不同规格怀牛膝不同极性部位HPLC指纹图谱

目的研究并比较不同规格怀牛膝不同极性部位的HPLC指纹图谱,探索其内部质量差异,为该药材的规格标准完善及临床用药提供参考。方法将怀牛膝用体积分数75%乙醇水浴回流提取,得体积分数75%乙醇回流提取物,用40 mL水溶解后,依次用石油醚、三氯甲烷、乙酸乙酯、正丁醇进行萃取,得到各萃取相及萃取后的水相,浓缩至浸膏,采集各部位HPLC指纹图谱;运用相似度、综合聚类法进行数据分析,同时对其不同部位的指纹图谱进行比对分析。结果石油醚、氯仿部位均标定了11个共有峰,乙酸乙酯部位标定了10个共有峰,正丁醇部位标定了19个共有峰,水部位标定8个共有峰;氯仿部位指纹图谱相似度差异较大,其他部位指纹图谱相似度差异较小,均在0.9以上;石油醚部位指纹图谱差异主要体现在峰高,乙酸乙酯部位、氯仿部位、水部位的化学成分种类及峰高均存在差异;综合聚类分析能将不同规格怀牛膝区分开。结论不同规格怀牛膝内部质量存在差异;实验中所建立的HPLC指纹图谱可以全面反映不同规格怀牛膝的化学成分分布,为不同规格怀牛膝的整体质量评价提供参考。

HPLC Fingerprints of Different Polarity Fractions of Achyranthes bidentata of Different Specifications

OBJECTIVE To study and compare the HPLC fingerprints of different polar parts of Achyranthes bidentata of different specifications,investigate their internal quality differences,and provide reference for the standard improvement and clinical use of the medicinal material.METHODS The Achyranthes bidentata samples were refluxed with 75%ethanol,the extract was dissolved with 40 mL water and then,extracted in turn with petroleum ether,chloroform,ethyl acetate and n-butanol,each extracted phase and aqueous phase were obtained and concentrated.The HPLC fingerprint of each part was collected.The data was analyzed by similarity and comprehensive clustering.Meanwhile,the fingerprints of different parts were compared and analyzed.RESULTS Eleven common peaks were determined in the HPLC fingerprints of petroleum ether fraction and chloroform extract fraction,10,19,and 8 common peaks were determined in the HPLC fingerprints of ethyl acetate fraction,n-butyl alcohol fraction,and water extract respectively.The similarity of the fingerprints of chloroform parts was quite low.The similarity of the fingerprints of other parts was higher,all above 0.9.The difference in the fingerprints of petroleum ether parts was mainly reflected in the peak height.The chemical compositions and peak heights of ethyl acetate,chloroform and water extracts were different.Comprehensive cluster analysis could distinguish different specifications of Achyranthes bidentata.CONCLUSION The internal qualities of different specifications of Achyranthes bidentata are different.HPLC fingerprint established in this experiment can comprehensively reflect the chemical components distribution of Achyranthes bidentata of different specifications.It provides reference for the whole quality evaluation of different specifications of Achyranthes bidentata.