Developing earthworm bioconcentration factors of nitrogen-based compounds for predicting environmentally significant parameters for new munition compounds in soil

We investigated the bioconcentration potential of nitrogen-based compounds 4-nitroanisole (4-NAN), 3,5-dinitro-o-toluamide (3,5-DNoTAME), and 2-methoxy-5 nitropyridine (2-M-5-NPYNE) using earthworm Eisenia andrei exposures in aqueous exposure media in sand. Separate toxicity studies were conducted prior to bioconcentration studies using a range of chemical concentrations to establish the sublethal exposure conditions for the earthworms. The objectives of the present studies were to: (1) develop an experimental test system for estimating bioconcentration potentials of new and emerging munition compounds that partition into earthworms, using aqueous exposure media; and (2) apply this experimental model to establish original bioconcentration data for 4-NAN, 3,5-DNoTAME, and 2-M-5-NPYNE. Experimental design includes earthworm exposures to chemicals for up to 14 days in aqueous media (Römbke medium; 0.08 mM KCl, 2 mM CaCl2, 0.5 mM MgSO4, and 0.8 mM NaHCO3) in the presence of water-washed coarse sand (0.5-1.0 mm) substrate. Concentrations of test chemicals in respective exposure media remained relatively stable during these independent studies. Tissue analyses revealed a rapid uptake of each chemical by the earthworms; a steady state was attained within 24 h from commencement of these exposures. Estimated steady-state bioconcentration factors (BCFss; mL/g dry tissue) were 47, 6, and 11 for 4-NAN, 3,5-DNoTAME, and 2-M-5-NPYNE, respectively. These results will contribute to the BCF database being developed for use in models aimed at predicting environmentally significant parameters for new munition compounds in soil.