Simple, Sensitive, High-Throughput Method for the Quantification of Mitragynine in Rat Plasma Using UPLC-MS and Its Application to an Intravenous Pharmacokinetic Study |
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Authors: | Pradeep K Vuppala Sai P Boddu Edward B Furr Christopher R McCurdy Bonnie A Avery |
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Institution: | 1. Department of Pharmaceutics, School of Pharmacy, The University of Mississippi, University, MS, 38677, USA 2. Department of Medicinal Chemistry, School of Pharmacy, The University of Mississippi, University, MS, 38677, USA 3. Department of Pharmacology, School of Pharmacy, The University of Mississippi, University, MS, 38677, USA
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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?Cliquid extraction using methyl t-butyl ether. Mitragynine was separated on an Acquity UPLC? 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-to-charge (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?C5,000 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?C93% 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. |
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