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Title:Enzyme-linked immunosorbent assay for herbicide detection
Author(s):Koppatschek, Fritz K.
Doctoral Committee Chair(s):Liebl, Rex A.
Department / Program:Crop Sciences
Discipline:Crop Sciences
Degree Granting Institution:University of Illinois at Urbana-Champaign
Degree:Ph.D.
Genre:Dissertation
Subject(s):Agriculture, Agronomy
Chemistry, Agricultural
Abstract:The purpose of this research was to develop polyclonal antibodies to the soybean herbicides clomazone (2- ((2 chlorophenyl)methyl) -4,4-dimethyl-3-isoxazolidinone), imazaquin (2- (4,5-dihydro-4-methyl-4-(1-methylethyl)-5-oxo-1H-imidazol-2-yl) -3-quinolinecarboxylic acid), and metribuzin (4-amino-6-(1,1-dimethylethyl)-3-(methylthio)-1,2,4-triazin-5(4H)-one). The herbicide-specific antibodies were to be used in an enzyme linked immunosorbent assay (ELISA) for herbicide detection in soil.
Antibodies, specific to clomazone, were produced in New Zealand white rabbits to a BSA-clomazone conjugate. Attempts at producing antibodies specific to imazaquin, via BSA-imazaquin and BSA-hexanoic acid-imazaquin conjugates, were not successful. Antibodies specific to metribuzin, via a porcine gamma globulin-metribuzin conjugate, were not successfully produced.
The antibodies to the BSA-clomazone conjugate were sensitive to clomazone having an I$\sb{50}$ of 12 ng/mL. The antibodies were also specific to clomazone and demonstrated no cross reactivity to the soybean herbicides metribuzin, metolachlor (2-chloro-N-(2-ethyl-6-methylphenyl)-N-(2-methoxy-1-methylethyl)acetamide) or trifluralin (2,6-dinitro-N,N-dipropyl-4-(trifluromethyl)benzenamine).
Soil extracts, from several different soil types, did not interfere with detection of clomazone in this ELISA. Soil extraction procedures could also be simplified without reducing the sensitivity of the assay.
The ELISA system compared favorably with conventional clomazone detection systems of wheat bioassay and gas chromatography. The low level of detection for both gas chromatography and ELISA was from a 10 ppb acetonitrile extract of clomazone treated soil.
Clomazone-treated soil was extracted with phosphate buffer and clomazone was quantified with ELISA. Detection of clomazone in the phosphate buffer extracts was influenced by soil type, with greater detection in low organic matter soil. This technique offers a novel method for quantifying a portion of the biologically available herbicide.
The ELISA system proved to be a fast, accurate and sensitive method of clomazone detection.
Issue Date:1989
Type:Text
Language:English
URI:http://hdl.handle.net/2142/21758
Rights Information:Copyright 1989 Koppatschek, Fritz K.
Date Available in IDEALS:2011-05-07
Identifier in Online Catalog:AAI9010922
OCLC Identifier:(UMI)AAI9010922


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