<p>The possibility of fluorescent detection of albumin through its reaction with Coomassie Brilliant Blue G (CBBG) in organized media of nonionic surfactants (n-Surf) has been investigated. Optimal conditions for generating the fluorescent signal of CBBG in solution have been determined and proposed. The optimal conditions for detecting the fluorescent signal of albumin in the aqueous-alcohol CBBG–n-Surf system have also been established. It has been shown that classical micelle-forming surfactants, particularly Triton X-100 (1.0–6.0)&#xa0;×&#xa0;10^–5&#xa0;M and Triton X-305 (1.0–2.0)&#xa0;×&#xa0;10⁻⁴&#xa0;M, are most suitable for constructing analytical systems for albumin determination. The presence of n-Surf in solutions enhances the signal intensity of both the dye and the analyte while stabilizing the colloid-chemical state of the solutions. A fluorescence-based technique for determining albumin content using the n-Surf-modified CBBG-ethanol system (<i>r</i>² = 0.99, LOD = 1.1&#xa0;mg/l) has been developed and tested for albumin determination in urine. The proposed approach can be applied to optimize and enhance the metrology of other reagent systems for the fluorescence determination of trace elements.</p> Graphical abstract <p></p>