title: Preparation of antibodies and development of an enzyme immunoassay for determination of atrazine in environmental samples.
abstract: An indirect competitive enzyme-linked immunosorbent assay (ELISA) has been developed and optimized for atrazine determination in soil at different depths (0-10, 10-20, and 20-30 cm) before and after 48 h of application, corn shoot and cow milk samples collected from Dina farm, Egypt. This assay was based on a specific polyclonal antibodies (PAb) raised by immunizing New Zealand rabbits with an immunogen prepared by coupling 3-{4-(ethylamino)-6-(isopropylamino)-1,3,5-triazine-2-yl} thiopropanoic acid to bovine serum albumin (BSA) via N-hydroxysuccinimide (NHS) active ester method. The sensitivity (estimated as IC₅₀ value) was 17.5 μg mL⁻¹ with a detection limit of 0.1 ng mL⁻¹. The maximum atrazine concentration was found in soil especially in the deepest layer (325 and 890 μg kg⁻¹ before and after application, respectively). Atrazine concentration in corn shoot was 333.28, μg kg⁻¹ dry plant, while there was no detectable amount in milk. All samples screened by ELISA were validated by gas chromatography mass spectrometer procedure (GC/MS). Good correlation was achieved between the two methods (r = 0.997 for soil and 0.9814 for plant). This study demonstrates the utility and convenience of the simple, practical and cost-effective ELISA method in the laboratory for analysis of environmental samples. The method is ideal for the rapid screening of large numbers of samples in laboratories where access to GC/MS facilities, is limited or lacking.
authors.affiliation: Department of Pesticide Chemistry, Faculty of Agriculture, Alexandria University, Alexandria, Egypt. kawther52@hotmail.com
authors.forename: Kawther S
authors.initials: KS
authors.lastname: El-Gendy
authors.forename: Nagat M
authors.initials: NM
authors.lastname: Aly
authors.forename: Eman M
authors.initials: EM
authors.lastname: Mosallam
authors.forename: Ahmed K
authors.initials: AK
authors.lastname: Salama
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D001280:Atrazine / Q000032:analysis*; D002417:Cattle; D004534:Egypt; D004784:Environmental Monitoring / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000191:economics / Q000379:methods*; D008401:Gas Chromatography-Mass Spectrometry; D006241:Haptens / Q000276:immunology; D006540:Herbicides / Q000032:analysis*; D020128:Inhibitory Concentration 50; D008892:Milk / Q000737:chemistry*; D011817:Rabbits; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000276:immunology; D012987:Soil / Q000737:chemistry*; D012989:Soil Pollutants / Q000032:analysis; D003313:Zea mays / Q000737:chemistry*
chemical_list: D000906:Antibodies; D006241:Haptens; D006540:Herbicides; D012987:Soil; D012989:Soil Pollutants; D012710:Serum Albumin, Bovine; D001280:Atrazine