Qualitative and Quantitative Phytochemical Screening and Free Radical Scavenging Activity of Different Parts of Rubus ellipticus Sm.

Abstract

Free radicals are molecules with reactive unpaired electrons which are produced during cell metabolism and create the oxidative stress inside tissue resulting in tissue damage. The aim of the present study was qualitative and quantitative phytochemical screening, determination of total phenolic and total flavonoid content, and free radical scavenging activity of different parts of ethnomedicinally used Rubus ellipticus Sm. plant. Root, stem, and leaves of Rubus ellipticus Sm. were collected from Annapurna Rural Municipality, Kaski, Western Nepal. Ethyl acetate and ethanol extracts of the plant parts were obtained by subsequent maceration process. The phytochemical screening of most of the extract showed the presence of phenols, carbohydrates, flavonoids, and glycosides. The ethanolic extract of stem showed the higher phenolic content with the value of 343.75 ± 2.21 µg GAE/mg. Ethanolic extract of stem had the highest amount of flavonoid content (1563.17 ± 10.79 µg QE/mg of extract), whereas all the ethyl acetate extracts of root, leaves and stem showed comparable flavonoid content. Ethanolic extracts of leaves showed potent DPPH free radical scavenging activity with IC50 value of 5.03µg/ml while ethyl acetate extract of stem showed the maximum free radical scavenging properties. The result depicted that the ethanolic extract of Rubus ellipticus Sm. showed the potent antioxidant activity by scavenging free radicals.

Keywords

• Antioxidant
• Ethnomedicine
• Free Radical Scavenging
• Phytochemicals
• Rubus ellipticus Sm.

References

1. Akter, M. S., Ahmed, M., & Eun, J. B., 2010. Solvent effects on antioxidant properties of persimmon (Diospyros kaki L. cv. Daebong) seeds. International Journal of Food Science & Technology, 45(11), 2258-2264. https://doi.org/10.1111/j.1365-2621.2010.02400.x
2. Anup, B. (2020/04/16). Lead acetate test (Lead sulfide test): Principle, Reaction, Reagents, Procedure and Result Interpretation. biocheminfo online library. Retrieved 2022/10/17 from http://biocheminfo.com/2020/04/16/lead-acetate-test-lead-sulfide-test-principle-reaction-reagents-procedure-and-result-interpretation/
3. Atanassova, M., Georgieva, S., & Ivancheva, K.,2011. Total phenolic and total flavonoid contents, antioxidant capacity and biological contaminants in medicinal herbs. Journal of the University of Chemical Technology & Metallurgy, 46(1).
4. Badhani, A., Rawat, S., Bhatt, I. D., & Rawal, R. S.,2015. Variation in Chemical Constituents and Antioxidant Activity in Y ellow H imalayan (R ubus ellipticus S mith) and Hill Raspberry (R ubus niveus T hunb.). Journal of Food Biochemistry, 39(6), 663-672. https://doi.org/10.1111/jfbc.12172
5. Bagewadi, Z. K., Siddanagouda, R., & Baligar, P. G.,2014. Phytoconstituents investigation by LC-MS and evaluation of anti-microbial and anti-pyretic properties of cynodon dactylon. International Journal of Pharmaceutical sciences and research, 5(7), 2874. http://dx.doi.org/10.13040/IJPSR.0975-8232.5(7).2874-89
6. Baliyan, S., Mukherjee, R., Priyadarshini, A., Vibhuti, A., Gupta, A., et al. 2022. Determination of Antioxidants by DPPH Radical Scavenging Activity and Quantitative Phytochemical Analysis of Ficus religiosa. Molecules, 27(4), 1326. https://doi.org/10.3390/molecules27041326
7. Baral, R., Subedi, L., Gurung, M., Ojha, S., Shrestha, B., et al. 2021. Phytochemical screening, free radical scavenging activity, In-vitro Alpha-amylase enzyme and glucose diffusion inhibition activity of ethyl acetate and water extracts of selected medicinal plants of Nepal. International journal of Herbal medicine, 9(3), 18-21. https://doi.org/10.38093/cupmap.1122429
8. Barros, L., Ferreira, M.-J., Queiros, B., Ferreira, I. C., & Baptista, P.,2007. Total phenols, ascorbic acid, β-carotene and lycopene in Portuguese wild edible mushrooms and their antioxidant activities. Food chemistry, 103(2), 413-419. https://doi.org/10.1016/j.foodchem.2006.07.038

