ISOLATION OF ACTIVE COMPOUND FROM RED POMEGRANATE (PUNICA GRANATUM L.) SEEDS ETHANOLIC EXTRACTS
Keywords:Active compound, Ethanolic extracts, Fraction, Isolation, Red pomegranate seeds
Objective: Pomegranate (Punica granatum L.) is rich in antioxidants and antibacterials, lending itself to beneficial effects on health improvement. The seeds contain a variety of active compounds. The purpose of this research was to isolate the active compound of red pomegranate seeds through ethanolic extraction.
Methods: Pomegranate seeds were extracted into 24 hexane fractions of the concentrated crude extract and left in open vials until all the solvent evaporated, leaving behind crystals. Crystals in vials 11, 12, and 13 were purified using n-hexane, ethyl acetate, methanol, chloroform, acetone and isopropyl alcohol as the solvents. The crystals were monitored with Thin Layer Chromatography (TLC) on a Kiesel gel 254 GF plate (e. Merck) using the developing system of hexane: ethyl acetate: formic acid (1:1:0.1) mixture and a UV light spot viewer with 366 wavelength emissions. Purification test with TLC two directions using hexane: ethyl acetate: formic acid (1:1:0.1) mixture as the eluents for the first development, continued with the second development using chloroform: methanol: acetate acid (9:0.5:0.5) mixture. The Retention Factor (Rf) value for the spot was measured.
Results: Chromatogram showed only 1 spot with Rf value 0.88.
Conclusion: There is one pure (active) isolate in red pomegranate seeds ethanolic extract.
2. Rahmani AH, Alsahli MA, Almatroodi SA, Rahmani AH, Alsahli MA. Active constituents of pomegranates (Punica granatum) as potential candidates in the management of health through modulation of biological activities. Pharmacogn J 2017;9:689-95.
3. Bhandari PR. Pomegranate (punica granatum L). Ancient seeds for modern cure? Review of potential therapeutic applications. Int J Nutr Pharmacol Neurol Dis 2012;2:171-84.
4. Venkata C, Prakash S, Prakash I. Bioactive chemical constituents from pomegranate (punica granatum) juice, seed and peel-a review. Int J Res Chem Environ Int J Res Chem Environ 2011;1:1-181.
5. Arun N, Road R, Pradesh U. Punica granatum: a review on pharmacological and therapeutic properties. IJSPR 2012;3:1240-5.
6. Setiadhi R, Sufiawati I, Zakiawati D, Nuraeny N, Hidayat W, Firman DR. Inhibition growth of pomegranate seeds extract against streptococcus sanguis: the cause of recurrent aphthous stomatitis. J Dentomaxillofac Sci 2017;2:7-11.
7. Harborne J. Phytochemical Methods. 3rd ed. Chapman and Hall; 1998.
8. Chromatography T. Thin layer chromatography. J Chromatogr A 1987;403:395.
9. Kannan R. Basics of chromatographic techniques course. J Neurochem 2016;1.
10. Levinson R. Chromatography. Mod Chem Tech; 2001. p. 116-59.
11. Chromatography TL. Thin Layer Chromatography; 2012. p. 1–11.
12. Lade BD, Patil AS, Paikrao HM, Kale AS, Hire KK. A comprehensive working, principles and applications of thin layer chromatography. Res J Pharm Biol Chem Sci 2014;5:486-503.
13. Shiga GPEA. Antibacterial activity of punica granatum peel extracts against shiga toxin-producing E Coli. IJLBPR 2012;1:164-72.
14. Feist P. Calculating retention factors (R. Chemistry (Easton); 2010. p. 3117–8.
15. Pinalia A. Penentuan metode rekristalisasi yang tepat untuk meningkatkan kemurnian kristal amonium perklorat (Ap). Maj Sains Dan Teknol Dirgant 2012;6:64-70.
16. Riani S, Irna S, Dewi Z, Nanan N, Wahyu H, Dani RF. Inhibition growth of pomegranate seeds extracts against streptococcus sanguis: the cause of recurrent aphthous stomatitis. J Dentomaxillofac Sci 2017;2:7-12.