EXTRACTION AND ISOLATION OF FLAVONOID QUERCETIN FROM THE LEAVES OF TRIGONELLA FOENUM-GRAECUM AND THEIR ANTI-OXIDANT ACTIVITY
Objective: The present study was designed for isolation of bioactive flavonoid molecule quercetin from the leaves of Trigonella foenum-graecum and their subsequent characterization.
Methods: Crude extracts of fenugreek were prepared using various solvents such as hexane, ethyl acetate, and ethanol. The plant extracts were subjected for photochemical analysis and total flavonoid content. The extracts were then subjected to column chromatography followed by TLC. The isolated compound was subjected to FT-IR, 1H NMR, 13C NMR, mass spectroscopy and their free radical scavenging activity was studied.
Results: The ethanol extract showed the presence of higher flavonoid content when compared with other solvent extracts. The ethanol extract was subjected to fractionalization by column chromatography. The eluted fractions were run in TLC mobile phase with the different solvent ratio. The fractions showed Rf value equal to standard quercetin in TLC were combined and crystallized. The characterization techniques confirmed that the isolated compound was found to be quercetin. The free radical scavenging activity suggests that the isolated compound quercetin could act as a potent source of antioxidants.
Conclusion: The flavonoid quercetin was isolated effectively from the leaves of Trigonella foenum-graecum and their antioxidant activity was studied.
Keywords: Antioxidant activity, DPPH, Flavonoids, NMR, Quercetin
2. Sharma R, Raghuram T, Rao N. Effects of fenugreek seeds on blood glucose and serum lipid in type I diabetes. Eur J Clin Nutr 1990;44:301-6.
3. Cowan MM. Plant product as antimicrobial agents. Clin Microbiol Rev 1999;12:564-82.
4. Jeevan KP, Chao CW, Stephen B. Mass spectrometric methods for the determination of flavonoids in biological samples. Free Radical Biol Med 2004;37:1324-50.
5. Su FC, Shin N, Chien HW, Chia LL, Yun CC, Shiang YL. Reappraisal of the anticancer efficacy of quercetin in oral cancer cells. J Chin Med Assoc 2013;76:146-52.
6. Shetty K. Biotechnology to harness the benefits of dietary phenolics: focus on Lamiaceae. Asia Pac J Clin Nutr 1997;21:79-102.
7. Sharma RD, Sarkar A, Hazra DK. Hypolipidaemic effect of fenugreek seeds: a chronic study in non-insulin dependent diabetic patients. Phytother Res 1996;10:332-4.
8. Ajabnoor M, Tilmisany A. Effect of Trigonella foenum-graecum on blood glucose levels in normal and alloxan-diabetic mice. J Ethnopharmacol 1998;22:45-9.
9. Pandian S, Anuradha C, Viswanathan P. Gastroprotective effect of fenugreek seeds (Trigonella foenum-graecum) on an experimental gastric ulcer in rats. J Ethnopharmacol 2002;81:393-7.
10. Chauhan G, Sharma M, Kharkwal H, Varma A. Pharmacognostic, preliminary phytochemical studies and anticancerous potential of Trigonella foenum-graecum. Pharm Sci Manitor 2010;2:350-9.
11. Han Y, Nishibe S, Noguchi Y, Jin Z. Flavonol glycosides from the stems of Trigonella foenum-graecum. Phytochemistry 2001;58:577-80.
12. Harborne JB. Phytochemical methods: a guide to modern techniques of plant analysis. 2nd ed. London: Chapman and Hall Publishers; 1973. p. 49-188.
13. Edeoga HO, Okwu DE, Mbaebie BO. Phytochemical constituents of some Nigerian medicinal plants. Afr J Biotechnol 2005;4:685-8.
14. Sakanaka S, Tachibana Y, Okada Y. Preparation and antioxidant properties of extracts of Japanese persimmon leaf tea (kakinoha-cha). Food Chem 2005;84:569-75.
15. Kumar N, Bhandari P, Singh B, Bari SS. Antioxidant activity and ultra-performance LC-electrospray ionization quadrupole time-of-flight mass spectrometry for phenolics-based fingerprinting of Rose species: Rosa damascena, Rosa bourboniana and Rosa brunonii. Food Chem Toxicol 2009;47:361-7.
16. Chourasiya A, Upadhayay A, Shukla RN. Isolation of quercetin from the leaves of Azadirachta indica and antidiabetic study of the crude extracts. J Pharm Biomed Sci 2012;25:179-81.
17. Selvaraj K, Chowdhury R, Bhattacharjee C. Isolation and structural elucidation of flavonoids from aquatic fern Azolla Microphylla and evaluation of free radical scavenging activity. Int J Pharm Pharm Sci 2013;5:743-9.
18. Benzie IFF, Strain JJ. The ferric reducing ability of plasma (FRAP) as a measure of â€œantioxidant powerâ€: the FRAP assay. Anal Biochem 1996;239:70-6.
19. Xu W, Chu K, Li H, Zhang Y, Zheng H, Chen R, et al. Ionic liquid-based microwave-assisted extraction of flavonoids from Bauhinia championii (Benth.). Benth Mol 2012;17:14323-35.
20. Sambandam B, Sathesh Kumar S, Ayyaswamy A, Nagarjuna Yadav BV, Thiyagarajan D. Synthesis and characterization of poly D-L Lactide (PLA) nanoparticles for the delivery of quercetin. Int J Pharm Pharm Sci 2015;7:1-8.
21. Bukhari SB, Bhanger MI, Memon S. Antioxidative activity of extracts from fenugreek seeds (Trigonella foenum-graecum). Pak J Anal Environ Chem 2008;9:78-83.