Capillary Gas Chromatographic Determination of Gamma Aminobutyric acid and Putrescine in Cerebrospinal Fluid using Trifluoroacetylacetone as Derivatizing Reagent

Abstract

determination of gamma aminobutyric acid (GABA) and putrescine (Put) using trifluoroacetylacetone (FAA) as derivatizing reagent from Cerebrospinal Fluid (CSF) samples prior to their gas chromatographic- flame ionization detector (GC-FID) analysis. GABA, Put, cadaverine (Cad) and tyramine (TY) as imitative of FAA extracted from the column HP-5 (30 m x 0.32 mm i.d) at temperature 110 oC for 1 min, tracked by heating rate 25 oC to 260 oC /min. The detection was carried out by FID with segregate ratio 10:1, v/v with whole run time 10 min. The proposed method showed linear calibration range between 2.5-50 µg/mL with low limit of detection 1.0 - 2.5 µg/mL analogous to 0.1 ng to 0.25 ng for selected Put, Cad, GABA, and TY. The method based on the pre-concentration was used for the determination of GABA and Put from CSF of human being and amounts found were 0.25- 0.56 µg/mL and 0.16 - 0.41 µg/mL with relative standard deviation (RSD) within 0.8 - 1.1 and 1.1 - 1.5 %, respectively. Many of aminoacids tested, separated completelyand did not variate the determinations of GABA and Put.