Phenolic and Flavonoid Content of Black Mulberry (Morus nigra L.) Stem and Their Evaluation Antioxidant and Cytotoxic Profile
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Submitted
19 July 2022
Published
30 November 2022
Keywords:
-
Cytotoxic, Flavonoid, Phenolic, Morus nigra L, Antioxidant
Abstract
The black mulberry (Morus nigra) plant has been widely developed for its bioactivity as natural medicine. This study evaluated plant extracts' total phenolic and flavonoid content and their correlation to M. nigra stem's antioxidant activity and toxicity. Dry powder from the M. nigra stem was extracted by maceration with 96% ethanol to obtain a thick extract (TE) and fractionated using hexane (HF), ethyl acetate (EAF), and ethanol-aqueous (EF). The samples were analyzed for a compound using reagents and the determination of total phenolic and flavonoid content. The samples were evaluated for antioxidant activity using several parameters and their cytotoxic effects using the BSLT method. Identification of compounds in EAF was confirmed to contain phenolic, flavonoids, alkaloids, saponins, tannins, steroids, and terpenoids. EAF showed higher phenolic and flavonoid content than others. The evaluation of antioxidant activity showed that extracts and fractions from M. nigra stems showed the ability to reduce ions and free radicals. EF sample has activity in reducing Mo (IV) ion by TAC method of 98.82±0.53 µM/mg, indicating substantial antioxidant capacity. In addition, EAF samples showed potential activity in reducing DPPH, hydroxyl, and peroxide radicals in the β-carotene bleaching method with IC50 values of 12.13, 42.06, and 57.6 µg/mL, respectively. Similar activity was also seen in the cytotoxic effect of a robust EAF sample with an LC50 value of 16.31 µg/mL. The results show that EAF can be developed as a raw material for traditional medicine as an antioxidant and anticancer candidate with a significant flavonoid and phenolics content.
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References
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26. Hidayati N, Kurniawan T, Kusumawardani NI, Sari RP. Comparison of Maceration and Ultrasonication Methods on Indigofera tinctoria Linn Leaf Extraction. J Bahan Alam Terbarukan. 2018;7(1):54–8. doi:10.15294/jbat.v7i1.11405
27. Chaves JO, de Souza MC, da Silva LC, Lachos-Perez D, Torres-Mayanga PC, Machado APdF, et al. Extraction of Flavonoids from Natural Sources Using Modern Techniques. Front Chem. 2020;8:507887. doi:10.3389/fchem.2020.507887
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31. Pham PP, Morales NP, Pitaksuteepong T, Hemstapat W. Antioxidant activity of mulberry stem extract: A potential used as supplement for oxidative stress-related diseases. Songklanakarin J Sci Technol. 2017;39(3):407–14.
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33. Apak R, Guclu K, Ozyurek M, Celik SE. Mechanism of antioxidant capacity assays and the CUPRAC (cupric ion reducing antioxidant capacity) assay. Microchim Acta. 2008;160:413–9. doi:10.1007/s00604-007-0777-0
34. Khan MA, Rahman AA, Islam S, Khandokhar P, Parvin S, Islam MB, et al. A comparative study on the antioxidant activity of methanolic extracts from different parts of Morus alba L. (Moraceae). BMC Res Notes. 2013;6:24. doi:10.1186/1756-0500-6-24
