A colorimetric platform using highly active Prussian blue composite nanocubes for the rapid determination  of ascorbic acid and acid phosphatase

Cobalt-doped Prussian blue composite nanocubes (Co-PB NCs) were synthesized, which can quickly convert O₂ to O₂^•− and ¹O₂. Due to the presence of cobalt and iron transition metal redox electron pairs, Co-PB NCs with high oxidase mimetic activity can rapidly oxidize the substrate 3,3′,5,5′-tetramethylbenzidine (TMB) to produce blue products (ox-TMB) without the assistance of unstable H₂O₂. Using ascorbic acid-2-phosphate trisodium salt (AAP) as a substrate, it can be converted to reduced ascorbic acid (AA) under acid phosphatase (ACP) hydrolysis, resulting in suppression of TMB oxidation. Therefore, an enzyme cascade signal amplification strategy for rapid colorimetric detection of AA/ACP was developed based on the high-efficiency oxidase-like activity of Co-PB NCs combined with the hydrolysis effect of ACP. The color changes at low concentrations of AA and ACP could be observed by the naked eye, and the detection limits of AA and ACP were 1.67 μM and 0.0266 U/L, respectively. The developed colorimetric method was applied to the determination of AA in beverages and ACP in human serum, and the RSDs were less than 3%, showing good reproducibility. This work provides a promising strategy for the use of metal-doped Prussian blue composite material for the construction of rapid colorimetric sensing platforms that avoid the use of unstable hydrogen peroxide.