A Validated HPLC Method for the Determination of GABA by Pre-Column Derivatization with 2,4-Dinitrofluorodinitrobenzene and Its Application to Plant GAD Activity Study |
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Abstract: | A validated, selective, and sensitive chromatographic method for determination of γ-aminobutyric acid (GABA) has been developed by precolumn derivative of the sample with 2,4-dinitrofluorodinitrobenzene (FDNB). The derivatives were separated with RP-HPLC on a C18 column and UV detection (360 nm). The calibration curve was linear over a range of 0.2–5.0 mg/mL GABA with a correlation coefficient of 0.9954. The precision of this method is high (RSD = 0.061%). This method also possessed high recovery rate (>99%) and good stability over a period of four weeks. The method was successfully used to determine the glutamate decarboxylase (GAD) activity in several plant tissues. |
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Keywords: | γ-aminobutyric acid 2,4-dinitrofluorodinitrobenzene Glutamate decarboxylase HPLC Pre-column derivatization reaction |
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