The derivative agent using in the determination ofγ-aminobutyric acid by HPLC lacked systematic comparative analysis. So this paper studied the comparison of three kinds of common derivative agent and chromatographic process. A gradient elution was used for the separation and quantification of GABA precolumn derivatization with OPA, Dansyl-Cl, AQC. The column were intertsil ODS-C18 (by the first two), waters AccQ·Tag for free amino acids. GABA was determined with UV detector at 332 nm, 386 nm, 250 nm, and linear equations were Y=9.0×106X+2062.5(R2=0.9989), Y=7.0×106X+11337(R2=0.9982), Y=7.1×106X+10025 (R2=0.9999), and detection limits were 0.005 mg/mL, 0.01 mg/mL, 0.001 mg/mL, and RSD of peak area of GABA measured in the same sample was 1.16%, 1.98%, 0.94%, and the recoveries were 98.5%-106.9%, 95.8%-110.6%, 99.4%-103.4%, GABA content measured in the same sample was 11.24, 10.67, 13.47 mg/ 100 g dry weight, respectively. We learned from the above results, that in three kinds of derivative agent determining GABA by HPLC, the most accurate one is AQC, followed by OPA and Dansyl-Cl, AQC, as a new derivative agent, has the broad prospect inγ-aminobutyric acid and even amino acid analysis.
Key words
germinated brown rice;γ- aminobutyric acid; HPLC; pre- column derivation; orthophthalaldehyde (OPA); dansyl chloride (dansyl-Cl)
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