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Quantitation of γ ‐ aminobutyric acid in equine plasma by hydrophilic interaction liquid chromatography with tandem mass spectrometry
Author(s) -
Yi Rong,
Zhao Sarah,
Kong Noel,
Zhang Julia,
Loganathan Devan,
Mérette Sandrine,
Morrissey Barbara
Publication year - 2017
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201700245
Subject(s) - aminobutyric acid , chemistry , chromatography , tandem mass spectrometry , bovine serum albumin , mass spectrometry , quantitative analysis (chemistry) , endogeny , liquid chromatography–mass spectrometry , neurotransmitter , biochemistry , receptor
γ ‐ Aminobutyric acid is the principal inhibitory neurotransmitter in the central nervous system and regulates the neuronal excitability. There has been anecdotal evidence that γ ‐ aminobutyric acid has been used within a few hours prior to competition in equine sports to calm down nervous horses. However, regulating the use of γ ‐ aminobutyric acid is challenging because it is an endogenous substance in the horse. γ‐Aminobutyric acid is usually present at low ng/mL levels in equine plasma; therefore, a sensitive method has to be developed to quantify these low background levels. Measuring low concentrations of endogenous γ ‐ aminobutyric acid is essential to establish a threshold that can be used to differentiate levels attributable to exogenous administrations of γ ‐ aminobutyric acid. A hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry method was developed and validated for the quantitation of γ ‐ aminobutyric acid in equine plasma. Calibrators were prepared in artificial surrogate matrix consisting of 35 mg/mL equine serum albumin in phosphate buffered saline. Samples were prepared by protein precipitation with acetonitrile. Utilizing this methodology, a total of 403 equine plasma samples collected post‐competition from horses participating in equestrian events in Canada were analyzed.