Thousands different chemicals are discharged into the environment from agriculture, industry, medical facilities, house-holds. Currently, there is an increasing concern for the environmental impact of mixture of compounds since the additive and eventual synergistic effects are unknown and could produce serious adverse effects. Recently, a document from the European Commission on combination effects of chemicals highlighted the need to ensure that risks associated with chemical mixtures are properly understood and assessed. To address this issue, a joint-effort of 16 European and associated research groups participated to an exercise to test a synthetic reference chemical mixture on the own routine bioassays to investigate the chemical mixtures effects. The reference material included class of pesticides, pharmaceuticals, industrial products, heavy metals and polyaromatic hydrocarbons. The mixures were prepared, each compound at Equivalent Quality Standard (EQS) value, the safety limit concentration allowed by the European Water Framework Directive, (WFD).The bioassays proposed by the groups could cover the entire ecosystem from bacteria to fish as well in vitro assays providing an unique scenario from ecological risk assessment perspective. The results showed that effects were observed at very low concentration on algal-bacteria composition in a marine microcosm, immobilization in crustacean, fish embryo toxicity and frog embryo development. We conclude that some precaution on the chemical mixture assessment should be taken even in case the individual compounds are present at EQS, the safety limit concentration under European legislation
