Simultaneous quantification of ten key Kratom alkaloids in Mitragyna speciosa leaf extracts and commercial products by ultra-performance liquid chromatography−tandem mass spectrometry
Abhisheak Sharma
Department of Pharmaceutics, College of Pharmacy, University of Florida, Gainesville, FL, USA
Search for more papers by this authorShyam H. Kamble
Department of Pharmaceutics, College of Pharmacy, University of Florida, Gainesville, FL, USA
Search for more papers by this authorFrancisco León
Department of Medicinal Chemistry, College of Pharmacy, University of Florida, Gainesville, FL, USA
Search for more papers by this authorNelson J.-Y. Chear
Centre for Drug Research, Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia
Search for more papers by this authorTamara I. King
Department of Pharmaceutics, College of Pharmacy, University of Florida, Gainesville, FL, USA
Search for more papers by this authorErin C. Berthold
Department of Pharmaceutics, College of Pharmacy, University of Florida, Gainesville, FL, USA
Search for more papers by this authorSurash Ramanathan
Centre for Drug Research, Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia
Search for more papers by this authorCorresponding Author
Christopher R. McCurdy
Department of Medicinal Chemistry, College of Pharmacy, University of Florida, Gainesville, FL, USA
Correspondence
Dr Bonnie A. Avery, Department of Pharmaceutics, University of Florida, Gainesville, FL, 32610, USA. Email: [email protected];
Dr Christopher R. McCurdy, Department of Medicinal Chemistry, University of Florida, Gainesville, FL 32610, USA.
Email: [email protected]
Search for more papers by this authorCorresponding Author
Bonnie A. Avery
Department of Pharmaceutics, College of Pharmacy, University of Florida, Gainesville, FL, USA
Correspondence
Dr Bonnie A. Avery, Department of Pharmaceutics, University of Florida, Gainesville, FL, 32610, USA. Email: [email protected];
Dr Christopher R. McCurdy, Department of Medicinal Chemistry, University of Florida, Gainesville, FL 32610, USA.
Email: [email protected]
Search for more papers by this authorAbhisheak Sharma
Department of Pharmaceutics, College of Pharmacy, University of Florida, Gainesville, FL, USA
Search for more papers by this authorShyam H. Kamble
Department of Pharmaceutics, College of Pharmacy, University of Florida, Gainesville, FL, USA
Search for more papers by this authorFrancisco León
Department of Medicinal Chemistry, College of Pharmacy, University of Florida, Gainesville, FL, USA
Search for more papers by this authorNelson J.-Y. Chear
Centre for Drug Research, Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia
Search for more papers by this authorTamara I. King
Department of Pharmaceutics, College of Pharmacy, University of Florida, Gainesville, FL, USA
Search for more papers by this authorErin C. Berthold
Department of Pharmaceutics, College of Pharmacy, University of Florida, Gainesville, FL, USA
Search for more papers by this authorSurash Ramanathan
Centre for Drug Research, Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia
Search for more papers by this authorCorresponding Author
Christopher R. McCurdy
Department of Medicinal Chemistry, College of Pharmacy, University of Florida, Gainesville, FL, USA
Correspondence
Dr Bonnie A. Avery, Department of Pharmaceutics, University of Florida, Gainesville, FL, 32610, USA. Email: [email protected];
Dr Christopher R. McCurdy, Department of Medicinal Chemistry, University of Florida, Gainesville, FL 32610, USA.
Email: [email protected]
Search for more papers by this authorCorresponding Author
Bonnie A. Avery
Department of Pharmaceutics, College of Pharmacy, University of Florida, Gainesville, FL, USA
Correspondence
Dr Bonnie A. Avery, Department of Pharmaceutics, University of Florida, Gainesville, FL, 32610, USA. Email: [email protected];
Dr Christopher R. McCurdy, Department of Medicinal Chemistry, University of Florida, Gainesville, FL 32610, USA.
Email: [email protected]
Search for more papers by this authorAbstract
Kratom (Mitragyna speciosa) is a psychoactive plant popular in the United States for the self-treatment of pain and opioid addiction. For standardization and quality control of raw and commercial kratom products, an ultra-performance liquid chromatography−tandem mass spectrometry (UPLC−MS/MS) method was developed and validated for the quantification of ten key alkaloids, namely: corynantheidine, corynoxine, corynoxine B, 7-hydroxymitragynine, isocorynantheidine, mitragynine, mitraphylline, paynantheine, speciociliatine, and speciogynine. Chromatographic separation of diastereomers, or alkaloids sharing same ion transitions, was achieved on an Acquity BEH C18 column with a gradient elution using a mobile phase containing acetonitrile and aqueous ammonium acetate buffer (10mM, pH 3.5). The developed method was linear over a concentration range of 1–200 ng/mL for each alkaloid. The total analysis time per sample was 22.5 minutes. The analytical method was validated for accuracy, precision, robustness, and stability. After successful validation, the method was applied for the quantification of kratom alkaloids in alkaloid-rich fractions, ethanolic extracts, lyophilized teas, and commercial products. Mitragynine (0.7%–38.7% w/w), paynantheine (0.3%–12.8% w/w), speciociliatine (0.4%–12.3% w/w), and speciogynine (0.1%–5.3% w/w) were the major alkaloids in the analyzed kratom products/extracts. Minor kratom alkaloids (corynantheidine, corynoxine, corynoxine B, 7-hydroxymitragynine, isocorynantheidine) were also quantified (0.01%–2.8% w/w) in the analyzed products; however mitraphylline was below the lower limit of quantification in all analyses.
