Detection of organic toxic pollutants in water and waste-water by liquid chromatography and in vitro cytotoxicity tests

In vitro toxicity tests with fish cell lines have proved to be correlated with in vivo toxicity tets on fish. A group of toxicity tests on RTG-2 cell line (a fibroblastic line derived from rainbow trout) has been standardized in order to enhance reproducibility and sensitivity. The liquid chromatographic (LC) separation of organic chemicals from industrial effluents and polluted waters and the use of in vitro toxicity tests on RTG-2 as a biological detector of toxicity in the eluted peaks are reported. Effluents and polluted waters were concentrated, if required, using Sep-Pak C₁₈ cartridges, and analysed by reversed-phase LC using a 30-cm C₁₈ column with an acetonitrile gradient from 10 to 100% in water in 60 min at a flow-rate of 1 ml min^?1 and UV detection at 254 and 280 nm. The cytotoxicity test was adapted to use 20-μl fractions of acetonitrile-water mixtures, allowing toxicity detection every 12 s with eight replicates per sample (or every 5 s with four replicates).