In vitro comparative study of enzyme inhibitory properties of herbal plants and their phenolic, tannin, and flavonoid content as natural inhibitors in public health
Main Article Content
Keywords
amylase, Cinnamomum verum, Eucalyptus camaldulensis, glucosidase, Laurus nobilis, Nerium oleander
Abstract
Medicinal herbs are used to treat numerous diseases all over the world. This study investigates the enzyme inhibitory potential, antioxidant properties, and phytochemical composition of methanolic extracts from Cinnamomum verum, Nerium oleander, Laurus nobilis, and Eucalyptus camaldulensis leaves. These plants, known for their healing properties, were tested to determine how effectively they inhibit α-amylase and α-glucosidase. These essential enzymes help break down carbohydrates, and their antioxidant capacities were evaluated using DPPH and ABTS assays. Phytochemical profiling revealed significant levels of phenolics (26.44–63.56 mg GAE/g), flavonoids (6.09–37.56 mg QE/g), and tannins (12.98–34.80 mg TAE/g), with E. camaldulensis and L. nobilis exhibiting the highest phenolic content. Methanolic extracts of L. nobilis and N. oleander demonstrated the strongest inhibition of α-amylase (0.586 ± 0.015 and 0.564 ± 0.0001 mmol acarbose/g) and α-glucosidase (7.570 ± 0.107 and 8.242 ± 0.113 mmol acarbose/g), outperforming synthetic controls. Antioxidant activity correlated with phenolic content, with E. camaldulensis showing the lowest IC50 in DPPH (8.83 µg/mL) and L. nobilis in ABTS (30.32 µg/mL). These findings highlight the therapeutic promise of these plants as natural sources of enzyme inhibitors and antioxidants, supporting their use in oxidative stress-related disorders. The study underscores the significance of plant-derived bioactive compounds in developing safer, cost-effective pharmaceuticals and functional foods.
References
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Rauf, A., Bawazeer, S., Naseer, M., Alhumaydhi, F. A., Aljohani, A. S., Habib, A., Khan, R., Jehan, U., Qureshi, M. N., & Khan, M. (2021). In vitro α-glucosidase and urease enzyme inhibition profile of some selected medicinal plants of Pakistan. Natural Product Research, 35(23), 5434–5440. 10.1080/14786419.2020.1779264
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Tungmunnithum, D., Thongboonyou, A., Pholboon, A., & Yangsabai, A. (2018). Flavonoids and other phenolic compounds from medicinal plants for pharmaceutical and medical aspects: An overview. Medicines, 5(3), 93. 10.3390/medicines5030093
Khodja, Y. K., Bachir-Bey, M., Belmouhoub, M., Ladjouzi, R., Dahmoune, F., & Khettal, B. (2023). The botanical study, phytochemical composition, and biological activities of Laurus nobilis L. leaves: A review. International Journal of Secondary Metabolite, 10(2), 269–296. 10.21448/ijsm.1171836
Visvanathan, R., Qader, M., Jayathilake, C., Jayawardana, B. C., Liyanage, R., & Sivakanesan, R. (2020). Critical review on conventional spectroscopic α-amylase activity detection methods: Merits, demerits, and future prospects. Journal of the Science of Food and Agriculture, 100(7), 2836–2847. 10.1002/jsfa.10315
Lu, H., Xie, T., Wu, Q., Hu, Z., Luo, Y., & Luo, F. (2023). Alpha-glucosidase inhibitory peptides: Sources, preparations, identifications, and action mechanisms. Nutrients, 15(19), 4267. 10.3390/nu15194267
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Ayouaz, S., Oliveira-Alves, S. C., Lefsih, K., Serra, A. T., Da Silva, A. B., Samah, M., Karczewski, J., Madani, K., & Bronze, M. R. (2020). Phenolic compounds from Nerium oleander leaves: Microwave-assisted extraction, characterization, antiproliferative and cytotoxic activities. Food & Function, 11(7), 6319. 10.1039/D0FO01180K
Al Dhaheri A. S., Alkhatib, D. H., Jaleel, A., Tariq, M. M., Feehan, J., Apostolopoulos, V., & Stojanovska, L. (2023). Proximate composition and mineral content of spices increasingly employed in the Mediterranean diet. Journal of Nutritional Science, 12, e79. 10.1017/jns.2023.52
Bourais, I., Elmarrkechy, S., Taha, D., Mourabit, Y., Bouyahya, A., El Yadini, M., & Iba, N. (2023). A review on medicinal uses, nutritional value, and antimicrobial, antioxidant, anti-inflammatory, antidiabetic, and anticancer potential related to bioactive compounds of J. regia. Food Reviews International, 39(9), 6199–6249. 10.1080/87559129.2022.2094401
Basak, S. S., & Candan, F. (2013). Effect of Laurus nobilis L. essential oil and its main components on α-glucosidase and reactive oxygen species scavenging activity. Iranian Journal of Pharmaceutical Research, 12(2), 367–379.
