Foldable paper-based analytical device for the detection of an acetylcholinesterase inhibitor using an angle-based readout

[Display omitted] •A foldable 3D paper device was designed for sequential enzyme reactions.•Semiquantitative analysis with the naked eye was achieved by an angle-based readout.•All reagents required for enzyme reactions were preadsorbed on the paper device.•Dimethyl methylphosphonate as one of the AChE inhibitors was analyzed. The rapid on-site detection of organophosphate compounds such as nerve agents and pesticides is essential due to their lethal effect, which irreversibly inhibits the activity of acetylcholinesterase (AChE). In this study, we demonstrate a foldable paper-based analytical device (PAD) that is capable of detecting an AChE inhibitor in a semiquantitative manner. An angle-based readout of the yellow area in the ring-type paper channel was used for the semiquantitative analysis with the naked eye. The color of the paper channel was changed to yellow as a result of oxidized cerium oxide nanoparticles by hydrogen peroxide produced from the enzymatic reactions. All reagents required for enzyme reactions were preadsorbed on the pop-up card shaped PAD to minimize the manual steps for dealing with multiple reagents for sequential enzyme reactions. As a proof of concept, we were able to semiquantitatively analyze a nerve agent simulant called dimethyl methylphosphonate (DMMP), one of the AChE inhibitors, by simply reading the angle of the yellow area in the paper channel. The foldable PAD proposed here would serve as a valuable tool that allows unskilled people to semiquantitatively detect AChE inhibitors in the field, including nerve agents and organophosphate pesticides.