In-Vitro Estimation of Total Phenolic, Flavonoid, Antioxidant Contents and Determination of Flavonoid, Amino Acid, and Vitamin Profiles of Ethanol Seed Extract of Annona squamosa Linn
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Abstract
The aim of this research is to analyse the flavonoid, amino acid, and vitamin profiles of ethanol seed extract of Annona squamosa using high performance liquid chromatography (HPLC). The fruits of Annona squamosa were collected from Takum, Taraba State, the seeds were removed from the fruit washed with water, dried in the shade and then pulverized into fine powder. For the crude extraction, the method adopted by Yakubu et al. (2020) was adopted for this protocol. The IC50 of the seed was found to be 3.87. The total flavonoid content was significantly higher (182.76±0.61) compared with the total phenolic compound (70.34±3.04). A reliable detection and peak resolution were obtained from the HPLC analysis. The result showed the presence of nine amino acids, which include; Threonine (9.6%), Phenyl Alanine (17.0%), Aspartic Acid (26.0 %), Glutamic acid (16.0%), Leucine (5.7%), Iso-Leucine (5.3%), Glycine (5.0 %), Arginine (2.3), Glutamine (13.2%), three vitamins; vitamin K (0.4%), vitamin B1(63.0 %) and B3 (36.6%), and three flavonoid compounds Trolox (0.2%), Quercetin (2.1%) and Coumaric acid (97.7%) in plant sample. The phytochemical study confirms that the seed of Annona squamosa rich sources of amino acids such as and these herbal raw materials can exhibit a wide range of pharmacological activities.
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Yakubu, O. E., Audu, J. P., & Adebisi, A. K. (2024). In-Vitro Estimation of Total Phenolic, Flavonoid, Antioxidant Contents and Determination of Flavonoid, Amino Acid, and Vitamin Profiles of Ethanol Seed Extract of Annona squamosa Linn. African Journal of Biochemistry and Molecular Biology Research, 1(1), 170-182. https://doi.org/10.58578/ajbmbr.v1i1.3470
Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
References
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29. Singleton, V. L., Orthofer, R. and Lamuela-Raventos, R. M. (2002). Analysis of total phenol and other oxidative substrates and antioxidants by means of Folin-ciocalteu reagent. Methods in Enzymology, 299, 152-178.
30. Triantis, T., Stelakis, A., Dimotikali, D., & Papadopoulos, K. (2005). Investigations on the antioxidant activity of fruit and vegetable aqueous extracts on superoxide radical anion using chemiluminescence techniques. Analytica chimica acta, 536(1-2), 101-105.
31. Trovato, M., Funck, D., Forlani, G., Okumoto, S., & Amir, R. (2021). Amino acids in plants: regulation and functions in development and stress defense. Frontiers in Plant Science, 12, 772810.
32. Ullah , A., Munir, S., Badshah, S. L., Khan, N., Ghani, L., Poulson, B. G., Emwas, A. H., & Jaremko, M. (2020). Important Flavonoids and Their Role as a Therapeutic Agent. Molecules (Basel, Switzerland), 25(22), 5243.
33. Vanitha, V., Umadevi, K. J., & Vijayalakshmi, K. (2010). In vitro assessment of alcoholic leaf extracts of Annona squamosa and Aegle marmelos. The Bioscan, An International Quarterly Journal of Life Sciences, 5(2), 2010, 225-229.
34. Vyas, K., Manda, H., Sharma, R. K., & Singhal, G. (2012). An update review on Annona Squamosa. IJPT, 3(2), 107-118.
35. Wu, G. (2009). Amino acids: metabolism, functions, and nutrition. Amino Acids, 37, 1-17.
36. Yakubu, O. E., Nwodo, O. F. C., Joshua, P. E., Ugwu, M. N., Odu, A. D., & Okwo, F. (2014). Fractionation and determination of total antioxidant capacity, total phenolic and total flavonoids contents of aqueous, ethanol and n-hexane extracts of Vitex doniana leaf. African Journal of Biotechnology, 13(5), 693-698.
37. Yakubu, O.E., Ojogbane, E., Nwaneri-Chidozie, V.O., Ibuzo, P.N., Ale, E.M., & Awudu, S.S. (2020). Ameliorative effects of ethanol extract of Ficus sur on diethylnitrosamine-induced hepatotoxicity in wistar rats. American Journal of Biochemistry and Molecular Biology, 10, 6-11.
38. Zibadi, S., Farid, R., Moriguchi, S., Lu, Y., Foo, L. Y., Tehrani, P. M., Ulreich, J.B. & Watson, R. R. (2007). Oral administration of purple passion fruit peel extract attenuates blood pressure in female spontaneously hypertensive rats and humans. Nutrition Research, 27(7), 408-416.
