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A simple and rapid ultra‐high‐performance liquid chromatography–tandem mass spectrometry method to determine plasma biotin in hemodialysis patients
Author(s) -
Yagi Shigeaki,
Nishizawa Manabu,
Ando Itiro,
Oguma Shiro,
Sato Emiko,
Imai Yutaka,
Fujiwara Masako
Publication year - 2016
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.3680
Subject(s) - chemistry , chromatography , protein precipitation , ammonium formate , biotin , electrospray ionization , mass spectrometry , tandem mass spectrometry , ammonium acetate , hydrophilic interaction chromatography , calibration curve , selected reaction monitoring , high performance liquid chromatography , detection limit , analytical chemistry (journal) , biochemistry
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 m m 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.