9. Bhakuni, R., Shukla, Y., & Thakur, R.,1987. Chemical examination of the roots of Rubus ellipticus. Indian Drugs, 24, 272.
10. Brown, G. C., & Borutaite, V.,2012. There is no evidence that mitochondria are the main source of reactive oxygen species in mammalian cells. Mitochondrion, 12(1), 1-4. https://doi.org/10.1016/j.mito.2011.02.001
11. Castleman, M.,1991. The Healing Herb: The Ultimate Guide to the Curative Power of Nature’s Medicines. Pensylvania: G. and C. Merriam, 128-135.
12. Cesquini, M., Torsoni, M., Stoppa, G. t.-., & Ogo, S. t.,.2003. t-BOOH-induced oxidative damage in sickle red blood cells and the role of flavonoids. Biomedicine & pharmacotherapy, 57(3-4), 124-129. https://doi.org/10.1016/S0753-3322(03)00018-0
13. Chang, C.-C., Yang, M.-H., Wen, H.-M., & Chern, J.-C.,2002. Estimation of total flavonoid content in propolis by two complementary colorimetric methods. Journal of food and drug analysis, 10(3). https://doi.org/10.38212/2224-6614.2748
14. Choudhary, G.,2015. Free radical scavenging properties of the ethanol extract of Cynodon dactylon. Asian Journal of Plant Science and Research, 5(6), 88-90.
15. Chun, O. K., Kim, D.-O., & Lee, C. Y.,2003. Superoxide radical scavenging activity of the major polyphenols in fresh plums. Journal of agricultural and food chemistry, 51(27), 8067-8072. https://doi.org/10.1021/jf034740d
16. Deweirdt, J., Quignard, J., Crobeddu, B., Baeza-Squiban, A., Sciare, J., et al. 2017. Involvement of oxidative stress and calcium signaling in airborne particulate matter-induced damages in human pulmonary artery endothelial cells. Toxicology in Vitro, 45, 340-350. https://doi.org/10.1016/j.tiv.2017.07.001
17. Duh, P.-D., Tu, Y.-Y., & Yen, G.-C.,1999. Antioxidant activity of water extract of Harng Jyur (Chrysanthemum morifolium Ramat). LWT-Food Science and Technology, 32(5), 269-277. https://doi.org/10.1006/fstl.1999.0548
18. Erdemoglu, N., Turan, N. N., Cakõcõ, I., Sener, B., & Aydõn, A.,2006. Antioxidant activities of some Lamiaceae plant extracts. Phytotherapy Research: An International Journal Devoted to Pharmacological and Toxicological Evaluation of Natural Product Derivatives, 20(1), 9-13. https://doi.org/10.1002/ptr.1816
19. Fattahi, S., Zabihi, E., Abedian, Z., Pourbagher, R., Ardekani, A. M., et al.,2014. Total phenolic and flavonoid contents of aqueous extract of stinging nettle and in vitro antiproliferative effect on hela and BT-474 Cell lines. International journal of molecular and cellular medicine, 3(2), 102.
20. Gutteridge, J. M.,1994. Biological origin of free radicals, and mechanisms of antioxidant protection. Chemico-biological interactions, 91(2-3), 133-140. https://doi.org/10.1016/0009-2797(94)90033-7
21. Halliwell, B.,1995. How to characterize an antioxidant: an update. Biochemical Society Symposium, https://doi.org/10.1042/bss0610073
22. Halliwell, B., & Gutteridge, J. M., 2015. Free radicals in biology and medicine. Oxford university press, USA.
23. Indradi, R. B., Fidrianny, I., & Wirasutisna, K. R.,2017. DPPH scavenging activities and phytochemical content of four Asteraceae plants. International Journal of Pharmacognosy and Phytochemical Research, 9(6), 755-759. https://doi.org/10.25258/phyto.v9i6.8173
24. Jabbari, M., & Jabbari, A.,2016. Antioxidant potential and DPPH radical scavenging kinetics of water-insoluble flavonoid naringenin in aqueous solution of micelles. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 489, 392-399. https://doi.org/10.1016/j.colsurfa.2015.11.022