Batiha, G. E.-S., Beshbishy, A. M., Guswanto, A., Nugraha, A., Munkhjargal, T., Abdel-Daim, M. M., Mosqueda, J., & Igarashi, I. (2020). Phytochemical characterization and chemotherapeutic potential of Cinnamomum verum extracts on the multiplication of protozoan parasites in vitro and in vivo. Molecules, 25(4), 996. 10.3390/molecules25040996
Chandorkar, N., Tambe, S., Amin, P., & Madankar, C. (2021). A systematic and comprehensive review on current understanding of the pharmacological actions, molecular mechanisms, and clinical implications of the genus Eucalyptus. Phytomedicine Plus, 1(4), 100089. 10.1016/j.phyplu.2021.100089
Kumar, M. D., Gowrishankar, S., Harikrishna, S. I., Nithish, S., Nidamanuri, B. S., Praharsh, K. M., & Jawahar, N. (2022). Mini review on chewing gum for oral hygiene. Journal of Pharmaceutical Sciences and Research, 14(6), 769–775.
Christoforidi, I., Kollaros, D., Manios, T., & Daliakopoulos, I. N. (2022). Drought-and salt-tolerant plants of the Mediterranean and their diverse applications: The case of Crete. Land, 11(11), 2038. 10.3390/land11112038
Dobroslavić, E., Repajić, M., Dragović-Uzelac, V., & Elez Garofulić, I. (2022). Isolation of Laurus nobilis leaf polyphenols: A review on current techniques and future perspectives. Foods, 11(2), 235. 10.3390/foods11020235
Hirsch, H., Allsopp, M. H., Canavan, S., Cheek, M., Geerts, S., Geldenhuys, C. J., Harding, G., Hurley, B. P., Jones, W., & Keet, J.-H. (2020). Eucalyptus camaldulensis in South Africa–Past, present, future. Transactions of the Royal Society of South Africa, 75(1), 1–16. 10.1080/0035919X.2019.1669732
Hussain, H. A., Islam, M., Saeed, H., Ahmad, A., Hussain, A., & Rafay, M. Z. (2025). Insight to phytochemical investigation and anti-hyperlipidemic effects of Eucalyptus camaldulensis leaf extract using in vitro, in vivo and in silico approach. Journal of Biomolecular Structure and Dynamics, 43(1), 325–347. 10.1080/07391102.2023.2280814
Huang, Z., Pang, D., Liao, S., Zou, Y., Zhou, P., Li, E., & Wang, W. (2021). Synergistic effects of cinnamaldehyde and cinnamic acid in cinnamon essential oil against S. pullorum. Industrial Crops and Products, 162, 113296. 10.1016/j.indcrop.2021.113296
Johnson, J., Collins, T., Walsh, K., & Naiker, M. (2020). Solvent extractions and spectrophotometric protocols for measuring the total anthocyanin, phenols and antioxidant content in plums. Chemical Papers, 74(12), 4481–4494. 10.1007/s11696-020-01261-8
Limam, H., Ezzine, Y., Tammar, S., Ksibi, N., Selmi, S., Del Re, G., Ksouri, R., & Msaada, K. (2021). Phenolic composition and antioxidant activities of thirteen Eucalyptus species cultivated in north east of Tunisia. Plant Biosystems-An International Journal Dealing with All Aspects of Plant Biology, 155(3), 587–597. 10.1080/11263504.2020.1762791
