Research on the Quality Evaluation of Chansu Medicinal Materials Based on Fingerprint Atlas and Multi component Quantitative Analysis. As a traditional precious traditional Chinese medicine in China, Chansu mainly contains various chemical components such as alkaloids and toxic esters, which exert pharmacological activities such as anti-tumor and anti muscle ischemia. However, it also has certain toxicity and requires strict control of dosage. Shandong, as a genuine production area for toad venom medicinal materials, is expensive in the market, with significant differences in quality. Some medicinal materials even contain toad skin and glands, and there is insufficient management of the collection and initial processing of toad venom, resulting in a lack of quality evaluation of the source medicinal materials. In recent years, HPLC fingerprinting and multi-component content determination have been widely used in the study of traditional Chinese medicine. The 2020 edition of the Chinese Pharmacopoeia stipulates that the total mass fraction of the three components, Huachansu toxin base, Zhichandu ligand, and Chandu Ling, should not be less than 7.0. However, existing research lacks systematic evaluation studies using multiple indicators and analytical methods, and so far, literature reports on the quality evaluation components of toad venom are not comprehensive enough. On this basis, this study added 8 new components and used 11 main components such as sand toad venom essence and toad venom spirit as comprehensive indicators to evaluate the quality of toad venom. 40 batches of toad venom samples were tested through fingerprint analysis, and their quality was comprehensively evaluated by combining similarity analysis, cluster analysis, principal component analysis, and partial least squares discriminant analysis results.

 

Shandong Province is a genuine production area for toads. According to surveys, there are as many as 127 toad farmers in Shandong Province, distributed in all cities except Dongying City. Due to differences in breeding scale, the quality of toad venom varies greatly. The combination of fingerprint spectrum and multi-component quantitative analysis can accurately distinguish the quality differences between toad venom samples, which can be used as a preliminary criterion for evaluating the quality of commercial medicinal materials. This study is based on the establishment of HPLC fingerprint spectra of 11 chemical components in toad venom samples, mainly targeting the cardiac glycosides bufadienolactones in toad venom medicinal materials. Modern pharmacological experiments have shown that these components have anti-tumor, immune regulatory activity, and the ability to attenuate cancer-related pain.

In addition, similarity analysis was conducted on the chromatographic peaks, and the results showed that the similarity between different batches of toad venom in Shandong Province was greater than 0.900, indicating that the similarity of characteristic components among different batches of toad venom in Shandong Province was good. However, the similarity between the six batches of samples outside the province and other batches was significantly different, especially the similarity between the six batches of samples in Hunan and Jiangsu was between 0.779 and 0.802.

To further analyze the differences in common peaks among different batches of Chansu medicinal materials, clustering, principal component analysis, and partial least squares discriminant analysis were conducted on 40 batches of Chansu to effectively distinguish the samples. The 40 batches of samples can be divided into three categories, with consistent classification results. There are differences in the quality of Chansu between different batches, and the VIP analysis of partial least squares discriminant analysis showed that the 12 components with the highest contribution degree include Sha Chan toxin essence, Bu Ta Li Ding, Zhi Chan toxin ligand, Hua Chan toxin base, Ri Chan toxin spirit, and Bu Tu spirit.

Through content determination of 40 batches of Bufonisu medicinal materials from different batches, it was found that the Bufonisu from Weishan, Jining had the highest content of Bufonisu Ling, Fat Bufonisin Formula, and Hua Bufonisu Formula according to the pharmacopoeia. In addition, the sample with the highest total content of all ingredients was Bufonisu from Juxian, Rizhao. The toad venom samples from within Shandong province have higher content than those from outside the province in terms of the sum of the three components specified in the pharmacopoeia and the sum of all measured components. However, the toad venom samples from Hunan and Jiangsu provinces do not meet the pharmacopoeia requirements, indicating that Shandong toad venom has certain advantages in terms of quality. The analysis results of the content of toad venom in Shandong Province show that the toad venom from Weishan in Jining and Juxian in Rizhao not only scored higher in principal component analysis, but also had a higher content of bufalin, lipid bufalin, and Chinese toad venom according to the pharmacopoeia. Moreover, the total amount of ingredients contained in this sample is also higher among all batches. Therefore, the author believes that the comprehensive quality of this batch of toad venom is the best, which may be related to the climate conditions and geographical location of these two places, providing a theoretical basis for the vigorous development of toad farming in this area.

In summary, this study not only established a fingerprint spectrum for the simultaneous determination of 11 components in Chansu, but also had a certain correlation with the quality identification and evaluation of Chansu, comprehensively evaluating the quality of Chansu and providing scientific basis for quality control in the production process of Chansu. In the process of raising toads, the living conditions of toads should be fully considered, scientific and rigorous quality control indicators should be adopted, more reasonable and scientific domestication bases should be established, and the economic benefits of the production area should be improved.