柱前荧光标记法结合HPLC-FLD测定石榴中四种三萜酸

Determination of 4 Tritepenoidic Acids in Pomegranate by Precolumn Fluorescent Derivatization Coupled with HPLC-FLD

本文利用2-（7H-二苯并[a,g]咔唑）乙基对甲苯磺酸酯（DBCETS）作为荧光标记试剂,建立了三萜酸的快速、高灵敏HPLC检测方法,利用响应面优化法优化了三萜酸的荧光标记条件（标记时间、标记温度及标记试剂用量）;建立的新方法具有很好的重现性及准确性,衍生物线性相关系数均大于0.9995,检测限为1.10~1.48ng/m L（S/N=3）;同时该方法首次应用于石榴中科罗索酸、山楂酸、齐墩果酸及熊果酸检测,结果令人满意。

In this study, a fast and sensitive HPLC method has been developed for the determination of triterpenic acids using 2-（7H-dibenzo[ a, g ] carbazol-7-yl） ethyl 4-methylbenzenesulfonate （ DBCETS ） as the fluorescent labeling rea- gent. A response surface methodology （RSM） based on a Box-Behnken design （BBD） was employed to optimize the main parameters affecting the fluorescence labeling including labeling time, reaction temperature and the concentration of labeling reagent. Under the optimized conditions ,the proposed method gave good reproducibility and accuracy. The linear correlation coefficients for all triterpenic acid derivatives were of 〉 0. 9995, and the limit of detections were in the range of 1.10 ~ 1.48 ng/mL （ S/N = 3）. The proposed method has been firstly applied for the determination of corosolic acid, maslinic acid, oleanolic acid and ursolic acid in pomegranate, and got satisfactory results.