Recently, free radical involving active oygen speceies and nitric oxide are widely investigated because of their role in physiological, pathological and toxicological phenomena. Electron spin resonance (ESR) spectrometer is used for detecting free radicals. More recently, in vivo ESR has been developed to analyse free radical reactions in living animals. In the present paper, ESR measurement of bioradicals is reviewed, including its principle, application of spin trapping technique, in vivo ESR system, and non-invasive analysis of free radical reaction in mice.

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Because the waste treatment is regarded as one of the major sources of environmental contaminations by hazardous substances, regulations against hazardous wastes were newly established and revised. The Waste Disposal and Public Cleansing Law and the criteria of the hazardous waste were revised, and then more volume of hazardous wastes were treated specially. Internationally, ocean dumpings of wastes will be prohibitted and transboundary movements of hazardous wastes were regulated. In Japan, environmental contaminations caused by the waste disposal have not been revealed. However, leachates from landfill sites were found to include various hazardous substances and they were proved to be toxic by bioassay tests.

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Aqueous anthracene/phenanthrene compound solutions were treated with chlorine under various experimental conditions. Diethyl ether extracts from the chlorinated waters were determined by gas chromatography for their product distributions, and were tested for their mutagenic effects on TA98 and TA100 strains with and without S9 mix. Aqueous chlorination not only generated mutagens from non-mutagenic compounds (anthracene, 2-methylanthracene, phenanthrene and 2-methylphenanthrene), but also enhanced the activity of mutagenic compounds (9, 10-dimethyl anthracene and 1-methylphenanthrene). The chlorination products of anthracene compounds were most mutagenic on TA98 strain with S9 mix, while these products of phenanthrene compounds were mutagenic without S9 mix. The parent compounds were shown to produce a variety of chlorination products, including chlorine-substituted, oxygenated, and hydroxylated compounds. The formation of chlorination products and mutagenic substances was dependent on the reaction pH and chlorine dose. The significance of these findings for water chlorination systems is discussed.

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Organic compounds in tap water trapped by microporous polyethylene hollow fiber (EHF) membrane were investigated. Samples of tap water (15 or 30 m³) obtained from Katsushika-ku (Tokyo), Shinjuku-ku (Tokyo), Osaka-shi and Shizuoka-shi were passed through EHF membrane cartridges. The cartridge was then washed with 3 l of methanol, each methanol extract was concentrated and a sticky dark brown material was obtained. Weight levels of the materials except that of Shizuoka-shi were similar (3000 mg) in spite of difference in the volume of water passed. Though the percentages of total organic carbons in the extracts except that of Shizuoka-shi ranged from 14 to 19% in 15 m³ of tap water and from 23 to 29% in 30 m³ of tap water. The mutagenic activity of the extracts to Salmonella typhimurium TA98 and TA100 without S9 was highest in the Osaka-shi tap water. Fractions from this extract with a Sep-Pak Silica cartridge were analyzed by gas chromatography/mass spectrometry (GC/MS). The eluates with CH₂Cl₂, which contained more than 100 organic compounds, were again fractionated with a Sep-Pak Florisil cartridge and by silica-gel thin layer chromatography. Three halogen compounds were identified as oxadiazon, butachlor and its chloro analogue. Oxadiazon and butachlor in the fractions were quantified by GC/MS in selected ion monitoring mode to be 2.18 and 1.58 ppt in the Osaka-shi tap water, respectively.

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Degradation of tetrachloroethylene (PCE) by a gram negative, anaerobic bacterium strain T isolated from sludge in drainage from a laundry was examined. The activity of degradation was obtained in the culture supernatant of strain T, but not in the cell-extract by sonication. The reaction was optimal between pH 7.0 and 8.0 and at a temperature of 37°C. The activity was unstable in oxygen, heat labile and inhibited not only by trypsin and chymotrypsin but also by thiol inhibitors such as N-ethylmaleimide and p-hydroxymercuribenzoic acid. The substance degrading PCE in the culture supernatant of a gram negative anaerobic bacterium strain T might be a protein with the thiol group as an active site.

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The natural coloring matter (blue color) produced from an aqueous extract of gardenia fruits (Gardenia augusta MERR.) and the decomposed protein was found to be of a mean molecular weight of 15600±400, based on the measurements of the osmotic pressure. The absorption from the gastrointestinal tract of this natural coloring matter was examined by testing the stability of the color in the gastrointestinal tract, by observing the absorption, both in vitro and in situ, and by conducting histopathological examinations of the gastrointestinal tract and other related organs in mice which had received the color for five months. The results showed that the color was very stable at the pH levels of the gastrointestinal tract and in the presence of digestive enzymes and enterococci. No evidence of the gastrointestinal absorption was obtained.

