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The main objective of technical protective clothing is to enhance people safety at work, which may save their life or keep them healthy away against some hazards. We developed a warning cotton fabric with a traffic safety warning photoluminescence character that continues emitting light for a long period of time after the removal of the illuminant source. Rare earth-doped strontium aluminate was dispersed in an aqueous medium of a polyacrylic-based binder to give a cross-linkable photoluminescent formula to be applied onto cotton substrate employing spray-coat approach. To introduce a transparent photoluminescent film, the Rare earth pigment must be fully dispersed to prevent aggregation. The long-persistent photoluminescent layer was deposited on cotton surface employing different concentrations of the rare earth pigment phosphor. The excitation wavelength maximum band of the spray-coated film on cotton fabric was found to occur at 365 nm, while the emission was recorded at 515 nm. Yellowish-green emissive color was monitored by CIE color data under the ultraviolet excitation source. The spray-coated fabric was characterized by wavelength dispersive X-ray fluorescence (WD-XRF), phosphorescence and excitation spectra, elements mapping, scanning electron microscopy (SEM) and energy-dispersive X-ray (EDX). The comfort measurements were studied by exploring both of fabric stiffness and air-permeability. Furthermore, the spray-coated textile substrates displayed good fastness properties and a reversible luminescent glow in the dark.