CONFLICT OF INTEREST
The authors declare no conflict of interest.
Supporting Information
| Filename | Description |
|---|---|
| dta2604-supp-0001-Figures_S1-S36.docxWord 2007 document , 8.5 MB |
Figure S1. 1H NMR spectrum of corynantheidine in DMSO-d6, 600 MHz Figure S2. 13C NMR spectrum of corynantheidine in DMSO-d6, 150 MHz Figure S3. 1H NMR spectrum of corynoxine in CD3OD, 600 MHz Figure S4. 13C NMR spectrum of corynoxine in CD3OD, 150 MHz Figure S5. 1H NMR spectrum of corynoxine B in CDCl3, 500 MHz Figure S6. 13C NMR spectrum of corynoxine B in CDCl3, 125 MHz Figure S7. 1H NMR spectrum of 7-hydroxy mitragynine in CDCl3, 500 MHz Figure S8. 13C NMR spectrum of 7-hydroxy mitragynine in CDCl3, 125 MHz Figure S9. 1H NMR spectrum of isocorynantheidine in CDCl3, 500 MHz Figure S10. 13C NMR spectrum of isocorynantheidine in CDCl3, 125 MHz Figure S11. 1H NMR spectrum mitragynine in CDCl3, 500 MHz Figure S12. 13C NMR spectrum of mitragynine in CDCl3, 125 MHz Figure S13. 1H NMR spectrum of mitragynine hydrochloride salt in CD3OD, 600 MHz Figure S14. 13C NMR spectrum of mitragynine hydrochloride salt in CD3OD, 150 MHz Figure S15. 1H NMR spectrum mitraphylline in CDCl3, 500 MHz Figure S16. 13C NMR spectrum of mitraphylline in CDCl3, 125 MHz Figure S17. 1H NMR spectrum of paynantheine in CD3OD, 600 MHz Figure S18. 13C NMR spectrum of paynantheine in CD3OD, 150 MHz Figure S19. 1H NMR spectrum speciociliatine in CDCl3, 500 MHz Figure S20. 13C NMR spectrum of speciociliatine in CDCl3, 125 MHz Figure S21. 1H NMR spectrum speciogynine in CDCl3, 500 MHz Figure S22. 13C NMR spectrum of speciogynine in CDCl3, 125 MHz Figure S23. Base peak chromatogram and corresponding MS spectrum of corynantheidine using UPLC-Q-TOF Figure S24. Base peak chromatogram and corresponding MS spectrum of corynoxine using UPLC-Q-TOF Figure S25. Base peak chromatogram and corresponding MS spectrum of corynoxine using UPLC-Q-TOF Figure S26. Base peak chromatogram and corresponding MS spectrum of 7-hydroxymitragynine using UPLC-Q-TOF, Water adduct of 7-hydroxymitragynine) Figure S27. Base peak chromatogram and corresponding MS spectrum of isocorynantheidine using UPLC-Q-TOF Figure S28. Base peak chromatogram and corresponding MS spectrum of mitragynine using UPLC-Q-TOF Figure S29. Base peak chromatogram and corresponding MS spectrum of mitraphylline using UPLC-Q-TOF Figure S30. Base peak chromatogram and corresponding MS spectrum of paynantheine using UPLC-Q-TOF Figure S31. Base peak chromatogram and corresponding MS spectrum of speciogynine using UPLC-Q-TOF Figure S32. Base peak chromatogram and corresponding MS spectrum of speciogynine using UPLC-Q-TOF Figure S33. Product ion spectra (MS/MS) of (A) corynantheidine, (B) corynoxine, (C) corynoxine B, (D) 7-hydroxymitragynine, (E) isocorynantheidine, (F) mitragynine, (G) mitraphylline, (H) paynantheine, (I) speciociliatine, and (J) speciogynine Figure S34. Representative calibration curves for corynantheidine, corynoxine, corynoxine B, 7-hydroxymitragynine, isocorynantheidine, mitragynine, mitraphylline, paynantheine, speciociliatine, and speciogynine Figure S35. Chromatograms of (1A) corynantheidine, (1B) corynoxine, (1C) corynoxine B, (1D) 7-hydroxymitragynine, (1E) isocorynantheidine, (1F) mitragynine, (1G) mitraphylline, (1H) paynantheine, (1I) speciociliatine, (1 J) speciogynine and (1 K) internal standard during the analysis of ethanolic extract of kratom Figure S36. Chromatograms of (1A) corynantheidine, (1B) corynoxine, (1C) corynoxine B, (1D) 7-hydroxymitragynine, (1E) isocorynantheidine, (1F) mitragynine, (1G) mitraphylline, (1H) paynantheine, (1I) speciociliatine, (1 J) speciogynine and (1 K) internal standard during the analysis of alkaloid fraction of kratom |
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