Combining untargeted and targeted metabolomics approaches for the standardization of polyherbal formulations through UPLC–MS/MS
Polyherbal formulations are an integral part of various indigenous medicinal systems such as Traditional Chinese Medicine (TCM) and Ayurveda. The presence of a very large number of compounds makes the quality control of polyherbal formulations very difficult.
To overcome this problem, we have developed a comprehensive strategy for the dereplication of natural products in polyherbal formulations by using Adhatoda vasica as a case study.
The strategy is based on five major steps: the collection of plant samples from different locations to observe the effects of environmental variables; LC–ESI–MS/MS-based untargeted metabolite profiling of the plant samples to identify marker compounds using extensive chemometric analysis of the obtained data; the identification of marker compounds in polyherbal products; the isolation, purification and characterization of the marker compounds; and MRM-based quantitative analysis of the isolated marker compounds using LC–ESI–MS/MS.
Using this strategy, we identified a total of 51 compounds in the methanolic extract of A. vasica plants from 14 accessions. Chemical fingerprinting of the plant led to the identification of characteristic peaks that were used to confirm the presence of A. vasica in complex polyherbal formulations. Four quinazoline alkaloids (marker compounds) were isolated, purified and quantified in various herbal formulations containing A. vasica.
This method demonstrates a comprehensive strategy based on untargeted and targeted metabolite analysis that can be used for the standardization of complex polyherbal formulations.
KeywordsChemical profiling Standardization Identification Quantification Polyherbal formulation
The authors express gratitude to Mr. Arsalan Tahir and Mr. Junaid Ul Haq for technical assistance in UHPLC–MS/MS analyses. Mr. Faraz Ul Haq would also like to acknowledge the Higher Education Commission (HEC), Pakistan for an Indigenous PhD Fellowship.
SGM proposed the subject, designed the study and actively participated in manuscript reviewing. SR, FUH, MNK and SMZS performed the experiments and actively involved in the write-up of the manuscript. AA, AA and HRES assisted in reviewing the manuscript. All authors reviewed the manuscript.
This work was supported by the Higher Education Commission (HEC), Pakistan under the Industry Academic & Government Linkages program (HEC/R&D/UITSP/03/2016).
Compliance with ethical standards
Conflict of interest
The authors declare no conflict of interest.
No animals were used/harmed in this study. This study did not involve any human participants.
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