TNT Concentration Research Articles

Bioremediation of 2,4,6-Trinitrotoluene by Bacterial Nitroreductase Expressing Transgenic Aspen

Trees belonging to the genus Populus are often used for phytoremediation due to their deep root formation, fast growth and high transpiration rates. Here, we study the capacity of transgenic hybrid aspen (Populus tremula x tremuloides var. Etropole) which expresses the bacterial nitroreductase gene, pnrA, to tolerate and take-up greater amounts of the toxic and recalcitrant explosive, 2,4,6-trinitrotoluene (TNT) from contaminated waters and soil. Transgenic aspen tolerate up to 57 mg TNT/L in hydroponic media and more than 1000 mg TNT/ kg soil, whereas the parental aspen could not endure in hydroponic culture with more than 11 mg TNT/L or soil with more than 500 mg TNT/kg. Likewise, the phytotoxicological limit for transgenic plants to a constant concentration of TNT was 20 mg TNT/L while wild-type plants only tolerated 10 mg TNT/L. Transgenic plants also showed improved uptake of TNT over wild-type plants when the original TNT concentration was above 35 mg TNT/L in liquid media or 750 mg TNT/kg in soil. Assays with 13C-labeled TNT show rapid adsorption of TNT to the root surface followed by a slower entrance rate into the plant. Most of the 13C-carbon from the labeled TNT taken up bythe plant (> 95%) remains in the root with little translocation to the stem. Altogether, transgenic aspen expressing pnrA are highly interesting for phytoremediation applications on contaminated soil and underground aquifers.

Characterization of wastewater from the Brazilian TNT industry

The objective of this work was to characterize the effluent originating from a Brazilian TNT production industry. Analyses were performed using physical, chemical, spectroscopic and ecotoxicological assays, which demonstrated that the effluent had a significant pollution potential, mainly due to the low pH and high concentration of TNT (156 ± 10 mg L −1). The results also demonstrated that the effluent presented significant acute toxicity, and could cause countless damages if released into the receiving body without being adequately treated first. The observed pollution potential justifies studies to evaluate treatment technologies or recover the residue generated in the TNT industry.

Denitration of 2,4,6-trinitrotoluene by Pseudomonas aeruginosa ESA-5 in the presence of ferrihydrite

Denitration of 2,4,6-trinitrotoluene (TNT) was evaluated in oxygen-depleted enrichment cultures. These cultures were established starting with an uncontaminated or a TNT-contaminated soil inoculum and contained TNT as sole nitrogen source. Incubations were carried out in the presence or absence of ferrihydrite. A significant release of nitrite was observed in the liquid culture containing TNT, ferrihydrite, and inoculum from a TNT-contaminated soil. Under these conditions, Pseudomonas aeruginosa was the predominant bacterium in the enrichment, leading to the isolation of P. aeruginosa ESA-5 as a pure strain. The isolate had TNT denitration capabilities as confirmed by nitrite release in oxygen-depleted cultures containing TNT and ferrihydrite. In addition to reduced derivatives of TNT, several unidentified metabolites were detected. Concomitant to a decrease of TNT concentration, a release of nitrite was observed. The concentration of nitrite peaked and then it slowly decreased. In the absence of TNT, the drop in the concentration of nitrite in oxygen-depleted cultures was lower when ferrihydrite was provided, suggesting that ferrihydrite inhibited the utilization of nitrite by P. aeruginosa ESA-5.

Photocatalytic Degradation of 2,4,6-Trinitrotoluene in Wastewater Using a Thin-Film TiO2Reactor

The photocatalytic treatment of water contaminated with 2,4,6-trinitrotoluene (TNT) was explored in bench-scale experiments in batch mode using a Pyrex tube coated with a thin film of TiO2 located inside a photoreactor. The reactor was aerated by purging it with compressed air before initiating the photocatalytic reaction. The rate of TNT degradation approximated first-order kinetics. The reaction rate constant decreased as the TNT concentration increased from 25 to 100 mg/L, while the first-order kinetics could be modeled using a Langmuir adsorption isotherm. The addition of the organic reductants methanol and EDTA significantly enhanced the rate of TNT degradation, with optimum results in the presence of 20% methanol by volume. EDTA increased the rate of TNT removal by enhancing the role of the reductants.

