In-vitro evaluation of antidiabetic and antioxidant activities of lime juice extract of Gossypium herbaceum leaves
Article Sidebar
Main Article Content
Management of diabetes mellitus increasingly focuses on controlling postprandial hyperglycemia through inhibition of carbohydrate-hydrolyzing enzymes responsible for gastrointestinal glucose release and absorption. This study evaluated the in vitro antidiabetic and antioxidant activities of a lime juice extract derived from Gossypium herbaceum leaves. Fresh leaves were extracted using lime juice (Citrus aurantifolia) as a natural solvent medium. Inhibitory activities against α-amylase and α-glucosidase were assessed alongside antioxidant capacity using DPPH, ABTS, and hydroxyl radical scavenging assays at concentrations of 7.8125–1000 μg/mL, conducted in triplicate using standard spectrophotometric protocols. Acarbose served as the reference enzyme inhibitor, while butylated hydroxytoluene (BHT) was used as the antioxidant standard. The extract exhibited concentration-dependent inhibition of both carbohydrate-digesting enzymes and significant free radical scavenging activity. Although reference standards demonstrated higher activity across most concentrations, ABTS radical scavenging increased progressively with dose, approaching BHT performance at higher concentrations. Notably, hydroxyl radical scavenging at 31.25 μg/mL showed no significant difference from BHT, indicating strong antioxidant potential at moderate dosage. In contrast, α-glucosidase inhibition remained significantly lower than acarbose (p < 0.05) at all tested concentrations, suggesting moderate regulation of carbohydrate digestion rather than complete enzyme suppression. These findings demonstrate that lime-mediated extraction of Gossypium herbaceum leaves yields appreciable hypoglycemic potential through partial enzyme inhibition combined with enhanced antioxidant defense mechanisms. Synergistic contributions from ascorbic acid and bioactive phytochemicals present in Citrus aurantifolia likely underpin the observed bioactivity. Overall, the extract represents a promising natural candidate for development as an oral antidiabetic phytomedicine aimed at complementary diabetes management.
Downloads
References
Amirqulova, G. M., & Xidirova, N. X. (2024). Vascular complications of diabetes. European Journal of Modern Medicine and Practice, 4(11), 180-185.
Apak, R., Özyürek, M., Güçlü, K. & Çapanoğlu, E. (2022). Antioxidant activity/capacity measurement: Recent trends and advances. Journal of Food Composition and Analysis, 103, 104171. https://doi.org/10.1016/j.jfca.2021.104171.
Apostolidis, E., Kwon, Y. I. & Setty, K. (2007). Inhibitory potential of herb, fruit, and fungal-enriched cheese against key enzymes linked to type-2 diabetes. Innov Food Sci Emerg Technol, 8, 46–54.
Bernfield, P. (1951). Enzymes of starch degradation and synthesis. Adv. Enzymol Relat Subj Biochem, 12, 379.
Bios, M. S. (1958). Antioxidant determination by the use of stable free radical. Nature, 81, 1199-2000.
Brand-Williams, W., Cuvelier, M. E., & Berset, C. L. W. T. (1995). Use of a free radical method to evaluate antioxidant activity. LWT-Food Science and Technology, 28(1), 25-30.
Catherine, A. E., Janvier, Y., Martin, F., Guy, T. G., Dupon, A. A., Vidal, N. J., ... & Julius, O. E. (2023). Anti-Arthritic Effect of the Aqueous Extracts of the Roots and Barks of Gossypium herbaceous in Complete Freund’s Adjuvant Induced Arthritis in Female Wistar Rats. Journal of Complementary and Alternative Medical Research, 22(3), 28-37.
Dash, R. P., Babu, R. J., & Srinivas, N. R. (2018). Reappraisal and perspectives of clinical drug–drug interaction potential of α-glucosidase inhibitors such as acarbose, voglibose and miglitol in the treatment of type 2 diabetes mellitus. Xenobiotica, 48(1), 89-108.
Ebulue, M. M. (2024). Inhibitory Properties of Polyphenolic Phytochemicals of Cola nitida on Carbohydrate Hydrolyzing Enzymes of Wistar Rat In-Vivo. Saudi J Biomed Res, 9(1), 22-27.
Fidyasari, A., Estiasih, T., Khatib, A., Wulan, S. N., & Raharjo, S. J. (2026). Molecular Dynamics Simulation of Bioactive Compounds from Moringa Oleifera Leaf Powder Extract as Antidiabetic by Inhibiting α-Amylase and α-Glucosidase Enzymes. Trends in Sciences, 23(1), 10964-10964.
Flieger, J., Flieger, W., Baj, J., & Maciejewski, R. (2021). Antioxidants: Classification, natural sources, activity/capacity measurements, and usefulness for the synthesis of nanoparticles. Materials, 14(15), 4135.
Franz, M. H., Birzoi, R., Maftei, C. V., Maftei, E., Kelter, G., Fiebig, H. H., & Neda, I. (2018). Studies on the constituents of Helleborus purpurascens: analysis and biological activity of the aqueous and organic extracts. Amino Acids, 50(1), 163-188.
Genitsaridi, I., Salpea, P., Salim, A., Sajjadi, S. F., Tomic, D., James, S., ... & Magliano, D. J. (2026). of the IDF Diabetes Atlas: global, regional, and national diabetes prevalence estimates for 2024 and projections for 2050. The Lancet Diabetes & Endocrinology, 14(2), 149-156.
