Development of an enzyme immunoassay for the detection of trinitrotoluene (TNT) in soil and water

Conference · Sun Dec 31 04:00:00 EST 1995

OSTI ID:276393

Larkin, K A; Ferguson, B S; Matt, J J; Fan, T S ^[1]

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1. ImmunoSystems Inc., Scarborough, ME (United States)

An enzyme immunoassay (EIA) has been developed which is capable of detecting as little as 0.5 ppb TNT in water and 0.25 ppm TNT in soil. Rabbit polyclonal antibodies were raised against a TNT-derivative coupled to bovine serum albumin. These antibodies are coated on both polystyrene tubes and 96-well microtiter plates. Water samples or soil extracts together with a TNT-horseradish peroxidase conjugate are then added and compete for antibody binding sites. Excess conjugate is washed away in a simple rinse step, and substrate (3,3{prime},5,5{prime}-tetramethylbenzidine) is added; the amount of color developed is inversely proportional to the concentration of TNT in the sample. This competitive ELA is relatively specific: 4-amino-2,6-DNT and 2,6-DNT are only 20% as reactive as TNT, and 2,4-DNT shows approximately 1% cross-reactivity. Other common explosives and important metabolites, such as RDX, HMX, nitroglycerin, and dichlorobenzoic acids, are essentially non-reactive in this assay. The microliter plate-based assay is complete in 90 minutes and is suitable for laboratory use. The tube-based test is run in less than 30 minutes and is field-portable. The soil extraction protocol employed for this assay utilizes methanol as the solvent and can be completed in 10 minutes. A simple dilution step is the only sample preparation that is required before running soil extracts in the immunoassay.

• OSTI ID: 276393
• Report Number(s): CONF-950209--
• Country of Publication: United States
• Language: English

Related Subjects

45 MILITARY TECHNOLOGY, WEAPONRY, AND NATIONAL DEFENSE
54 ENVIRONMENTAL SCIENCES
ENZYME IMMUNOASSAY
PORTABLE EQUIPMENT
REMEDIAL ACTION
SITE CHARACTERIZATION
SOILS
TECHNOLOGY ASSESSMENT
TNT
WATER