No abstract

To analyze the fate of 2,4,6-trinitrotoluene (TNT) and its reduction products, a TNT-contaminated soil (350 mg TNT/ kg dry soil) was spiked with [ring-UL-14.C]TNT and treated in a laboratory slurry reactor. During an anaerobic/aerobic treatment, the total radioactivity measured in the supernatant and methanolic soil extracts decreased to 2 per cent. The decrease corresponded to an increase of strongly bound radioactivity to the soil. Throughout the whole treatment process, mineralization of TNT was not observed. During the reductive process, unidentified polar substances increased to a maximum amount of 23.2 per cent of the total radioactivity on the day after the start of the experiment. After the end of the anaerobic phase, still 9.7 per cent of the radioactivity was found in this fraction. Only during the aerobic phase did the polar substances disappear completely. The irreversible character of the binding of the reduced metabolites of TNT to the soil was indicated by the failure of desorption even under rigorous and longterm extraction conditions. A significant release of radioactivity could be measured only by using high concentrations of HCl (5.N) or EDTA (12.5 per cent and 5.9 per cent, respectively). However, in none of the extracts were TNT or any reduced metabolites detected by HPLC and 14.C-radiocounting. Size-exclusion chromatography of humic acids extracted from the treated soil indicated that the metabolites of TNT were evenly bound to the complete range of molecular size of the humic acids

Evidence is presented for the covalent binding of biologically reduced metabolites of 2,4,6-15N3-trinitrotoluene (TNT) to different soil fractions (humic acids, fulvic acids, and humin) using liquid 15N NMR spectroscopy. A silylation procedure was used to release soil organic matter from humin and whole soil for spectroscopic measurements. TNT-contaminated soil was spiked with 2,4,6-15N3-trinitrotoluene and 14C-ring labeled TNT, before treatment in a soil slurry reactor. During the anaerobic/aerobic incubation the amount of radioactivity detected in the fulvic and humic acid fractions did not change significantly (11−16%), whereas the radioactivity bound to humin increased to 71%. The 15N NMR spectra of the fulvic acid samples were dominated by a large peak that corresponded to aliphatic amines or ammonia. In the early stages of incubation, 15N NMR analysis of the humic acids indicated bound azoxy compounds. The signals arising from nitro and azoxy groups disappeared with further anaerobic treatment. At the end of incubation, the NMR shifts showed that nitrogen was covalently bound to humic acid as substituted amines and amides. The NMR spectra of the silylated humin suggest formation of azoxy compounds and imine linkages. Bound metabolites possessing nitro groups were also detected. Primary amines formed during the anaerobic incubation disappeared during the aerobic treatment. Simultaneously, the amount of amides and tertiary amines increased. Nitro and azoxy groups of bound molecules were still present in humin at the end of the incubation period. Formation of azoxy compounds from partially reduced TNT followed by binding and further reduction appears to be an important mechanism for the immobilization of metabolites of TNT to soil.