Phytochemical Screening, Total Phenolic and Flavonoid Contents, Antioxidant, Tyrosinase Inhibitory, and HaCaT Cell Cytotoxic Activities of Leaf Extracts  from Bouea oppositifolia (Roxb.) Meisn.

Natthaphong  Lamkhwan; Aritsara  Sriraksa; Nuengruethai  Sinakkharanan; Aranya  Jutiviboonsuk; Suthira  Yanasob; Kanokporn  Sawasdee; Pattawat Seekhaw

Authors

Natthaphong Lamkhwan Department of Chemistry, Faculty of Science and Technology, Pibulsongkram Rajabhat University, Phitsanulok, 65000 Thailand
Aritsara Sriraksa Department of Chemistry, Faculty of Science and Technology, Pibulsongkram Rajabhat University, Phitsanulok, 65000 Thailand
Nuengruethai Sinakkharanan Department of Chemistry, Faculty of Science and Technology, Pibulsongkram Rajabhat University, Phitsanulok, 65000 Thailand
Aranya Jutiviboonsuk Faculty of Pharmaceutical Sciences, Huachiew Chalermprakiet University, Samut Prakan, 10540 Thailand
Suthira Yanasob Faculty of Pharmaceutical Sciences, Huachiew Chalermprakiet University, Samut Prakan, 10540 Thailand
Kanokporn Sawasdee Faculty of Pharmaceutical Sciences, Huachiew Chalermprakiet University, Samut Prakan, 10540 Thailand
Pattawat Seekhaw Department of Chemistry, Faculty of Science and Technology, Pibulsongkram Rajabhat University, Phitsanulok, 65000 Thailand

Keywords:

Bouea oppositifolia (Roxb.) Meisn., Antioxidant activity, Tyrosinase inhibitory, Cytotoxicity

Abstract

Natural medicines from plants have proved to display effective activity, especially through phytochemicals that can prevent or protect against various chronic diseases caused by free radicals. Bouea oppositifolia (Roxb.) Meisn., commonly known as Marian plum or Ma-Yong-Chid in Thai, is a perennial tree belonging to the family Anacardiaceae. It is closely related to plum mango that has been reported to possess significant biological activities, notably, with strong antioxidant activity in its leaf extract. In this study, the researchers aimed to investigate the phytochemical screening, total phenolic, and flavonoid contents, as well as the antioxidant, tyrosinase inhibitory, and HaCaT cytotoxic activities of leaf extracts from B. oppositifolia. Successive extraction of the dried leaf powder was carried out using three different solvents (hexane, ethyl acetate, and methanol) via the maceration technique. The preliminary phytochemical screening of B. oppositifolia extracts revealed the presence of phenolics, saponins, flavonoids, phytosterols, glycosides, anthraquinones, diterpenes, triterpenoids, and tannins. Among the three extracts (hexane, ethyl acetate, and methanol), the methanol extract had the highest total phenolic and flavonoid contents (74.25±3.31 mg GAE/g, dried leaf and 71.48±1.94 mg QE/g, dried leaf, respectively.), along with the strongest antioxidant capacity with IC₅₀ values of 1.39±0.40 µg/mL and 21.97±0.31 µg/mL for DPPH and ABTS assays, respectively, and a FRAP value of 505.55±24.32 mg FeSO₄/g, of dried leaf. On the other hand, the ethyl acetate extract exhibited the strongest tyrosinase inhibitory activity, with an IC₅₀ value of 242.90±7.48 µg/mL, and showed the lowest cytotoxicity toward HaCaT cells (%cell viability of 59.04±0.15%, at the concentration of 1,000 mg/mL). This study suggests that leaf extracts from B. oppositifolia are abundant in potential antioxidant and biological activities and could be further applied in pharmaceutical and cosmetic industries.

References

Akyilmaz, E., Yorganci, E., & Asav, E. (2010). Do copper ions activate tyrosinase enzyme? A biosensor model for the solution. Bioelectrochemistry, 78(2), 155–160.

Barbosa, A.D.P. (2014). An overview on the biological and pharmacological activities of saponins. International Journal of Pharmacy and Pharmaceutical Sciences, 6(8), 47–50.

Benzie, I.F., & Strain, J.J. (1996). The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: The FRAP assay. Analytical Biochemistry, 239(1), 70–76.

Bhatti, H.N., & Khera, R.A. (2014). Biotransformations of diterpenoids and triterpenoids: A review. Journal of Asian Natural Products Research, 16(1), 70–104.

