Establishment of a LC-MS/MS method for detecting gamma-aminobutyric acid in plasma and its clinical application
10.3969/j.issn.1672-8467.2025.05.012
- VernacularTitle:液相色谱-串联质谱检测血浆γ-氨基丁酸方法的建立和临床应用
- Author:
Jia-qian QIN
1
;
Lei CAO
;
Ying-fei PENG
;
Fang-jun CHEN
;
Bai-shen PAN
;
Bei-li WANG
;
Wei GUO
Author Information
1. 复旦大学附属中山医院检验科 上海 200032
- Publication Type:Journal Article
- Keywords:
gamma-aminobutyric acid(GABA);
liquid chromotography-tandem mass spectrometry(LC-MS/MS);
sleep disorders;
performance validation;
reference range
- From:
Fudan University Journal of Medical Sciences
2025;52(5):708-716
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a stable liquid chromatography-tandem mass spectrometry(LC-MS/MS)method for detecting gamma-aminobutyric acid(GABA)in plasma,and to evaluate the value of GABA detection in the diagnosis of sleep disorders.Methods GABA was detected using a UPLC Xevo TQs system.The method was pre-validated and its performance was verified to establish a reference range for healthy individuals.The difference in plasma GABA levels between apparently healthy individuals and patients with sleep disorders was compared.Results We employed deuterated compounds as isotopic internal standards and utilized an Amide chromatographic column for separation.The mobile phase was 0.050%formic acid in water and 90%acetonitrile in water containing 0.175%formic acid and 5 mmol/L ammonium acetate with gradient elution in the column temperature of 35℃.The linear range for the detection of GABA by LC-MS/MS was 0.05-10.00 μmol/L,with a lower limit of quantification of 0.02 μmol/L,the inter-day CV<3.00%and intra assay CV<4.00%,respectively,and the recovery rate was 101.06%-109.02%.The reference ranges for plasma GABA were established by analyzing 300 healthy controls stratified by age:18-34 years(0.08-0.15 μmol/L),35-49 years(0.10-0.20 μmol/L),and≥50 years(0.12-0.23 μmol/L).Then plasma GABA was used as a biomarker for auxiliary diagnosis of sleep disorders in analyzing 221 patients and 300 healthy controls,which revealed that AUC values were 0.510(P=0.850),0.686(P=0.002),and 0.890(P<0.001)in the groups of 18-34 years,35-49 years,and≥50 years,respectively,with optimal cut-off values of 0.09,0.10 and 0.11 μmol/L.Conclusion A reliable LC-MS/MS method for detecting GABA has been established,which can detect plasma GABA levels sensitively and accurately and can be used in assisting the clinical diagnosis of sleep disorders.
