An underlying assumption of laboratory-based toxicity tests is that the sensitivity and exposure of organisms in the laboratory is comparable to that in the field. We sought to make a comparison between field-based and laboratory-based sediment toxicity tests using a recently developed rapid amphipod reproduction test that encompasses gametogenesis, fertilization, and embryo development before hatching. The test species, Melita plumulosa, is an epibenthic, detritivorous amphipod native to Eastern Australia. Test sediments were sourced from Lake Macquarie, a large saltwater lagoon located 100 km north of Sydney (New South Wales, Australia) that has received heavy-metal pollution over many decades, primarily from a Pb/Zn smelter but also from collieries, coal-fired power stations, and urban areas. This has led to a north-south trace-metal concentration gradient, including Pb, Zn, Cd and Cu, in the sediments of Lake Macquarie. Sediments from these northern bays were demonstrated to reduce amphipod fecundity in laboratory-based tests. For the current study, the amphipod reproduction test has been modified for use in situ. In situ test chambers were deployed at the mouth of Cockle Creek, Lake Macquarie. Sediments that were demonstrated to reduce fecundity of M. plumulosa in the laboratory reproduction test were not similarly toxic when amphipods were exposed to the same sediments in situ. Factors related to the regular tidal renewal of overlying water likely altered exposure profiles in situ, including the provision of additional or alternative nutrition that obviated the need for amphipods to interact with the contaminated sediments, and a washout effect that prevented the accretion of dissolved zinc in the overlying water.