Development and validation of a high-performance liquid chromatographic method for the determination of gamma-hydroxybutyric acid in rat plasma.
A method for the determination of gamma-hydroxybutyric acid (GHB) in rat plasma was developed using solid-phase extraction (SPE) and high-performance liquid chromatography (HPLC) with UV detection. GHB was isolated from plasma using strong anion-exchange SPE columns. The chromatographic separation was performed on a C18 Aqua column. The lower limit of quantification was 10 microg/ml using 60 microl of plasma. The linearity of the calibration curves was satisfactory as indicated by correlation coefficients of >0.990. The within-day and between-day precision were <10% (n=24), the accuracy was nearly 101%. Plasma concentrations in rats after GHB infusion determined by HPLC-UV were compared with the corresponding concentrations determined with a validated gas chromatographic-mass spectrometric method by orthogonal distance regression. A good correlation was observed and a t-test indicated no significant differences from 0 and 1 for the intercept and slope, respectively.
Heymans Institute of Pharmacology, Ghent University, Faculty of Medicine and Health Sciences, Belgium.
C A
CA
de Vriendt
D K
DK
van Sassenbroeck
M T
MT
Rosseel
E J
EJ
van de Velde
A G
AG
Verstraete
Y
Y
Vander Heyden
F M
FM
Belpaire
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008401:Gas Chromatography-Mass Spectrometry; D051381:Rats; D013056:Spectrophotometry, Ultraviolet; D005680:gamma-Aminobutyric Acid / Q000097:blood*
D005680:gamma-Aminobutyric Acid