Biofunctional activity and phytochemical composition by LC-QToF-MS of Mitragyna speciosa Korth leaves extracted in Northern Thailand
Main Article Content
Keywords
Mitragyna speciosa Korth; Antioxidant activity; Anti-inflammatory; Flavonoid; Phenolic; LC–QToF-MS
Abstract
Mitragyna speciosa Korth, a traditional Thai herb, is known for its medicinal properties, including enhancing physical strength and promoting relaxation. However, its misuse can lead to addiction. This study evaluated the antioxidant capacity, anti-inflammatory activity, and chemical composition of M. speciosa using liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-QToF-MS). Leaves were extracted with ethanol, methanol, and water. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay showed that ethanol extraction had the best IC50 value (0.23 ± 0.01 mg/mL). Methanol extraction excelled in the 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay (2.24 ± 0.33 mg/mL), while water extraction displayed the highest antioxidant capacity in the fluorescence recovery after photobleaching (FRAP) assay (231.87 ± 11.51 mg ascorbic acid equivalent [AAE]/100 g). The water extract exhibited the highest total phenolic content (153.41 ± 0.06 mg gallic acid equivalent [GAE]/g) and total flavonoid content (6.24 ± 0.83 mg QE/g). It also provided the maximum anti-inflammatory protection, with a protection rate of 96.35% and a hemolysis of 3.65%. LC-QToF-MS analysis identified key bioactive compounds, such as stigmatellin Y (99.17%) and mitragynine (98.58%), along with other compounds with antioxidant and anti-inflammatory activities. These findings highlight M. speciosa’s potential of being a therapeutic agent. Future research should focus on elucidating the molecular mechanisms of its bioactive compounds, optimizing extraction techniques, and conducting in vivo and clinical studies to validate its safety and therapeutic efficacy.
References
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Khalil, S., Abdullah, S.A.J. and Ahmad, R. 2020. Enforcement status of the Poison Act 1952 against offences related to kratom (Mitragyna speciosa Korth) misuse in Malaysia. UUM Journal of Legal Studies (UUMJLS) 11(1): 75–93. https://doi.org/10.32890/uumjls.11.1.2020.6928
Matsumoto, K., Hatori, Y., Murayama, T., Tashima, K., Wongseripipatana, S., Misawa, K. and Horie, S. 2006. Involvement of μ-opioid receptors in antinociception and inhibition of gastrointestinal transit induced by 7-hydroxymitragynine, isolated from Thai herbal medicine Mitragyna speciosa. European Journal of Pharmacology 549(1–3): 63–70. https://doi.org/10.1016/j.ejphar.2006.08.013
Nakaphan, T., Teerachaisakul, M., Puttum, S., Sompimai, K. and Nootim, P. 2016. Traditional uses of kratom (Mitragyna speciosa Korth) among folk healers in southern Thailand. Journal of Traditional Thai and Alternative Medicine 14(3): 274–285.
Parthasarathy, S., Azizi, J.B., Ramanathan, S., Ismail, S., Sasidharan, S., Said, M.I.M. and Mansor, S.M. 2009. Evaluation of antioxidant and antibacterial activities of aqueous, methanolic and alkaloid extracts from Mitragyna speciosa (Rubiaceae family) leaves. Molecules 14(10): 3964–3974. https://doi.org/10.3390/molecules14103964
Ponglux, D., Wongseripipatana, S., Takayama, H., Kikuchi, M., Kurihara, M., Kitajima, M., ... & Sakai, S.I. 1994. A new indole alkaloid, 7 α-hydroxy-7H-mitragynine from Mitragyna speciosa in Thailand. Planta Medica, 60(06): 580–581. https://doi.org/10.1055/s-2006-959578
Racho, M.M.M., Narceda, R.J.A. and Uy Jr, S.P. 2022, Oct. Hierarchical cluster analysis potential application on suspected plant-based new psychoactive substances (NPS) found in the Philippines using FT-IR/ATR spectral data. Forensic Asia 12: 11–16.