35. Nur S, Wierson Y, Yulia, Sami FJ, Megawati M, Andi NA, et al. Characterization, Antioxidant and α-Glucosidase Inhibitory Activity of Collagen Hydrolysate from Lamuru (Caranx ignobilis) Fishbone. Sains Malays. 2021;50(8):2329–41. doi:10.17576/jsm-2021-5008-16
36. Treml J, Šmejkal K. Flavonoids as Potent Scavengers of Hydroxyl Radicals. Compr Rev Food Sci Food Saf. 201;15(4):720–38. doi:10.1111/1541-4337.12204
37. Bogacz-Radomska L, Harasym J. β-Carotene—properties and production methods. Food Qual Saf. 2018;2(2):69–74. doi:10.1093/fqsafe/fyy004
38. Nerdy N, Lestari P, Sinaga JP, Ginting S, Zebua NF, Mierza V, et al. Brine Shrimp (Artemia salina Leach.) Lethality Test of Ethanolic Extract from Green Betel (Piper betle Linn.) and Red Betel (Piper crocatum Ruiz and Pav.) through the Soxhletation Method for Cytotoxicity Test. Open Access Maced J Med Sci. 2021;9(A):407–12. doi:10.3889/oamjms.2021.6171
2. Erarslan ZB, Karagöz S, Kültür Ş. Comparative Morphological and Anatomical Studies on Morus Species (Moraceae) in Turkey. Turk J Pharm Sci. 2021;18(2):157-66. doi:10.4274/tjps.galenos.2020.02779
3. Lim S, Choi CI. Pharmacological Properties of Morus nigra L. (Black Mulberry) as A Promising Nutraceutical Resource. Nutrients. 2019;11(2):437. doi:10.3390/nu11020437
4. Lúcio KdP, Rabelo ACS, Araújo CM, Brandão GC, de Souza GHB, da Silva RG, et al. Anti-Inflammatory and Antioxidant Properties of Black Mulberry (Morus nigra L.) in a Model of LPS-Induced Sepsis. Oxid Med Cell Longev. 2018;2018:5048031. doi:10.1155/2018/5048031
5. Budiman A, Aulifa DL. Antibacterial Activity and Mode of Action of Black Mulberry (Morus nigra) Fruits Extract Against Streptococcus mutans. Pharmacogn J. 2020;12(6s):1722–6. doi:10.5530/pj.2020.12.233
6. Souza GR, Oliveira-Junior RG, Diniz TC, Branco A, Lima-Saraiva SRG, Guimarães AL, et al. Assessment of the antibacterial, cytotoxic and antioxidant activities of Morus nigra L. (Moraceae). Braz J Biol. 2018;78(2):248–54. doi:10.1590/1519-6984.05316
7. Kostiæ D, Dimitrijeviæ D, Mitiæ S, Mitiæ M, Stojanoviæ G, Živanoviæ A. Phenolic Content and Antioxidant Activities of Fruit Extracts of Morus nigra L (Moraceae) from Southeast Serbia. Trop J Pharm Res. 2013;12(1):105–10. doi:10.4314/tjpr.v12i1.17
8. Aulifa DL, Fitriansyah SN, Ardiansyah SA, Wibowo DP, Julata YA, Christy DS. Phytochemical Screening, Antibacterial Activity, and Mode of Action on Morus nigra. Pharmacogn J. 2018;10(1):167–71. doi:10.5530/pj.2018.1.28
9. Sánchez-Salcedo EM, Mena P, García-Viguera C, Martínez JJ, Hernández F. Phytochemical evaluation of white (Morus alba L.) and black (Morus nigra L.) mulberry fruits, a starting point for the assessment of their beneficial properties. J Funct Foods. 2015;12:399–408. doi:10.1016/j.jff.2014.12.010
10. Tungmunnithum D, Thongboonyou A, Pholboon A, Yangsabai A. Flavonoids and Other Phenolic Compounds from Medicinal Plants for Pharmaceutical and Medical Aspects: An Overview. Medicines. 2018;5(3):93. doi:10.3390/medicines5030093
11. Galano A, Castañeda-Arriaga R, Pérez-González A, Tan DX, Reiter RJ. Phenolic Melatonin-Related Compounds: Their Role as Chemical Protectors against Oxidative Stress. Molecules. 2016;21(11):1442. doi:10.3390/molecules21111442
