title: Simultaneous quantification of methylene blue and its major metabolite, azure B, in plasma by LC-MS/MS and its application for a pharmacokinetic study.
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.
authors.affiliation: Translational Medicine, Life Science R&D Park, SK Biopharmaceuticals, Daejeon, 305-712, Korea.
authors.forename: Soo-Jin
authors.initials: SJ
authors.lastname: Kim
authors.forename: Dong-Jin
authors.initials: DJ
authors.lastname: Ha
authors.forename: Tae-Sung
authors.initials: TS
authors.lastname: Koo
mesh_terms: D000284:Administration, Oral; D000818:Animals; D001399:Azure Stains / Q000032:analysis* / Q000737:chemistry / Q000493:pharmacokinetics; D001774:Blood Chemical Analysis; D002853:Chromatography, Liquid / Q000379:methods*; D016014:Linear Models; D008297:Male; D008751:Methylene Blue / Q000032:analysis* / Q000737:chemistry / Q000493:pharmacokinetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D001399:Azure Stains; C025819:Azure B; D008751:Methylene Blue
keywords: LC/MS/MS; azure B; methylene blue; pharmacokinetics; rat plasma