【摘要】 在HypersilC18色谱柱上,利用新型荧光试剂苯并[b]吖啶酮-5-乙基对甲苯磺酸酯(BAETS)作柱前衍生化试剂,采用梯度洗脱对10种胆汁酸衍生物进行了优化分离.在二甲亚砜溶剂中,以碳酸钾作催化剂,95℃,45min后获得稳定的荧光产物.衍生物在稳态荧光条件下,在乙腈和甲醇水溶液中的百分离子化δ值在0%～88.83%和0%～89.15%范围内.最大激发和发射波长为λex/λem=272/505nm.采用大气压化学电离源(APCI)正离子模式实现了胆汁中胆汁酸的定性定量测定.线性回归系数均在0.9995以上,线性范围宽,检出限为0.76～1.62ng/mL.更多还原

【Abstract】 On a reversed-phased Hypersil C18 column, 10 bile acid derivatives were separated using 2-(12-oxobenzo[b]acridin-5(12H)-yl)ethyl 4-toluenesulfonate (BAETS) 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 BAETS at 95 ℃ for 30 min in the presence of basic catalyst potassium carbonate in dimethylsulfoxide solvent. The optimum excitation and emission wavelengths were at λex/λem=272 nm/505 nm. The percent ionization δ values in aqueous acetonitrile and aqueous methanol were in the range of 0%～88.83% and 0%～89.15% respectively. The identification of bile acid derivatives was obtained on the basis of an atmosphere-pressure chemical ionization source at positive model detection. Correlation coefficients are >0.9995, and detection limits at signal-to-noise of 3 are 0.76～1.62 ng/mL for the labeled bile acids.更多还原