Gulcin, İ. (2020). Antioxidants and antioxidant methods: An updated overview. Archives of Toxicology, 94(3), 651-715.
Huang, Q., Li, Y., Yu, M., Lv, Z., Lu, F., Xu, N., ... & Jiang, H. (2025). Global burden and risk factors of type 2 diabetes mellitus from 1990 to 2021, with forecasts to 2050. Frontiers in Endocrinology, 16, 1538143.
Indriyani, N. N., Al Anshori, J., Permadi, N. & Julaeha, E. (2023). Bioactive components and biological activities of Citrus aurantifolia for food and health applications. Foods, 12(10), 2036. https://doi.org/10.3390/foods12102036.
Karki, N., Achhami, H., Pachhai, B. B., Bhattarai, S., Shahi, D. K., Bhatt, L. R., & Joshi, M. K. (2024). Evaluating citrus juice: A comparative study of physicochemical, nutraceutical, antioxidant, and antimicrobial properties of citrus juices from Nepal. Heliyon, 10(23).
Kawami, M., Yumoto, R., Junyaprasert, V. B., Soonthornchareonnon, N., Patanasethanont, D., Sripanidkulchai, B., & Takano, M. (2026). Disaccharidase Inhibitory Activity of Thai Plant Extracts. Nutrients, 18(3), 456.
Kyada, A., Rahamathulla, M., Santani, D., Vadia, N., Baldaniya, L., Rana, M., ... & Pasha, I. (2023). Phytochemical analysis, in vitro free radical scavenging, and LDL protective effects of different solvent fractions of Calotropis procera (R.) Br. root bark extract. Journal of Food Biochemistry, 2023(1), 6689595.
Larayetan, R. A., Ayeni, G., Yahaya, A., Ajayi, A., Omale, S., Ishaq, U., ... & Enyioma-Alozie, S. (2021). Chemical composition of Gossypium herbaceum Linn and its antioxidant, antibacterial, cytotoxic and antimalarial activities. Clinical Complementary Medicine and Pharmacology, 1(1), 100008.
McKeirnan, K. C., & Rodin, N. M. (2023). α-Glucosidase Inhibitors. Guide to Medications for the Treatment of Diabetes Mellitus, 24.
Mili, S., Konwar, A., Singh, Y. R., & Kalita, J. C. (2025). Comprehensive Phytochemical Analysis and Antifertility Assessment of Medicinal Plants Traditionally Used in Contraception across Different Cultures. Pharmacological Research-Natural Products, 100403.
Ogunyinka, B. I., Oboh, G., Ademiluyi, A. O., & Boligon, A. A. (2016). Inhibitory effect of aqueous extract of different parts of Gossypium herbaceum on key enzymes linked with type 2 diabetes and oxidative stress in rat pancreas in vitro. Journal of Basic and Applied Sciences, 5(2), 180–186. https://doi.org/10.1016/j.bjbas.2016.05.003
Okunlola, A., Adewoyin, A. G. & Odeku, O. A. (2021). Chemical composition of Gossypium herbaceum Linn and its antioxidant, antibacterial, cytotoxic and antimalarial activities. Clinical Complementary Medicine and Pharmacology, 1(1), 100008. https://doi.org/10.1016/j.ccmp.2021.100008.
Olanrewaju, A. A., Ogunlakin, A. D., Ogundele, D. O., Oyebamiji, A. K., Ojo, O. A., Akinola, O. T., ... & Oke, D. G. (2025). Antioxidant and antidiabetic activities of Gossypium barbadense L. leaves using in vitro, and in silico methods. Vegetos, 1-12.
Parameswari, R., Kumar, P. M., Pavithra, S. A., Iswarya, S. J., Yogesh, T., & Babujanarthanam, R. (2025). Diabetes: Secondary Complications. In Algae in Diabetes Management: Therapeutic Properties and Applications (pp. 35-88). Singapore: Springer Nature Singapore.
Pathak, S. R., Senwar, K. R., & Sharma, K. N. (2025). Alpha-glucosidase in diabetes mellitus. In Diabetes Mellitus (pp. 63-78). Academic Press.
Re, R., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M., & Rice-Evans, C. (1999). Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biology and Medicine, 26(9-10), 1231-1237.
Saeedi, P., Petersohn, I. & Salpea, P., et al. (2019). Global and regional diabetes prevalence estimates for 2019 and projections for 2030 and 2045. Diabetes Research and Clinical Practice, 157, 107843. https://doi.org/10.1016/j.diabres.2019.107843.
Tumilaar, S. G., Hardianto, A., Dohi, H., & Kurnia, D. (2024). A comprehensive review of free radicals, oxidative stress, and antioxidants: Overview, clinical applications, global perspectives, future directions, and mechanisms of antioxidant activity of flavonoid compounds. Journal of Chemistry, 2024(1), 5594386.
Vig, H., Wal, A., Sharma, P., Dwivedi, J., Saxena, B., Singh, N., ... & Gasmi, A. (2026). Phytochemical Composition and Mechanistic Pharmacology of the Citrus Genus: Focus on Organic Constituents. Current Organic Chemistry.
This work is licensed under a Creative Commons Attribution 4.0 International License.
All articles published in our journal are licensed under CC-BY 4.0, which permits authors to retain copyright of their work. This license allows for unrestricted use, sharing, and reproduction of the articles, provided that proper credit is given to the original authors and the source.