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CC BY-NC-ND 4.0 · Planta Med 2022; 88(09/10): 838-857
DOI: 10.1055/a-1795-5876
Natural Product Chemistry and Analytical Studies
Original Papers

Kratom (Mitragyna speciosa) Validation: Quantitative Analysis of Indole and Oxindole Alkaloids Reveals Chemotypes of Plants and Products[ # ]

Preston K. Manwill
Department of Chemistry and Biochemistry, University of North Carolina at Greensboro, Greensboro, NC, USA
,
Laura Flores-Bocanegra
Department of Chemistry and Biochemistry, University of North Carolina at Greensboro, Greensboro, NC, USA
,
Manead Khin
Department of Chemistry and Biochemistry, University of North Carolina at Greensboro, Greensboro, NC, USA
,
Huzefa A. Raja
Department of Chemistry and Biochemistry, University of North Carolina at Greensboro, Greensboro, NC, USA
,
Nadja B. Cech 
Department of Chemistry and Biochemistry, University of North Carolina at Greensboro, Greensboro, NC, USA
,
Nicholas H. Oberlies 
Department of Chemistry and Biochemistry, University of North Carolina at Greensboro, Greensboro, NC, USA
,
Daniel A. Todd
Department of Chemistry and Biochemistry, University of North Carolina at Greensboro, Greensboro, NC, USA
› Author AffiliationsSupported by: National Center for Complementary and Integrative Health U54 AT008909
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Abstract

Many consumers are turning to kratom (Mitragyna speciosa) to self-manage pain and opioid addiction. In the United States, an array of capsules, powders, and loose-leaf kratom products are readily available. Additionally, several online sites supply live kratom plants. A prerequisite to establishing quality control and quality assurance standards for the kratom industry, or understanding how alkaloid levels effect clinical outcomes, is the identification and quantitation of major and minor alkaloid constituents within available products and preparations. To this end, an ultra-high performance liquid chromatography-high resolution mass spectrometry method was developed for the analysis of 8 indole alkaloids (7-hydroxymitragynine, ajmalicine, paynantheine, mitragynine, speciogynine, isopaynantheine, speciociliatine, and mitraciliatine) and 6 oxindole alkaloids (isomitraphylline, isospeciofoleine, speciofoline, corynoxine A, corynoxeine, and rhynchophylline) in US-grown kratom plants and commercial products. These commercial products shared a qualitatively similar alkaloid profile, with 12 – 13 detected alkaloids and high levels of the indole alkaloid mitragynine (13.9 ± 1.1 – 270 ± 24 mg/g). The levels of the other major alkaloids (paynantheine, speciociliatine, speciogynine, mitraciliatine, and isopaynantheine) and the minor alkaloids varied in concentration from product to product. The alkaloid profile of US-grown M. speciosa “Rifat” showed high levels of the indole alkaloid speciogynine (7.94 ± 0.83 – 11.55 ± 0.18 mg/g) and quantifiable levels of isomitraphylline (0.943 ± 0.033 – 1.47 ± 0.18 mg/g). Notably, the alkaloid profile of a US-grown M. speciosa seedling was comparable to the commercial products with a high level of mitragynine (15.01 ± 0.20 mg/g). This work suggests that there are several M. speciosa chemotypes.

Key words

kratom - Mitragyna speciosa - Rubiaceae - indole alkaloid - oxindole alkaloid - chemotype - UPLC-HRMS

# Dedicated to Professor Dr. A. Douglas Kinghorn on the occasion of his 75th birthday.


Supporting Information

    The following are available in the Supporting Information: List of publications covering the analytical analysis of kratom plants and products (Table 1S), LC-MS chromatograms comparing a generic screening method using a BEH C18 column with the developed method using a Kinetex F5 column (Fig. 1S), precision and accuracy of the method for kratom alkaloid quantification (Table 2S), concentration and percent recovery of internal standard (Table 3S), base peak MS chromatograms representing kratom materials quantified in [Tables 2], [3] and [4] (Fig. 2S, 3S, and 4S, respectively), literature references for purchased indole and oxindole alkaloid standards (Table 4S), the commercial kratom products and M. speciosa specimens analyzed in this study (Table 5S), graphical overview of BLAST results in NCBI GenBank database using the Internal Transcribed Spacer (ITS) region for K64 (Fig. 5S) and K68 (Fig. 6S), phylogram showing that K64 and K68 group with other M. speciosa (Fig. 7S), the precursor ion inclusion list used for MS2 analysis (Table 6S), and the constrain peak width settings used for peak integration (Table 7S).

  • Supporting Information


Publication History

Received: 08 October 2021

Accepted after revision: 23 February 2022

Article published online:
25 April 2022

© 2022. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commecial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/)

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