Rech, M.A., Donahey, E., Cappiello Dziedzic, J.M., Oh, L. and Greenhalgh, E. 2015. New drugs of abuse. Pharmacotherapy: The Journal of Human Pharmacology and Drug Therapy 35(2): 189–197. https://doi.org/10.1002/phar.1522
Salim, H.M., Puspitarini, M.D., Setiwati, Y. and Shimabukuro, M. 2021. Antibacterial mechanism of Kratom (Mitragyna speciosa) methanol extract on Streptococcus pneumoniae and Eschericia coli bacteria. Biomolecular and Health Science Journal 4(2): 99–103. https://doi.org/10.20473/bhsj.v4i2.28933
Sengnon, N., Vonghirundecha, P., Chaichan, W., Juengwatanatrakul, T., Onthong, J., Kitprasong, P. and Wungsintaweekul, J. 2023. Seasonal and geographic variation in alkaloid content of kratom (Mitragyna speciosa (Korth.) Havil.) from Thailand. Plants, 12(4): 949. https://doi.org/10.3390/plants12040949
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Tagrida, M. and Benjakul, S. 2021. Betel (Piper betle L.) leaf ethanolic extracts dechlorophyllized using different methods: antioxidant and antibacterial activities, and application for shelf-life extension of Nile tilapia (Oreochromis niloticus) fillets. RSC Advances, 11(29): 17630–17641. https://doi.org/10.1039/D1RA02464G
Veltri, C. and Grundmann, O. 2019. Current perspectives on the impact of Kratom use. Substance Abuse and Rehabilitation 10: 23–31. https://doi.org/10.2147/SAR.S164261
Zakaria, F., Anuar, N.N.M., Hisam, N.S.N., Tan, J.K., Zakaria, F., Fauzi, S.M.M. and Ashari, S.E. 2023. An investigation of the in vitro wound healing potential of Mitragyna speciosa (Korth.) Havil leaf ultrasound-assisted methanol crude extract and fractions. Biocatalysis and Agricultural Biotechnology 50: 102707. https://doi.org/10.1016/j.bcab.2023.102707
Azizi, J., Ismail, S., Mordi, M.N., Ramanathan, S., Said, M.I.M. and Mansor, S.M. 2010. In vitro and in vivo effects of three different Mitragyna speciosa Korth leaf extracts on phase II drug metabolizing enzymes—glutathione transferases (GSTs). Molecules 15(1): 432–441. https://doi.org/10.3390/molecules15010432
Benjakul, S., Kittiphattanabawon, P., Sumpavapol, P. and Maqsood, S. 2014. Antioxidant activities of lead (Leucaena leucocephala) seed as affected by extraction solvent, prior dechlorophyllisation and drying methods. Journal of Food Science and Technology 51: 3026–3037. https://doi.org/10.1007/s13197-012-0846-1
Chear, N.J.Y., León, F., Sharma, A., Kanumuri, S.R.R., Zwolinski, G., Abboud, K.A., ... & McCurdy, C.R. 2021; Hassan, Z., Muzaimi, M., Navaratnam, V., Yusoff, N.H., Suhaimi, F.W., Vadivelu, R., ... and Müller, C.P. 2013. Exploring the chemistry of alkaloids from Malaysian Mitragyna speciosa (Kratom) and the role of oxindoles on human opioid receptors. Journal of Natural Products 84(4): 1034–1043. https://doi.org/10.1021/acs.jnatprod.0c01055
Chippada, S.C., Volluri, S.S., Bammidi, S.R. and Vangalapati, M. 2011. In vitro anti-inflammatory activity of methanolic extract of Centella asiatica by HRBC membrane stabilisation. Rasayan Journal of Chemistry 4(2): 457–460.
