Blass K G
Department of Chemistry, University of Regina, Saskatchewan, Canada.
Clin Biochem. 1995 Apr;28(2):107-11. doi: 10.1016/0009-9120(95)00004-s.
To develop a new quantitative fluorometric assay for creatinine with enhanced specificity and inherent fluorescence sensitivity.
Creatinine is reacted with 3,5-dinitrobenzoate under alkaline reaction conditions to produce a fluorophore. Alternatively, 3,5-dinitrobenzoyl chloride or methyl-3,5-dinitrobenzoate may be employed as reagents. Fluorophore development was evaluated for Group 1A bases under aqueous and mixed solvent conditions.
The chemical reaction between creatinine and 3,5-dinitrobenzoate performed under highly alkaline conditions produced a fluorescent product with excitation and emission maxima near 410 and 475 nm, respectively. The reaction proceeds rapidly in the presence of group IA bases under both aqueous and mixed solvent conditions, in particular with water/dimethylsulfoxide and water/1,4-butanediol solvent systems containing 1.0 M LiOH. A linear calibration curve was obtained when the fluorescence intensity measured in microamperes was plotted versus creatinine concentration in the range of 5-50 mumol/L. The detection limit for creatinine was well below 1 mumol/L.
A simple, sensitive and highly specific fluorometric method for the determination creatinine has been developed.
开发一种用于肌酐的新型定量荧光测定法，具有更高的特异性和内在荧光灵敏度。
肌酐在碱性反应条件下与3,5-二硝基苯甲酸反应生成荧光团。另外，也可使用3,5-二硝基苯甲酰氯或3,5-二硝基苯甲酸甲酯作为试剂。在水性和混合溶剂条件下，对第1A族碱的荧光团生成情况进行了评估。
肌酐与3,5-二硝基苯甲酸在高碱性条件下进行的化学反应产生了一种荧光产物，其激发和发射最大值分别接近410和475nm。在水性和混合溶剂条件下，特别是在含有1.0M氢氧化锂的水/二甲基亚砜和水/1,4-丁二醇溶剂体系中，该反应在第1A族碱存在下迅速进行。当以微安为单位测量的荧光强度与5-50μmol/L范围内的肌酐浓度作图时，得到了一条线性校准曲线。肌酐的检测限远低于1μmol/L。
已开发出一种简单、灵敏且高度特异的荧光法用于测定肌酐。