Validation of an Antibody-Based Biosensor for Rapid Quantification of 2,4,6-Trinitrotoluene (TNT) Contamination in Ground Water and River Water

Nitroaromatics are common pollutants of soil and groundwater at military installations because of their manufacture, storage, and use at these sites. Long-term monitoring of these pollutants comprise a significant percentage of restoration costs. Further, remediation activities often have to be delayed, while the samples are processed via traditional chemical assessment protocols. Here we describe a rapid (<5 min), cost-effective, accurate method using a KinExA Inline Biosensor for monitoring of 2,4,6-trinitrotoluene (TNT) in field water samples. The biosensor, which is based on KinExA technology, accurately estimated the concentration of TNT in double-blind comparisons with similar accuracy to traditional high-performance liquid chromatography(HPLC). In the assessment of field samples, the biosensor accurately predicted the concentration of TNT over the range of 1-30,000 microg/L when compared to either HPLC or quantitative gas chromatography-mass spectrometry (GC-MS). Various pre-assessment techniques were explored to examine whether field samples could be assessed untreated, without the removal of particulates or the use of solvents. In most cases, the KinExA Inline Biosensor gave a uniform assessment of TNT concentration independent of pretreatment method. This indicates that this sensor possesses significant promise for rapid, on-site assessment of TNT pollution in environmental water samples.

Fenton-like oxidation of 2,4,6-trinitrotoluene using different iron minerals

Degradation of 2,4,6-trinitrotoluene (TNT) was investigated in presence of different oxidants (Fenton's reagent, sodium persulfate, peroxymonosulfate and potassium permanganate) and different iron minerals (ferrihydrite, hematite, goethite, lepidocrocite, magnetite and pyrite) either in aqueous solution or in soil slurry systems. Fenton's reagent was the only oxidant able to degrade TNT in solution (k(app)=0.0348 min(-1)). When using iron oxide as heterogeneous catalyst at pH 3, specific reaction rate constants per surface area were k(surf)=1.47.10(-3) L min(-1) m(-2) and k(surf)=0.177 L min(-1) m(-2) for magnetite and pyrite, respectively while ferric iron minerals were inefficient for TNT degradation. The major asset of iron mineral catalyzed Fenton-like treatment has been the complete oxidation of the pollutant avoiding the accumulation of possible toxic by-products. In soil slurry systems, 38% abatement of the initial TNT concentration (2 g/kg) was reached after 24 h treatment time at neutral pH. Rate limiting steps were the availability of soluble iron at neutral pH together with desorption of the TNT fraction sorbed on the clay mineral surfaces.

Diagnostic value of D-dimer measurement in patients referred to the emergency department with suspected myocardial ischemia

The accurate identification of patients with acute myocardial infarction (AMI) remains one of the most difficult challenges facing emergency physicians. The introduction of early and reliable biomarkers of AMI should hence be acknowledged, since they would increase the efficiency of the diagnostic process. A total of 2,276 consecutive patients referred to the emergency department for clinical symptoms suggestive for AMI underwent cardiac troponin T (cTnT) and D-dimer testing between January and December 2006. Patients sample were eligible for inclusion in this investigation if they had been collected prior to medication or intervention and 12-24 h after the time of patient's arrival to the emergency department, when the diagnostic efficiency of cTnT is the highest. cTnT was assayed on the Elecsys 2010 and test results were stratified according to the decisional threshold corresponding to the lowest TnT concentration associated with a 10% total imprecision in the assay (>0.03 microg/l). Plasma D-dimer was measured employing Vidas DD. The results of 741 patient's samples fulfilled the above criteria and were included in the study, 252 (34%) of whom had cTnT values>0.03 microg/l. The D-dimer value distribution (median and 95% C.I.) was significantly different in patients with cTnT values>0.03 microg/l than in those with cTnT values<0.03 microg/l (2,227 microg/l, 431-10,000 microg/l versus 1,039 microg/l, 143-6,338 microg/l; P<0.001). The area under the receiver operating characteristic (ROC) curve, was 0.734 (95% confidence interval: 0.715-0.753; P<0.001). At the 500 microg/l diagnostic threshold estimated by the ROC curve analysis, corresponding to the cut-off for the diagnosis of VTE, sensitivity and specificity of Vidas D-dimer were 95% and 27%, respectively. The positive and negative predictive values were estimated as 92% and 41%, respectively. In linear regression analysis, no significant association (r=0.090; P=0.077) was observed between D-dimer and cTnT in patients with cTnT levels exceeding the decisional threshold of the assay. Results of the present investigation on patients with AMI established by accepted diagnostic criteria (cTnT values above the decisional threshold of the assay associated with suggestive clinical symptoms), testify that D-dimer testing would not add clinically meaningful information to the sole determination of the cardiospecific troponins 12-24 h after patient's admission at the emergency department, when the cumulative data indicate that the diagnostic efficiency of cTnT is the highest.

