柱前衍生-HPLC荧光法测定人血清中的饱和与不饱和脂肪酸  

DETERMINATION OF FREE FATTY ACIDS IN HUMAN SERUMS BY PRE-COLUMN DERIVATIZATION HPLC FLUOROMETRIC METHOD

作　　者：李谦[1] 张晓萍[2] 邱宗荫[2] 

机构地区：[1]四川省自贡市疾病预防控制中心,自贡643000 [2]重庆医科大学

出　　处：《现代预防医学》2007年第24期4714-4717,共4页Modern Preventive Medicine

摘　　要：[目的]建立人血清中饱和及多不饱和脂肪酸高效液相色谱分离及测定方法。[方法]3-溴甲基-6,7-二甲氧基-1-甲基-2(1H)喹喔啉酮(Br-DMEQ)作荧光标记试剂,74%乙腈-水溶液作流动相,岛津shim-pack CLC-C8(150mm×6.0mmΦ)为色谱柱,HPLC柱前衍生化分离测定脂肪酸。[结果]得到该衍生化试剂与脂肪酸的衍生化反应条件及脂肪酸生成的酯衍生物的荧光光谱,衍生物荧光强度衰减情况、最佳色谱分离条件及C12:0-C22:6的11种脂肪酸的分离图谱。2.0ng/ml～8.0μg/ml浓度范围内线性关系良好,回收率在91%～105%;本方法最低检测限0.2pg/ml,首次在国内报道了以该法测定人血清中C18:3,C20:4,C20:5,C22:6脂肪酸的结果。[结论]方法重现性好,灵敏度高,快速、简便,准确可靠,是一种可用于常规检测脂肪酸的荧光-HPLC法。[Objective]To establish a method to separate and determine free long chain,polyunsaturated fatty acids in human serum by high-performance liquid chromatography（HPLC）.[Methods]3-bromomethyl-6,7-diomethoxy-1-methyl-2（1H）-quinoxaline（Br-DMEQ）served as fluorescent labeled reagent,acetonitrile solution（74%）as mobile phase,shim-pack CLC-C8（150 mm×6.0 mmΦ）as chromatographic column,then the fatty acids were separated and determined by the corresponding by pre-column derivatization HPLC.[Results]The fluorescence spectra of the reactive condition for this derivatization reagents reacted with fatty acid derivatization and the ester derivatives of fatty acid,the attenuation of fluorescence intensity derivatives,the optimum separated condition and chromatogram of 11 fatty acids（ C12：0-C22：6 ）were found.Linear relationship were found within the range of 2.0 ng/ml-8.0 μg/ml.The rate of recovery of the spiked samples was 91%-105%;The detection limit of this method was 0.2pg/ml.The fatty acids in human serum were firstly determined by this method in our country including a-linolenic C18：3,arachidonic C20：4,eicosapentaenoic C20：5,docosah-exaenoic C22：6[Conclusion]HPLC fluorometry is a method with fine reproducibility,high sensitivity,rapid,convenience,reliability and accuracy,which can be used as a routine method for detecting the fatty acids.

关 键 词：Br-DMEQ 脂肪酸 荧光检测-HPLC 

分 类 号：Q547[生物学—生物化学]