Long-Term Ambient Storage and Retained Hypnotic–Sedative Activity of a Confiscated Mitragyna speciosa Specimen: A Forensic Chemical and Pharmacological Evaluation

Supiyani; Sarah Salsabil

doi:10.32734/jcnar.v7i2.25976

Authors

Supiyani Department of Pharmacy, Faculty of Pharmacy, Universitas Muslim Nusantara Al-Washliyah, Jl. Garu II No. 93 Medan 20147, Indonesia; Forensic Laboratory Unit, North Sumatra Region, Indonesian National Police, Jl. Sisingamangaraja Km.10.5 No. 60 Medan 20148, Indonesia https://orcid.org/0009-0008-6942-6698
Sarah Salsabil Department of Biology, Faculty of Mathematics and Sciences, Universitas Syiah Kuala, Banda Aceh 23111, Aceh, Indonesia https://orcid.org/0009-0006-6854-1905

DOI:

https://doi.org/10.32734/jcnar.v7i2.25976

Keywords:

Forensic Pharmacognosy, Hypnotic–Sedative, Mitragynine, Mitragyna speciosa, Long-Term Stability

Abstract

Mitragyna speciosa (kratom) is a Southeast Asian medicinal plant whose principal alkaloid, mitragynine, exhibits stimulant and sedative properties. Long-term preservation of bioactive constituents in stored plant material is a quality control concern. This study investigated alkaloid stability and pharmacological recoverability of M. speciosa leaf powder stored under ambient, uncontrolled conditions for over ten years following confiscation in 2016. Three extraction solvents, ethanol, acetone, and water–methanol, were employed and their phytochemical profiles compared by Thin-Layer Chromatography with iodoplatinate visualization and Gas Chromatography–Mass Spectrometry (GC–MS). Phytochemical screening revealed consistent presence of alkaloids, sterols, triterpenoids, fatty acids, terpenoids, phenolics, and nitrogen-containing heterocycles across all solvent systems. Acetone yielded the highest relative mitragynine content (8.52%) compared with ethanol (7.01%) and water–methanol (3.64%). Preserved pharmacological activity of the acetone extract was confirmed in vivo using the glass cylinder escape latency test in male mice (Mus musculus), demonstrating a statistically significant, concentration-dependent hypnotic–sedative effect across doses of 4.5–114 mg/kg. One-way ANOVA with Tukey's HSD and Duncan's post hoc tests revealed statistically significant differences among treatment groups (p < 0.001). Within the limitations of this single-specimen evaluation, findings indicate that a detectable alkaloid profile and measurable hypnotic–sedative activity of M. speciosa are retained over extended ambient storage, with solvent selection influencing relative mitragynine recovery; absolute quantification and comparison with a fresh reference specimen were not possible and are recommended. This study provides reference data for forensic evaluation of aged kratom specimens and underscores the need for standardized storage and quality control protocols for kratom preparations.

Downloads

Download data is not yet available.

References

[1] Edinoff AN, Kaufman SE, Mahoney TC, Upshaw WC, Gong J, Cornett EM, Murnane KS, Kaye AM, Varrassi G, Shekoohi S, Kaye AD. Kratom: A Narrative Review of the Possible Clinical Uses and Dangers of This Opioid-Like Plant. Cureus 2024;16:e73058. https://doi.org/10.7759/CUREUS.73058.

[2] Alford AS, Moreno HL, Benjamin MM, Dickinson CF, Hamann MT. Exploring the Therapeutic Potential of Mitragynine and Corynoxeine: Kratom-Derived Indole and Oxindole Alkaloids for Pain Management. Pharmaceuticals 2025;18:222. https://doi.org/10.3390/PH18020222.

[3] Manwill PK, Flores-Bocanegra L, Khin M, Raja HA, Cech NB, Oberlies NH, Todd DA. Kratom (Mitragyna speciosa) Validation: Quantitative Analysis of Indole and Oxindole Alkaloids Reveals Chemotypes of Plants and Products. Planta Med 2022;88:838. https://doi.org/10.1055/A-1795-5876.

[4] Veeramohan R, Zamani AI, Azizan KA, Goh HH, Aizat WM, Razak MFA, Yusof NSM, Mansor SM, Baharum SN, Ng CL. Comparative metabolomics analysis reveals alkaloid repertoires in young and mature Mitragyna speciosa (Korth.) Havil. Leaves. PLoS One 2023;18:e0283147. https://doi.org/10.1371/JOURNAL.PONE.0283147.

[5] Kowalska T, Sajewicz M. Thin-Layer Chromatography (TLC) in the Screening of Botanicals–Its Versatile Potential and Selected Applications. Molecules 2022;27:6607. https://doi.org/10.3390/MOLECULES27196607.

[6] Chatepa LEC, Mwamatope B, Chikowe I, Masamba KG. Effects of solvent extraction on the phytoconstituents and in vitro antioxidant activity properties of leaf extracts of the two selected medicinal plants from Malawi. BMC Complement Med Ther 2024;24:317. https://doi.org/10.1186/S12906-024-04619-7.

[7] Al Muqarrabun LMR, Amrianto A. Mitragynine: A review of its extraction, identification, and purification methods. Current Research in Biosciences and Biotechnology 2021;3:165–71. https://doi.org/10.5614/crbb.2021.3.1/TMPNSA4H.

[8] Lee JE, Jayakody JTM, Kim J Il, Jeong JW, Choi KM, Kim TS, Seo C, Azimi I, Hyun JM, Ryu BM. The Influence of Solvent Choice on the Extraction of Bioactive Compounds from Asteraceae: A Comparative Review. Foods 2024;13:3151. https://doi.org/10.3390/FOODS13193151/S1.

