苦丁茶主物质提取及其HPLC指纹图谱分析

Extraction and HPLC fingerprint analysis of the main substance of Ilex latifolia Thunb.

通过对苦丁茶成分的提取方式和条件初步优化,选取最优的色谱分离条件,运用高效液相色谱技术(HPLC),针对4个不同产地的10个批次苦丁茶建立指纹图谱.从图谱中所含主成分的种类和化学成分的提取率选择超声提取的方式,确定75%的乙醇为提取溶剂和30 min的超声时间.在对HPLC条件的优化过程中,根据最终实验结果的优越性选择流动相是甲醇—0.5%醋酸,等梯度洗脱的方式能更好达到分离效果,260 nm作为检测波长,流速为6 mL·min-1.多重方法学的考察结果表明提取剂具有良好稳定性,HPLC法检测后不同批次共有峰保留时间的基本重合,即RSD均<1%,相似度均>0.9,符合要求.

Through the preliminary optimization of the extraction method and conditions of the components of Ilex latifolia Thunb.,the optimum chromatographic separation conditions were selected by using high performance liquid chromatography(HPLC).The ultrasonic extraction method was selected from the types of principal components and the extraction rate of chemical components in the map,and 75%ethanol was determined as the extraction solvent with the ultrasonic time of 30 min.In the optimization of HPLC conditions,the mobile phase was selected as methanol-0.5%acetic acid according to the superiority of the final experimental results.The separation effect would be better achieved by gradient elution.260 nm was used as the detection wavelength and the flow rate was 0.6 mL·min-1.The results of multiple methods showed that the extract solvent had good stability,and the common peak retention time of different batches was basically coincident after HPLC detection(RSD was<1%and similarity>0.9),which meets the requirements.