A tiered framework provides a structured approach to assess and manage risk and underpins much of the legislation concerning chemicals and environmental management. Management decisions regarding appropriate controls can have high cost implications to the regulated community. The risk framework provides an evidence-based approach to reduce uncertainty in decision making. Traditional assessment is heavily dependent on laboratory-generated toxicity test data and estimations of exposure and effect. Despite many well documented demonstrations of in situ methodologies, they are rarely used by regulators to help improve assessment or to validate risk. Emerging legislation puts greater emphasis on environmental outcomes and represents a significant shift from the reliance on chemical measures alone toward biological responses that improve assessment and demonstrate ecological benefit. Diagnostic methods, that could include in situ–based measures, will help assess and manage environments failing to achieve good status and it is likely that a weight of evidence approach will be needed to help inform management decisions. The potential application of such measures in the risk framework is reviewed in the context of current and emerging legislation concerning chemicals. Effect measures on the basis of in situ methods provide an alternative line of evidence and can help reduce uncertainty in decision making. Criteria are presented to help select appropriate methods in a multiple-line, weight of evidence approach.