25. Jigna, P., Rathish, N., & Sumitra, C.,2005. Preliminary screening of some folklore medicinal plants from western India for potential antimicrobial activity. Indian journal of pharmacology, 37(6), 408-409. Karuppusamy, S., Muthuraja, G., & Rajasekaran, K.,2011. Antioxidant activity of selected lesser known edible fruits from Western Ghats of India.
26. Kaur, C., & Kapoor, H. C.,2002. Anti‐oxidant activity and total phenolic content of some Asian vegetables. International Journal of Food Science & Technology, 37(2), 153-161. https://doi.org/10.1046/j.1365-2621.2002.00552.x Kirtikar, K., Basu, B., & CS, I.,2001. Indian medicinal plants, oriental enterprises. Dehradun, 6, 2029-2035.
27. Kunwar, R. M., & Bussmann, R. W.,2008. Ethnobotany in the nepal himalaya. Journal of ethnobiology and ethnomedicine, 4(1), 1-8. https://doi.org/10.1186/1746-4269-4-24
28. Majidaee, E., Hosseyni Talei, S. R., Gholamnezhad, S., & Ebrahimzadeh, M. A.,2020. Comparing the Effect of Different Extraction Methods and the Role of Solvent Polarity on Total Phenolic and Flavonoid Contents and Antioxidant Activities of Ferula persica. Journal of Mazandaran University of Medical Sciences, 30(188), 26-39.
29. Maxwell, S. R.,1995. Prospects for the use of antioxidant therapies. Drugs, 49(3), 345-361. https://doi.org/10.2165/00003495-199549030-00003
30. Mukherjee, A. K., Basu, S., Sarkar, N., & Ghosh, A. C.,2001. Advances in cancer therapy with plant based natural products. Current medicinal chemistry, 8(12), 1467-1486. https://doi.org/10.2174/0929867013372094
31. Okoduwa, S. I. R., Umar, I. A., James, D. B., Inuwa, H. M., & Habila, J. D.,2016. Evaluation of extraction protocols for anti-diabetic phytochemical substances from medicinal plants. World journal of diabetes, 7(20), 605. https://doi.org/10.4239%2Fwjd.v7.i20.605
32. Öztürk, M., Aydoğmuş-Öztürk, F., Duru, M. E., & Topçu, G.,2007. Antioxidant activity of stem and root extracts of Rhubarb (Rheum ribes): An edible medicinal plant. Food chemistry, 103(2), 623-630. https://doi.org/10.1016/j.foodchem.2006.09.005
33. Pandey, A., & Tripathi, S.,2014. Concept of standardization, extraction and pre phytochemical screening strategies for herbal drug. Journal of Pharmacognosy and Phytochemistry, 2(5).
34. Patel, A., Rojas-Vera, J., & Dacke, C.,2004. Therapeutic constituents and actions of Rubus species. Current medicinal chemistry, 11(11), 1501-1512. https://doi.org/10.2174/0929867043365143
35. Pfoze, N. L., Kumar, Y., & Myrboh, B.,2012. Survey and assessment of ethnomedicinal plants used in Senapati District of Manipur State, Northeast India. Phytopharmacology, 2(2), 285-311.
36. Polidori, M. C., & Mecocci, P.,2022. Modeling the dynamics of energy imbalance: The free radical theory of aging and frailty revisited. Free Radical Biology and Medicine. https://doi.org/10.1016/j.freeradbiomed.2022.02.009
37. Raj, S., Berthomier, M., Babu, M. A., Karthikeyan, S., Sivakumar, A., et al. 2013. Antioxidant, Antibacterial and Anti-Proliferative Activity and Phytochemical Analysis of Selected Medicinal Plants from Dasapushpam of Kerala. International Journal of Pharmaceutical Sciences Review and Research,23(1), 172-179. https://doi.org/10.1016/j.genrep.2022.101651
38. Raman, G., Cho, M., Brodbelt, J. S., & Patil, B. S.,2005. Isolation and purification of closely related Citrus limonoid glucosides by flash chromatography. Phytochemical Analysis: An International Journal of Plant Chemical and Biochemical Techniques, 16(3), 155-160. https://doi.org/10.1002/pca.835
