A strategy for the metabolomics-based screening of active constituents and quality consistency control for natural medicinal substance toad venom

Natural medicinal substances (NMS) have great inconsistency due to chemical variability, seriously limiting their development as therapeutics. Here, we chose toad venom with anti-tumor activities as a model and developed a pipeline of metabolomics-based screening and quality consistency control (MSQCC) as one potential solution to this long-standing problem. Our study firstly exemplified the power of the co-correlation screen of metabolomic and biological profiles of 180 fractions prepared from natural heterogeneous venom samples, to identify a series of bufadinolides as quality control markers for cancer cell inhibition. The next quantitative monitoring of these markers revealed great batch-to-batch variation of toad venoms. Finally, we developed a marker-based blending program (Markers-NMBT) to normalize heterogeneity of NMS. It created the blends for the conversion of the unqualified venoms with high variation in the contents of bufadienolides, into qualified products consistent with the reference. Thus, this work provides a strategy for rapid, large-scale discovery, quantification and application of quality control markers to ensure batch-to-batch consistency, and can be a crucial technology in the development of modern NMS preparations.