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We investigated the ecotoxicity of the chemical-warfare agent (CWA) HD (Mustard) using Collembola reproduction test with numbers of adults and juveniles as measurement endpoints. Toxicity tests were conducted using soils with contrasting parameters to investigate the effects of soil properties on chemical toxicity. These included standard artificial soil (SAS; 10% OM; 6 pH), O’Neill-Hall sandy loam (OHSL; natural soil with 4.3% OM; 5.1 pH), and Sassafras sandy loam (SSL; natural soil with 2% OM; 4.9 pH). Soils were individually spiked with HD concentrations ranging from 1 to 125 mg kg –1 . Lowest observed effect concentrations (LOECs) for adult mortality were 25, 50, and 6.97 mg kg –1 for SAS, OHSL, and SSL, respectively. The LOECs for reproduction were 6.97, 6.25, and 1.9 mg kg –1 for SAS, OHSL, and SSL, respectively. HD toxicity to both adults and juveniles was greater in SSL. These results show that soil toxicity testing should not rely solely on the adult acute endpoints using artificial soils, but should include assays with reproductive endpoints using natural soils with varying physical and chemical parameters to adequately assess toxicity to test species.