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

Abstract

In present work, a new capillary gas chromatographic procedure was established and validated for the 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 amino-acids tested, separated completely and did not variate the determinations of GABA and Put.

Keywords

• Trifluoroacetylacetone
• Gamma aminobutyric acid
• Putrescine
• Amino-acids
• Cerebrospinal Fluid