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Abstract
Kratom is illegally used in traditional medicine and health supplement products. Thus, the analytical method development is needed to identify its presence in final products. Currently, the availability of its markers is limited due to it belongs to restricted substance in Indonesia. The research was conducted to produce alternative analytical method without the use of reference standard. In addition, the research was developed by biomarker profiling approach using mass spectrometer. Mass spectrometry was performed using either gas chromatography or liquid chromatography-high resolution mass spectrometry (LC-HRMS). Both systems were developed to confirm the presence of kratom. A mass spectrometry test was carried out after the marker compounds were extracted from the leaves. The gas chromatography test was carried out using selected ion monitoring (SIM) mode at m/z 214, 397, 383, and 269. The LC-HRMS chromatography system used 0.1 % formic acid as an ion modifier. The analysis was carried out using a quadrupole time of flight (QToF) mass spectrometer as a mass analyzer with electrospray ionization (ESI) in positive mode, running in scan mode analysis. Qualitative analysis was carried out using retention time (Rt) data and ionic fragments of kratom marker compounds. There were three marker compounds obtained, namely speciociliatine, speciogynine, and mitragynine. Specificity test was carried out by identifying their retention time, followed by the determination of their Retention Index (RI) value by GCMS. In addition, m/z value of the marker compounds were confirmed by LC-HRMS QToF. The results showed that three main marker compounds can be used in the identification of kratom adulteration in traditional medicines. This method can be applied as a confirmation test of kratom adulteration in traditional medicine where the LC-HRMS QToF method is more sensitive than GCMS and TLC with LOD values of 149.2 µg/g and 67.6 mg/g respectively after extracted and isolated by TLC.
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References
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References
Avula, Bharathi et al. 2015. “Identification and Characterization of Indole and Oxindole Alkaloids from Leaves of Mitragyna Speciosa Korth Using Liquid Chromatography - Accurate QToF Mass Spectrometry.” Journal of AOAC International 98(1): 13–21.
Boffa, Luisa et al. 2018. “Alkaloid Profiles and Activity in Different Mitragyna Speciosa Strains.” Natural Product Communications 13(9): 1111–16.
BPOM. 2016. “Surat Edaran Pelarangan Penggunaan Mitragyna Speciosa (Kratom) Dalam Obat Tradisional Dan Suplemen Kesehatan Tahun 2016.” Badan Pengawas Obat dan Makanan.
Casey, Christine R et al. 2015. “Quantitative and Qualitative Analysis of Mitragynine in Kratom ( Mitragyna Speciosa ) by GC-MS , LC-MS/MS and UPLC-PDA.” Journal of Regulatory Science 02: 1–14.
Cheng, Sy Chyi, Suhail Muzaffar Bhat, Chi Wei Lee, and Jentaie Shiea. 2018. “Thin Layer Chromatography Combined with Electrospray Ionization Mass Spectrometry for Characterizing Herbal Compounds.” International Journal of Mass Spectrometry 434: 264–71. https://doi.org/10.1016/j.ijms.2018.09.024.
E. Adkins, Jessica, Edward W. Boyer, and Christopher R. McCurdy. 2011. “Mitragyna Speciosa, A Psychoactive Tree from Southeast Asia with Opioid Activity.” Current Topics in Medicinal Chemistry 11(9): 1165–75.
Elsa, Livia. 2016. “Pengembangan Metode Isolasi Dan Identifikasi Mitragynine Dalam Daun Kratom (Mitragyna Speciosa).” Jurnal Biosains Pascasarjana 18(3): 191.
Flores-Bocanegra, Laura et al. 2020. “The Chemistry of Kratom [ Mitragyna Speciosa]: Updated Characterization Data and Methods to Elucidate Indole and Oxindole Alkaloids.” Journal of Natural Products 83(7): 2165–77.
Fu, Hanzhuo et al. 2015. “Screening and Identification of Mitragynine and 7-Hydroxymitragynine in Human Urine by LC-MS/MS.” Chromatography 2(2): 253–64.
Istyawan, M, A Fadholi, … D Surtikanthi - JURNAL, and undefined 2022. “Legalitas Narkotika Jenis Baru (Kratom): Antara Ancaman Dan Peluang Bagi Ketahanan Nasional Indonesia.” Jurnal.Politap.Ac.Id: 69–80. https://jurnal.politap.ac.id/index.php/literasi/article/view/170.
Kruegel, Andrew C., and Oliver Grundmann. 2018. 134 Neuropharmacology The Medicinal Chemistry and Neuropharmacology of Kratom: A Preliminary Discussion of a Promising Medicinal Plant and Analysis of Its Potential for Abuse. Elsevier Ltd. http://dx.doi.org/10.1016/j.neuropharm.2017.08.026.
Raini, Mariana. 2017. “Kratom (Mitragyna Speciosa Korth): Manfaat, Efek Samping Dan Legalitas.” Media Penelitian dan Pengembangan Kesehatan 27(3): 175–84.
Reich, Eike. 2019. Encyclopedia of Analytical Science Thin-Layer Chromatography | Overview. Elsevier Inc. http://dx.doi.org/10.1016/B978-0-12-409547-2.00540-0.
“Retention Index in Column Chromatography, I.” 2008. The IUPAC Compendium of Chemical Terminology 819: 5360.
Rivier, Laurent. 2003. “Criteria for the Identification of Compounds by Liquid Chromatography – Mass Spectrometry and Liquid Chromatography – Multiple Mass Spectrometry in Forensic Toxicology and Doping Analysis.” 492: 69–82.
Sardjono, Ratnaningsih E. 2015. “Modul 2 Isomeri.” Isomeri. http://file.upi.edu/Direktori/FPMIPA/JUR._PEND._KIMIA/196904191992032-RATNANINGSIH_EKO_SARDJONO/MODUL_2_isomer_20_6_08_revisi.pdf.
Todd, D. A. et al. 2020. “Chemical Composition and Biological Effects of Kratom (Mitragyna Speciosa): In Vitro Studies with Implications for Efficacy and Drug Interactions.” Scientific Reports 10(1): 1–13. https://doi.org/10.1038/s41598-020-76119-w.