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The simultaneous determination of phenylurea and carbamate herbicides (diuron, linuron, siduron, methyldymron, chlorpropham, and swep) in water was studied using FRIT/fast atom bombardment (FAB)-liquid chromatography/mass spectrometry (LC/MS). These herbicides were extracted from water by the use of an octadecyl-silica solid phase extraction (SPE) cartridge, and then eluted from the SPE cartridge with acetonitrile. Ethyl-N-phenylcarbamate was added as an internal standard to the eluates and these herbicides were determined by the FRIT/FAB-LC/MS-selected ion monitoring (SIM) method. By this method, these herbicides were analyzed in river water, ground water and tap water to which the standard solution was added. These recoveries were more than approximately 90% and these standard deviations were less than 10%, respectively.

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To search indicator organisms for environmental pollution by volatile chlorinated hydrocarbons (VCHs), 1, 1, 1-trichloroethane (MC), carbon tetrachloride (CTC), trichloethylene (TCE) and tetrachloroethylene (PCE) were tested for their ability of bioconcentration in blue mussel, Mytilus edulis inhabiting chiefly seawater, and killifish, Oryzias latipes inhabiting chiefly freshwater. Extraction of VCHs from blue mussel was carried out by the purge-trap method using a Dean-stark apparatus. In the case of killifish, we developed a new method using and improved purge-trap apparatus because it needs small scale analysis due to the little livestock. The levels of VCHs were determined by ECD-GC. Recoveries of VCHs by this method were 81.1% (MC), 72.7% (CTC), 80.1% (TCE) and 75.8% (PCE). Blue mussel and killifish were exposed to VCHs (5-100 ppb) for 8 d and 21 d, respectively. The bioaccumulation factors for VCHs were 2.95 (MC), 4.52 (TCE) and 25.7 (PCE) for blue mussel, and 2.54 (MC), 3.49 (CTC), 2.71 (TCE) and 13.42 (PCE) for killifish. These results showed that these two aquatic organisms, blue mussel and killifish, have a large bioaccumulation potency of PCE. The monitor of VCHs levels using these organisms may offer an easy way to get some information of water pollution by VCHs.

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Multiple copper chlorophyll analogues were detected in the chewing gum product containing the copper chlorophyll dye using a HPLC apparatus with photodiode array detection. The main components among these analogues were assumed to be 10-hydroxy copper chlorophyll a and b by IR, FAB/mass and NMR spectrometry. Since these copper chlorophyll analogues were main components in the commercial food additive dye, "copper chlorophyll", it was clear that these analogues in the chewing gum product were carried over from this food additive. The 10-hydroxy pheophytins, which are precursors of these copper chlorophyll analogues, were detected from the extract of silkworm excreta, the original material of this dye. These copper chlorophyll analogues were possibly produced by the metalation reaction between the copper and the 10-hydroxy pheophytins which might be generated from the oxidation of pheophytins, like "allomerization reaction", during the process of the synthesis of the original material and its storage. It is possible that these analogues have already been the main components in the original dye.

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The simultaneous determination of residual 13 synthetic antibacterials in edible viscera by HPLC was studied and the clean up method was developed by Sep-Pak Alumina N and also tC₁₈ cartridges. This method was applicable to sulfamerazine, sulfadimidine, sulfamonomethoxine, sulfadimethoxine, sulfaquinoxaline, ormethoprim, trimethoprim, pyrimethamine difulazon, clopidol, morantel, carbadox and nicarbazin in the bovine liver and swine kidney. The recoveries of the addition of 0.4μg/g of these antibacterials to the bovine liver and to the swine kidney were in the range of 54.0 to 74.9%, and their detection limits were in the range of 30 to 50 ppb.

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The application of ethyl acetate as the elution solvent for solid-phase extraction (SPE) of 34 organic pesticides in drinking water was evaluated using octadecyl (C18) bonded porous silica, octyl (C8) bonded porous silica and styrene divinyl benzene copolymer (PS-1) cartridges. A 500 ml volume of drinking water spiked with 0.5 μg of each pesticide was passed through each cartridge, absorbed target pesticides were eluted with 2 ml of ethyl acetate and analyzed using capillary gas chromatography-mass spectrometry. Target pesticides except for captan could be detected at 0.2-0.52μg/l level from each cartridge, it was confirmed that ethyl acetate was a suitable solvent as the eluent.

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Volatile halogenated organic compounds in tonics, waters for injection and infusion preparations were determined by the headspace gaschromatograph method. Chloroform and 1, 1, 1-trichloroethane were detected in tonics. The highest concentration of chloroform was 0.011 mg/l and that of 1, 1, 1-trichloroethane was 0.010 mg/l. The contents of these compounds in water for injection and infusion preparations were very low.

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