2. Al-Nemari, R., Al-Senaidy, A., Semlali, A., Ismael, M., Badjah-Hadj-Ahmed, A. Y., & Ben Bacha, A. (2020). GC-MS profiling and assessment of antioxidant, antibacterial, and anticancer properties of extracts of Annona squamosa L. leaf. BMC Complementary Tedicine and Therapies, 20(1), 296.
3. Alu’datt, M. H., Rababah, T., Alhamad, M. N., Al-Mahasneh, M. A., Almajwal, A., Gammoh, S., Ereifej, K., Johargy, A. & Alli, I. (2017). A review of phenolic compounds in oil-bearing plants: Distribution, identification and occurrence of phenolic compounds. Food Chemistry, 218, 99-106.
4. Bhat R. and Paliyath G., (2016). Fruits of tropical climates: dietary importance and health benefits, Editor(s): Benjamin Caballero, Paul M. Finglas, Fidel Toldrá, Encyclopedia of Food and Health. Academic Press, 144-149.
5. Chang, C. C., Yang, M. H., Wen, H. M., & Chern, J. C. (2002). Estimation of total flavonoid content in propolis by two complementary colometric methods. Journal of Food and Drug Analysis, 10(3), 178-182.
6. Cylwik, D., Mogielnicki, A., & Buczko, W. (2005). L-arginine and cardiovascular system. Pharmacol Rep, 57(1), 14-22.
7. Egbuna, C., & Dable-Tupas, G. (2020). Functional foods and nutraceuticals. Springer Nature Switzerland AG, 1, 1-632.
8. Egydio, A. P. M., Santa Catarina, C., Floh, E. I. S., & dos Santos, D. Y. A. C. (2013). Free amino acid composition of Annona (Annonaceae) fruit species of economic interest. Industrial Crops and Products, 45, 373-376.
9. Feliciano, R. P., Pritzel, S., Heiss, C., & Rodriguez-Mateos, A. (2015). Flavonoid intake and cardiovascular disease risk. Current Opinion in Food Science, 2, 92-99.
10. Kumar, M., Changan, S., Tomar, M., Prajapati, U., Saurabh, V., Hasan, M., Sasi, M., Maheshwari, C., Singh, S., Dhumal, S., Radha, Thakur, M., Punia, S., Satankar, V., Amarowicz, R., & Mekhemar, M. (2021). Custard Apple (Annona squamosa L.) Leaf: Nutritional Composition, Phytochemical Profile, and Health-Promoting Biological Activities. Biomolecules, 11(5): 614
11. Lachman, J., Hamouz, K., Orsák, M., Pivec, V., & Dvořák, P. (2000). The influence of flesh colour and growing locality on polyphenolic content and antioxidant activity in potatoes. Scientia Horticulturae, 117(2), 109-114.
12. Lin, D., Xiao, M., Zhao, J., Li, Z., Xing, B., Li, X., Kong, M., Li, L., Zhang, Q., Liu, Y., Chen, H., Qin, W., Wu, H., & Chen, S. (2016). An Overview of Plant Phenolic Compounds and Their Importance in Human Nutrition and Management of Type 2 Diabetes. Molecules (Basel, Switzerland), 21(10), 1374.
13. Ling, Z. N., Jiang, Y. F., Ru, J. N., Lu, J. H., Ding, B., & Wu, J. (2023). Amino acid metabolism in health and disease. Signal Transduction and Targeted Therapy, 8(1), 345.
14. Loizzo, M. R., Tundis, R., Bonesi, M., Menichini, F., Mastellone, V., Avallone, L., & Menichini, F. (2012). Radical scavenging, antioxidant and metal chelating activities of Annona cherimola Mill.(cherimoya) peel and pulp in relation to their total phenolic and total flavonoid contents. Journal of Food Composition and Analysis, 25(2), 179-184.
15. Mannino, G., Gentile, C., Porcu, A., Agliassa, C., Caradonna, F., & Bertea, C. M. (2020). Chemical Profile and Biological Activity of Cherimoya (Annona cherimola Mill.) and Atemoya (Annona atemoya) Leaf. Molecules, 25(11): 2612
16. Mateo, R. D., Wu, G., Bazer, F. W., Park, J. C., Shinzato, I., & Kim, S. W. (2007). Dietary L-arginine supplementation enhances the reproductive performance of gilts. The Journal of Nutrition, 137(3), 652-656.
17. Nonfon, M., Lieb, F., Moeschler, H., & Wendisch, D. (1990). Four annonins from Annona squamosa. Phytochemistry, 29(6), 1951-1954.
18. Patel, J. D., & Kumar, V. (2008). Annona squamosa L.: phytochemical analysis and antimicrobial screening. Journal of Pharmacy Research, 1(1), 34-38.
19. Pietta, P. G. (2000). Flavonoids as antioxidants. Journal of Natural Products, 63(7), 1035-1042.
20. Rahman, M. M., Parvin, S., Haque, M. E., Islam, M. E., & Mosaddik, M. A. (2005). Antimicrobial and cytotoxic constituents from the seeds of Annona squamosa. Fitoterapia, 76(5), 484-489.