12. Mathew S, Abraham TE, Zakaria ZA. Reactivity of phenolic compounds towards free radicals under in vitro conditions. J Food Sci Technol. 2015;52(9):5790-8. doi:10.1007/s13197-014-1704-0
13. Kumar S, Pandey AK. Chemistry and Biological Activities of Flavonoids: An Overview. ScientificWorldJournal. 2013;2013:162750. doi:10.1155/2013/162750
14. Kopustinskiene DM, Jakstas V, Savickas A, Bernatoniene J. Flavonoids as Anticancer Agents. Nutrients. 2020;12(2):457. doi:10.3390/nu12020457
15. Nur S, Mus S, Marwati, Fadri A, Nursamsiar, Sami FJ. Determination of Total Phenolic and Flavonoid Levels of Mangkokan Leaf Extract (Polyscias scutellaria). J Pharm Med Sci. 2020;5(1):24–7. doi:10.32814/jpms.v5i1.116
16. Harborne JB. Phytochemical Methods: A Guide to Modern Techniques of Plant Analysis. Berlin, Germany: Springer Dordrecht; 1984. doi:10.1007/978-94-009-5570-7
17. Nur S, Aswad M, Yulianti R, Burhan A, Patabang WJD, Fadri A, et al. Profil Aktivitas Antioksidan dari Ekstrak Buah Kersen (Muntingia calabura L.) dengan Metode TAC dan CUPRAC. JPSCR J Pharm Sci Clin Res. 2022;7(1):79–88. doi:10.20961/jpscr.v7i1.56653
18. Rahman MDM, Islam MdB, Biswas M, Alam AHMK. In vitro antioxidant and free radical scavenging activity of different parts of Tabebuia pallida growing in Bangladesh. BMC Res Notes. 2015;8:621. doi:10.1186/s13104-015-1618-6
19. Sami FJ, Nur S, Ramli N, Sutrisno B. Uji Aktivitas Antioksidan Daun Kersen (Muntingia calabura L.) Dengan Metode DPPH (1,1-difenil-2-pikrilhidrazil) dan FRAP (Ferric Reducing Antioxidant Power). As-Syifaa J Farmasi. 2017;9(2):106–11. doi:10.56711/jifa.v9i2.258
20. Kutlu T, Kasim T, Çeken B, Murat K. DNA damage protecting activity and in vitro antioxidant potential of the methanol extract of Cherry (Prunus avium L). J Med Plants Res. 2014;8(19):715–26. doi:10.5897/JMPR2013.5350
21. Nur S, Aswad M, Yulianti R, Sami FJ, Burhan A, Fadri A, et al. Antioxidant activity profile of extract and fraction of kersen (Muntingia calabura L.) fruits prepared by different methods. IOP Conf Ser Earth Environ Sci. 2022;976(1):012066. doi:10.1088/1755-1315/976/1/012066
22. Olugbami JO, Gbadegesin MA, Odunola OA. In vitro evaluation of the antioxidant potential, phenolic and flavonoid contents of the stem bark ethanol extract of Anogeissus leiocarpus. Afr J Med Med Sci. 2014;43(Suppl 1):101–9.
23. Nur S, Mubarak F, Jannah C, Winarni DA, Rahman DA, Hamdayani LA, et al. Total phenolic and flavonoid compounds, antioxidant and toxicity profile of extract and fractions of paku atai tuber (Angiopteris ferox Copel). Food Res. 2019;3(6):734–40. doi:10.26656/fr.2017.3(6).135
24. Nguyen VT, Nguyen MT, Tran QT, Thinh PV, Bui LM, Le THN, et al. Effect of extraction solvent on total polyphenol content, total flavonoid content, and antioxidant activity of soursop seeds (Annona muricata L.). IOP Conf Ser Mater Sci Eng. 2020;736:022063. doi:10.1088/1757-899X/736/2/022063
25. Sultana B, Anwar F, Ashraf M. Effect of Extraction Solvent/Technique on the Antioxidant Activity of Selected Medicinal Plant Extracts. Molecules. 2009;14(6):2167–80. doi:10.3390/molecules14062167
