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bioRxiv · 10.1101/2022.01.25.477015

High performance liquid chromatography-based metabolomics of Psidium guajava Linn. leaf extracts

Gholkar, M. S.·Daswani, P. G.·Iyer, V. V.·Birdi, T. J.
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Abstract

BackgroundOver the years, a number of methods have been introduced in the field of herbal medicine. Amongst these, metabolomics has rapidly emerged as a method of choice due to its wide applicability and versatility. High performance liquid chromatography (HPLC) is a preferred method for fingerprinting with advantages such as easy and wide availability, and relatively low maintenance cost. The current study used HPLC profiling and attempted to correlate it to anti-diarrhoeal activity of Psidium guajava (guava) extracts. MethodsNinety samples of guava leaves were collected from three locations in Maharashtra, India over three seasons. Hydroalcoholic extracts (50:50, ethanol:water) were prepared and the HPLC chromatogram of all 90 extracts obtained. A total of eight bioassays representing the important features of diarrhoeal pathogenesis were performed and the extracts were differentiated as good or poor according to cut-offs for each assay. The numerical data of the bioassays and the HPLC chromatogram was correlated using suitable mathematical models comprising Principal component analysis (PCA), orthogonal partial least squares (OPLS, regression) and orthogonal partial least squares discriminant analysis (OPLS-DA). ResultsOPLS-DA showed good seasonal and regional segregation of extracts. For most of the bioassays, PCA was unsuccessful in showing significant discrimination. Hence, OPLS plots were further developed; differentiation of good and poor extracts among the 90 extracts was successfully demonstrated for antibacterial activity against Shigella flexneri and Vibrio cholerae, inhibition of invasion of enteropathogenic Escherichia coli into HEp-2 epithelial cells, and cholera toxin (CT) production by V. cholerae. However, for other assays, namely, inhibition of adherence of E. coli, invasion of S. flexneri into HEp-2 cells, and inhibition of E. coli labile toxin (LT), subsets of 90 extracts were selected to demonstrate a significant correlation. ConclusionHPLC-based metabolomics has the potential to differentiate good and poor activities of guava leaf extracts. This approach can be extended for identifying phytoconstituents responsible for the anti-diarrhoeal activity of guava leaf and help standardization of crude extracts.

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Gholkar, M. S., Daswani, P. G., Iyer, V. V., Birdi, T. J.. 2022-01-28. High performance liquid chromatography-based metabolomics of Psidium guajava Linn. leaf extracts. https://doi.org/10.1101/2022.01.25.477015

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