Combination of HPLC Fingerprint and QAMS as a New Analytical Approach for Determination of Bufadienolides in &amp;lt;i&amp;gt;Bufonis Venenum&amp;lt;/i&amp;gt;

Yi Luo,Fei He,Shengci Fan,Zhaoqing Meng,Cejia Liu

doi:10.11648/j.ijcm.20200401.11

Abstract

The present study presents an effective and comprehensive evaluation method for assessing the quality of Bufonis Venenum, based on analysis of high performance liquid chromatography (HPLC) fingerprints in combination with similarity analysis, hierarchical cluster analysis (HCA), principal component analysis (PCA) and a quantitative analysis multi-components by single marker (QAMS) method. Nine common peaks identified by fingerprint detection results of 10 batches of samples were collected and used for the similarity analysis, HCA, PCA and QAMS analysis. These methods drew a similar conclusion that the quantitative analysis multi-components by single marker (QAMS) method 41 Bufonis Venenum samples were categorized into four main groups by HCA and PCA, and the majority of the samples with similar ingredients were mainly concentrated in Shandong area. When QAMS method was compared with the external standard method (ESM), it was feasible to evaluate the quality of Bufonis Venenum by the values of relative correction factors (RCFs) from cinobufagin (the internal reference) versus arenobufagin, bufalin, bufotalin, resibufogenin and telocinobufogin. In conclusion, these methods were successfully applied to identification of the origin and evaluation the quality of Bufonis Venenum. Therefore, these evaluation methods are promising to be widely applied in the quality control of Traditional Chinese Medicines (TCMs).

Highlights

Bufonis Venenum is a dry secretion derived from the parotoid glands, and skin of two species of toads, Bufo bufo gargarizans Cantor and B. melanostrictus Schneider [1, 2]
The results showed that relative retention time (RRT) was stable and could be used to identify the other five compounds referring to cinobufotalin in spite of the differences in columns or high performance liquid chromatography (HPLC) instruments, or those arising from analysis being performed in different laboratories
Cinobufagin and resibufogenin are detailed as the appropriate control markers to be used for Bufonis Venenum analysis in the Chinese Pharmacopoeia, while bufalin, cinobufagin and resibufogenin are mandated by the Japanese and Korean Pharmacopoeia

Summary

Introduction

Bufonis Venenum is a dry secretion derived from the parotoid glands, and skin of two species of toads, Bufo bufo gargarizans Cantor and B. melanostrictus Schneider [1, 2]. It is known as Chansu in China and Senso in Japan. To date, according to the references reported, 71 kinds of bufadienolide have been described, of which cinobufagin, resibufogenin, telocinobufogin, arenobufagin, bufalin and bufotalin are typically the most abundant These six compounds are known to possess cardiac function and antitumor activities [7,8,9]. Bufalin is known to possess anti-inflammatory and anesthetic properties [6, 10]

Methods

Results

Conclusion