In this study, a novel fluorescent labeling reagent 2-(9-acridone)-ethyl chloroformate (AEC-Cl) was designed, synthesized and applied for the determination of free amino acids by high-performance liquid chromatography with a fluorescence detector (HPLC-FLD). The free amino acids were rapidly and efficiently labeled by AEC-Cl in the presence of basic catalyst (pH 9.0) within 5 min at room temperature (25 °C). The derivatives exhibited excellent stability and fluorescence properties, with maximum excitation and emission wavelengths at 268 nm and 438 nm, respectively. Derivatives of 22 kinds of natural amino acids were completely separated by gradient elution on a Hypersil ODS C18 column. Under the optimal conditions, the calibration curves exhibited excellent linear responses, with correlation coefficients of R² > 0.9994. The detection and quantification limits were in the range of 0.61–2.67 μg kg⁻¹ and 2.07–8.35 μg kg⁻¹, respectively. Therefore, AEC-Cl was successfully applied for the detection of trace levels of free amino acids in honey samples.

在这项研究中，设计，合成了新型荧光标记试剂2-（9-ac啶酮）-氯甲酸乙酯（AEC-Cl），并用于高效液相色谱和荧光检测器（HPLC- FLD）。在室温（25°C）下5分钟内，在碱性催化剂（pH 9.0）存在下，通过AEC-Cl快速有效地标记游离氨基酸。衍生物表现出优异的稳定性和荧光性质，最大激发和发射波长分别在268 nm和438 nm。在Hypersil ODS C18色谱柱上通过梯度洗脱将22种天然氨基酸的衍生物完全分离。在最佳条件下，校准曲线表现出出色的线性响应，相关系数为R ² > 0.9994。检出限和定量限分别在0.61–2.67μgkg ^-1和2.07–8.35μgkg ^-1之间。因此，AEC-Cl已成功应用于蜂蜜样品中痕量游离氨基酸的检测。