The real time monitoring of some organophosphorus based pesticides is of great concern to environmentalists because the widespread use of pesticides is causing severe health hazards to all living beings and also hampering our ecological balance. The traditional methods of measurement of pesticide residues are time consuming, need sample pre-treatment, and lack desired specificity and accuracy. We have developed an amperometric biosensor for indirect measurement of the pesticide concentration precisely in ppb level. The method is based on the action of two enzymes namely acetylcholine esterase and choline oxidase which are uniquely immobilized in a polymeric porous network directly on the working electrode of a screen-printed sensor. Polyacrylamide matrix has been prepared by copolymerisation of acrylamide and N,N'-methylenebisacrylamide using Potassium peroxodisulphate (K2S2O8) as initiator. A linear relationship was obtained between the range of 0 to 10 ppb. © Springer Science+Business Media, Inc. 2006.