Application of a cholinesterase biosensor to screen for organophosphorus pesticides extracted from soil

Based on the principle of enzyme inactivation, a butyrylcholinesterase (EC 3.1.1.8.) biosensor, to determine some organophosphorus (ORP) pesticides (Fenitrothion, Diazinon, Parathion ethyl, Mevinphos and Heptenophos) in soil extracts, is presented. The enzyme was immobilized on pre-activated Pall Biodyne(TM) transfer membranes, which were physically attached to the sensitive ends of glass pH electrodes. Contact of the enzyme with pesticide samples results in specific inhibition of enzyme activity. Sensor calibration was possible by correlating the inhibition of enzyme activity (monitored by observing reduction in electrode potential changes with substrate additions) with varying concentrations of pesticide compounds in a buffer solution. A simple procedure was designed to extract ORP pesticides from spiked soil samples using a mixture of dichloromethane and acetone as the extraction solvent mixture. The sensor was successfully used to determine pesticide concentrations ranging from a low of 35 ppb (Diazinon) to 21 ppm (Fenitrothion) in soil, with resultant relative standard deviations of percentage enzyme inactivation less than 12%. The complete extraction and analytical procedure is simple, inexpensive and rapid. Mass production of the enzyme membranes and their easy attachment to the electrodes, render them disposable after a single use. The biosensor is seen as a potential analytical instrument for early warning against pesticide contaminations in soil.