Regulatory authorities involved in safety and risk assessments have a primary duty to protect humans, animals and the environment. Safety evaluation studies, to assess the potential hazards and risks posed by products and substances, currently depend predominantly on animal testing as a limited number of non-animal methods have been developed and received regulatory acceptance. The ways in which regulatory authorities can assist in replacing animal procedures and in reducing the suffering caused to animals in safety testing were highlighted in 1997, when a set of principles on animal welfare was drawn up by them in collaboration with the Home Office. The principles are based on the replacement, reduction and refinement of animal experiments, known as the Three Rs. Focus on Alternatives (FOA), an umbrella group of organisations working together to advance the replacement of animal experiments, wrote to each of the UK regulatory authorities asking them how, and to what extent, they have implemented these principles. This paper reviews how well the various authorities are implementing the five principles, and also presents FOA's recommendations on where further effort is needed. These animal welfare principles and FOA recommendations may also apply to similar regulatory authorities worldwide.