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An in-capillary derivatization and stacking capillary electrophoresis (CE) technique has been applied to redeem the detection of dilute analytes in the analysis of gamma-aminobutyric acid (GABA) and alanine (Ala) in tea samples. Extracts from samples were diluted to eliminate matrix interference before introduction into the CE system. GABA and Ala in the diluted sample zone were derivatized with o-phthaldialdehyde/2-mercaptoethanol (OPA/2-ME) to form fluorescence-labeled products in the stacking process, and the labeled derivatives were then enriched by online stacking. Optimal conditions for the stacking, such as the concentration of the background buffer solution, the matrix of the sample zone (sample solution), and the volume of the sample injection, were investigated and then applied to real sample analysis. Under optimum conditions, the detections were linear in the range of 5.0 nM-2.5 microM with the square of correlation coefficients (R2) of 0.9995 and 0.9992 for GABA and Ala, respectively. Detection limits were found to be 0.7 and 0.8 nM for GABA and Ala, respectively. Tea samples were analyzed with recoveries between 92.33 and 97.87% and between 94.36 and 96.46% for GABA and Ala, respectively. This method is a rapid, convenient, and sensitive process for determining GABA and Ala in complicated matrix samples such as tea samples.

The aim of this study was to investigate an in-capillary derivatization capillary electrophoresis (CE) technique that was performed to determine the concentration of gamma-aminobutyric acid (GABA) and alanine (Ala) in tea after being derivatized with o-phthaldialdehyde/2-mercaptoethanol (OPA/2-ME) to form fluorescence-labeled products. The conditions of labeled derivatization and CE separation were optimized and then applied to real sample analysis. The labeled derivatization with 20 mM OPA and 26.67 mM 2-ME (mol ratio=0.75) at pH 10 offered the most sensitive detection, and the separation with 30 mM sodium tetraborate buffer (pH 10.0) under 21 kV achieved good selectivity within 14 min. The detections were linear in the range of 0.05-5 microM with correlation coefficients (R2) of 0.9995 and 0.9964 and with detection limits of 0.004 and 0.02 microM for GABA and Ala, respectively. The recoveries were 94.22% (3.58% RSD) and 93.54% (6.46% RSD) for five determinations of GABA and Ala, respectively. This method is a fast, convenient, sensitive, and eco-friendly way to determine the GABA and Ala in tea samples from different manufacturing processes.