高效液相色谱柱前衍生荧光检测法测定游离脂肪醇

Determination of Aliphatic Alcohols by HPLC/MS with Fluorescence and Atmospheric Pressure Chemical Ionization(APCI) Detection

本文利用新型荧光试剂1,2-苯并-3,4-二氢咔唑-9-乙酸(BCAA)作为柱前衍生试剂建立了测定游离脂肪醇的方法,实验以1-乙基-3-(3-二甲氨基丙基)环己碳二亚胺(EDAC)作为缩合剂,4-二甲氨基吡啶(DMAP)为催化剂,55℃下衍生反应25min后获得稳定的荧光产物。在Eclipse XDB-C8色谱柱上,通过梯度洗脱对12种游离脂肪醇进行了分离和在线质谱定性。采用大气压化学电离源(APCI)正离子模式,对抗静电剂(十二烷基磷酸酯钾盐)中游离脂肪醇进行定性及相应含量测定。脂肪醇的线性回归系数大于0.9997,检测限在9.40-25.32fmol。

A simple and highly sensitive method based on the derivatization of aliphatic alcohols utilizing 1,2-benzo-3,4-dihydrocarbazole-9-acetic acid （BCAA）as the novel pre-column derivatization reagent followed by high-performance liquid chromatography with fluorescence detection and online mass spectrometric identification was developed .Optimum derivatization was obtained by the reaction of aliphatic alcohols with BCAA at 55℃ for 25min in the presence of EDAC and DMAP in acetonitrile solvent. The condition of derivation was mildness. Derivatives were sufficiently stable to be efficiently analyzed by high-performance chromatography without pretreatment. On a reversed-phase Eclipse XDB-C8 column,twelve kinds of aliphatic alcohols （C1-C12）derivatives were separated in conjunction with a gradient elution with a good baseline resolution. The precise determining aliphatic alcohols from antistat was obtained by post-column online mass spectrometric identification with atmospheric pressure chemical ionization （APCI）source under positive mode detection. The established method exhibited excellent reproducibility and recovery. Linear responses were observed with coefficients of 0.9997,detection limits（at signal-to-noise of 3：1）9.40-25.32fmol.