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血清中游离脂肪酸的液相色谱荧光测定及质谱鉴定
Determination of Free Fatty Acids in Serum by Liquid Chromatography Mass Spectrum with Fluorescence Detector
【摘要】 利用新型荧光试剂1,2 苯并 3,4 二氢咔唑 9 乙基对甲苯磺酸酯(BDETS)对19种游离脂肪酸(FFAs)进行柱前衍生,在EclipseXDB C8反相色谱柱上,采用梯度洗脱优化分离.90℃下在DMF溶剂中以K2CO3作催化剂,衍生反应30min获得稳定的荧光产物.激发和发射波长分别为λex=333nm,λem=390nm,采用大气压化学电离源(APCI)正离子模式进行柱后在线质谱定性.多数脂肪酸的线性回归系数大于0.9989,检测限为24.80~80.37fmol.实现了人体血清中长链脂肪酸的定性及相应含量测定.
【Abstract】 On a reversed-phase Eclipse XDB-C8 column, 19 free fatty acids (FFA) derivatized using 1,2-benzo-3,4-dihydrocarbazole -9-ethyl-toluene-4-sulfonate (BDETS) as pre-column derivatization reagent were separated in conjunction with a gradient elution. Optimum derivatization, giving the corresponding stable fluorescent derivatives, were done by reacting of fatty acids with BDETS at 90℃ for 30 min in the presence of K2CO3 as catalyst in DMF solvent. The fluorescence excitation and emission wavelengths were set at 333 nm λex and 390 nm λem, respectively. The identification of long-chain FFA derivatives was obtained on the basis of APCI detection at positive mode. Correlation coefficients for FFA derivatives are >0.998 9, and detection limits at signal-to-noise of 3:1 are 24.80~80.37 fmol for the labeled FFAs. The established method is sensitive and reproducible for the determination of FFAs in extracted serum with satisfactory results.
【Key words】 high-performance liquid chromatography; mass spectrum; fluorescence detection; pre-column derivatization; fatty acids;
- 【文献出处】 化学研究 ,Chemical Researches , 编辑部邮箱 ,2004年04期
- 【分类号】R446.1
- 【被引频次】25
- 【下载频次】394