[9] Heywood J, Smallets S, Paustenbach D. Beneficial and adverse health effects of kratom (Mitragyna speciosa): A critical review of the literature. Food and Chemical Toxicology 2024;192:114913. https://doi.org/10.1016/J.FCT.2024.114913.

[10] Chungsiriporn J, Pongyeela P, Chairerk N, Iewkittayakorn J, Duangsuwan W. Drying Characteristics and Mitragynine Content of Kratom Leaves. Chiang Mai Journal of Science 2023;50. https://doi.org/10.12982/CMJS.2023.059.

[11] Mustafa RR, Sukor R, Mariam S, Nor M, Saari N, Azri FA. Enhancing Extraction Yield And Purity Of Mitragynine From Mitragyna speciosa Through Sequential Solvent Extraction And Characterisation Using Nmr Technique 2020.

[12] Isnaeni N, Saefumillah A, Cahyana AH. Preliminary Study of Isolation and Purification Mitragynine from Kratom Leaves. Materials Science Forum, vol. 1061, Trans Tech Publications Ltd; 2022, p. 173–9. https://doi.org/10.4028/p-74ggl5.

[13] Hasanuddin S, Jumarniati, Dewi C. Uji Aktivitas Hipnotik-Sedatif Ekstrak Daun Kirinyuh (Eupatorium odoratum) Pada Mencit (Mus musculus) Jantan. Jurnal Pharmacia Mandala Waluya 2023;2:175–86. https://doi.org/10.54883/jpmw.v2i4.20.

[14] McConnell EL, Basit AW, Murdan S. Measurements of rat and mouse gastrointestinal pH, fluid and lymphoid tissue, and implications for in-vivo experiments. J Pharm Pharmacol 2008;60:63–70. https://doi.org/10.1211/JPP.60.1.0008.

[15] Friatna GN, Khumaisah LL, Anwar DI. Uji Efektivitas Sedatif-Hipnotik Ekstrak Daun Keji Beling (Strobilanthes crispa Blume) terhadap Mencit Putih Jantan Sedative-Hypnotic Effectiveness of the Extract of Strobilanthes crispa Blume in Male Mice. vol. 20. 2023.

[16] Shovkova O V, Klimenko LY, Shpychak O, Shovkova Z V, Ulanova VA, Shpychak OS. Application of thin layer chromatography in analysis of secnidazole, ornidazole, tinidazole and nimorazole. 2018.

[17] Melo T, Figueiredo ARP, da Costa E, Couto D, Silva J, Domingues MR, Domingues P. Ethanol extraction of polar lipids from nannochloropsis oceanica for food, feed, and biotechnology applications evaluated using lipidomic approaches. Mar Drugs 2021;19:593. https://doi.org/10.3390/MD19110593/S1.

[18] Alhusain F, Alshalhoub M, Homaid M Bin, Esba LCA, Alghafees M, Al Deeb M. Clinical presentation and management of methanol poisoning outbreaks in Riyadh, Saudi Arabia: a retrospective analysis. BMC Emerg Med 2024;24:64. https://doi.org/10.1186/S12873-024-00976-1.

[19] Chakraborty S, Uprety R, Slocum ST, Irie T, Le Rouzic V, Li X, Wilson LL, Scouller B, Alder AF, Kruegel AC, Ansonoff M, Varadi A, Eans SO, Hunkele A, Allaoa A, Kalra S, Xu J, Pan YX, Pintar J, Kivell BM, Pasternak GW, Cameron MD, McLaughlin JP, Sames D, Majumdar S. Oxidative Metabolism as a Modulator of Kratom’s Biological Actions. J Med Chem 2021;64:16553–72. https://doi.org/10.1021/ACS.JMEDCHEM.1C01111.

[20] Flores-Bocanegra L, Raja HA, Graf TN, Augustinović M, Wallace ED, Hematian S, Kellogg JJ, Todd DA, Cech NB, Oberlies NH. The Chemistry of Kratom [Mitragyna speciosa]: Updated Characterization Data and Methods to Elucidate Indole and Oxindole Alkaloids. J Nat Prod 2020;83:2165–77. https://doi.org/10.1021/ACS.JNATPROD.0C00257/ASSET/IMAGES/LARGE/NP0C00257_0004.JPEG.

[21] Annuar NAK, Azlan UK, Mediani A, Tong X, Han R, Al-Olayan E, Baharum SN, Bunawan H, Sarian MN, Hamezah HS, Jantan I. An insight review on the neuropharmacological effects, mechanisms of action, pharmacokinetics and toxicity of mitragynine. Biomedicine & Pharmacotherapy 2024;171:116134. https://doi.org/10.1016/J.BIOPHA.2024.116134.

[22] Sears SMS, Hewett SJ. Influence of glutamate and GABA transport on brain excitatory/inhibitory balance. Exp Biol Med 2021;246:1069. https://doi.org/10.1177/1535370221989263.

[23] Hossain R, Sultana A, Nuinoon M, Noonong K, Tangpong J, Hossain KH, Rahman MA. A Critical Review of the Neuropharmacological Effects of Kratom: An Insight from the Functional Array of Identified Natural Compounds. Molecules 2023, Vol 28, Page 7372 2023;28:7372. https://doi.org/10.3390/MOLECULES28217372.

[24] Agbangba CE, Sacla Aide E, Honfo H, Glèlè Kakai R. On the use of post-hoc tests in environmental and biological sciences: A critical review. Heliyon 2024;10:e25131. https://doi.org/10.1016/J.HELIYON.2024.E25131.