Adduct supported analysis of γ-hydroxybutyrate in human serum with LC-MS/MS

Abstract

To avoid the detection of small fragmentation products of γ-hydroxybutyrate (GHB), a liquid chromatography–tandem mass spectrometry GHB quantification method in human serum supported by adduct formation was developed and validated. The continuous infusion of GHB/GHB-D6 made the identification of two adducts possible and GHB/GHB-D6 sodium acetate adduct fragmentation was used as target mass transition. A Luna 5 μm C18 (2) 100 A, 150 mm × 2 mm analytical column and elution with a programmed flow of the mobile phase consisting of 10 % A (H₂O/methanol = 95/5, v/v) and 90 % B (H₂O/methanol = 3/97, v/v), both with 10 mM ammonium acetate and 0.1 % acetic acid (pH = 3.2), were used. Protein precipitation with 1 mL of the mobile phase B was used as the sample preparation. The calculated limit of detection/quantification was 1 μg/mL. The presented study shows that the fragmentation of GHB sodium acetate adducts is an effective way of quantification of this small molecule and is an interesting alternative to other methods based on the detection of ions smaller than 85 Da. This fact together with the short analysis time of 3 min and the fast sample preparation make this method very attractive for forensic/clinical application.