ASE-HPLC法快速测定决明子中大黄酚和橙黄决明素

摘要

目的: 建立快速溶剂萃取(ASE)及HPLC测定决明子中大黄酚和橙黄决明素的分析方法.方法: 利用正交试验对ASE350快速溶剂萃取系统进行提取方法的研究,采用HPLC法同时测定决明子中大黄酚和橙黄决明素的含量.色谱柱为ACE Excel C18-PFP柱(75 mm×2.1 mm,2.5 μm),流动相为乙腈-0.1%磷酸溶液梯度洗脱,流速为0.4 ml·min-1,检测波长为284 nm,柱温为40℃.结果: 采用甲醇为萃取溶剂、萃取温度为120℃、静态萃取时间为5 min以及循环萃取3次的方法最优,可以很好地提取决明子中大黄酚和橙黄决明素,耗时仅为药典提取方法的1/9.大黄酚和橙黄决明素线性范围为0.73~58.57 μg·ml-1(r=0.999 7)和1.09~87.29 μg·ml-1(r=0.999 6),平均回收率分别为102.7%(RSD=0.8%)和98.2%(RSD=1.5%). 结论: 该方法能简便、快速、准确的测定决明子中的大黄酚和橙黄决明素.%Objective: To establish an accelerated solvent extraction(ASE)-HPLC method to determine chrysophanol and aurantio-obtusin in Cassia obtusifolia L.Methods: The optimal extraction conditions were defined by orthogonal tests using ASE.The method was carried out on an ACE Excel C18-PFP column (75 mm×2.1 mm,2.5 μm) with the mobile phase consisting of 0.1% phosphoric acid solution-acetonitrile with gradient elution.The column temperature was 40 ℃,the flow rate was 0.4 ml·min-1, and the detection wavelength was 284 nm. Results: The best process parameters of ASE were as follows:the extraction solvent was methanol, the extraction temperature was 120 ℃ and the static extraction duration was 5 minutes for three cycles.The ASE method needed only 1/9 of the time as the pharmacopoeia method,while the extraction efficiency of the ASE method was higher.The linear ranges of cassia obtusifolia L.and Chrysophanol were at 0.73~58.57 μg·ml-1(r=0.999 7) and 1.09~87.29 μg·ml-1(r=0.999 6).The average recoveries were 102.7%(RSD=0.8%) and 98.2%(RSD=1.5%).Conclusion: The method is simple, rapid and sensitive, which can be used for the rapid determination of aurantio obtusin and chrysophanol in Cassia obtusifolia L.