ISOLATION AND CHARACTERIZATION OF CHEMICAL COMPOUNDS FROM THE STEM BARK FRACTIONS OF CITRUS GRANDIS AND EVALUATION OF THEIR ANTIMICROBIAL, CYTOTOXIC, AND ANTIOXIDANT ACTIVITIES

Sheikh Adnan Sakib; Zimam Mahmud; Md. Rejaul Karim; Tania Rahman; Md. Enamul Haque

doi:10.3329/brc.v9i2.67089

Authors

Sheikh Adnan Sakib Department of Pharmacy, State University of Bangladesh, Dhaka-1205, Bangladesh
Zimam Mahmud Department of Biochemistry and Molecular Biology, University of Dhaka, Dhaka-1000, Bangladesh
Md. Rejaul Karim Department of Biochemistry and Molecular Biology, University of Dhaka, Dhaka-1000, Bangladesh
Tania Rahman Department of Biochemistry and Molecular Biology, University of Dhaka, Dhaka-1000, Bangladesh
Md. Enamul Haque Department of Biochemistry and Molecular Biology, University of Dhaka, Dhaka-1000, Bangladesh

DOI:

https://doi.org/10.3329/brc.v9i2.67089

Keywords:

Citrus grandis, Medicinal plant, Zeylenol, Limonin, Antimicrobial activities, Cytotoxic activities, Antioxidant activities

Abstract

Citrus grandis belongs to the Rutaceae family and is a naturally occurring citrus plant. The fractionated crude extracts of C. grandis have been studied for the investigation of its phytochemicals and allowed for the evaluation of biological activities, especially antimicrobial, cytotoxic, and antioxidant activities. At first, we assessed the antibacterial and antifungal properties using the disc diffusion method and conducted cytotoxicity screening using the brine shrimp lethality bioassay. Among the raw extracts, we isolated two compounds named CG-F3-B1 and CG-F3-B2 through column chromatographic separation of the chloroform extract. By comparing their proton NMR spectra with published spectra, we identified the chemical structures of these compounds as Zeylenol and Limonin, respectively. Regarding antibacterial testing, we observed that the methanol soluble fractions and petroleum ether fractions exhibited mild to moderate activity against bacteria. Additionally, the ethyl acetate soluble fraction showed moderate antibacterial activity. In addition, the crude extract displayed moderate antifungal activity against specific fungi. Furthermore, the cytotoxic assay revealed a relevant cytotoxic effect of a crude extract with an LC₅₀ value of 37 µg/ml. The antioxidant activity was determined using the DPPH assay, and the methanol soluble extract exhibited modest antioxidant activity with an IC₅₀ value of 89 µg/ml. Our findings demonstrated that the stem bark fractions and isolated compounds from C. grandis exhibited various promising biological activities, including antibacterial, antifungal, cytotoxic, and antioxidant properties.

References

Abudayeh, Z. H., I. I. Al Khalifa, S. M. Mohammed and A. A. J. P. R. Ahmad (2019). "Phytochemical content and antioxidant activities of pomelo peel extract." 11(3).

Ali, M. Y., N. N. Rumpa, S. Paul, M. S. Hossen, E. M. Tanvir, T. Hossan et al. (2019). "Antioxidant Potential, Subacute Toxicity, and Beneficiary Effects of Methanolic Extract of Pomelo (Citrus grandis L. Osbeck) in Long Evan Rats." J Toxicol 2019: 2529569.

Anmol, R. J., S. Marium, F. T. Hiew, W. C. Han, L. K. Kwan, A. K. Y. Wong et al. (2021). "Phytochemical and Therapeutic Potential of Citrus grandis (L.) Osbeck: A Review." J Evid Based Integr Med 26: 2515690x211043741.

Bauer, A. W., W. M. Kirby, J. C. Sherris and M. Turck (1966). "Antibiotic susceptibility testing by a standardized single disk method." Am J Clin Pathol 45(4): 493-496.

