高效液相色谱荧光测定及质谱鉴定分析血清中胆汁酸  

Analyses of bile acids extracted from serum by high-performance liquid chromatography mass spectrum (HPLC/MS) with fluorescence detection

作　　者：石运伟[1] 王彩萍[1] 薛霞[1] 邓英霞[1] 尤进茂[1] 

机构地区：[1]曲阜师范大学化学科学学院

出　　处：《分析试验室》2005年第9期1-5,共5页Chinese Journal of Analysis Laboratory

基　　金：国家自然科学基金资助项目(20075016)

摘　　要：利用新型荧光试剂1,2-苯并-3,4-二氢咔唑-9-乙基苯磺酸酯(BDEBS)作柱前衍生化试剂, 在Eclipse XDB-C8色谱柱上,采用梯度洗脱对10种胆汁酸(BAs)荧光衍生物进行了优化分离.95 ℃下在二甲基亚砜(DMSO)溶剂中以柠檬酸钾作催化剂,衍生反应35 min后获得稳定的荧光产物,衍生反应完全.荧光激发和发射波长分别为λex=333 nm,λem=390 nm.采用大气压化学电离源(APCI Source)正离子模式,实现了血清中胆汁酸的定性测定.分析物的定量测定采用荧光法进行.线性回归系数均在0.9996以上,检出限为22.36～44.57×10-15 mol.Using 1, 2-benzo - 3, 4-dihydrocarbazole-9- ethyl-benzenesulfonate （BDEBS） as pre-column derivatization reagent, on a reversed-phased Eclipse XDB-C8 column, 10 bile acid derivatives were separated by high- performance liquid chromatography in conjunction with a gradient elution. Optimal derivatization was obtained by the reaction of bile acids with BDEBS in DMSO solvent at 95 ℃ for 35 min in the presence of mild basic catalyst potassium citrate. The fluorescence excitation and emission wavelengths were set at 333 and 390 nm, respectively. The qualitative analysis of derivatives in serum samples was accomplished by post-column mass spectrum with atmospheric pressure chemical ionization （APCI） source detection under positive mode. Quantitative determination of the analytes were achieved by means of fluorescence detection. Correlation coefficients for bile acid derivatives are 〉 0.9996, and detection limits（at signal- to-noise of 3 ： 1 ） are 22.36 - 44.57 fmol for the labeled bile acids. The established method is sensitive and reproducible for the determination of bile acids from real samples with satisfactory results.

关 键 词：高效液相色谱-质谱 荧光检测 柱前衍生 血清 胆汁酸 

分 类 号：O658.1[理学—分析化学]