Colucci, S., Culbreth, S., Alsarraf, E. and Fanikos, J. 2019. Why does the food and drug administration need to ban kratom. Current Emergency and Hospital Medicine Reports 7: 169–174. https://doi.org/10.1007/s40138-019-00201-5
Dewanto, V., Wu, X., Adom, K.K. and Liu, R.H. 2002. Thermal processing enhances the nutritional value of tomatoes by increasing total antioxidant activity. Journal of Agricultural and Food Chemistry, 50(10): 3010–3014. https://doi.org/10.1021/jf0115589
Eastlack, S.C., Cornett, E.M. and Kaye, A.D. 2020. Kratom—pharmacology, clinical implications, and outlook: a comprehensive review. Pain and Therapy 9: 55–69. https://doi.org/10.1007/s40122-020-00151-x
Firmansyah, A., Sundalian, M. and Taufiq, M. 2021. Kratom (Mitragyna speciosa Korth) for a new medicinal: a review of pharmacological and compound analysis. Biointerface Research in Applied Chemistry, 11(2): 9704–9718. https://doi.org/10.33263/BRIAC112.97049718
Goh, Y.S., Karunakaran, T., Murugaiyah, V., Santhanam, R., Abu Bakar, M.H. and Ramanathan, S. 2021. Accelerated solvent extractions (ASE) of Mitragyna speciosa Korth (Kratom) leaves: evaluation of its cytotoxicity and antinociceptive activity. Molecules 26(12): 3704. https://doi.org/10.3390/molecules26123704
Handa, S.S., Khanuja, S.P.S., Longo, G. and Rakesh, D.D. 2008. “Extraction Technologies for Medicinal and Aromatic Plants,” No. 66. United Nations Industrial Development Organization, Vienna, Austria, and International Centre for Science and High Technology. Trieste, Italy, pp. 21–25.
Hassan, Z., Muzaimi, M., Navaratnam, V., Yusoff, N.H., Suhaimi, F.W., Vadivelu, R., ... and Müller, C.P. 2013. From Kratom to mitragynine and its derivatives: physiological and behavioural effects related to use, abuse, and addiction. Neuroscience & Biobehavioral Reviews 37(2): 138–151. https://doi.org/10.1016/j.neubiorev.2012.11.012
Jeeno, P., Tongban, S., Yana, P., Wongta, A., Sutan, K., Yadoung, S. and Hongsibsong, S. 2022. Tentative identification of phytochemicals from Smilax glabra and Smilax corbularia extracts by LC-QToF-MS and their bioactive potential. Plants 11(16): 2089. https://doi.org/10.3390/plants11162089
Kang, W.Y., Li, C.F. and Liu, Y.X. 2010. Antioxidant phenolic compounds and flavonoids of Mitragyna rotundifolia (Roxb.) Kuntze in vitro. Medicinal Chemistry Research 19: 1222–1232. https://doi.org/10.1007/s00044-009-9265-x
Kek, S.P., Chin, N.L., Yusof, Y.A., Tan, S.W. and Chua, L.S. 2017. Classification of entomological origin of honey based on its physicochemical and antioxidant properties. International Journal of Food Properties 20(Sup 3): S2723–S2738. https://doi.org/10.1080/10942912.2017.1359185
Khalil, S., Abdullah, S.A.J. and Ahmad, R. 2020. Enforcement status of the Poison Act 1952 against offences related to kratom (Mitragyna speciosa Korth) misuse in Malaysia. UUM Journal of Legal Studies (UUMJLS) 11(1): 75–93. https://doi.org/10.32890/uumjls.11.1.2020.6928
Matsumoto, K., Hatori, Y., Murayama, T., Tashima, K., Wongseripipatana, S., Misawa, K. and Horie, S. 2006. Involvement of μ-opioid receptors in antinociception and inhibition of gastrointestinal transit induced by 7-hydroxymitragynine, isolated from Thai herbal medicine Mitragyna speciosa. European Journal of Pharmacology 549(1–3): 63–70. https://doi.org/10.1016/j.ejphar.2006.08.013
Nakaphan, T., Teerachaisakul, M., Puttum, S., Sompimai, K. and Nootim, P. 2016. Traditional uses of kratom (Mitragyna speciosa Korth) among folk healers in southern Thailand. Journal of Traditional Thai and Alternative Medicine 14(3): 274–285.
