不同产地酒制蜂胶的HPLC指纹图谱研究

Study on the HPLC Fingerprint of Propolis Processed with Ethanol Collected from Different Place of Origin

目的应用高效液相色谱(HPLC)法建立酒制蜂胶指纹图谱,对蜂胶进行质量控制,为产地的选择提供理论依据。方法采用Phenomenex Kinetex 5μm F5 100A(4.6 mm×250 mm)色谱柱,以甲醇(A)-0.3%醋酸水溶液(B)作为流动相进行梯度洗脱:0～15 min,42%～52%A; 15～30 min,52%～55%A; 30～60 min,55%～59%A; 60～80 min,59%～83%A;柱温35℃;流速:0.8 m L·min^(-1)。测定22批不同产地的酒制蜂胶,通过"中药色谱指纹图谱相似度评价系统"生成指纹图谱,并进行相似度评价。结果该方法可使蜂胶中各成分得到较好的分离,指纹图谱确定了25个共有峰,并指认了其中6个化学成分。不同产地间样品相似度差异较大。结论该HPLC指纹图谱研究方法简便、准确、重复性好,可用于蜂胶的质量控制,为产地的选择提供理论依据。

OBJECTIVE To establish the fingerprint of propolis processed with ethanol by HPLC method,and then to provide theoretical basis for controlling its quality and choosing place of origin. METHODS We used Phenomenex Kinetex 5 μm F5 100 A( 4.6 mm×250 mm) column,with methanol as mobile phase A while 0.3% aqueous acetic acid solution as B,which were eluted gradiently:0-15 min,42%-52%A;15-30 min,52%-55%A;30-60 min,55%-59%A;60-80 min,59%-83%A;column temperature was set at35 ℃ and the flow rate was 0. 8 m L·min-1. Twenty-two propolis processed with ethanol taken from three different place of origin were analyzed,and fingerprint was established by using the'Chinese medicine chromatographic fingerprint similarity evaluation system',and then evaluated their similarities. RESULTS Most chemical composition of propolis processed with ethanol could be separated and the fingerprint retained 25 common peaks,and 6 of these were confirmed. There was obvious discrimination in similarity of fingerprint between different places of origin. CONCLUSION The research method about HPLC fingerprint of propolis processed with ethanol were convenient,accurate as well as repeatable,which could be used to control quality and choose place of origin of propolis.