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A simple and rapid ultra‐high‐performance liquid chromatography–tandem mass spectrometry method to determine plasma biotin in hemodialysis patients
Authors:Shigeaki Yagi  Manabu Nishizawa  Itiro Ando  Shiro Oguma  Emiko Sato  Yutaka Imai  Masako Fujiwara
Affiliation:1. Fuso Pharmaceutical Industries Ltd Research and Development Center, Osaka, Japan;2. Laboratory of Clinical Pharmacology and Therapeutics, Tohoku University Graduate School of Pharmaceutical Sciences, Sendai, Japan;3. Koujinkai Nagamachi Clinic, Sendai, Japan;4. Department of Planning for Drug Development and Clinical Evaluation, Tohoku University Graduate School of Pharmaceutical Sciences, Sendai, Japan
Abstract:A simple, rapid, and selective method for determination of plasma biotin was developed using ultra‐high‐performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS). After single‐step protein precipitation with methanol, biotin and stable isotope‐labeled biotin as an internal standard (IS) were chromatographed on a pentafluorophenyl stationary‐phase column (2.1 × 100 mm, 2.7 μm) under isocratic conditions using 10 mm ammonium formate–acetonitrile (93:7, v/v) at a flow rate of 0.6 mL/min. The total chromatographic runtime was 5 min for each injection. Detection was performed in a positive electrospray ionization mode by monitoring selected ion transitions at m/z 245.1/227.0 and 249.1/231.0 for biotin and the IS, respectively. The calibration curve was linear in the range of 0.05–2 ng/mL using 300 μL of plasma. The intra‐ and inter‐day precisions were all <7.1%. The accuracy varied from ?0.7 to 8.2%. The developed UHPLC–MS/MS method was successfully applied to determine plasma biotin concentrations in hemodialysis patients. Copyright © 2016 John Wiley & Sons, Ltd.
Keywords:biotin  UHPLC–  MS/MS  human plasma  hemodialysis
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