Development and validation of volumetric absorptive microsampling coupled with UHPLC–MS/MS for the analysis of gamma-hydroxybutyric acid in human blood

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Abstract

A volumetric microsampling (VAMS) device (20 μl) was evaluated and validated for the analysis of γ-hydroxybutyric acid (GHB) in venous blood using a simple ultra-high-pressure liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) method. GHB was extracted from VAMS device by acetonitrile, after a re-hydration step in a temperature-controlled ultrasonic bath at 60°C for 10 min. Chromatographic analysis was carried out on a Kinetex C18 column using 0.1% formic acid in water and acetonitrile as binary gradient mobile phase (from 5 to 95% of acetonitrile from 1 to 2.5 min) at a flow rate of 0.3 ml/min. The VAMS method was fully validated according to current guidelines with satisfactory results in terms of linearity, selectivity, precision, absolute recovery, matrix effect and stability. The linearity was determined from 0.5 to 200 μg/ml and the lower limit of quantitation was 0.5 μg/ml. The novel VAMS–UHPLC–MS/MS method was successfully compared with plasma-based method in a GHB-treated patient as a proof of concept.

Original languageEnglish
Article numbere4781
JournalBiomedical Chromatography
Volume34
Issue number3
DOIs
Publication statusPublished - Mar 2020

Keywords

  • ultra-high-performance liquid chromatography–tandem mass spectrometry
  • VAMS
  • γ-hydroxybutyric acid

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry

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