Reflectometric cutinase assay for rapid screening of contaminants and residues of insecticidal organophosphates and carbamates

Abstract

Because of the extensive use of insecticides in agriculture, there is an increasing demand for rapid analytical methods for residues in food and feed control. To meet this need, a completely new application of the reflectometric lipase test (Reflectoquant®, Merck) was developed. By using the cutinase-induced reaction of the substrate 5-bromo-4-chloro-3-indoxyl caprylate on the test strips, residues of organophosphates and carbamates can be determined on the basis of enzyme inhibition in a fast and inexpensive way. With this technique, we investigated the inhibition effects of representative insecticides, i.e., chlorpyrifos oxon, paraoxon, and carbaryl. The bimolecular inhibitory rate constants (ki) were found to agree well with those obtained by a previously described spectrophotometric cutinase assay in the microtiter-plate format. Recoveries determined with the strip test from spiked samples compared well with those obtained by both the cutinase microtiter-plate assay and liquid chromatography/mass spectrometry.