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

Biomed Chromatogr. 2020 Mar;34(3):e4781. doi: 10.1002/bmc.4781. Epub 2020 Jan 14.

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.

Keywords: VAMS; ultra-high-performance liquid chromatography-tandem mass spectrometry; γ-hydroxybutyric acid.

MeSH terms

  • Blood Specimen Collection
  • Chromatography, High Pressure Liquid / methods*
  • Humans
  • Hydroxybutyrates / blood*
  • Hydroxybutyrates / pharmacokinetics
  • Hydroxybutyrates / therapeutic use
  • Limit of Detection
  • Linear Models
  • Narcolepsy / drug therapy
  • Reproducibility of Results
  • Tandem Mass Spectrometry / methods*

Substances

  • Hydroxybutyrates
  • 4-hydroxybutyric acid