Fabrication of a novel immunosensor using functionalized self-assembled monolayer for trace level detection of TNT by surface plasmon resonance

We have developed a new immunosensor based on self-assembly chemistry for highly sensitive and label-free detection of 2,4,6-trinitrotoluene (TNT) using surface plasmon resonance (SPR). A monolayer of amine terminated poly(ethylene glycol) hydrazinehydrochloride (PEG-NH 2) thiolate was constructed on an activated gold surface and immobilized with trinitrophenyl-β-alanine (TNPh-β-alanine) by amide coupling method. The binding interaction of a monoclonal anti-TNT Ab (M-TNT Ab) with TNPh-β-alanine immobilized thiolate monolayer surface was monitored and evaluated for detection of TNT based on the principle of indirect competitive immunoreaction. Here, the competition between the self-assembled TNT derivative and the TNT in solution for binding with antibody yields in the response signal that is inversely proportional to the concentration of TNT in the linear detection range. With the present immunoassay format, TNT could be detected in the concentration range from 0.008 ng/ml (8 ppt) to 30 ng/ml (30 ppb). The response time for an immunoreaction was 2 min and one immunocycle could be done with in 4 min including surface regeneration. Bound antibodies could be easily eluted from the self-assembled immunosurface at high recoveries (more than 100 cycles) using pepsin solution without any damage to the TNT derivatives immobilized on the surface. The compact self-assembled monolayer was highly stable and prevented the non-specific adsorption of proteins on the surface favoring error free measurement.

The involvement of cytochrome P450 system in the fate of 2,4,6-trinitrotoluene (TNT) in European eel [Anguilla anguilla (Linnaeus, 1758)

TNT (2,4,6-trinitrotoluene) was the most common nitro aromatic explosive available in World War II ammunitions. The presence of ordnance dumped at sea might represent a great concern for marine species living close to dumping sites and the toxicological properties of the chemicals released into the marine environments need to be evaluated. The aim of the present study is to investigate the involvement of CYP (cytochrome P450) system in the metabolism of TNT in marine organisms by using the European eel [Anguilla anguilla (Linnaeus, 1758)] as model species. In vivo exposure to sublethal concentration of TNT (0.5, 1 and 2.5 mg/l) leads to a significant decrease in the phase I CYP1A catalytic activities such as EROD (7-ethoxyresorufin-O-de-ethylase) and MROD (7-methoxyresorufin-O-de-ethylase). On the opposite, a significant increase in NADPH cytochrome c reductase activity as well as phase II UDP-glucuronosyltransferase activity is observed. An inhibition at enzyme level is hypothesized for both CYP1A enzymes, also confirmed by a similar decrease observed after in vitro exposure. An active role of NADPH cytochrome c reductase and phase II enzymes in the TNT metabolism may also be hypothesized.

Phototransformation of 2,4,6-trinitrotoluene: Sensitized by riboflavin under different irradiation spectral range

Riboflavin-sensitized phototransformation of 2,4,6-trinitrotoluene (TNT) under natural sunlight was investigated with reverse-phase high performance liquid chromatography/mass spectrometry (HPLC/MS) and gas chromatography/mass spectrometry (GC/MS). The effect of different spectral region of sunlight on TNT phototransformation in the absence or presence of riboflavin was also investigated by using optical filters with cut-off at 400 or 455 nm. The concentration of riboflavin in the phototransformation of TNT was optimized. Concentration of riboflavin and TNT was 1.0 and 50 μM, respectively. The rates of phototransformation of TNT under natural sunlight in the presence or absence of riboflavin were conformed to initial pseudo-first-order rate equation. The photolysis half life of TNT in the presence of riboflavin was 21.87 min, compared to 39 min in the absence of riboflavin under natural sunlight. Two major phototransformation products of TNT, 3,5-dinitroaniline (3,5-DNA) and 1,3,5-trinitrobenzene (1,3,5-TNB), were detected in the samples in the presence of riboflavin receiving irradiation at full wavelength or wavelength >400 nm. The results indicate that riboflavin mediates TNT sensitized-phototransfomation under natural sunlight or near-UV–vis light.

