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Multi-component identification and target cell-based screening of potential bioactive compounds in toad venom by UPLC coupled with high-resolution LTQ-Orbitrap MS and high-sensitivity Qtrap MS

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Abstract

Traditional Chinese medicines (TCMs) are undoubtedly treasured natural resources for discovering effective medicines in treating and preventing various diseases. However, it is still extremely difficult for screening the bioactive compounds due to the tremendous constituents in TCMs. In this work, the chemical composition of toad venom was comprehensively analyzed using ultra-high performance liquid chromatography (UPLC) coupled with high-resolution LTQ-Orbitrap mass spectrometry and 93 compounds were detected. Among them, 17 constituents were confirmed by standard substances and 8 constituents were detected in toad venom for the first time. Further, a compound database of toad venom containing the fullest compounds was further constructed using UPLC coupled with high-sensitivity Qtrap MS. Then a target cell-based approach for screening potential bioactive compounds from toad venom was developed by analyzing the target cell extracts. The reliability of this method was validated by negative controls and positive controls. In total, 17 components in toad venom were discovered to interact with the target cancer cells. Further, in vitro pharmacological trials were performed to confirm the anti-cancer activity of four of them. The results showed that the six bufogenins and seven bufotoxins detected in our research represented a promising resource to explore bufogenins/bufotoxins-based anticancer agents with low cardiotoxic effect. The target cell-based screening method coupled with the compound database of toad venom constructed by UPLC-Qtrap-MS with high sensitivity provide us a new strategy to rapidly screen and identify the potential bioactive constituents with low content in natural products, which was beneficial for drug discovery from other TCMs.

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Acknowledgements

This work was supported by National Natural Science Foundation of China (Grant No. 21575146, 21635008, and 21621062), the Fundamental Research Funds for the Central public welfare research institutes (Grant No. ZZ10-007), and National Standardlization Project of Chinese Medicine (Grant No. ZYBZH-C-AH-01).

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Authors and Affiliations

  1. Beijing National Laboratory for Molecular Sciences, CAS Research/Education Center for Excellence in Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry Chinese Academy of Sciences, Beijing Mass Spectrum Center, North First Street 2, Beijing, 100190, China

    Wei Ren, Ming Guan, Hui Yang, Chao Han, Na Li, Tuo Li, Shilei Li, Yangyang Zhang & Zhenwen Zhao

  2. Graduate School, University of Chinese Academy of Sciences, No.19(A) Yuquan Road, Shijingshan District, Beijing, 100049, China

    Wei Ren, Ming Guan, Hui Yang, Chao Han, Na Li, Tuo Li, Shilei Li, Yangyang Zhang & Zhenwen Zhao

  3. Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, No. 16 Nanxiaojie, Dongzhimennei Ave, Beijing, 100700, China

    Lingyu Han, Mengyi Luo, Baolin Bian & Haiyu Zhao

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  1. Wei Ren
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  2. Lingyu Han
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  3. Mengyi Luo
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  4. Baolin Bian
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  5. Ming Guan
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  7. Chao Han
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  8. Na Li
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  9. Tuo Li
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  10. Shilei Li
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  11. Yangyang Zhang
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  12. Zhenwen Zhao
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  13. Haiyu Zhao
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Correspondence to Zhenwen Zhao or Haiyu Zhao.

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Animal experiments were conducted with the formal approval of the ethics committee of the China Academy of Chinese Medical Sciences (Beijing, China).

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Ren, W., Han, L., Luo, M. et al. Multi-component identification and target cell-based screening of potential bioactive compounds in toad venom by UPLC coupled with high-resolution LTQ-Orbitrap MS and high-sensitivity Qtrap MS. Anal Bioanal Chem 410, 4419–4435 (2018). https://doi.org/10.1007/s00216-018-1097-4

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  • DOI: https://doi.org/10.1007/s00216-018-1097-4

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