HPLC Coupled with Chemical Fingerprinting for Multi-Pattern Recognition for Identifying the Authenticity of Clematidis Armandii Caulis

Fangqing Wang; Zengkai Qian; Guolu Liao; Jing Zeng; Dan Huang; Qiquan Liu; Xingliang Xie

doi:10.3791/64690

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Method Article

HPLC Coupled with Chemical Fingerprinting for Multi-Pattern Recognition for Identifying the Authenticity of Clematidis Armandii Caulis

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DOI:

10.3791/64690

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November 11th, 2022

Fangqing Wang^*¹ , Zengkai Qian^*¹ , Guolu Liao^*² , Jing Zeng² , Dan Huang² , Qiquan Liu² , Xingliang Xie¹

¹School of Pharmacy, The Second Class Laboratory of Traditional Chinese Medicine Pharmaceutics, National Administration of Traditional Chinese Medicine, Chengdu Medical College, ²Pengzhou Second People's Hospital

^* These authors contributed equally

Summary

Here, we present a protocol to establish high-performance liquid chromatography (HPLC), coupled with chemical fingerprint multi-pattern recognition, which provides a new strategy for effectively identifying the genuine varieties of Clematidis Armandii Caulis and its adulterants.

Abstract

A method for identifying Chinese medicinal materials and their related adulterants was constructed by taking Clematidis Armandii Caulis (Chuanmutong, a universally used traditional Chinese medicine) as an example. Ten batches of genuine Chuanmutong varieties and five batches of related adulterants were analyzed and compared based on the high-performance liquid chromatography (HPLC) fingerprints combined with chemometrics, including cluster analysis (CA), principal component analysis (PCA), and orthogonal partial least-squares discrimination analysis (OPLS-DA). In addition, the content of β-sitosterol was determined. The control chemical fingerprint of Chuanmutong was established, and 12 common peaks were identified. The similarity between the fingerprint of 10 batches of genuine Chuanmutong varieties and the control fingerprint was 0.910-0.989, while the similarity of five batches of adulterants was only 0.133-0.720. Based on the common peaks in the chromatogram, 15 batches of samples were classified into three content levels by PCA, and were aggregated into four categories by CA, achieving a clear distinction between authentic Chuanmutong and adulterants of Chuanmutong. Further, seven differential components that can effectively identify authentic Chuanmutong and adulterants of Chuanmutong were found through OPLS-DA. The β-sitosterol content of 10 batches of genuine Chuanmutong varieties was 97.53-161.56 µg/g, while the β-sitosterol content of the five batches of adulterants varied greatly, among which the β-sitosterol content of Clematis peterae Hand.-Mazz. and Clematis gouriana Roxb. Var. finetii Rehd. et Wils. was significantly lower than that of authentic varieties of Chuanmutong. The HPLC index component content and chemical fingerprint multi-pattern recognition method established in this study provide a new strategy for effectively identifying authentic Chinese medicinal materials and related adulterants.

Introduction

Chuanmutong, dry Caulis of Clematis armandii Franch. or Clematis montana Buch.-Ham., is a traditional Chinese medicine commonly used in clinics¹^,²^,³. It is used for treating urinary problems, edema, sores on the tongue and mouth, decreased milk secretion, joint stiffness, and muscle pain caused by damp heat⁴. Chuanmutong has always been obtained from wild varieties, mainly distributed in southwest China, especially in Sichuan, where the best quality can be found⁵^,⁶. It is diff....