Photo–Fenton treatment of TNT contaminated soil extract solutions obtained by soil flushing with cyclodextrin

The technical feasibility and performances of coupling flushing abilities of cyclodextrin solutions for 2,4,6-trinitrotoluene (TNT) removal from contaminated soil and the ability of Photo–Fenton treatment for final disposal of soil extract solutions containing high TNT loads have been investigated at laboratory scale. Methylated-β-cyclodextrin (MCD) has shown better ability than hydroxypropyl-β-cyclodextrin (HPCD) to complex TNT. The MCD solution increased the aqueous concentration of TNT in soil extract effluents as much as 2.1 times the concentrations obtained during the water flush of the soil. TNT in soil extract solution has been treated by Photo–Fenton. Our results indicate that MCD has a beneficial effect on the degradation rates of TNT. This relative improvement of TNT degradation rate (1.3 time) in presence of high amounts of hydroxyl radical scavengers can be ascribed to the formation of a ternary complex (TNT-cyclodextrin-iron) which can direct hydroxyl radical reaction toward TNT. Complete mineralization of soil extraction solutions was not achieved and TNT degradation pathway has been elucidated in order to ensure that no potential toxic intermediate is left at the end of the treatment time. After successive TNT hydroxylations, oxidative opening of the TNT aromatic ring quickly occurred, leading to the accumulation of short chain carboxylic acids such as oxalic acid and formic acid.

2,4,6-Trinitrotoluene Transformation Using Spinacia Oleracea: Saturation Kinetics of the Nitrate Reductase Enzyme

A series of aqueous phase and soil-slurry phase microcosm studies were conducted on 2,4,6-trinitrotoluene (TNT) to obtain kinetic data for optimizing a treatment protocol using an enzyme extract from spinach (Spinacia oleracea). Crude extract was obtained by homogenization of fresh leaves with a buffered protease inhibitor, and employed as phytoremediation agent. Aqueous phase microcosms containing 20 mg∕L TNT and soil–slurry microcosms containing 1 g of a characterized sandy loam soil contaminated with 2,500 mg∕kg TNT and 1,000 mg∕kg hexahydro-1,3,5-trinitro-1,3,5-triazine were dosed with fixed aliquots of extract and analyzed for TNT transformation over time. The TNT concentration was monitored using a colorimetric method for nitroaromatic compounds based on EPA Method 8515. Nitrate reductase activity of the applied crude extract was simultaneously quantified. The transformation of TNT was described by a pseudofirst-order reaction. Coupling kinetic rate information with enzyme activity allowed for estim...

Assessment of a 2,4,6-Trinitrotoluene–Contaminated Site Using Aporrectodea rosea and Eisenia andrei in Mesocosms

Polynitro-organic compounds such as 2,4,6-trinitrotoluene (TNT) can be released into the environment from production and processing facilities and military firing ranges as well as through field use and disposal practices. Based on laboratory toxicity data, TNT has lethal (at >/=260 mg TNT/kg dry soil) and sublethal effects (at >/=59 mg TNT/kg dry soil) to the earthworm. However, field studies are needed to relate exposure of organisms to explosives in mixed-contaminated soil under field conditions and to define effects-based ecotoxicologic benchmarks for TNT-contaminated soil. In the present study, the lethal and sublethal effects of a 10-day in situ exposure at a TNT-contaminated field site using mesh-bag mesocosms were assessed. In addition to the survival end point, the biomarkers of earthworm exposure and effect-including tissue residues, lysosomal neutral red retention time (NRRT), and total immune activity (TIA)-were measured. Concentrations of TNT in soil mesocosms ranged from 25 to 17,063 mg/kg. Experiments indicated a trend toward decreasing survival of caged Aporrectodea rosea and Eisenia andrei as the concentration of TNT and total nitroaromatic compounds increased. E. andrei tolerated higher concentrations of TNT (up to 4050 mg/kg dry soil) in mesocosms than did indigenous earthworms, who survived only at </=1146 mg TNT/kg. Earthworms E. andrei and A. rose survived in 67% and 75% of TNT-contaminated mesocosms, respectively, compared with references groups. NRRT was significantly decreased in surviving earthworms from the contaminated areas compared with those from the reference site. TIA was not affected by field exposure to TNT. Earthworm tissue concentrations of TNT metabolites 2-amino-4,6-dinitrotoluene and 4-amino-2,6-dinitrotoluene were not correlated with TNT soil concentrations. In addition, higher tissue concentrations of TNT metabolites were observed at concentrations ranging from 116 to 130 mg TNT/kg soil. The results showed that earthworm exposure in TNT-contaminated soil produced both lethal and sublethal effects in the field. The results of study indicated that mesocosm experiments would be useful to assess the toxicity of a site and to characterize the overall effects of contaminants. However, mesocosm experiments present special considerations (e.g., abiotic factors, exposure period) when used at heterogenous sites, and data must be interpreted with caution.

