Algal growth inhibition test in filled, closed bottles for volatile and sorptive materials

Exposure concentrations of many hydrophobic substances are difficult to maintain in algal growth inhibition tests performed in open agitated flasks. This is partly because such compounds tend to volatilize from aqueous solution and partly because of sorption to the algal biomass as well as to the test container. A simple filled closed bottle test with low algal densities and bicarbonate enrichment is described here as an approach to minimize the loss of test material from solution. The algal medium was enriched with 300 mg NaHCO3/L, the pH was adjusted to 7.0 by addition of HCl, and the resulting dissolved CO2 concentration supported maximum algal growth rates without pH drift for algal densities up to 4 mg dry weight/L. Two-day toxicity tests with kerosene were performed with this new test design and compared with an open bottle test and with a closed bottle test with headspace. Exposure concentrations of the volatile fraction of kerosene decreased by 99% in the open test, by 77% in the closed flask test with headspace, and by 16% in the filled closed bottle test. Algal growth inhibition was observed at much lower additions of kerosene in the new test design because of the improved maintenance of a constant exposure.