Study on the Quality Standard and Fingerprint of Achyranthes bidentata Decoction Pieces

WANG Xiaoyan; CHANG Junmin; GUO Changrun; CHEN Li

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Study on the Quality Standard and Fingerprint of Achyranthes bidentata Decoction Pieces

更新时间：2022-06-21

- Study on the Quality Standard and Fingerprint of Achyranthes bidentata Decoction Pieces
- China Pharmacy Vol. 31, Issue 24, (2020)
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WANG Xiaoyan, CHANG Junmin, GUO Changrun, et al. Study on the Quality Standard and Fingerprint of Achyranthes bidentata Decoction Pieces. [J]. China Pharmacy 31(24).(2020)
DOI：

WANG Xiaoyan, CHANG Junmin, GUO Changrun, et al. Study on the Quality Standard and Fingerprint of Achyranthes bidentata Decoction Pieces. [J]. China Pharmacy 31(24).(2020) DOI：

摘要

目的：建立怀牛膝饮片的质量评价标准并建立不同产地饮片的指纹图谱。方法：采用薄层色谱法（TLC）鉴别怀牛膝饮片；按2015年版《中国药典》（四部）方法检查怀牛膝饮片的水分、总灰分、醇溶性浸出物的含量；采用紫外分光光度法测定总甾酮的含量；采用高效液相色谱法（HPLC）测定β-蜕皮甾酮的含量；采用HPLC法建立10批怀牛膝饮片的指纹图谱，以β-蜕皮甾酮（10号峰）为参照，采用《中药色谱指纹图谱相似度评价系统（2012版）》进行相似度评价，确定共有峰；采用SPSS21.0软件进行聚类分析、主成分分析以评价怀牛膝饮片的综合质量。结果：TLC鉴别结果显示，在供试品色谱中，与β-蜕皮甾酮、人参皂苷Ro对照品色谱相应位置上有相同颜色的斑点。10批怀牛膝饮片的平均水分含量为4.07％～6.33％，总灰分含量为5.04％～6.43％，醇溶性浸出物含量为6.57％～11.12％。总甾酮（以β-蜕皮甾酮计）、β-蜕皮甾酮检测质量浓度的线性范围分别为0.01～0.08、68.5～479.5µg/mL（R2均大于0.999），精密度、稳定性、重复性试验的RSD均小于3％，平均加样回收率分别为98.85％（RSD＝1.89％，n＝6）、100.34％（RSD＝2.12％，n＝9），平均含量分别为0.34％～0.56％、0.07％～0.09％。10批怀牛膝饮片共有24个共有峰，相似度均大于0.900。聚类分析结果显示，10批怀牛膝饮片可聚为4类，其中S1、S3、S6、S10为一类，S2、S7、S8为一类，S4、S9为一类，S5为一类。主成分分析结果显示，前4个主成分因子的累积方差贡献率为86.774％。综合质量高低依次为S8＞S5＞S9＞S4＞S3＞S7＞S6＞S10＞S2＞S1。结论：所建质量标准、含量测定方法和HPLC指纹图谱稳定、准确性好，可用于全面评价怀牛膝饮片的质量。

Abstract

OBJECTIVE：The establ ish the qu ality standard of Achyranthes bidentata decoction pieces and establish the fingerprint of decoction pieces of different origins. METHODS ：TLC method was used to identify A. bidentata decoction pieces. The contents of water ，total ash and ethanol-soluble extract in A. bidentata decoction pieces were determined according to the method in 2015 edition of Chinese Pharmacopoeia （part Ⅳ）. UV spectrophotometry was used to determine the content of total steroid. The content of β-ecdysterone in A. bidentata decoction pieces was determined by HPLC. HPLC method was used to establish the fingerprint of 10 batches of A. bidentata decoction pieces. Using β-ecdysterone（No. 10 peak）as reference ，similarity evaluation was conducted by using Similarity Evaluation System for TCM Chromatographic Fingerprints （2012 edition），and the common peaks were determined. SPSS 21.0 software was used for cluster analysis and principal component analysis so as to evaluate the comprehensive quality of A. bidentata decoction pieces. RESULTS ：Results of TLC identification showed that the spots with the same color on the corresponding positions of β-ecdysterone and ginsenoside Ro control in chromatogram of test sample. The average water content of 10 batches of A. bidentata decoction pieces was 4.07％ -6.33％ . The total ash content was 5.04％ -6.43％ . The ethanol-soluble extract was 6.57％ -11.12％ . The linear range of total sterone （by β-ecdysterone） and β-ecdysterone were 0.01-0.08 mg/mL and 68.5-479.5 µg/mL （R2＞0.999）， respectively. RSDs of precision ，stability and repeatability tests were all less than 3％. The average recovery rates were 98.85％（RSD＝1.89％，n＝6）and 100.34％（RSD＝ 2.12％，n＝9），respectively. The average contents were 0.34％-0.56％ and 0.07％-0.09％，respectively. There were 24 common peaks in 10 batches of A. bidentata decoction pieces ，and the similarity was all over 0.900. Cluster analysis results showed that 10 batches of A. bidentata decoction pieces could be grouped into 4 categories，among which S 1，S3，S6 and S 10 were one category ， S2，S7 and S 8 were one category ，S4 and S 9 were one category ，and S 5 was one category. The results of principal component analysis showed that the cumulative variance contribution rate of the first four principal components was 86.774％. The order of comprehensive quality is S 8＞S5＞S9＞S4＞S3＞S7＞S6＞S10＞S2＞S1. CONCLUSIONS ：The established quality standard ， content deter mination method and HPLC fingerprint are stable and accurate ，and can be used for the quality evaluation of A. bidentata decoction pieces. Δ 基金项目：国家科技部重点研发计划（No.2018YFC1707105）＊硕士研究生。研究方向：中药质量控制。E-mail：995354085@

关键词

怀牛膝饮片薄层色谱法高效液相色谱法含量测定指纹图谱聚类分析主成分分析

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College of Pharmacy， Hubei University of Chinese Medicine

Dept. of Pharmacy， Hubei Provincial Hospital of Traditional Chinese Medicine/the Affiliated Hospital of Hubei University of Chinese Medicine/Hubei Province Academy of Traditional Chinese Medicine

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