De Silva, G.O., Abeysundara, A.T., & Aponso, M.M.W. (2017). Extraction methods, qualitative and quantitative techniques for screening of phytochemicals from plants. American Journal of Essential Oils and Natural Products, 5(2), 29–32.

Dirar, A.I., Alsaadi, D.H.M., Wada, M., Mohamed, M.A., Watanabe, T., & Devkota, H.P. (2019). Effects of extraction solvents on total phenolic and flavonoid contents and biological activities of extracts from Sudanese medicinal plants. South African Journal of Botany, 120, 261–267.

Evers, A.W.M., Duller, P., Van De Kerkhof, P.C.M., Van der Valk, P.G.M., De Jong, E.M.G.J., Gerritsen, M.J.P., ... Kraaimaat, F.W. (2008). The impact of chronic skin disease on daily life (ISDL): A generic and dermatology‐specific health instrument. British Journal of Dermatology, 158(1), 101–108.

Fathi Hafshejani, S., Lotfi, S., Rezvannejad, E., Mortazavi, M., & Riahi‐Madvar, A. (2023). Correlation between total phenolic and flavonoid contents and biological activities of 12 ethanolic extracts of Iranian propolis. Food Science & Nutrition, 11(7), 4308–4325.

Forni, C., Facchiano, F., Bartoli, M., Pieretti, S., Facchiano, A., D’Arcangelo, D., ... Jadeja, R.N. (2019). Beneficial role of phytochemicals on oxidative stress and age‐related diseases. BioMed Research International, 2019(1), 8748253.

Fraga-Corral, M., Otero, P., Echave, J., Garcia-Oliveira, P., Carpena, M., Jarboui, A., ... Prieto, M.A. (2021). By-products of agri-food industry as tannin-rich sources: A review of tannins’ biological activities and their potential for valorization. Foods, 10(1), 137.

Hajighasemi, F., & Tajic, S. (2017). Assessment of cytotoxicity of dimethyl sulfoxide in human hematopoietic tumor cell lines. Iranian Journal of Blood and Cancer, 9(2), 48–53.

Huyut, Z., Beydemir, Ş., & Gülçin, İ. (2017). Antioxidant and antiradical properties of selected flavonoids and phenolic compounds. Biochemistry Research International, 2017(1), 7616791.

Iloki-Assanga, S.B., Lewis-Luján, L.M., Lara-Espinoza, C.L., Gil-Salido, A.A., Fernandez-Angulo, D., Rubio-Pino, J.L., & Haines, D.D. (2015). Solvent effects on phytochemical constituent profiles and antioxidant activities, using four different extraction formulations for analysis of Bucida buceras L. and Phoradendron californicum. BMC Research Notes, 8, 1–14.

International Organization for Standardization (ISO). (2009). Biological evaluation of medical devices – Part 5: Tests for in vitro cytotoxicity (3rd ed.). https://www.iso.org/standard/36406.html

Itsarasook, K., Prompamorn, P., Tanghiranrat, J., Chantree, K., & Itsarasuk, I. (2023). Antioxidant activities and cytotoxicity effect on normal human dermal fibroblasts of roselle (Hibiscus sabdariffa L.) calyces extracts and W/O/W emulsion loaded extract for cosmetic applications. Journal of Food Health and Bioenvironmental Science, 16(2), 38–48.

Jaengklang, C., Muangman, T., Pathtubtim, I., & Jaroennon, P. (2024). Development of dosage form of Ya Kae Fok Buam Mueai Khop, a Thai traditional formula, and its phytochemical contents and anti-inflammatory activity. Journal of Food Health and Bioenvironmental Science, 17(2), 33–43.

Jin, W., Stehbens, S.J., Barnard, R.T., Blaskovich, M.A., & Ziora, Z.M. (2024). Dysregulation of tyrosinase activity: A potential link between skin disorders and neurodegeneration. Journal of Pharmacy and Pharmacology, 76(1), 13–22.

Kaur, M., Asthir, B., & Mahajan, G. (2017). Variation in antioxidants, bioactive compounds and antioxidant capacity in germinated and ungerminated grains of ten rice cultivars. Rice Science, 24(6), 349–359.

Khan, H., Pervaiz, A., Intagliata, S., Das, N., Nagulapalli Venkata, K.C., Atanasov, A.G., ... Bishayee, A. (2020). The analgesic potential of glycosides derived from medicinal plants. DARU Journal of Pharmaceutical Sciences, 28(1), 387–401.

