Abstract
Research on natural anti-oxidant agents increased due to their wide range of applications on various diseases. HPTLC-DPPH is the novel technique where anti-oxidant potential of complex herbal extract can be detected on TLC plate. In thin-layer chromatography, silica gel plates (60 F-254) were used as a stationary phase along with THF/Toluene/Acetic Acid/Water [16:8:2:1 (v/v)] as a mobile phase. Cytochrome P-450 inhibitors were predicted by in-silico method as anti-oxidant agent. HPTLC method development and validation were performed on CAMAG HPTLC system along with quantitative estimation of epigallocatechin in hydroalcoholic extract of Acacia suma. HPTLC method had ensured no change in RF value (0.944) at wavelength 269 nm and revealed presence of epigallocatechin in Acacia suma is 404.2 mg/g. Yellow coloured bands on TLC plate were observed with DPPH reagent to confirm radical scavenging property. The in vitro DPPH radical scavenging assay, hydrogen peroxide assay, nitric oxide assay, total anti-oxidant assay and lipid peroxidation assay were performed, and IC50 values are 81.46 ± 2.72, 61.39 ± 1.85, 21.30 ± 2.26, 55.13 ± 2.86 and 77.03 ± 2.47 µg/ml, respectively. Additionally, in-silico data predicted ‘Diinsininol’ as cytochrome P450 inhibitor with docking score -13.51 kcal/mol. The study findings evident that hydroalcoholic extract of Acacia suma had significant anti-oxidant potential which was confirmed by in vitro assays, molecular docking study and HPTLC-DPPH anti-oxidant approaches.
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Acknowledgements
Authors are thankful to Late Dr. Sanjay Mishra for his suggestions in refining this draft. Authors are also thankful to the director of Analytical laboratory at KAHER’s BSRC, Belagavi 590010, Karnataka, India for providing facilities to complete this study.
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NK, SM, SN contributed to conceptualization. NK contributed to writing. SM, SN, SA contributed to review. NK, SN contributed to data analysis. NK, SM contributed to additional works. All authors agreed position and publication.
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Kanbarkar, N., Mishra, S., Nandanwadkar, S. et al. Assessment of anti-oxidant activity and quantification of epigallocatechin in Acacia suma heartwood by HPTLC-DPPH fingerprinting method. Chem. Pap. 76, 5865–5878 (2022). https://doi.org/10.1007/s11696-022-02295-w
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DOI: https://doi.org/10.1007/s11696-022-02295-w