Bocco, A., M.-E. Cuvelier, H. Richard and C. Berset (1998). "Antioxidant Activity and Phenolic Composition of Citrus Peel and Seed Extracts." Journal of Agricultural and Food Chemistry 46(6): 2123-2129.

Carvalho, J. C. S., J. R. Pirani and M. J. P. Ferreira (2022). "Esenbeckia (Pilocarpinae, Rutaceae): chemical constituents and biological activities." Brazilian Journal of Botany 45(1): 41-65.

Chang, S. and A. Azrina (2016). Antioxidant content and activity in different parts of pomelo [Citrus grandis (L.) Osbeck] by-products. III International Conference on Agricultural and Food Engineering 1152.

Choi, H. S., H. S. Song, H. Ukeda and M. Sawamura (2000). "Radical-scavenging activities of citrus essential oils and their components: detection using 1,1-diphenyl-2-picrylhydrazyl." J Agric Food Chem 48(9): 4156-4161.

Chung S. K., K. S. H., Choi Y.H., Song EY, Kim S.H. (2000). "Status of citrus fruit production and view of utilization in Cheju." Food Industry and Nutrition 5: 42-52.

Cowan, M. M. (1999). "Plant products as antimicrobial agents." Clin Microbiol Rev 12(4): 564-582.

Franke, A. A., L. J. Custer, C. Arakaki and S. P. Murphy (2004). "Vitamin C and flavonoid levels of fruits and vegetables consumed in Hawaii." Journal of Food Composition and Analysis 17(1): 1-35.

Ghani, A. (1998). Medicinal plants of Bangladesh: Chemical constituents and uses.

Haque, M. E., Z. Mahmud, A. K. M. M. Hasan, R. Sardar, M. L. Kabir and S. T. Haque (2022). "Betulin-3-caffeate and Amyrin from the Stem Bark of Barringtonia acutangula (L)." Bioresearch Communications - (BRC) 1(2): 121-123.

Heim, K. E., A. R. Tagliaferro and D. J. Bobilya (2002). "Flavonoid antioxidants: chemistry, metabolism and structure-activity relationships." The Journal of Nutritional Biochemistry 13(10): 572-584.

Hou, Y. C., S. L. Hsiu, C. W. Tsao, Y. H. Wang and P. D. Chao (2000). "Acute intoxication of cyclosporin caused by coadministration of decoctions of the fruits of Citrus aurantium and the Pericarps of Citrus grandis." Planta Med 66(7): 653-655.

Islam, L. N., A. Rahman, Z. Mahmud, A. H. M. N. Nabi, M. Hossain and M. Mohasin (2015). "Assessment of Physicochemical and Biochemical Qualities of Tannery Effluents of Hazaribagh, Dhaka, and Comparison with Non-Tannery Water Samples." International Journal of Environment 4(1): 68-81.

Jayaprakasha, G. K., R. P. Singh, J. Pereira and K. K. Sakariah (1997). "Limonoids from Citrus reticulata and their moult inhibiting activity in mosquito Culex quinquefasciatus larvae." Phytochemistry 44(5): 843-846.

Kedare, S. B. and R. P. Singh (2011). "Genesis and development of DPPH method of antioxidant assay." J Food Sci Technol 48(4): 412-422.

Kumar, D., M. S. Ladaniya and M. Gurjar (2019). "Underutilized Citrus sp. Pomelo (Citrus grandis) and Kachai lemon (Citrus jambhiri) exhale in phytochemicals and antioxidant potential." J Food Sci Technol 56(1): 217-223.

Lim, H.-K., J. Y. Moon, H. Kim, M. Cho and S. K. Cho (2009). "Induction of apoptosis in U937 human leukaemia cells by the hexane fraction of an extract of immature Citrus grandis Osbeck fruits." Food Chemistry 114(4): 1245-1250.

Lu, Q., N. Huang, Y. Peng, C. Zhu and S. Pan (2019). "Peel oils from three Citrus species: volatile constituents, antioxidant activities and related contributions of individual components." J Food Sci Technol 56(10): 4492-4502.