39. Rojas‐Vera, J., Patel, A. V., & Dacke, C. G.,2002. Relaxant activity of raspberry (Rubus idaeus) leaf extract in guinea‐pig ileum in vitro. Phytotherapy Research: An International Journal Devoted to Pharmacological and Toxicological Evaluation of Natural Product Derivatives, 16(7), 665-668. https://doi.org/10.1002/ptr.1040
40. Saini, R., Dangwal, K., Singh, H., & Garg, V.,2014. Antioxidant and antiproliferative activities of phenolics isolated from fruits of Himalayan yellow raspberry (Rubus ellipticus). Journal of food science and technology, 51(11), 3369-3375. https://doi.org/10.1007/s13197-012-0836-3
41. Saxena, M., Saxena, J., Nema, R., Singh, D., & Gupta, A.,2013. Phytochemistry of medicinal plants. Journal of Pharmacognosy and Phytochemistry, 1(6).
42. Schulz, M., Seraglio, S. K. T., Della Betta, F., Nehring, P., Valese, A. C., et al. 2019. Blackberry (Rubus ulmifolius Schott): Chemical composition, phenolic compounds and antioxidant capacity in two edible stages. Food research international, 122, 627-634. https://doi.org/10.1016/j.foodres.2019.01.034
43. Sharma, U. S., & Kumar, A.,2011. In vitro antioxidant activity of Rubus ellipticus fruits. Journal of advanced pharmaceutical technology & research, 2(1), 47. https://doi.org/10.4103%2F2231-4040.79805
44. Shikha, D., & Kashyap, P.,2020. Yellow Himalayan Berry. In Antioxidants in Fruits: Properties and Health Benefits (pp. 67-81). Springer. https://doi.org/10.1007/978-981-15-7285-2_4
45. Sies, H., & Cadenas, E.,1985. Oxidative stress: damage to intact cells and organs. Philosophical Transactions of the Royal Society of London. B, Biological Sciences, 311(1152), 617-631. https://doi.org/10.1098/rstb.1985.0168
46. Srivastava, J., Lambert, J., & Vietmeyer, N.,1996. Medicinal plants: An expanding role in development (Vol. 320). World Bank Publications.
47. Subba, B., Gaire, S., & Sharma, K. R.,2019. Analysis of phyto-constituents, antioxidant, and alpha amylase inhibitory activities of persea americana Mill., Rhododendron arboretum Sm. Rubus ellipticus Sm. from Arghakhanchi district Nepal. Asian Journal of Pharmaceutical and Clinical Research, 12(1), 301. http://dx.doi.org/10.22159/ajpcr.2019.v12i1.29679
48. Surabhi, S., & Leelavathi, S.,2010. Anti-oxidant property of ethanolic extract of Catunaregam spinosa Thunb. Int J Drug Dev Res, 2, 399-403.
49. Uniyal, S. K., Singh, K., Jamwal, P., & Lal, B.,2006. Traditional use of medicinal plants among the tribal communities of Chhota Bhangal, Western Himalaya. Journal of ethnobiology and ethnomedicine, 2(1), 1-8. https://doi.org/10.1186/1746-4269-2-14
50. Vadivelan, R., Bhadra, S., Ravi, A., Shanish, K. S. A., Elango, K.,et al. 2009. Evaluation of anti-inflammatory and membrane stabilizing property of ethanol root extract of Rubus ellipticus Smith in Albino rats. Journal of Natural Remedies, 9(1), 74-78.
51. Valko, M., Leibfritz, D., Moncol, J., Cronin, M. T., Mazur, M., et al.2007. Free radicals and antioxidants in normal physiological functions and human disease. The international journal of biochemistry & cell biology, 39(1), 44-84. https://doi.org/10.1016/j.biocel.2006.07.001
52. Yadav, N., Tyagi, G., Jangir, D. K., & Mehrotra, R.,2011. Rapid determination of polyphenol, vitamins, organic acids and sugars in Aegle marmelos using reverse phase-high performance liquid chromatography. J Pharm Res, 4(3), 717-719.
53. Yadav, R., & Agarwala, M.,2011. Phytochemical analysis of some medicinal plants. Journal of phytology, 3(12).