Analytical isolation, separation and identification of mutagens from nonvolatile organics of drinking water

A general procedure has been developed for the concentration/fractionation of mutagenic residue organics from small, less than 50L, and large, to 1200L, volumes of drinking water obtained from a variety of sources. This procedure features concentration of the residue organics chromatographically by passage of the water through XAD-2 and XAD-7 resins in specially designed columns, details of which are given. The residue organics are eluted from the resins via organic solvents, followed by solvent removal and subsequent bioassay for mutagenicity. Then the residue organics are fractionated via a coupled bioassay/analytical fractionation method which progressively focuses to the bioactive constituents of the complex mixture of residue organics. In this report, results for the optimal operation and validation of the concentration system are given, using drinking water derived from an industrially polluted river system, a wilderness river system and a major aquifer system. The predominant type of mutagenesis observed for the residue organics isolated from these samples was direct-acting to the Salmonella tester strain, TA98, which was decreased by the addition of the metabolic activation system from the livers of rats previously treated with Arochlor 1254. Some TA100 direct-acting mutagenesis was observed for all samples. Fractionation of the residue organics indicated the mutagens to be nonpolar. Samples of residue organics collected over a period of a year from each type of drinking water showed no discernable pattern of mutagenesis versus season. The methodologies described in this paper provide a comprehensive approach for the concentration/isolation of residue organics from drinking water for studies to identify biohazardous compounds and to characterize these compounds biologically.