深海鱼油中脂肪酸的柱前衍生-高效液相色谱串联质谱分析

用1-2-(对甲苯磺酸酯)乙基-2-苯基咪唑4,5-f9,10-菲(TSEPIP)作为柱前荧光衍生试剂,在EclipseXDB-C8(4.6×150mm,5μm,Agilent)反相色谱柱上,采用梯度洗脱在检测波长为380nm(激发波长为260nm)的条件下,实现了阿拉斯加深海鱼油中饱和脂肪酸含量的外标法定量测定。26种饱和脂肪酸的线性范围是200.0pmol～48.83fmol,线性相关系数均大于0.9996,检测限为3.824～47.13fmol(信噪比为3:1测得,S/N3:1)。经柱后串联质谱大气压化学电离源(APCI)正离子模式实现了各种饱和与不饱和脂肪酸衍生物的质谱鉴定,进而通过峰面积归一化法得出了所有饱和与不饱和脂肪酸的相对含量。结果表明,深海鱼油主要含有C12～C22的脂肪酸,共鉴定出25种脂肪酸,其中不饱和脂肪酸含量占69.71%(峰面积百分比,下同),特别是具有重要生理作用的多不饱和脂肪酸,如C20:5:5,8,11,14,17-二十碳五烯酸(5,8,11,14,17-eicosapentaenoicacid,EPA,16.62%),C22:6:2,5,8,11,14,17-二十二碳六烯酸(2,5,8,11,14,17-docosahexenoicacid,DHA,12.31%)。

Pre-column Derivatization High-Performance Liquid Chromatography Tandem Mass Spectrometry Determination of Fatty Acids from Deep-sea Fish Oil

A simple and sensitive method for the determination of fatty acids in Alaska deep-sea fish oil using 1-[2-(p-toluenesulfonate)-ethyl]-2-phenylimidazole-[4,5-f]-9,10-phenanthrene (TSEPIP) as derivatization reagent on a reversed phase Eclipse XDB-C8 (4.6×150mm, 5μm, Agilent) column in conjunction with a gradient elution followed by reversed phase high-performance liquid chromatography with fluorescence detection at 380 nm(excitation wavelength 260 nm) and tandem mass spectrometric identification has been developed. The contents of 26 free fatty acids in deep-sea fish oil were determined by external standard method, and the linear range of them was 200.0 pmol～48.83 fmol, all the correlation coefficients>0.9996, detection limits were 3.824～47.13 fmol (at signal to noise 3:1, S/N 3:1). The identification of all saturated and unsaturated fatty acid derivatives was carried out by post-column tandem mass spectrometry with atmospheric pressure chemical ionization (APCI) source in positive-ion mode, and the relative content of all saturated and unsaturated fatty acids was contrasted by peak area. According to the analysis of HPLC/MS/APCI, 25 fatty acids were identified, the main fatty acids in deep-sea fish oil are C12～C22 and 69.71% (peak area percent) of them are unsaturated fatty acids, especially polyunsaturated fatty acids which have important physiological function, such as 5, 8, 11, 14, 17-eicosapentaenoic acid(EPA, 16.62%), 2, 5, 8, 11, 14, 17-docosahexenoic acid (DHA, 12.31%).