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Comprehensive analysis using DNA metabarcoding, SCAR marker based PCR assay, and HPLC unveils the adulteration in Brahmi herbal products

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Abstract

Background

Brahmi is one of the important nootropic botanicals, widely sold in the market, with the name “Brahmi’’ being used to describe both Bacopa monnieri and Centella asiatica species. The Brahmi herbal products market is expanding; hence, economically motivated adulteration is highly prevalent.

Methods and results

This study aimed to develop DNA-based methods, including SCAR marker-based PCR and metabarcoding, to authenticate Brahmi herbal products and compare these methods with HPLC. These methods have been validated using mock controls (in-house blended formulations). All targeted plant species in mock controls were detected successfully with all three methods, whereas, in market samples, only 22.2%, 55.6%, and 50.0% were found positive for Brahmi by PCR assay, DNA metabarcoding, and HPLC, respectively. Metabarcoding can detect the presence of non-labeled plants together with targeted species, which is an advantage over PCR assay or HPLC.

Conclusion

SCAR marker-based PCR is a rapid and cost-effective method for detecting the presence of B. monnieri and C. asiatica. However, in this study, the success rate of PCR amplification was relatively low because the primers targeted either RAPD or ITS-based SCAR markers. HPLC assay, although an alternative, was unable to detect the presence of other botanicals, just like the SCAR marker-based PCR assay. On the other hand, metabarcoding can be utilized to identify the target plants, even in very small quantities, while also providing simulated identification of other botanicals. This study successfully addressed the need for quality control of Brahmi herbal products and provided the first-time report of DNA metabarcoding for such products.

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Data availability

This article and its supplementary information files include all data generated or analyzed during this study.

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Acknowledgements

The authors would like to thank Prof. Padamnabhi S. Nagar, The Maharaja Sayajirao University of Baroda, India, for helping us in plant collection and authentication, Mr. Dipesh Parikh, Technical assistance, GBRC for carrying out HPLC, Mr. Nitin Savaliya, Technical assistance from Thermo Fisher Scientific, for NGS instrument handling and run setup.

Funding

Gujarat State Biotechnology Mission (GSBTM), Gandhinagar, Gujarat, India, has financially supported the project under the Research Support Scheme, Grant ID GSBTM/JDRD/584/2018/204.

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Author notes

  1. Abhi P. Shah and Tasnim Travadi have contributed equally to this work.

Authors and Affiliations

  1. Gujarat Biotechnology Research Centre (GBRC), Department of Science and Technology, Government of Gujarat, Gandhinagar, India

    Abhi P. Shah, Tasnim Travadi, Sonal Sharma, Ramesh Pandit, Chaitanya Joshi & Madhvi Joshi

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Contributions

APS: Performed experiments, Data Analysis, Writing, and Editing manuscript; TT: Performed experiments, Data Analysis, Writing, and Validation of manuscript; RP: Designed primers for metabarcoding, established metabarcoding data analysis pipeline, Manuscript editing; SS: Designed primers for CA, performed experiments, Manuscript editing; CJ: Project administration, Methodology, Supervision, and Review & Editing; MJ: Principal Investigator, Conceptualization, Methodology, Supervision, and Review & Editing.

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Correspondence to Madhvi Joshi.

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The sequence of CA primers and the PCR conditions have been applied for an Indian patent (Application Number-202221035088).

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The sequence of CA primers and the PCR conditions have been applied for an Indian patent (Application Number-202221035088).

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Shah, A.P., Travadi, T., Sharma, S. et al. Comprehensive analysis using DNA metabarcoding, SCAR marker based PCR assay, and HPLC unveils the adulteration in Brahmi herbal products. Mol Biol Rep 50, 7605–7618 (2023). https://doi.org/10.1007/s11033-023-08653-5

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