A Phytochemical Screening and FTIR Analysis for Fractions of Ethanol Extract from Mitragyna speciosa leaves
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Abstract
Introduction: Mitragyna speciosa, also referred to as kratom, has long been utilised for its stimulant and analgesic effects, which are mostly due to its high alkaloid content. Most of the study of M. speciosa utilised methanol as the solvent of extraction. However, ethanol is a more environmentally friendly and nontoxic solvent that can also be used for the extraction process. The purpose of this study was to use ethanol to extract phytochemicals from M. speciosa leaves and separate the bioactive compounds by using Vacuum Liquid Chromatography (VLC). Methods: M. speciosa leaves underwent ethanol maceration followed by phytochemical screening with Dragendorffs, Shinoda, Salkowski, FeCl?, foam tests and VLC fractionation. The resulting five fractions were analysed by TLC and screened for alkaloids, phenolic, unsaturated compounds and flavonoids along with analysation of the functional groups by using the Fourier Transform Infrared Spectroscopy (FTIR). Results: Screening of the crude ethanol extract exhibits positive reactions for alkaloids, flavonoids, terpenoids, phenolics, tannins and saponin. Meanwhile, the TLC analysis of the fractions using solvent system n-hexane: ethyl acetate (30:20) shows presence of alkaloid compound with Rf value of 0.80 for fraction 3 and 0.83 for fraction 2,4 and 5 which aligns with the literature for mitragynine. FTIR results exhibit presence of important peaks of mitragynine such as C=O, N-H and C-H for the fractions while C=C, can only be found in fraction 3. Conclusion: This study supports the use of ethanol as solvent of extraction and proved that VLC is highly efficient in its ability to fractionate the ethanol extract of M. speciosa.
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References
Annuar, N. A. K., Azlan, U. K., Mediani, A., Tong, X., Han, R., Al-Olayan, E., Baharum, S. N., Bunawan, H., Sarian, M. N., Hamezah, H. S., & Jantan, I. (2024). An insight review on the neuropharmacological effects, mechanisms of action, pharmacokinetics and toxicity of mitragynine. Biomedicine & Pharmacotherapy, 171, 116-134. https://doi.org/10.1016/j.biopha.2024.116134
Azman, A. S., & Mat So’ad, S. Z. (2024). The Investigation of Phytochemicals and Antioxidant Properties of Champereia manillana(Blume) Merr Stem Bark. Journal of Pharmacy, 4(2), 151–164. https://doi.org/10.31436/jop.v4i2.303
Begum, T., Ahmed, Q. U., Arzmi, M. H., Khatib, A., Abbas, S. A., & Aziz, Z. H. A. (2025). Phytochemical Screening and Evaluation of in vitro Antioxidant, Antibacterial and Antibiofilm Activities of Mitragyna speciosa Leaf Extract. Malaysian Journal of Chemistry, 27(5), 80-94.
Beng, G. T., Hamdan, M. R., Siddiqui, M. J., Mordi, M. N., & Mansor, S. M. (2011). A Simple and CostEffective Isolation and Purification Protocol of Mitragynine From Mitragyna speciosa Korth (Ketum) Leaves. Malaysian Journal of Analytical Sciences, 15(1), 54-60.
Devmurari, V., Pandey, S., Goyani, M., & Jivani, N. (2010). Phytochemical Screening of Ethanol Extract of Artemisia Nilagirica. International Journal of Chemical Sciences, 8(4), 2099–2104. https://www.tsijournals.com/articles/phytochemical-screening-of-ethanolic-extract-of-artemisia-nilagirica.pdf
Dhanani, T., Shah, S., Gajbhiye, N. A., & Kumar, S. (2017). Effect of extraction methods on yield, phytochemical constituents and antioxidant activity of Withania somnifera. Arabian Journal of Chemistry, 10, S1193–S1199. https://doi.org/10.1016/j.arabjc.2013.02.015
Dragendorff reagent | Alkaloids Detection Agent | MedChemExpress. (n.d.). MedchemExpress.Com. Retrieved January 7, 2026, from https://www.medchemexpress.com/dragendorff-reagent.html
Eastlack, S. C., Cornett, E. M., & Kaye, A. D. (2020). Kratom—Pharmacology, Clinical Implications, and Outlook: A Comprehensive Review. Pain and Therapy, 9(1), 55–69. https://doi.org/10.1007/s40122-020-00151-x
Ernawati, T., Hermawan, F., & Kusumaningrum, S. (2024). A Review on Isolation, Characterization, Biosynthesis, Synthesis, Modification, Pharmacological Activities and Toxicology of Mitragynine. Gazi University Journal of Science, 37(4), 1654–1672. https://doi.org/10.35378/gujs.1395354
Garg, J., Ghoshal, G., Bhadada, S. K., & Katare, O. P. (2024). Derivatisation mechanistic-guided identification of phytoconstituents of different extracts of Cissus quadrangularis by TLC and standardization by HPTLC. Phytomedicine Plus, 4(3), 100601. https://doi.org/10.1016/j.phyplu.2024.100601
Haspi, N. F. Z., Mat So’Ad, S. Z., Arzmi, M. H., & Khatib, A. (2025). Sequential Extraction and Phytochemical Study of the Extracts of Mitragyna speciosa. Malaysian Journal of Chemistry,27(1),84-91.
