Smartphone-assisted colorimetric sensing of enzyme-substrate system using pH-responsive gold nanoparticle assembly

Highlights: • A pH-responsive gold nanoparticle assembly probe for sensing subtle pH changes is developed. • The developed probe shows a reversible and highly sensitive response to pH variation between 6.20 and 7.40. • The pH-responsive probe can detect a slight pH variation in the enzyme-catalyzed reactions. • The proposed strategy shows great potential for POCT of blood glucose and on-site determination of urea. Gold nanoparticle (AuNP)-based colorimetric biosensors have been widely used for pH sensing and monitoring its changes. However, few AuNP-based pH sensors have been developed through the manipulation of the aggregation states of AuNPs via i-motif DNA. We herein report i-motif DNA-assisted pH-responsive gold nanoparticle assembly (termed RGA), which shows a reversible and highly sensitive response to pH variation between 6.20 and 7.40. The acidic pH triggers the disassembly of the RGA, thus converting the AuNPs from aggregation state to disperse state, which leads to a color transition from blue-purple to red. Therefore, the pH value can be estimated by naked-eye determination or UV–vis spectroscopy analysis. More significantly, the visually detectable color change is monitored using the built-in camera of a smartphone. The RGB (red, green, blue) values of the RGA solution are measured by a smartphone application (APP). Following data processing, the RGB values can be converted into pH value, providing a new strategy for the on-site and real-time pH sensing. Furthermore, the pH-induced conformation change of i-motif DNA allows the RGA to detect a slight pH fluctuation in the catalytic oxidation of glucose by glucose oxidase and the hydrolysis of urea by urease.