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Simultaneous quantification of methylene blue and its major metabolite, azure B, in plasma by LC‐MS/MS and its application for a pharmacokinetic study
Author(s) -
Kim SooJin,
Ha DongJin,
Koo TaeSung
Publication year - 2014
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.3063
Subject(s) - chemistry , chromatography , protein precipitation , ammonium acetate , metabolite , methylene blue , detection limit , hydrophilic interaction chromatography , mass spectrometry , selected reaction monitoring , pharmacokinetics , acetonitrile , methanol , tandem mass spectrometry , high performance liquid chromatography , organic chemistry , medicine , biochemistry , photocatalysis , catalysis
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.