C18）、亚油酸（C18∶2）的含量.以1-乙基-3-（3-二甲胺丙基）碳酰亚胺（EDC）为缩合试剂,IPEF与羧酸在50℃反应40min即可较完全衍生.采用C18柱分离脂肪酸,色谱柱采用pH 6.5的甲醇-水（65/35,V/V）流动相平衡,进样后,流动相流速为0.8mL.min-1,检测波长λex为504nm,λem为525nm.该方法检出限可达0.2～0.8nmol.L-1. A simple and highly sensitive method for the determination of 12 fatty acids n-valeric acid（C5）,n-hexanoic acid（C6）,n-heptanoic acid（C7）,n-octanoic acid（C8）,n-nonanoic acid（C9）,n-decanoic acid（C10）,lauric acid（C12）,myristic acid（C14）,palmitic acid（C16）,stearic acid（C18）,oleic acid（C18∶1） and linoleic acid（C18∶2） is described.The method is based on the separation of the 6-Oxy-（isopropionyl ethylenediamine） fluorescein（IPEF） derivatives of 12 fatty acids by high performance liquid chromatography with fluorescence detection.The derivatization reaction with 12 fatty acids was completed at 50 ℃ for 40 min using 1-ethyl-3-（3-dimethylamino-propyl） carbodiimide（EDC） as the condensing reagent successfully.The chromatographic separation is performed using a mobile phase consisting of methanol-water（65/35,V/V） adjusted to pH 6.5 with Na2HPO4-H3Cit3 buffer on a Kromasil 100 C18 analytical column with flow rate of 0.8 mL/min and fluorescence detection at λex/λem=504 nm/525 nm.The detection limits were in the range of 0.2-0.8 nmol·L-1. 6-O-异丙酰乙二胺荧光素 衍生 高效液相色谱（HPLC） 荧光检测 脂肪酸 6-oxy-（isopropionyl ethylenediamine） fluorescein derivative high performance liquid chromatography（HPLC） fluorescence detection fatty acids 10.14188/j.1671-8836.2011.01.011 TS227 国家自然科学基金（20575047）资助项目 2011-01-01 2021-04-01 1 57