Parthasarathy, S., Azizi, J.B., Ramanathan, S., Ismail, S., Sasidharan, S., Said, M.I.M. and Mansor, S.M. 2009. Evaluation of antioxidant and antibacterial activities of aqueous, methanolic and alkaloid extracts from Mitragyna speciosa (Rubiaceae family) leaves. Molecules 14(10): 3964–3974. https://doi.org/10.3390/molecules14103964
Ponglux, D., Wongseripipatana, S., Takayama, H., Kikuchi, M., Kurihara, M., Kitajima, M., ... & Sakai, S.I. 1994. A new indole alkaloid, 7 α-hydroxy-7H-mitragynine from Mitragyna speciosa in Thailand. Planta Medica, 60(06): 580–581. https://doi.org/10.1055/s-2006-959578
Racho, M.M.M., Narceda, R.J.A. and Uy Jr, S.P. 2022, Oct. Hierarchical cluster analysis potential application on suspected plant-based new psychoactive substances (NPS) found in the Philippines using FT-IR/ATR spectral data. Forensic Asia 12: 11–16.
Rech, M.A., Donahey, E., Cappiello Dziedzic, J.M., Oh, L. and Greenhalgh, E. 2015. New drugs of abuse. Pharmacotherapy: The Journal of Human Pharmacology and Drug Therapy 35(2): 189–197. https://doi.org/10.1002/phar.1522
Salim, H.M., Puspitarini, M.D., Setiwati, Y. and Shimabukuro, M. 2021. Antibacterial mechanism of Kratom (Mitragyna speciosa) methanol extract on Streptococcus pneumoniae and Eschericia coli bacteria. Biomolecular and Health Science Journal 4(2): 99–103. https://doi.org/10.20473/bhsj.v4i2.28933
Sengnon, N., Vonghirundecha, P., Chaichan, W., Juengwatanatrakul, T., Onthong, J., Kitprasong, P. and Wungsintaweekul, J. 2023. Seasonal and geographic variation in alkaloid content of kratom (Mitragyna speciosa (Korth.) Havil.) from Thailand. Plants, 12(4): 949. https://doi.org/10.3390/plants12040949
Shaik Mossadeq, W.M., Sulaiman, M.R., Tengku Mohamad, T.A., Chiong, H.S., Zakaria, Z.A., Jabit, M.L., ... and Israf, D.A. 2009. Anti-inflammatory and antinociceptive effects of Mitragyna speciosa Korth methanolic extract. Medical Principles and Practice 18(5): 378–384. https://doi.org/10.1159/000226292
Tagrida, M. and Benjakul, S. 2021. Betel (Piper betle L.) leaf ethanolic extracts dechlorophyllized using different methods: antioxidant and antibacterial activities, and application for shelf-life extension of Nile tilapia (Oreochromis niloticus) fillets. RSC Advances, 11(29): 17630–17641. https://doi.org/10.1039/D1RA02464G
Veltri, C. and Grundmann, O. 2019. Current perspectives on the impact of Kratom use. Substance Abuse and Rehabilitation 10: 23–31. https://doi.org/10.2147/SAR.S164261
Zakaria, F., Anuar, N.N.M., Hisam, N.S.N., Tan, J.K., Zakaria, F., Fauzi, S.M.M. and Ashari, S.E. 2023. An investigation of the in vitro wound healing potential of Mitragyna speciosa (Korth.) Havil leaf ultrasound-assisted methanol crude extract and fractions. Biocatalysis and Agricultural Biotechnology 50: 102707. https://doi.org/10.1016/j.bcab.2023.102707
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