26. Hidayati N, Kurniawan T, Kusumawardani NI, Sari RP. Comparison of Maceration and Ultrasonication Methods on Indigofera tinctoria Linn Leaf Extraction. J Bahan Alam Terbarukan. 2018;7(1):54–8. doi:10.15294/jbat.v7i1.11405
27. Chaves JO, de Souza MC, da Silva LC, Lachos-Perez D, Torres-Mayanga PC, Machado APdF, et al. Extraction of Flavonoids from Natural Sources Using Modern Techniques. Front Chem. 2020;8:507887. doi:10.3389/fchem.2020.507887
28. Agbor GA, Vinson JA, Donnelly PE. Folin-Ciocalteau Reagent for Polyphenolic Assay. Int J Food Sci Nutr Diet. 2014;3(8):147–56. doi:10.19070/2326-3350-1400028
29. Blainski A, Lopes G, de Mello J. Application and Analysis of the Folin Ciocalteu Method for the Determination of the Total Phenolic Content from Limonium Brasiliense L. Molecules. 2013;18(6):6852–65. doi:10.3390/molecules18066852
30. Januarti IB, Taufiq H, Sulistyaningsih S. The Correlation of Total Flavonoid and Total Phenolic with Antioxidant Activity of Single Bulb Garlic (Allium sativum) from Tawangmangu and Magetan. J Farmasi Sains Komunitas J Pharm Sci Commun. 2020;16(2):96–103. doi:10.24071/jpsc.001798
31. Pham PP, Morales NP, Pitaksuteepong T, Hemstapat W. Antioxidant activity of mulberry stem extract: A potential used as supplement for oxidative stress-related diseases. Songklanakarin J Sci Technol. 2017;39(3):407–14.
32. Apak R, Güçlü K, Demirata B, Özyürek M, Çelik S, Bektaşoğlu B, et al. Comparative Evaluation of Various Total Antioxidant Capacity Assays Applied to Phenolic Compounds with the CUPRAC Assay. Molecules. 2007;12(7):1496–547. doi:10.3390/12071496
33. Apak R, Guclu K, Ozyurek M, Celik SE. Mechanism of antioxidant capacity assays and the CUPRAC (cupric ion reducing antioxidant capacity) assay. Microchim Acta. 2008;160:413–9. doi:10.1007/s00604-007-0777-0
34. Khan MA, Rahman AA, Islam S, Khandokhar P, Parvin S, Islam MB, et al. A comparative study on the antioxidant activity of methanolic extracts from different parts of Morus alba L. (Moraceae). BMC Res Notes. 2013;6:24. doi:10.1186/1756-0500-6-24
35. Nur S, Wierson Y, Yulia, Sami FJ, Megawati M, Andi NA, et al. Characterization, Antioxidant and α-Glucosidase Inhibitory Activity of Collagen Hydrolysate from Lamuru (Caranx ignobilis) Fishbone. Sains Malays. 2021;50(8):2329–41. doi:10.17576/jsm-2021-5008-16
36. Treml J, Šmejkal K. Flavonoids as Potent Scavengers of Hydroxyl Radicals. Compr Rev Food Sci Food Saf. 201;15(4):720–38. doi:10.1111/1541-4337.12204
37. Bogacz-Radomska L, Harasym J. β-Carotene—properties and production methods. Food Qual Saf. 2018;2(2):69–74. doi:10.1093/fqsafe/fyy004
38. Nerdy N, Lestari P, Sinaga JP, Ginting S, Zebua NF, Mierza V, et al. Brine Shrimp (Artemia salina Leach.) Lethality Test of Ethanolic Extract from Green Betel (Piper betle Linn.) and Red Betel (Piper crocatum Ruiz and Pav.) through the Soxhletation Method for Cytotoxicity Test. Open Access Maced J Med Sci. 2021;9(A):407–12. doi:10.3889/oamjms.2021.6171
Authors
1.
Nur S, Sami FJ, Marwati M, Nursamsiar N, Fadri A, Khairuddin K. Phenolic and Flavonoid Content of Black Mulberry (Morus nigra L.) Stem and Their Evaluation Antioxidant and Cytotoxic Profile. Borneo J Pharm [Internet]. 2022Nov.30 [cited 2024Dec.21];5(4):384-95. Available from: https://journal.umpr.ac.id/index.php/bjop/article/view/3760
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