sarin (c4h10fo2p,o-isopropyl methylphosphonofluoridate) is a colourless, odourless and highly toxic phosphonate that acts as a cholinesterase inhibitor and disrupts neuromuscular transmission. sarin and related phosphonates are chemical warfare agents, and there is a possibility of their application in a military or terrorist attack. this paper reports a lab-on-a-chip device for detecting a trace amount of sarin in a small volume of blood. the device should allow early detection of sarin exposure during medical triage to differentiate between those requiring medical treatment from mass psychogenic illness cases. the device is based on continuous-flow microfluidics with sequential stages for lysis of whole blood, regeneration of free nerve agent from its complex with blood cholinesterase, protein precipitation, filtration, enzyme-assisted reaction and optical detection. whole blood was first mixed with a nerve gas regeneration agent, followed by a protein precipitation step. subsequently, the lysed product was filtered on the chip in two steps to remove particulates and fluoride ions. the filtered blood sample was then tested for trace levels of regenerated sarin using immobilised cholinesterase on the chip. activity of immobilized cholinesterase was monitored by the enzyme-assisted reaction of a substrate and reaction of the end-product with a chromophore. resultant changes in chromophore-induced absorbance were recorded on the chip using a z-shaped optical window. loss of enzyme activity obtained prior and after passage of the treated blood sample, as shown by a decrease in recorded absorbance values, indicates the presence of either free or regenerated sarin in the blood sample. the device was fabricated in pmma (polymethylmethacrylate) using co2-laser micromachining. this paper reports the testing results of the different stages, as well as the whole device with all stages in the required assay sequence. the results demonstrate the potential use of a field-deployable hand-held device for point-of-care triage of suspected nerve agent casualties.
in recent years, the growing concern of terrorist attacks has resulted in a demand for point-of-care diagnostic devices that allow fast and effective triage of casualties exposed to chemical warfare agents from the “worried-well” masses. sarin (c4h10fo2p,o-isopropyl methylphosphonofluoridate) is a phosphonate that is highly toxic to humans because it acts as an acetylcholinesterase (ache) inhibitor to disrupt neuromuscular transmission. sarin belongs to the g-type nerve agents and is colourless and odourless. the lethal concentration-time dose of sarin for 50% of exposed individuals (humans) is 75 to 100 mg min?1 m?3.1 sarin and related phosphonates are chemical warfare agents and there is a possibility of their application in a military or terrorist attack.2 thus, there is a need for its detection in the prevention of an attack and for early treatment after exposure. fluorescence is frequently used for enzyme activity detection in the presence of a nerve agent. the enzyme can be immobilised on packed microbeads to increase the contact surface area.3 zhang and swager developed fluorescence-based functional group specific ratiometric chemosensors.4 simonian et al. developed a detection scheme based on a competitive inhibitor, whose change in fluorescence is generated by gold nanoparticles attached to the enzyme.5 viveros et al. used an opticalwave guidewith immobilised enzyme to detect the change in fluorescence.6 bencic-nagale et al. used nerve-agent-reactive microbead probes with fluorescence as the detector.7 all of these reported approaches did not use a microfluidic platform for sample analysis, and none of the papers reported above worked with whole blood as the sample.