Simultaneous quantification of methylene blue and its major metabolite,azure B,in plasma by LC‐MS/MS and its application for a pharmacokinetic study |
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| Authors: | Soo‐Jin Kim Dong‐Jin Ha Tae‐Sung Koo |
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| Affiliation: | 1. Translational Medicine, Life Science R&D Park, SK Biopharmaceuticals, Daejeon, Korea;2. Graduate School of Drug Discovery and Development, Chungnam National University, Daejeon, Korea |
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| Abstract: | A simple and sensitive liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) method was developed for the quantification of methylene blue (MB) and its major metabolite, azure B (AZB), in rat plasma. A simple protein precipitation using acetonitrile was followed by injection of the supernatant on to a Zorbax HILIC Plus column (3.5 µm, 2.1 × 100 mm) with isocratic mobile phase consisting of 5 mM ammonium acetate in 10:90 (v/v) water:methanol at a flow rate of 0.3 mL/min and detection in positive ionization mode. The standard curve was linear over the concentration range from 1 to 1000 ng/mL for MB and AZB with coefficient of determination above 0.9930. The lower limit of quantification was 1 ng/mL using 20 μL of rat plasma sample. The intra‐ and inter‐assay precision and accuracy were <12%. The developed analytical method was successfully applied to the pharmacokinetic study of MB and AZB in rats. Copyright © 2013 John Wiley & Sons, Ltd. |
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| Keywords: | methylene blue azure B pharmacokinetics LC/MS/MS rat plasma |
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