色氨酸与2,6-二氯醌氯亚胺荷移荧光光谱法的研究

Study on the Charge –Transfer Fluorescence Spectroscopy of Trytophan with 2,6- Dichloroquinone-4-Chloroimide

本文用荧光猝灭法研究了色氨酸与2,6-二氯醌氯亚胺(DBQ)的最佳反应条件。实验得知,DBQ在一定的条件下对色氨酸的自体荧光有显著的猝灭作用,在p H为11.0的三酸缓冲介质中,在19.8℃下反应15min,可生成稳定的荷移络合物。该荧光猝灭法的线性回归方程为△F=0.9251+37.647c(单位为μg/m L),相关系数R=0.9990。色氨酸的浓度在0.51~1.33μg/m L的范围内与荧光强度的猝灭程度呈良好的线性关系,检测限为0.02μg/m L。平行测定含0.51μg/m L的色氨酸15次,方法的相对标准偏差为1.3%,回收率在97.5~100.6%的范围内。

A new fluorescence quenching method has been developed for the determination of trytophan. It is based on the fluorescence quenching by charge transfer reaction of trytophan with 2,6-Dichloroquinone-4-Chloroimide in Britton-Robinson buffer solution. Experiment indicated that the reaction was carried in p H=11.0 BR buffer solution at room temperature for 15 min. The linear equation of the fluorescence quenching is △ F =0.9251+37.647c(μg/m L)(R=0.9990). The calibration curve is linear in the concentration range of 0.51 ~ 1.33 μg/m L, the detection limit is 0.02μg/m L. Fifteen replicate determinations of solutions containing 0.51μg/m L tryptophan show a relative standard deviation of 1.3%. This proposed method has been applied to the determination of trytophan in serum. The recovery is 97.5% ~ 100.6%.