PHYTOCHEMICAL ANALYSIS, HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY, AND EX VIVO ANTIOXIDANT POTENTIAL OF A HERBAL FRUIT EXTRACT
Objective: The objective of the present investigation was to determine the presence of various phytochemicals in an ethanolic herbal fruit extract (HFE) made up by a blend of fruits of Phyllanthus emblica (PE), Ananas comosus (AC), Punica granatum (PG), and Terminalia chebula (TC). Followed by a qualitative high-performance liquid chromatography (HPLC) analysis for standardization of the extract using ascorbic acid and gallic acid as standards, this study was also aimed at the determination of ex vivo antioxidant potential and calculation of inhibitory concentration (IC50) value of HFE.
Methods: The analysis of phytochemicals present in HFE was done qualitatively by methods as described by various researchers. Reversed-phase HPLC was carried out on a C-18 column on gradient mode, using methanol and acetic acid as mobile phase. Ex vivo antioxidant potential was evaluated by inhibition of lipid peroxidation (LPO) method by the preparation of packed cell volume.
Results: The results revealed the presence of multiple phytoconstituents in the HFE, and through the chromatographic analysis, the presence of gallic and ascorbic acid as marker compounds was achieved. IC50 values calculated after LPO inhibition method was found to be 21.87 μg/ml and 25.63 μg/ml for ascorbic acid (as standard) and ethanolic HFE, respectively.
Conclusion: The use of herbal antioxidants can provide protection to the human body by fighting against multiple diseases caused as a result of the accumulation of free radicals, generated in the normal metabolic processes going on inside the body. Nowadays, researchers are amused to see the long-term positive effects of using a combination of antioxidants, for example, a polyherbal formulation, rather than single entities.
2. Balunas MJ, Kinghorn AD. Drug discovery from medicinal plants. Life Sci 2005;78:431-41.
3. Kidd P. Th1/Th2 balance: The hypothesis, its limitations, and implications for health and disease. Altern Med Rev 2003;8:223-46.
4. Sato M, Ramarathnam N, Suzuki Y, Ohkubo T, Takeuchi M, Ochi H. Varietal differences in the phenolic content and superoxide radical scavenging potential of wines from different sources. J Agric Food Chem 1996;44:37-41.
5. Meyer AS, Yi OS, Pearson DA, Waterhouse AL, Frankel EN. Inhibition of human low density lipoprotein oxidation in relation to composition of phenolic antioxidants in grapes (Vitis vinifera). J Agric Food Chem 1997;45:1638-43.
6. Zhang YJ, Gan RY, Li S, Zhou Y, Li AN, Xu DP, et al. Antioxidant phytochemicals for the prevention and treatment of chronic diseases. Molecules 2015;20:21138-56.
7. Sharma G, Sharma V, Mishra T. Enhancement of in vitro antioxidant potential of Terminalia chebula by various fruit extracts and optimization of concentration by response surface methodology. Asian J Pharm Clin Res 2018;11:228-33.
8. Duraiswamy B, Singanan M, Varadarajan V. Physicochemical, phytochemicals and antioxidant evaluation of Guazuma ulmifolia fruit. Int J Pharm Pharm Sci 2018;10:87-91.
9. Chanda SV, Parekh J, Karathia N. Evaluation of antibacterial activity and phytochemical analysis of Bauhinia variegate. L bark. Afr J Biomed Res 2006;9:53-6.
10. Trease GE, Evans WC. Pharmacognosy: A Physicians Guide to Herbal Medicine. 13th ed. London, UK: Bailliere Tindall; 1989.
11. Kumar GS, Jayaveera KN, Kumar CK, Sanjay UP, Swamy BM, Kumar DV. Antimicrobial effects of Indian medicinal plants against acne-inducing bacteria. Trop J Pharm Res 2007;6:717-23.
12. Onwukaeme DN, Ikuegbvweha TB, Asonye CC. Evaluation of phytochemical constituents, antibacterial activities and effect of exudates of Pycanthus angolensis Weld Warb (Myristicaceae) on corneal ulcers in rabbits. Trop J Pharm Res 2007;6:725-30.
13. Edeoga HO, Okwu DE, Mbaebie BO. Phytochemical constituents of some Nigerian medicinal plants. Afr J Biotechnol 2005;4:685-8.
14. Sawant L, Prabhakar B, Pandita N. Quantitative HPLC analysis of ascorbic acid and gallic acid in Phyllanthus emblica. J Anal Bioanal Tech 2010;1:1-4.
15. Deep A, Rana P, Gosal SS, Soni G. Antioxidant potential of tissue cultured Mentha spicata. BioMedRx 2013;1:90-6.
16. Stocks J, Dormandy TL. The autoxidation of human red cell lipids induced by hydrogen peroxide. Br J Haematol 1971;20:95-111.
17. Menon A, Priya V, Gayathri R. Preliminary phytochemical analysis and cytotoxicity potential of pineapple extract on oral cancer cell lines. Asian J Pharm Clin Res 2016;9:140-3.
18. Babajide JM, Olaluwoye AA, Taofik ST, Adebisi MA. Physicochemical properties and phytochemical components of spiced cucumber-pineapple fruit drink. Niger Food J 2013;31:40-52.
19. Seal T. HPLC Determination of phenolic acids, flavonoids and ascorbic acid in four different solvent extracts of Zanthoxylum acanthopodium, a wild edible plant of Meghalaya state of India. Int J Pharm Pharm Sci 2016;8:103-9.
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