21. Rama, D. B., Malleswari, K., Naveen K. E. (2023). Phytochemicals and Pharmacological Activities of Annona Squamosa. International Journal for Multidisciplinary Research, 5(1), 1-9
22. Rehman, S. U., Ali, R., Zhang, H., Zafar, M. H., & Wang, M. (2023). Research progress in the role and mechanism of Leucine in regulating animal growth and development. Frontiers in physiology, 14, 1252089.
23. Roesler, R., Catharino, R. R., Malta, L. G., Eberlin, M. N., & Pastore, G. (2007). Antioxidant activity of Annona crassiflora: Characterization of major components by electrospray ionization mass spectrometry. Food Chemistry, 104(3), 1048-1054.
24. Saha, R. (2011). Pharmacognosy and pharmacology of Annona squamosa. International Journal of Pharmacy and Life Sciences, 2(10), 1183-1189.
25. Shan, X., Cheng, J., Chen, K., Liu, Y. M., & Juan, L. (2017). Comparison of lipoxygenase, cyclooxygenase, xanthine oxidase inhibitory effects and cytotoxic activities of selected flavonoids. DEStech Transactions on Environment, Energy and Earth Sciences, 10.
26. Shukla, S.K. Kumar, V. (2013). Chapter 36 - Bioactive foods and supplements for protection against liver diseases, Editor(s): Ronald Ross Watson, Victor R. Preedy, Bioactive Food as Dietary Interventions for Liver and Gastrointestinal Disease, Academic Press, 557-567
27. Singh, Y., Bhatnagar, P., & Thakur, N. (2019). A review on insight of immense nutraceutical and medicinal potential of custard apple (Annona squamosa Linn.). International Journal of Chemical Studies, 7(2), 1237-1245.
28. Singh, Y., Bhatnagar, P., & Thakur, N. (2019). A review on insight of immense nutraceutical and medicinal potential of custard apple (Annona squamosa Linn.). International Journal of Chemical Studies, 7(2), 1237-1245.
29. Singleton, V. L., Orthofer, R. and Lamuela-Raventos, R. M. (2002). Analysis of total phenol and other oxidative substrates and antioxidants by means of Folin-ciocalteu reagent. Methods in Enzymology, 299, 152-178.
30. Triantis, T., Stelakis, A., Dimotikali, D., & Papadopoulos, K. (2005). Investigations on the antioxidant activity of fruit and vegetable aqueous extracts on superoxide radical anion using chemiluminescence techniques. Analytica chimica acta, 536(1-2), 101-105.
31. Trovato, M., Funck, D., Forlani, G., Okumoto, S., & Amir, R. (2021). Amino acids in plants: regulation and functions in development and stress defense. Frontiers in Plant Science, 12, 772810.
32. Ullah , A., Munir, S., Badshah, S. L., Khan, N., Ghani, L., Poulson, B. G., Emwas, A. H., & Jaremko, M. (2020). Important Flavonoids and Their Role as a Therapeutic Agent. Molecules (Basel, Switzerland), 25(22), 5243.
33. Vanitha, V., Umadevi, K. J., & Vijayalakshmi, K. (2010). In vitro assessment of alcoholic leaf extracts of Annona squamosa and Aegle marmelos. The Bioscan, An International Quarterly Journal of Life Sciences, 5(2), 2010, 225-229.
34. Vyas, K., Manda, H., Sharma, R. K., & Singhal, G. (2012). An update review on Annona Squamosa. IJPT, 3(2), 107-118.
35. Wu, G. (2009). Amino acids: metabolism, functions, and nutrition. Amino Acids, 37, 1-17.
36. Yakubu, O. E., Nwodo, O. F. C., Joshua, P. E., Ugwu, M. N., Odu, A. D., & Okwo, F. (2014). Fractionation and determination of total antioxidant capacity, total phenolic and total flavonoids contents of aqueous, ethanol and n-hexane extracts of Vitex doniana leaf. African Journal of Biotechnology, 13(5), 693-698.
37. Yakubu, O.E., Ojogbane, E., Nwaneri-Chidozie, V.O., Ibuzo, P.N., Ale, E.M., & Awudu, S.S. (2020). Ameliorative effects of ethanol extract of Ficus sur on diethylnitrosamine-induced hepatotoxicity in wistar rats. American Journal of Biochemistry and Molecular Biology, 10, 6-11.
38. Zibadi, S., Farid, R., Moriguchi, S., Lu, Y., Foo, L. Y., Tehrani, P. M., Ulreich, J.B. & Watson, R. R. (2007). Oral administration of purple passion fruit peel extract attenuates blood pressure in female spontaneously hypertensive rats and humans. Nutrition Research, 27(7), 408-416.
