基于HPLC分析柴胡黄芩配伍的主要化学成分研究

Study on Main Chemical Components from Compatibility of Chaihu and Huangqin by HPLC

目的:基于高效液相色谱法(High Performance Liquid Chromatography,HPLC)分析柴胡黄芩配伍的主要化学成分,阐释中药合理配伍的科学内涵。方法:采用HPLC指纹图谱分析进行精密度试验测试HPLC仪器的精密度;重复性试验测试HPLC指纹图谱方法的重复性;稳定性试验测试样品的稳定性。HPLC指纹图谱:取10批柴胡黄芩,精密吸取5μL按照供试品溶液制备方法制备供试品溶液,根据色谱条件依次测定并记录10批柴胡黄芩配伍的色谱图。结果:1)柴胡黄芩精密度试验中峰面积大于总峰面积5%的峰有22、30、36和47号峰。2)重复试验和稳定性试验中各主要色峰的相似度均大于0.99,显示良好的重复性和稳定性。3)10批柴胡黄芩HPLC指纹图谱中大于总峰面积5%的共有峰面积为22、30、36、47号色谱峰,且10批柴胡黄芩配伍的药液相似度均>0.99,显示相似度良好。4)柴胡黄芩配伍的水煎液中,大多数物质归属柴胡,45 min以后的色谱峰是柴胡的特征峰,而15~45 min多集中了黄芩的特征峰;主要化学成分为柴胡皂苷a和黄芩苷。结论:柴胡黄芩配伍的水煎液中主要化学成分为柴胡皂苷a和黄芩苷。

Objective： To analyze the main chemical components from compatibility of Chaihu and Huangqin by HPLC and to explain the scientific connotation of rational compatibility. Methods： High performance liquid chromatography（ HPLC） fingerprint was used for test precision of HPLC instrument. Repeatability of HPLC fingerprint method was test and sample stability was also tested. HPLC fingerprint： Ten batches of Chaihu and Huangqin were taken and 5 μL solution was prepared according to the conditions of chromatography and the chromatogram. Fingerprint of compatibility of Chaihu and Huangqin was determined and recorded.Results： 1） The peak areas in the precision test over 5% of the total peak area were No. 22,30,36 and 47. 2） The similarity of the main color peaks in the repeatability test and the stability test was greater than 0. 99,which showed good repeatability and stability. 3） The total peak areas of the HPLC fingerprint of the ten batches of Chaihu and Huangqin greater than the total peak area were No. 22,30,36,and 47 and the similarity of the chemical compatibility of the ten batches of Chaihu and Huangqin was above0. 99,showing a good similarity. 4） Most substances in the decoction of Chaihu with Huangqin came from Chaihu and chromatographic peak after 45 min was the characteristic peak of Chaihu,while characteristic peak of Huangqin mainly appeared during 15~ 45 min. The main chemical components were saikosaponin a and baicalin. Conclusion： The main components in decoction of Chaihu and Huangqin were saikosaponin a and baicalin.