Citation
Norliana Izzati M. R., . and Lee, C. T. and Fauziah A., . and Zunoliza A., . and Ling, S. K. and Mohd Hafidz Hadi A., . and Ng, K. K. S. and Ng, C. H. and Tnah, L. H. and Ng, C. L. and Goh, H. H and Lee, S. L. (2024) HPLC-PDA method for the quantification of mitragynine in fresh kratom (Mitragyna speciosa) leaf. Journal of Tropical Forest Science (JTFS) (Malaysia), 36 (4). pp. 434-443. ISSN 0128-1283
Abstract
Kratom (Mitragyna speciosa) is a psychotropic tropical plant that is used as a traditional remedy in Southeast Asia. It has many medicinal properties and has great potential for clinical applications. Mitragynine and 7-hydroxymitragynine (7-OH) are the important pharmacologically active compounds found in kratom with mitragynine as the major alkaloid found in the leaf. We report the development and validation of a high-performance liquid chromatography-photodiode array method for the determination and quantification of mitragynine from M. speciosa fresh leaf extract. Fresh leaf samples were cryogenically ground and freeze-dried before methanol extraction. Gradient elution was conducted using acetonitrile and ammonium bicarbonate buffer (pH 9.5) at a flow rate of 1 mL min⁻¹. Chromatogram separation was achieved using Kinetex EVO C18 column in a duration of 25 min with MG eluted at 15.75 min. The calibration curves were in a linear relationship with determination coefficients (R²) ≥ 0.99. Mitragynine quantification was calculated based on the calibration curve equation. The precision value (RSD) was less than 2% and accuracy range from 93%–100%. The values for limit of detection (LOD) and limit of quantification (LOQ) were 0.67 μg mL⁻¹ and 1.51 μg mL⁻¹, respectively. This method has been validated and found suitable for the quantification of mitragynine in fresh leaf. We found that freeze-drying during sample preparation from fresh leaf yielded more reliable mitragynine quantification, compared with that of conventional oven-drying method.
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Official URL: https://jtfs.frim.gov.my/jtfs/article/view/2967/19...
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Abstract
Kratom (Mitragyna speciosa) is a psychotropic tropical plant that is used as a traditional remedy in Southeast Asia. It has many medicinal properties and has great potential for clinical applications. Mitragynine and 7-hydroxymitragynine (7-OH) are the important pharmacologically active compounds found in kratom with mitragynine as the major alkaloid found in the leaf. We report the development and validation of a high-performance liquid chromatography-photodiode array method for the determination and quantification of mitragynine from M. speciosa fresh leaf extract. Fresh leaf samples were cryogenically ground and freeze-dried before methanol extraction. Gradient elution was conducted using acetonitrile and ammonium bicarbonate buffer (pH 9.5) at a flow rate of 1 mL min⁻¹. Chromatogram separation was achieved using Kinetex EVO C18 column in a duration of 25 min with MG eluted at 15.75 min. The calibration curves were in a linear relationship with determination coefficients (R²) ≥ 0.99. Mitragynine quantification was calculated based on the calibration curve equation. The precision value (RSD) was less than 2% and accuracy range from 93%–100%. The values for limit of detection (LOD) and limit of quantification (LOQ) were 0.67 μg mL⁻¹ and 1.51 μg mL⁻¹, respectively. This method has been validated and found suitable for the quantification of mitragynine in fresh leaf. We found that freeze-drying during sample preparation from fresh leaf yielded more reliable mitragynine quantification, compared with that of conventional oven-drying method.
Additional Metadata
| Item Type: | Article |
|---|---|
| AGROVOC Term: | alkaloids |
| AGROVOC Term: | plant extracts |
| AGROVOC Term: | extraction |
| AGROVOC Term: | quantitative analysis |
| AGROVOC Term: | chromatography |
| AGROVOC Term: | medicinal plants |
| AGROVOC Term: | bioactive compounds |
| AGROVOC Term: | accuracy |
| AGROVOC Term: | precision agriculture |
| Geographical Term: | Malaysia |
| Uncontrolled Keywords: | Mitragyna speciosa, medicinal plant, bioactive compounds, method validation, freeze-drying |
| Depositing User: | Ms. Azariah Hashim |
| Date Deposited: | 16 Jun 2026 01:37 |
| Last Modified: | 16 Jun 2026 01:37 |
| URI: | http://webagris.upm.edu.my/id/eprint/3752 |
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