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Integrated post-acquisition data processing strategy for rapid steroid sulfate characterization in Toad gall-bladder.

Wei Chen1, Wenhui Zhao1, Lijuan Wu1

  • 1Modern Research Center for Traditional Chinese Medicine, Beijing Research Institute of Chinese Medicine, Beijing University of Chinese Medicine, Beijing, China; School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, China.

Journal of Pharmaceutical and Biomedical Analysis
|January 10, 2024
PubMed
Summary

This study introduces an integrated strategy for rapid chemical filtering of Chinese medicinal materials using liquid chromatography-mass spectrometry (LC-MS). The method efficiently profiles steroid sulfates in Toad gall-bladder, aiding in the characterization of complex natural products.

Keywords:
(34)S isotopically molecular ionBilichol sulfatePost-acquisition data processing strategySteroid sulfateToad gall-bladder

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Area of Science:

  • Analytical Chemistry
  • Natural Product Chemistry
  • Pharmacognosy

Background:

  • Liquid chromatography-mass spectrometry (LC-MS) is crucial for characterizing Chinese medicinal materials (CMMs) due to its rich MS/MS spectral data.
  • Processing large datasets from LC-MS to identify specific compounds is challenging and time-consuming, hindering efficient analysis of CMMs.
  • Steroid sulfates, including bufogenin, bile acid, and bilichol sulfates, are important bioactive compounds found in CMMs like Toad gall-bladder.

Purpose of the Study:

  • To develop and validate an integrated data filtering strategy for rapid, homologs-focused chemical characterization of CMMs using LC-MS.
  • To apply this strategy for in-depth profiling of steroid sulfates in Toad gall-bladder (Chandan).
  • To demonstrate the strategy's utility for submetabolome characterization in complex natural product matrices.

Main Methods:

  • An integrated strategy combining molecular weight imprinting (MWI), diagnostic fragment ion filtering (DFIF), neutral loss filtering (NLF), and isotope pattern filtering (IPF) was developed.
  • In vitro metabolism using recombinant human SULT2A1 generated bufogenin sulfates to establish MS/MS spectral behaviors.
  • LC-MS/MS data from Toad gall-bladder were filtered using MWI, IPF, DFIF (targeting SO3-· and HSO4-), and NLF (targeting SO3) for sulfate identification.

Main Results:

  • The integrated strategy successfully filtered and identified 71 compounds from Toad gall-bladder, including 7 bufogenin sulfates, 17 bile acid sulfates, and 13 bilichol sulfates.
  • Thirty-nine of the detected compounds were assigned plausible identities based on empirical mass fragmentation rules and spectral data.
  • The method enabled a comprehensive profile of steroid sulfates in Toad gall-bladder, showcasing its effectiveness for complex natural product analysis.

Conclusions:

  • The proposed integrated data filtering strategy significantly accelerates the homologs-focused chemical characterization of CMMs using LC-MS/MS.
  • This approach provides a valuable tool for the in-depth profiling of specific compound classes, such as steroid sulfates, within complex natural product samples.
  • The strategy offers a meaningful option for submetabolome characterization in CMMs and potentially other complex biological matrices.