【摘要】 在HypersilC18色谱柱上,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)作柱前衍生化试剂,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℃下在二甲基亚砜溶剂中以柠檬酸钾作催化剂,衍生反应30min后获得稳定的荧光产物,衍生反应完全。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,实现了血清中胆汁酸的定性和定量测定。线性回归系数均在0.9996以上,线性范围宽,检出限为12.94~21.94fmol。更多还原

【Abstract】 On a reversed-phased Hypersil C₁₈ column , 10 bile acids derivatives were separated using 1,2-benzo-3,4-dihydrocarbazole-9-ethyl-p-toluenesulfonate （BDETS） as pre-column derivatization reagent by high-performance liquid chromatography with fluorescence detection and mass spectrum identification in conjunction with a gradient elution. Optimum derivatization, giving the corresponding sensitively fluorescent derivatives, was obtained by the reacting of bile acids with BDETS at 95℃ for 30 min in the presence of mild basic catalyst potassium citrate in dimethyl sulfoxide solvent . The fluorescence excitation and emission wavelengths were set at λ_ex 333 and λ_em 390 nm, respectively. The identification of bile acids derivatives was obtained on the basis of atmospheric-pressure chemical ionization source at positive model detection. Correlation coefficients for the peak areasand bile acid derivative concentrations are >0.9996, and detection limits at signal-to-noise of （3∶1） are 12.94～21.94 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.更多还原