Simple, Sensitive, High-Throughput Method for the Quantification of Mitragynine in Rat Plasma Using UPLC-MS and Its Application to an Intravenous Pharmacokinetic Study

Abstract

A simple, sensitive and rapid ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) method was developed and validated for the quantification of mitragynine in rat plasma using amitriptyline hydrochloride as an internal standard. Sample preparation involved a one-step liquid-liquid extraction using methyl t-butyl ether. Mitragynine was separated on an Acquity UPLC TM BEH HILIC column using isocratic elution with a mobile phase of 10 mM ammonium formate buffer containing 0.1% formic acid:acetonitrile (15:85, v/v). At a flow rate of 0.2 mL min -1 , the retention time of mitragynine was found to be 1.3 min. Ionization was performed in the positive ion electrospray mode. The selected mass-tocharge (m/z) ratio transition of mitragynine ion [M ? H] ? used in the selected ion recording (SIR) was 399.1. The calibration curve was found to be linear over a concentration range of 1-5,000 ng mL -1 (r = 0.999) with a lower limit of quantification (LLOQ) of 1 ng mL -1 . Intra-and inter-day assay variations were found to be less than 15%. The extraction recoveries ranged from 85-93% at the three concentrations (2, 400 and 4,000 ng mL -1 ) in rat plasma.This method was successfully used to quantify mitragynine in rat plasma following intravenous administration of the compound.

Simultaneous quantification of ten key Kratom alkaloids in Mitragyna speciosa leaf extracts and commercial products by ultra‐performance liquid chromatography−tandem mass spectrometry

“…It is necessary to separate alkaloids sharing the same ion transitions chromatographically to assure their peak purity; otherwise, samples may be inappropriately over quantified for certain alkaloids. For example, mitragynine might have been over quantified along with its three other co‐eluting diastereomers by earlier reported isocratic or short gradient methods . Therefore, various mobile phase compositions (mobile phase A: aqueous ammonium acetate, water containing acetic acid, aqueous ammonium formate, or water containing formic acid; pH 3.0–6.8, mobile phase B: acetonitrile and methanol) and columns (Acquity UPLC BEH C18, CSH C18, Cyano, and HILIC) were tested to achieve at least a resolution of ≥2 between alkaloids sharing the same ion transitions with Gaussian peak shapes.…”

“…The first step would be a proper way to identify and quantify the multiple kratom alkaloids that are present in each product and traditional preparations. There are multiple analytical and bioanalytical methods available in the literature for the quantification of kratom alkaloids . Most of the reported methods were only able to quantify mitragynine and/or 7‐hydroxymitragynine except the method reported by Kikura‐Hanajiri et al which is able to quantify five kratom alkaloids (mitragynine and 7‐hydroxymitragynine along with speciogynine, speciociliatine, and paynantheine), but there is no method available which can quantify 10 kratom alkaloids simultaneously.…”

“…There are multiple analytical and bioanalytical methods available in the literature for the quantification of kratom alkaloids. 3,[7][8][9][10][11][12][13][14] Most of the reported methods were only able to quantify mitragynine and/or 7-hydroxymitragynine except the method reported by Kikura-Hanajiri et al 9 which is able to quantify five kratom alkaloids (mitragynine and 7-hydroxymitragynine along with speciogynine, speciociliatine, and paynantheine), but there is no method available which can quantify 10 kratom alkaloids simultaneously. For this purpose, an ultra-performance liquid chromatography−mass spectrometry (UPLC−MS/MS) method was developed for the quantification of ten alkaloids in kratom products: corynantheidine, corynoxine, corynoxine B, 7-hydroxymitragynine, isocorynantheidine, mitragynine, mitraphylline, paynantheine, speciociliatine, and speciogynine ( Figure 1).…”

Here Today, Gone Tomorrow…and Back Again? A Review of Herbal Marijuana Alternatives (K2, Spice), Synthetic Cathinones (Bath Salts), Kratom, Salvia divinorum, Methoxetamine, and Piperazines

“…In another study, LC-ESI-MS analysis of 13 samples of commercial products advertising Kratom on their packaging confirmed the presence of the following compounds: mitragynine, 7-OH-mitragynine, paynantheine, speciogynine, and speciociliatine [150]. An ultraperformance liquid chromatography (UPLC) technique has also been described to identify mitragynine [151]. Recently, investigators in Japan have developed a PCR-based method to test different plant products in order to determine whether or not they contain Kratom [135].…”

Novel Treatment Strategies for Cocaine and Opioid Abuse

“…The leaves of the plant have traditionally been used by natives as a substitute for opium or to treat withdrawal, as well as for their stimulant effects [38][39][40]. Furthermore, the leaves have also been used by southern Thai villagers as a medicine to treat coughing, diarrhea, muscle pain and hypertension.…”