Kinetics and mechanism of TNT degradation in TiO 2 photocatalysis

The photocatalytic degradation of TNT in a circular photocatalytic reactor, using a UV lamp as a light source and TiO 2 as a photocatalyst, was investigated. The effects of various parameters such as the initial TNT concentration, and the initial pH on the TNT degradation rate of TiO 2 photocatalysis were examined. In the presence of both UV light illumination and TiO 2 catalyst, TNT was more effectively degraded than with either UV or TiO 2 alone. The reaction rate was found to obey pseudo first-order kinetics represented by the Langmuir–Hinshelwood model. In the mineralization study, TNT (30 mg/l) photocatalytic degradation resulted in an approximately 80% TOC decrease after 150 min, and 10% of acetate and 57% of formate were produced as the organic intermediates, and were further degraded. NO − 3 NO − 2, and NH + 4 were detected as the nitrogen byproducts from photocatalysis and photolysis, and more than 50% of the total nitrogen was converted mainly to NO − 3in the photocatalysis. However, NO − 3 did not adsorbed on the TiO 2 surface. TNT showed higher photocatalytic degradation efficiency at neutral and basic pH.

Solute transport and extraction by a single root in unsaturated soils: model development and experiment

A contaminant transport model was developed to simulate the fate and transport of organic compounds such as TNT (2,4,6-trinitrotoluene), using the single-root system. Onions were planted for this system with 50-ml plastic tubes. Mass in the soil, soil solution, root and leaf was monitored using 14C-TNT. Model parameters were acquired from the experiments in the single-root system and were used to simulate total TNT concentration in soil, providing the average concentrations in the rhizosphere and bulk soil as well as root and leaf compartments. Because the existing RCF (root concentration factor) and TSCF (transpiration stream concentration factor) equations based on log K ow (octanol–water partition coefficient) were not correlated to TNT uptake, a new term, root uptake rate ( R ur), and a new T scf equation, based on the experimental data, were introduced in the proposed model. The results from both modeling and experimental studies showed higher concentrations of TNT in the rhizosphere than in the bulk soil, because mass transported from the surrounding soil into the rhizosphere was higher than that by root uptake.

In vitro Degradation of 2,4,6‐Trinitrotoluene Using Plant Tissue Cultures of Solanum aviculare and Rheum palmatum

AbstractThe degradation of TNT was tested in suspension cultures (Rheum palmatum and Solanum aviculare) and was followed by the identification of degradation products and the determination of the phytotoxicity of TNT to both cultures. The concentration of TNT inhibited the growth of cell cultures by 50 %, i.e., 37.8 mg/ and 38.1 mg/L for Rheum palmatum and Solanum aviculare, respectively. The TNT uptake was studied by determining the concentration of TNT and its degradation products, such as aminodinitrotoluenes and diaminonitrotoluenes, in the cultivation medium as well as in plant cells. The kinetics of the degradation showed that TNT was mostly taken up within 10 hours and 6 hours for S. aviculare and R. palmatum, respectively. Aminodinitrotoluenes were preferentially produced by cultures of S. aviculare, whereas diaminonitrotoluenes and aminodinitrotoluenes were revealed in cultures of R. palmatum. The final concentrations of identified degradation products did not stoichiometrically correspond to the decreased concentration of TNT in the medium. The different concentrations of degradation products in each culture were an indication that the metabolism of TNT is controlled by different enzymatic systems. Therefore, it was concluded that studying different species for TNT degradation is necessary for the search of most suitable candidates for TNT phytoremediation.