Mahmud, Z., S. A. Shabnam, I. D. Mishu, F.-T. Johura, S. B. Mannan, A. Sadique et al. (2021). "Virotyping, genotyping, and molecular characterization of multidrug resistant Escherichia coli isolated from diarrheal patients of Bangladesh." Gene Reports 23: 101182.

Mäkynen, K., S. Jitsaardkul, P. Tachasamran, N. Sakai, S. Puranachoti, N. Nirojsinlapachai et al. (2013). "Cultivar variations in antioxidant and antihyperlipidemic properties of pomelo pulp (Citrus grandis [L.] Osbeck) in Thailand." Food Chem 139(1-4): 735-743.

Mandal, M., L. Kabir, I. Khan, S. T. Haque and M. E. Haque (2022). "Isolation of Bioactive Principles and Studies of Antimicrobial, Cytotoxic and Antioxidant Activities of the stem bark of Baccaurea ramiflora (Euphorbiaceae)." Bioresearch Communications - (BRC) 4(2): 565-571.

Meyer, B. N., N. R. Ferrigni, J. E. Putnam, L. B. Jacobsen, D. E. Nichols and J. L. McLaughlin (1982). "Brine shrimp: a convenient general bioassay for active plant constituents." Planta Med 45(5): 31-34.

Mokbel, M. S. and F. Hashinaga (2006). "Evaluation of the antioxidant activity of extracts from buntan (Citrus grandis Osbeck) fruit tissues." Food Chemistry 94(4): 529-534.

Njoroge, S. M., H. Koaze, P. N. Karanja and M. Sawamura (2005). "Volatile constituents of redblush grapefruit (Citrus paradisi) and pummelo (Citrus grandis) peel essential oils from Kenya." J Agric Food Chem 53(25): 9790-9794.

Palasuwan, A., S. Soogarun, T. Lertlum, P. Pradniwat and V. Wiwanitkit (2005). "Inhibition of Heinz body induction in an in vitro model and total antioxidant activity of medicinal Thai plants." Asian Pac J Cancer Prev 6(4): 458-463.

Shao, S. Y., W. J. Xu, J. Tao, J. H. Zhang, X. R. Zhou, G. Yuan et al. (2017). "Glycemic index, glycemic load, and glycemic response to pomelo in patients with type 2 diabetes." J Huazhong Univ Sci Technolog Med Sci 37(5): 711-718.

Shethi, K. J. (2022). "Evaluation of Antioxidant Activity of Four Locally Grown Aromatic Herbs Commonly Consumed in Bangladesh." Bioresearch Communications - (BRC) 3(2): 391-396.

Tao, N., G. Yumei, L. YueJin and G. Fei (2010). "Carotenoids from the peel of Shatian pummelo (Citrus grandis Osbeck) and its antimicrobial activity." Am. Eurasian J. Agric. Environ. Sci. 7: 110-115.

Tian, D., F. Wang, M. Duan, L. Cao, Y. Zhang, X. Yao et al. (2019). "Coumarin Analogues from the Citrus grandis (L.) Osbeck and Their Hepatoprotective Activity." J Agric Food Chem 67(7): 1937-1947.

Tsai, H. L., S. K. Chang and S. J. Chang (2007). "Antioxidant content and free radical scavenging ability of fresh red pummelo [Citrus grandis (L.) Osbeck] juice and freeze-dried products." J Agric Food Chem 55(8): 2867-2872.

Wu, T.-S., S.-C. Huang, T.-T. Jong, J.-S. Lai and C.-S. Kuoh (1988). "Coumarins, acridone alkaloids and a flavone from Citrus grandis." Phytochemistry 27(2): 585-587.

Zhang, M., C. Duan, Y. Zang, Z. Huang and G. Liu (2011). "The flavonoid composition of flavedo and juice from the pummelo cultivar (Citrus grandis (L.) Osbeck) and the grapefruit cultivar (Citrus paradisi) from China." Food Chemistry 129(4): 1530-1536.