https://doi.org/10.55373/mjchem.v27i1.84
Karunakaran, T., Ngew, K. Z., Zailan, A. A. D., Mian Jong, V. Y., & Abu Bakar, M. H. (2022). The Chemical and Pharmacological Properties of Mitragynine and Its Diastereomers: An Insight Review. Frontiers in Pharmacology, 13, 805-986. https://doi.org/10.3389/fphar.2022.805986
Phoonket, S., Yisarakun, W., Srikoon, P., & Poolpol, K. (2025). Anti-migration and Anti-invasion Effects of Mitragyna speciosa Polar Extract on A549 Cell. Trends in Sciences, 22(12), 10150–10150. https://doi.org/10.48048/tis.2025.10150
Prasetya, F., Indriyanti, N., Mus, N. M., Bafadal, M., Fadilla, R., Widiyastuti, Y., Chaidir, C., Kuncoro, H., Fajriah, S., Heryanto, R., Narsa, A. C., Fricillia, O. Z., Sastyarina, Y., Fitriani, V. Y., Rouchmana, S., Sobah, N., Bahar, Z., Nisaa, N. R. K., Helmi, H., & Anshory, H. (2026). Kratom (Mitragyna speciosa) as a Phytochemical-Based Natural Product Exhibiting Opioid-like Analgesic Effects with Reduced Tolerance and Dependence Liability via TLR4-Associated Neuroimmune Modulation. Molecules, 31(9), 1428. https://doi.org/10.3390/molecules31091428
Ramanathan, S., & McCurdy, C. R. (2020). Kratom (Mitragyna speciosa): Worldwide issues. Current Opinion in Psychiatry, 33(4), 312–318. https://doi.org/10.1097/YCO.0000000000000621
Sokup, B., & Pippin, M. M. (2025). Kratom. In StatPearls. StatPearls Publishing. http://www.ncbi.nlm.nih.gov/books/NBK585120/
León, F., Habib, E., Adkins, J. E., Furr, E. B., McCurdy, C. R., & Cutler, S. J. (2009). Phytochemical Characterization of the Leaves of Mitragyna speciosa Grown in USA. Natural Product Communications, 4(7), 907–910.
Masriani, M., Muharini, R., Wijayanti, D. K., Melanie, P., & Widiansari, M. L. (2023). Phytochemical Screening of Ethanol Extracts from Three Variants of Kratom Leaves (Mitragyna speciosa Korth.). Hydrogen. Jurnal Kependidikan Kimia, 11(2), 192. https://doi.org/10.33394/hjkk.v11i2.7122
Mohd Zubri, N. S., Amirah Azhar, N. A., Amran, N. A., Weng, S. I. S., Awal, A., & Jusoh, S. A. (2025). Interactions of Kratom Alkaloids–Mitragynine,7-Hydroxymitragynine, and Mitragynine Pseudoindoxyl with Lipid Bilayers: A Molecular Dynamics Study. International Journal of Pharmaceuticals, Nutraceuticals and Cosmetic Science, 8(1), 124–139. https://doi.org/10.24191/IJPNaCS.v8i1.09
Mustafa, R., Sukor, R., Mariam, S., Mohd Nor, S. M., Saari, N., & Azri, F. (2020). Enhancing Extraction Yield and Purity of Mitragynine From Mitragyna speciosa Through Sequential Solvent Extraction and characterisation Using NMR Technique. International Journal of Scientific & Technology Research, 9, 3846–3854.
Ramanathan, S., & McCurdy, C. R. (2020). Kratom (Mitragyna speciosa): Worldwide issues. Current Opinion in Psychiatry, 33(4), 312–318. https://doi.org/10.1097/YCO.0000000000000621
Setyawati, H. (2020). Uji Aktivitas Antioksidan Ekstrak etanol Daun Kratom (Mitragyna Speciosa) Dengan Metode 1, 1 Difenil-2-Pikrihidrazil (DPPH). Jurnal Farmasi Udayana, 204. https://doi.org/10.24843/jfu.2020.v09.i03.p09
Sokup, B., & Pippin, M. M. (2025). Kratom. In StatPearls. StatPearls Publishing. http://www.ncbi.nlm.nih.gov/books/NBK585120/
Tuntiyasawasdikul, S., Junlatat, J., Tabboon, P., Limpongsa, E., & Jaipakdee, N. (2024). Mitragyna speciosa ethanolic extract: Extraction, anti-inflammatory, cytotoxicity, and transdermal delivery assessments. Industrial Crops and Products, 208, 117909. https://doi.org/10.1016/j.indcrop.2023.117909