Application of a Homogenous Assay for the Detection of 2,4,6-Trinitrotoluene to Environmental Water Samples

Ellen R Goldman,Michael E Lassman,Igor L Medintz,George P Anderson,Adrienne L Egge

doi:10.1100/tsw.2005.58

Ellen R Goldman, Michael E Lassman + Show 3 more

Open Access

https://doi.org/10.1100/tsw.2005.58

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Abstract

A homogeneous assay was used to detect 2,4,6-trinitrotoluene (TNT) spiked into environmental water samples. This assay is based on changes in fluorescence emission intensity when TNT competitively displaces a fluorescently labeled, TNT analog bound to an anti-TNT antibody. The effectiveness of the assay was highly dependent on the source of the sample being tested. As no correlation between pH and assay performance was observed, ionic strength was assumed to be the reason for variation in assay results. Addition of 10x phosphate-buffered saline to samples to increase their ionic strength to that of our standard laboratory buffer (about 0.17 M) significantly improved the range over which the assay functioned in several river water samples.

Highlights

Munitions storage and processing has led to the contamination of soil and groundwater with the explosives, including 2,4,6-trinitrotoluene (TNT)[1,2]
TNT in the test samples competes with the fluorescent TNT analog for binding by the antibody. This assay has the advantages of being fast, simple, and sensitive, all qualities essential for adaptation to a high-throughput format. We demonstrate that this assay format works with an alternative dye-labeled analog and apply it to TNT spiked into environmental water samples
The two river waters that had performed the poorest in the assays were retested with the addition of 10× phosphate-buffered saline (PBS) to bring the final ionic strength of the sample to at least that of our laboratory PBS buffer

Summary

Introduction

Munitions storage and processing has led to the contamination of soil and groundwater with the explosives, including 2,4,6-trinitrotoluene (TNT)[1,2]. TNT in the test samples competes with the fluorescent TNT analog for binding by the antibody. We demonstrate that this assay format works with an alternative dye-labeled analog and apply it to TNT spiked into environmental water samples.

Results

Conclusion