Metabolomics method based on ultra high performance liquid chromatography with time-of-flight mass spectrometry to analyze toxins in fresh and dried toad venom

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• 作者：Hongyue Ma ; Huixia Niu ; Qin Cao ; Jing Zhou ; Yan Gong ; Zhenhua Zhu ; Xiang Lv ; Liuqing Di ; Dawei Qian ; Qinan Wu and Jin'ao Duan
• 刊名：Journal of Separation Science
• 出版年：2016
• 出版时间：December 2016
• 年：2016
• 卷：39
• 期：24
• 页码：4681-4687
• 全文大小：3527K
• ISSN：1615-9314

文摘

Drying is a critical step to prolong the storage time in natural medicine processing but it changes the chemical characteristics of the product. In this study, research was performed to characterize the metabolomic changes in toad venom induced by vacuum-drying at 60°C and air-drying at room temperature by ultra high performance liquid chromatography coupled with pattern recognition approaches. In total 52 metabolites, down-regulated or up-regulated, were identified as potential chemical markers. Compared with fresh toad venom, vacuum-drying at 60°C succeeded in raising the conjugated-type bufadienolide content significantly, while the content of free-type bufadienolides were slightly reduced. On the other hand, toad venom air-dried at room temperature presented a relatively low amount of bufadienolides compared with fresh venom. For example, the content of several known anti-tumor components (gamabufotalin, bufotalin, cinobufagin, etc.) were significantly reduced. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide bioassay further showed that venom air-dried at room temperature had weaker anti-tumor activity on human hepatocellular carcinoma SMMC-7721 proliferation in vitro than samples vacuum-dried at 60°C. These results showed that the great metabolomic changes of toad venom occurred during the drying process, suggesting that a proper drying procedure is important for sustaining the chemical quality of natural medicines.