Mahmoud Dogara, A., Ahmed Al-Zahrani, A., Bradosty, S. W., Hamad, S. W., Bapir, S. H., & Anwar, T. K. (2024). Antioxidant, α-glucosidase, antimicrobial activities chemical composition and in silico analysis of Eucalyptus camaldulensis Dehnh. AIMS Biophysics, 11(3), 255–280. 10.3934/biophy.2024015
Mouhcine, M., Amin, L., Saaid, A., Khalil, H., & Laila, B. (2019). Cytotoxic, antioxidant and antimicrobial activities of Nerium oleander collected in Morocco. Asian Pacific Journal of Tropical Medicine, 12(1), 32–38. 10.4103/1995-7645.250342
Mutlu-Ingok, A., Devecioglu, D., Dikmetas, D. N., & Karbancioglu-Guler, F. (2022). Advances in biological activities of essential oils. In Studies in Natural Products Chemistry (Vol. 74, pp. 331–362). 10.1016/B978-0-323-91099-6.00010-4
Omar, N., Ismail, C. N., & Long, I. (2022). Tannins in the treatment of diabetic neuropathic pain: Research progress and future challenges. Frontiers in Pharmacology, 12, 805854. 10.3389/fphar.2021.805854
Rauf, A., Bawazeer, S., Naseer, M., Alhumaydhi, F. A., Aljohani, A. S., Habib, A., Khan, R., Jehan, U., Qureshi, M. N., & Khan, M. (2021). In vitro α-glucosidase and urease enzyme inhibition profile of some selected medicinal plants of Pakistan. Natural Product Research, 35(23), 5434–5440. 10.1080/14786419.2020.1779264
Redha, A. A. (2020). Phytochemical investigations of Nerium oleander L. leaves and flowers. International Journal of Scientific Research in Chemical Sciences, 7(4).
Sharma, T., Gamit, R., Acharya, R., & Shukla, V. J. (2021). Quantitative estimation of total tannin, alkaloid, phenolic, and flavonoid content of the root, leaf, and whole plant of Byttneria herbacea Roxb. International Quarterly Journal of Research in Ayurveda, 42(3), 143–147. 10.4103/ayu.AYU_25_19
Sanna, G., Madeddu, S., Serra, A., Collu, D., Efferth, T., Hakkim, F. L., & Rashan, L. (2021). Anti-poliovirus activity of Nerium oleander aqueous extract. Natural Product Research, 35(4), 633–640. 10.1080/14786419.2019.1582046
Singh, P., Dhole, B., Choudhury, J., Tuli, A., Pandey, D., Velpandian, T., Gupta, S., & Chaturvedi, P. K. (2024). Calotropis procera extract inhibits prostate cancer through regulation of autophagy. Journal of Cellular and Molecular Medicine, 28(6), e18050. 10.1111/jcmm.18050
Chebabe, M., Elkhoudri, N., Chahboune, M., Laamiri, F. Z., Marc, I., & Mochhoury, L. (2025). Study of chemical characterization of medicinal plants used for traditional medicine: A review. Biomolecules and Pharmacology Journal, 18(1), 535–546. 10.13005/bpj/3106
Kurek, M., Benaida-Debbache, N., Elez Garofulić, I., Galić, K., Avallone, S., Voilley, A., & Waché, Y. (2022). Antioxidants and bioactive compounds in food: Critical review of issues and prospects. Antioxidants, 11(4), 742. 10.3390/antiox11040742
Mohammed, W. H., Soud, S. A., Hameed, A. H., & Hussein, N. N. (2021). Assessment of the cytotoxic activity of alcoholic extract of Eucalyptus camaldulensis on breast cancer cell line. Archives of Razi Institute, 76(5), 1307–1314.