Kiattisin, K., Intasai, N., Nitthikan, N., Nantarat, T., Lee, K., Lin, W., Lue, S. & Leelapornpisid, P. (2019). Antioxidant, anti-tyrosinase, anti-aging potentials and safety of arabica coffee cherry extract. Chiang Mai Journal of Science, 46(5), 930–945.

Kumar, A.P.N., Kumar, M., Jose, A., Tomer, V., Oz, E., ... Oz, F. (2023). Major phytochemicals: Recent advances in health benefits and extraction method. Molecules, 28(2), 887.

Kumar, M., Saurabh, V., Tomar, M., Hasan, M., Changan, S., Sasi, M., ... Mekhemar, M. (2021). Mango (Mangifera indica L.) leaves: Nutritional composition, phytochemical profile, and health-promoting bioactivities. Antioxidants, 10(2), 299.

Kumar, S., & Pandey, A.K. (2013). Chemistry and biological activities of flavonoids: An overview. The Scientific World Journal, 2013(1), 162750.

Lai, X., Wichers, H.J., Soler‐Lopez, M., & Dijkstra, B.W. (2018). Structure and function of human tyrosinase and tyrosinase‐related proteins. Chemistry–A European Journal, 24(1), 47–55.

Lamkhwan, N., & Seekhaw, P. (2025). Phytochemical screening, total phenolic and flavonoid contents, antioxidant and cytotoxic activities from twig crude extracts of Bouea burmanica Griff. Thai Science and Technology Journal, 33(2), 1–14.

Lee, S.Y., Baek, N., & Nam, T.G. (2016). Natural, semisynthetic and synthetic tyrosinase inhibitors. Journal of Enzyme Inhibition and Medicinal Chemistry, 31(1), 1–13.

Majhenič, L., Škerget, M., & Knez, Ž. (2007). Antioxidant and antimicrobial activity of guarana seed extracts. Food Chemistry, 104(3), 1258–1268.

Mamoona, Nosheen, S., Riaz, S., Shah, S.I., & Shahid, S. (2024). Optimizing extraction methods: The role of solvent polarity in enhancing phenolic content and antioxidant activity in biowaste. Biomass Conversion and Biorefinery, 15, 16721–16736.

Masuda, T., Yamashita, D., Takeda, Y., & Yonemori, S. (2005). Screening for tyrosinase inhibitors among extracts of seashore plants and identification of potent inhibitors from Garcinia subelliptica. Bioscience, Biotechnology, and Biochemistry, 69(1), 197–201.

Mavuso, V.L., & Yapwattanaphun, C. (2017). Effect of environmental conditions on flower induction of marian plum (Bouea burmanica Griff). Agriculture and Natural Resources, 51(4), 243–246.

Mehmood, A., Javid, S., Khan, M.F., Ahmad, K.S., & Mustafa, A. (2022). In vitro total phenolics, total flavonoids, antioxidant and antibacterial activities of selected medicinal plants using different solvent systems. BMC chemistry, 16(1), 64.

Moghadamtousi, S.Z., Goh, B.H., Chan, C.K., Shabab, T., & Kadir, H.A. (2013). Biological activities and phytochemicals of Swietenia macrophylla King. Molecules, 18(9), 10465–10483.

Muflihah, Y.M., Gollavelli, G., & Ling, Y.C. (2021). Correlation study of antioxidant activity with phenolic and flavonoid compounds in 12 Indonesian indigenous herbs. Antioxidants, 10(10), 1530.

Mutha, R.E., Tatiya, A.U., & Surana, S.J. (2021). Flavonoids as natural phenolic compounds and their role in therapeutics: An overview. Future journal of pharmaceutical sciences, 7, 1–13.

Nguyen, N.H., Nguyen, T.T., Ma, P.C., Ta, Q.T.H., Duong, T.H., & Vo, V.G. (2020). Potential antimicrobial and anticancer activities of an ethanol extract from Bouea macrophylla. Molecules, 25(8), 1996.

Onar, H.C., Yusufoglu, A., Turker, G., & Yanardag, R. (2012). Elastase, tyrosinase and lipoxygenase inhibition and antioxidant activity of an aqueous extract from Epilobium angustifolium L. leaves. Journal of Medicinal Plants Research, 6(5), 716–726.

