A high-throughput dual parameter assay for assessing drug-induced mitochondrial dysfunction provides additional predictivity over two established mitochondrial toxicity assays

► A 384-well dual parameter assay for assessing drug-induced mitochondrial dysfunction in cells was validated. ► The assay measures cellular oxygen consumption and extracellular acidification in both suspension and adherent cells. ► Two hundred drugs were tested in this assay and in two previously established mitochondrial toxicity assays. ► The dual parameter assay identified more compounds that cause mitochondrial impairment than the other two assays. ► Novel information was obtained on drugs not previously reported to cause mitochondrial impairment.