Ouattara, B., Semay, I., Ouédraogo, J. C. W., Gerbaux, P., & Ouédraogo, I. W. (2024). Optimization of the extraction of phenolic compounds from Eucalyptus camaldulensis Dehnh leaves using response surface methodology. Chemistry Africa, 7(3), 1251–1267. 10.1007/s42250-023-00821-1
Pathak, R., & Sharma, H. (2021). A review on medicinal uses of Cinnamomum verum (Cinnamon). Journal of Drug Delivery and Therapeutics, 11(6-S), 161–166. 10.22270/jddt.v11i6-S.5145
El Faqer, O., Elkoraichi, I., Latif, M., Debierre-Grockiego, F., Ouadghiri, Z., Rais, S., Dimier-Poisson, I., & Mtairag, E. M. (2024). Pharmacological insights into Laurus nobilis: HPLC profiling and evaluation of its anti-toxoplasma, antioxidant, and anti-hemolytic properties. Biochemical Systematics and Ecology, 117, 104891. 10.1016/j.bse.2024.104891
Chaachouay, N., & Zidane, L. (2024). Plant-derived natural products: A source for drug discovery and development. Drugs and Drug Candidates, 3(1), 184–207. 10.3390/ddc3010011
Rahman, M. H., Bajgai, J., Fadriquela, A., Sharma, S., Trinh, T. T., Akter, R., … Lee, K. J. (2021). Therapeutic potential of natural products in treating neurodegenerative disorders and their future prospects and challenges. Molecules, 26(17), 5327. 10.3390/molecules26175327
Akter, R., Afrose, A., Rahman, M. R., Chowdhury, R., Nirzhor, S. S. R., Khan, R. I., & Kabir, M. T. (2021). A comprehensive analysis into the therapeutic application of natural products as Sirt6 modulators in Alzheimer’s disease, aging, cancer, inflammation, and diabetes. International Journal of Molecular Sciences, 22(8), 4180. 10.3390/ijms22084180
Chaachouay, N., & Zidane, L. (2024). Plant-derived natural products: A source for drug discovery and development. Drugs and Drug Candidates, 3(1), 184–207. 10.3390/ddc3010011
Tungmunnithum, D., Thongboonyou, A., Pholboon, A., & Yangsabai, A. (2018). Flavonoids and other phenolic compounds from medicinal plants for pharmaceutical and medical aspects: An overview. Medicines, 5(3), 93. 10.3390/medicines5030093
Khodja, Y. K., Bachir-Bey, M., Belmouhoub, M., Ladjouzi, R., Dahmoune, F., & Khettal, B. (2023). The botanical study, phytochemical composition, and biological activities of Laurus nobilis L. leaves: A review. International Journal of Secondary Metabolite, 10(2), 269–296. 10.21448/ijsm.1171836
Visvanathan, R., Qader, M., Jayathilake, C., Jayawardana, B. C., Liyanage, R., & Sivakanesan, R. (2020). Critical review on conventional spectroscopic α-amylase activity detection methods: Merits, demerits, and future prospects. Journal of the Science of Food and Agriculture, 100(7), 2836–2847. 10.1002/jsfa.10315
Lu, H., Xie, T., Wu, Q., Hu, Z., Luo, Y., & Luo, F. (2023). Alpha-glucosidase inhibitory peptides: Sources, preparations, identifications, and action mechanisms. Nutrients, 15(19), 4267. 10.3390/nu15194267
Mohd Zaid, N. A., Sekar, M., Bonam, S. R., Gan, S. H., Lum, P. T., Begum, M. Y., … Fuloria, S. (2023). Promising natural products in new drug design, development, and therapy for skin disorders: An overview of scientific evidence and understanding their mechanism of action. Drug Design, Development and Therapy, 23–66. 10.2147/DDDT.S326332
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