Abstract
Objective: This study details the determination of mitragynine in various kratom samples using the thin-layer chromatography (TLC) technique and validation of analytical methods for quantifying the concentration of mitragynine in various kratom samples using liquid chromatography with photodiode array detector (LC-PDA). Methods: TLC technique using n-hexane: ethyl acetate: ammonia 25% (30: 15: 1 v/v/v) was applied to isolate mitragynine from kratom samples. Several interesting spots obtained were visualized under UV light at 254 nm. Samples were also prepared with organic solvent extraction directly prior to LC analysis (non-isolation method of preparation) to quantify the concentration of mitragynine. Mobile phases used were acetonitrile (MP A) and 0.1% formic acid in water (MP B). Samples and standards were run by gradient elution with a flow rate of 0.3 ml/min, detection using PDA detector at 254 nm. Results: Mitragynine was successfully isolated from kratom samples in Rf 0.50 by TLC system applied. The validation of analytical methods for mitragynine passed the acceptance criteria as described by UNODC Guidance. The concentration of mitragynine in various kratom samples seized in Indonesia ranged from 0.37%-1.70% (%w/w). Conclusion: Both TLC and LC analytical methods could be applied to determine and quantify the concentration of mitragynine in each examined sample, respectively.
Highlights
Kratom (Mitragyna speciosa Korth.) is widely cultivated in Indonesia especially in Kalimantan region
For the best result of analysis, both isocratic and gradient elution were compared in this study
Henceforward, the gradient elution was applied in the quantitative analysis of mitragynine using liquid chromatography with photodiode array detector (LC-PDA)
Summary
Kratom (Mitragyna speciosa Korth.) is widely cultivated in Indonesia especially in Kalimantan region. The plants are known as ‘purik’ or ‘ketum’ in West Kalimantan, ‘kedamba/kedemba’ in East Kalimantan and ‘kayu sapat/sepat’ in Middle and South Kalimantan. West Kalimantan is the largest center of kratom plants in Indonesia. Kratom plants grow abundantly along the Kapuas river in Kapuas Hulu district. Products of kratom cultivated in Indonesia, 90% in the form of kratom powders, are mostly exported to other countries. The society in Kalimantan traditionally used kratom to combat fatigue and enhance stamina by drinking the brewed fresh kratom leaves or powder, sometimes by adding other ingredients such as honey or oranges [1]
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