Portable Paper-Based Colorimetric Biosensor for Rapid Screening of Organophosphate Exposure via Acetylcholinesterase Activity and RGB Image Analysis.

BACKGROUND

Rapid screening of acetylcholinesterase (AChE) inhibition is essential for monitoring exposure to organophosphate compounds, particularly in field settings where access to laboratory infrastructure is limited. This study aimed to develop a portable paper-based colorimetric biosensor for the semi-quantitative detection of AChE activity in blood samples.

METHODS

The biosensor was based on a pH-dependent color change adapted from the modified Edson method. The platform was first optimized using experimental models and then evaluated in human capillary blood samples collected under real field conditions. Reference serum cholinesterase activity was determined by a certified clinical laboratory and used for comparison with the colorimetric response of the biosensor. RGB (red, green, and blue) image analysis was performed in a subset of samples to digitally characterize the chromatic response of the device.

RESULTS

Samples with preserved AChE activity showed a visible color shift associated with substrate hydrolysis, whereas samples with reduced or inhibited activity maintained darker blue-dominant tones. RGB analysis of human samples revealed significant associations between AChE activity and the R and G channels, supporting the ability of the platform to distinguish between chromatic patterns associated with different ranges of enzymatic activity.

CONCLUSIONS

The proposed platform demonstrated the feasibility of translating a pH-based AChE assay into a portable paper-based format for semi-quantitative visual and RGB-assisted screening. Its evaluation in human samples demonstrated the feasibility of its use under field conditions as a proof-of-concept. Further studies are needed to optimize its analytical performance and validate its application in larger and more diverse populations.