Pang, Y., Ahmed, S., Xu, Y., Beta, T., Zhu, Z., Shao, Y., & Bao, J. (2018). Bound phenolic compounds and antioxidant properties of whole grain and bran of white, red and black rice. Food chemistry, 240, 212–221.

Patel, O.P., Beteck, R.M., & Legoabe, L.J. (2021). Antimalarial application of quinones: A recent update. European Journal of Medicinal Chemistry, 210, 113084.

Phaniendra, A., Jestadi, D.B., & Periyasamy, L. (2015). Free radicals: properties, sources, targets, and their implication in various diseases. Indian Journal of Clinical Biochemistry, 30, 11–26.

Phong, H.X., Viet, N.T., Quyen, N.T.N., Van Thinh, P., Trung, N.M., & Ngan, T.T.K. (2022). Phytochemical screening, total phenolic, flavonoid contents, and antioxidant activities of four spices commonly used in Vietnamese traditional medicine. Materials Today: Proceedings, 56, A1–A5.

Rafi, M., Meitary, N., Septaningsih, D.A., & Bintang, M. (2020). Phytochemical profile and antioxidant activity of Guazuma ulmifolia leaves extracts using different solvent extraction. Indonesian Journal of Pharmacy, 31(3), 171–180.

Rey-Blanes, C., Pérez-Portero, Y., Morris-Quevedo, H.J., Casas, V., Abdala, R., Quesada, A.R., ... Medina, M.Á. (2020). In vitro evaluation of the antitumoral and antiangiogenic effects of extracts from Spondias mombin L. leaves. Biomedicine & Pharmacotherapy, 131, 110716.

Seekhaw, P., Nearmayhom, N., Promla, N., Na-Wichian, K., & Jutiviboonsuk, A. (2022). Phytochemical screening, acute toxicity against skin irritation and corrosion in rabbits of aqueous fabric dye from Bouea burmanica Griff. leaf extract. PSRU Journal of Science and Technology, 7(1), 117–128.

Shaikh, J.R., & Patil, M. (2020). Qualitative tests for preliminary phytochemical screening: An overview. International journal of chemical studies, 8(2), 603–608.

Sisein, E.A. (2014). Biochemistry of free radicals and antioxidants. Scholars Academic Journal of Biosciences, 2(2), 110–118.

Song, Y., Chen, S., Li, L., Zeng, Y., & Hu, X. (2022). The hypopigmentation mechanism of tyrosinase inhibitory peptides derived from food proteins: An overview. Molecules, 27(9), 2710.

Sriraksa, A., & Seekhaw, P. (2024). Comparison of phytochemicals and antioxidant capacities of ethanol extracts from different parts of Elsholtzia beddomei C.B.Clarke ex Hook. f. Thai Science and Technology Journal, 32(2), 1–12.

Tada, M., Kohno, M., & Niwano, Y. (2014). Alleviation effect of arbutin on oxidative stress generated through tyrosinase reaction with L-tyrosine and L-DOPA. BMC Biochemistry, 15, 23.

Widyawati, P.S., Budianta, T.D.W., Kusuma, F.A., & Wijaya, E.L. (2014). Difference of solvent polarity to phytochemical content and antioxidant activity of Pluchea indicia less leaves extracts. International Journal of Pharmacognosy and Phytochemical Research, 6(4), 850–855.

Xiao, F., Farag, M.A., Xiao, J., Yang, X., Liu, Y., Shen, J., & Lu, B. (2022). The influence of phytochemicals on cell heterogeneity in chronic inflammation-associated diseases: The prospects of single cell sequencing. The Journal of Nutritional Biochemistry, 108, 109091.

Yoo, S., Kim, K., Nam, H., & Lee, D. (2018). Discovering health benefits of phytochemicals with integrated analysis of the molecular network, chemical properties and ethnopharmacological evidence. Nutrients, 10(8), 1042.

Zagórska-Dziok, M., Ziemlewska, A., Nizioł-Łukaszewska, Z., & Bujak, T. (2020). Antioxidant activity and cytotoxicity of Medicago sativa L. seeds and herb extract on skin cells. BioResearch Open Access, 9(1), 229–242.

Zengin, G., Uysal, S., Ceylan, R., & Aktumsek, A. (2015). Phenolic constituent, antioxidative and tyrosinase inhibitory activity of Ornithogalum narbonense L. from Turkey: A phytochemical study. Industrial crops and products, 70, 1–6.