In vitro anaerobic trinitrotoluene (TNT) degradation with rumen fluid ...

throughout the run. The mass spectrometer was operated using an ionizing voltage of
70 eV and an ionizing current of 300 mA.
2.4 Results
Incubation of the tubes at 38°C containing 13.8% of 14C-TNT and non-radio
labelled TNT in 127 mg/1 of total concentration was terminated by freezing the tubes.
One ml tube samples were centrifuged (18000 xg) for six minutes and analyzed by
HPLC. The chromatogram of the analysis is illustrated in Figure 2.1 (minutes
intervals) and Figure 2.2 (day intervals). The 14C-TNT completely disappeared within
one hour, leaving MADNT and DAMNT as intermediates. After five days incubation,
14 C-TNT added to the supernatant was transformed into unidentified products which
were retained at 2, 15, and 16.5 minutes, respectively, in the HPLC system.
To identify the transformation products of TNT degradation by the ruminal
organism, only non-radio labeled TNT was added to a 10% rumen fluid with glucose
as a carbon source, and incubated at 30°C for two days. The supernant was extracted
with CHC13(1:1). The condensed extract was injected into GC/MS.
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