Enzyme-based microassay for accurate determination of soman in blood samples

Successful medical therapy for nerve agent intoxication requires early diagnosis and treatment. Current clinical diagnostic methods do not permit early or definitive confirmation of intoxication. To improve the chances of successful medical therapy against nerve agent intoxication, a sensitive enzyme-based microassay for rapid and accurate quantification of residual soman levels in blood was developed. The new analytical technique is based on the linear correlation between residual eel acetylcholinesterase activities and the inhibitor concentration. Blood samples were deproteinized with perchloric acid, followed by immediate neutralization after deproteinization. The mixtures were centrifuged at 3000g and the supernatant was directly assayed for soman. The sensitivity of the technique (18-1820 pg/ml blood) is comparable to that attained by GC-FID analysis (250 pg/ml blood). To facilitate routine analysis, the linear range of the assay was optimized to span over a factor of 100 (0.1-10 nM), with a typical correlation factor of at least 0.999 (six standards). The assay accuracy, checked with four different concentrations of soman, was within +/- 10%. The assay capability in monitoring the pharmacokinetic of soman was validated using both in vitro and in vivo rat models.