The chemical and biogenotoxic characterization of organic xenobiotics in aquatic sediment materials 1. The application and comparison of chemically non-specific and biogenotoxic methods

The aim of this work was to evaluate the Ames assay and mixed function oxidase (MFO)-Induct Test used in parallel with chemical group tests (ECD fingerprint and PAH estimation) for the characterization of the organic pollution of water sediment materials. Sediment materials were collected from “clean” and relatively heavily polluted locations in the Middle Adriatic Sea, and from some locations in continental Croatia polluted with wastewaters from different enterprises. Characterization of the organic extracts of the sediment materials investigated was performed chemically using UV spectrofluorometry for the determination polyaromatic hydrocarbons (PAH) and gas chromatography for the determination of volatile EC detector sensitive materials. Genotoxic analysis of the extracts was performed using the MFO-Induct Test and mutagenicity testing using the Standard Plate Incorporation Test as described by Maron and Ames with Salmonella typhimurium TA 98. Measurement of the BaPMO enzyme activity in the livers of carp treated i.p. with total extracts of the sediment investigated confirmed that the methanol extracts generally contained more inducing matter than the petroleum ether extracts. Ames assay showed that for all the samples following the elimination of the sulfur, there was an increase in the number of revertants in comparison to the control number, which indicates that the samples contained mutagenic substances. The larger doses of extracts generally demonstrated cytotoxicity, as evidenced by a reduced number of spontaneous revertants in the Salmonella/Microsome Test. Investigation of the correlation of the chemical parameters with the biological parameter showed that the induction of BaPMO exhibited a statistically significant correlation with the level of the ECD fingerprint of the petroleum ether sediment extract.     

Minimization of blank values of the miniature silica gel column

Several methods involving silica gel separations of polychlorinated biphenyls from chlorinated insecticides are already described in literature. Our intention was to find a system of solvent and silica gel of the lowest blank values. The first step was to find a solvent mixture which will keep the homogenity of a silica gel column. PCBs elution was performed with pentane, while the chlorinated insecticides eluated with diethylether, acetone, methanol, and benzene. The elution of pesticides gave the best results for the following solvent systems: diethylether + hexane (1:1), acetone + hexane (1:4), methanole + pentane (1:8) and benzene.Separation of pesticides from polychlorinated biphenyls found in a synthetic mixture was also examined. It was found that all solvent systems used gave similar results of separation. However, only if benzene is used as eluant, the silica gel column can be used for four successive separations after the “on column” regeneration. In order to obtain the lowest blank values silica gel was cleaned with several solvents. The eluates after the silica gel separation were also cleaned up by means of deactivation alumina and concentrated sulphuric acid.     

Investigation of the production of lipophilic volatile organohalogen materials during laboratory chlorination of some ground water samples from Yugoslavia

Water chlorination leads to the generation of various halogenated products of natural and waste water organic matter. In natural surface and ground water humic matter usually makes the greatest part of the total organic matter. However, in the water polluted with various organic wastes, other organic matter is also present in significant concentrations. The investigation of the relationship between the concentration of organic matter in the samples of ground water in Yugoslavia on one side, and the production of the lipophilic volatile organohalogen materials during the process of their chlorination on the other side is described. The production of the halomethanes and other ECD response materials during the chlorination of water samples in laboratory conditions is compared with the concentrations of organic materials in water samples. From the obtained linear correlation coefficients, possibilities of predicting amounts of organohalides during the chlorination process by using several very simple methods for the organic materials estimation in the investigated water samples are discussed and compared.     

Evaluation of modifications of the simple spectrofluorometry method for estimating petroleum hydrocarbon levels in fresh and waste water samples

The monitoring of petroleum hydrocarbons in the water environment is very often performed using complicated methods. There are many difficulties encountered in analyzing a long number of samples in the monitoring programme. In such cases, a simpler though less precise and specific technique using inexpensive equipment to monitor a large number of water samples, might be more desirable.

This idea was realized in the assessment of the state of pollution of the Mediterranean Sea by petroleum hydrocarbons where the pollution of sea water with oil (dissolved and/or dispersed in sea water) was measured by spectrofluorometry. The paper describes an analytical evaluation of three modifications of simple the spectrofluorometry method, applied on numerous river and waste water samples collected from various areas of Croatia.