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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。
On a Hypersil C18 column, a new fluorescent reagent, 1,2-benzo-3,4-dihydrocarbazole-9-ethyl p-toluenesulfonate (BDETS) was used as a pre-column derivatization reagent, Ten bile acid (BAs) fluorescent derivatives were optimized. 95 ° C in dimethyl sulfoxide solvent with potassium citrate as a catalyst derived derivative reaction after 30min to obtain a stable fluorescent product, the derivatization reaction was complete. Excitation and emission wavelengths were λex = 333 nm, λem = 390 nm, respectively. The qualitative and quantitative determination of bile acid in serum was achieved by adopting the positive ion mode of atmospheric chemical ionization source (APCI). The linear regression coefficients were all above 0.9996 with wide linear range and the detection limit was 12.94 ~ 21.94fmol.