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A survey of the presence of mutagenic activity in drinking water of 18 cities in the Netherlands revealed that in drinking water of 13 cities mutagenic activity could be demonstrated. The activity was detected in the Ames test after concentrating the organic mutagens with a XAD4/8 procedure. Dose-related responses were observed with concentrates corresponding to 0.5 to 3.0 liters of drinking water.A study of the changes in mutagenic activity during the preparation of drinking water in a few waterworks showed that breakpoint chlorination, transport chlorination and post chlorination increased the mutagenic activity, while ozonation only reduced the activity with metabolic activation. When adsorption on activated carbon powder was used, a certain reduction of mutagenic activity was observed. The use of activated carbon filters, however, removed the activity completely. The majority of organic mutagens present in drinking water concentrates were shown to be nonvolatile and relatively stable and probably consist of compounds with a molecular weight in the order of 200. These mutagens are not identical to the organics identified up till now in drinking water by standard gas chromatography/mass spectrometry analysis. Finally, a group of organic mutagens, which adsorbs only at pH 2-3 on XAD-4/8 (acid fraction), could be demonstrated in Ames-positive drinking waters. IntroductionIn the Netherlands, groundwater, surface water, and a mixture of both are used as raw water sources for drinking water. In particular, the rivers Rhine and Meuse serve as important sources for the drinking water supply; the two rivers provide the potable water for about 5 million people in the Netherlands (1).In the Rhine and Meuse and in drinking water prepared from these rivers, several recognized mutagenic and carcinogenic organic compounds have been identified by Leer et al. (2) Thin Layer Chromatography (TLC)For the TLC fractionation of acetone concentrates of drinking water, preparative plates precoated with 2.0 mm of Silicagel-G (PSG Merck, Fertigplatten) were used. A volume of 2.0 ml acetone concentrate was applied to the plate with an automatic spraying device in the form of a small band (4.0 x 0.3 cm). The chromatograms were developed in one direction using ethyl acetate:isooctane (1:1) as the first solvent system. The plates were air-dried overnight and a second solvent system, benzene:methanol (4:1), was used. The plates were air-dried again prior to further investigation.The developed plates were examined under ultraviolet light (366 nm) in order to mark the separate bands. The marked fractions (six) were collected by scraping off and collecting the adsorbents with a Pasteur pipet connected to a vacuum pump. The outlet of the pipet was fitted with a plug of glass wool. The organic material was recovered by eluting the pipet with 5 ml DMSO. The eluates were stored at -20°C prior to mutagenicity testing. KOOL ET AL. Gel Filtration on Sephadex LH20The glass column (height 40 cm, 1 cm) was packed with Sephadex LH20 in dioxane-water ...
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