A kinetic model of aqueous-phase alkali hydrolysis of 2,4,6-trinitrotoluene

Alkali hydrolysis of 2,4,6-trinitrotoluene (TNT) was studied using batch experiments with starting pH values 11 and 12 in glass conical flasks covered with aluminum foil. Isothermal (25 and 40 °C) as well as non-isothermal experiments were conducted. Experiments starting at pH 12 resulted in >95% reduction in the concentration of TNT; those starting at pH 11 had a maximum reduction of 20–25% in TNT concentration. In all the experiments, one major stable intermediate was produced but it could not be identified. The experimental data were used to determine the overall stoichiometry of TNT and hydroxyl ions. Approximately 100 mol (standard deviation 11.4) of hydroxyl ions were consistently consumed per mole of TNT. However, control experiments showed that all but 13 mol (standard deviation 2) of hydroxyl ions were consumed in reactions not related to TNT. A simple kinetic model involving formation of the intermediate was proposed to account for changes in concentrations of TNT and hydroxyl ions. The rate constants and activation energies of the reactions were identified using isothermal data and the kinetic model was then used to predict the experimental data in the non-isothermal experiments reasonably well.

Characterization and performance evaluation of in vivo and in vitro produced monoclonal anti-TNT antibodies for the detection of TNT

In this paper are the experimental results used to characterize four distinct monoclonal anti-TNT antibodies (in vivo and in vitro cloned) for potential use in a field-portable immunosensor. Direct and competitive enzyme-linked immunosorbent assays (ELISA) were performed to determine their affinity for TNT and a fluorescently labeled analog of TNT for use in an immunosensor. Effective concentrations (EC 50), inhibition concentration (IC 50) and cross-reactivity measurements to related nitroaromatics (e.g., 2,4,6-trinitrobenzene [TNB], methyl-2,4,6-trinitrophenyl nitramine [tetryl], 2-amino-4,6-dinitrotoluene [2A-4,6-DNT], 2,4-dinitrotoluene [2,4-DNT] and 1,3-dinitrotoluene [1,3-DNT]) were measured. Final characterization of the monoclonal antibodies was based on performance (measured by fluorescence dose response) using a fluorescence-based microcapillary displacement assay. Analytical techniques showed a high degree of affinity for TNT and varying degrees of cross-reactivity with each respective monoclonal antibody. Microcapillary displacement immunoassays with each of the antibodies resulted in detection capabilities at the lowest applied TNT concentration (10 ng/ml).

Non-aerosol detection of explosives with a continuous flow immunosensor.

Contamination of groundwater, soil, and the marine environment by explosives is a global issue. Identification, characterization and remediation are all required for a site recognized as contaminated with 2,4,6-trinitrotoluene (TNT) or hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX). For each step, a method to accurately measure the contaminant level is needed. This paper reviews some of the current methods with emphasis on a single biosensor developed in our laboratory. Current regulatory methods require samples to be sent off-site to a certified laboratory resulting in time delays up to a month. A continuous flow biosensor for detection of explosives has been developed and tested for the rapid field screening of environmental samples. The detection system is based on a displacement immunoassay in which monoclonal antibodies to (TNT) and RDX are immobilized on solid substrates, allowed to bind fluorescently labeled antigens, and then exposed to explosives in aqueous samples. Explosive compounds present in the sample displace proportional amounts of the fluorescent label, which can then be measured to determine the original TNT or RDX concentration. The system can accurately detect ppb to ppt levels of explosives in groundwater or seawater samples and in extracts of contaminated soil. The biosensor has applications in environmental monitoring at remediation sites or in the location of underwater unexploded ordnance.

Electrochemiluminescence enzyme immunoassay for TNT.

Details of an electrochemiluminescence (ECL) enzyme immunoassay for TNT (2,4,6-trinitrotoluene) are reported. The design and construction of a computer controlled flow injection electrochemiluminometer in which the immunoassays are carried out is described. This system is used to select and pump solutions through a flow cell, which contains a gold working electrode as part of a three-electrode arrangement. The deposition of a re-usable immunosorbent dextran surface anchored to a gold surface in the flow cell by chemiadsorbed thiol groups is described. Antibodies are labeled with the enzyme glucose oxidase and used in competitive immunoassays in which the separation step is carried out by concentrating unbound antibodies on the immunosorbent surface. Hydrogen peroxide generated by the enzyme label when glucose is pumped through the flow cell is detected using luminol ECL. Light intensity was inversely proportional to the concentration of TNT in the sample in the range 0-100 ppb. The results are compared with colorimetric ELISA's carried out using the same reagents, and potential for developing a portable instrument for use in the field is discussed.