Application of 4-Diethylaminobenzoic Acid N-Succinimidyl Ester and Its Deuterated Isotopologue as Derivatization Reagents to Quantitative Analysis of γ-Aminobutyric Acid in Serum by LC/ESI-MS/MS

gamma-Aminobutyric acid (GABA) is an inhibitory neurotransmitter having anti-anxiety, stress-reducing and sleep-enhancing effects. The blood GABA concentration has the potential to become a diagnostic index for depression and schizophrenia. In this study, we developed a method for the quantification of GABA in human serum by liquid chromatography/electrospray ionization-tandem mass spectrometry (LC/ESI-MS/MS) combined with derivatization using a pair of reagents, 4-diethylaminobenzoic acid N-succinimidyl ester (DEABAS) and its deuterated isotopologue (dDEABAS). The deproteinized serum samples were derivatized with DEABAS, to which the dDEABAS-derivatized standard GABA of known amount was then added. The DEABAS-derivatization enhanced not only the ESI-MS/MS sensitivity but also the LC retention, which enabled the separation of GABA from the interfering isomers. Satisfactory assay precision and accuracy were also acquired by adding dDEABAS-derivatized GABA, which served as the internal standard and worked well for correcting the matrix effect and instrument drift. The developed method had satisfactory applicability to the real sample analysis because it was successfully used to quantify the serum GABA of the healthy subjects.