ELISA test, a new method to detect and quantify isoproturon in soil

Abstract

An indirect competitive Enzyme-Linked ImmunoSorbent Assay (ELISA) has been developed to quantify isoproturon (a commonly used phenylurea herbicide) in soil samples. To produce the specific antibodies needed for detection of isoproturon, an isopropylanilin analogue was covalently linked to a proteic carrier and the resulting conjugate used as immunogen. Hybridomas were obtained from spleen cells of mice immunized with this conjugate. One specific monoclonal antibody (MAb) was selected and used to the ELISA allowing the detection of small amounts of herbicide (20-250 micrograms l-1). Cross-reactivities with other current phenylureas appear to be insignificant. Samples of six soils were mixed with several quantities of herbicide (to reach final concentrations ranging between 0.15 and 1 microgram g-1) and extracted either with methanol to determine the total residue or with a solution of CaCl2, to establish the herbicide available from the soil solution. The extracts were analyzed simultaneously by gas chromatography and ELISA in these conditions, correlation coefficients between both methods were never lower than 93.5%. Organic matter and ionic strength appeared to be the most important parameters interfering the ELISA test. To avoid these drawbacks, standard curves were established for the ELISA by spiking isoproturonless soils extracts. In these conditions, correlations between both methods were considered as sufficient. ELISA of untreated samples were carried out to verify the isoproturon availability curves established previously.