Activated carbon-chloroform (CCE), diethyl ether (CEE), and acetone extracts (CAE) of drinking water taken from Jan. to Nov. 1986 were tested for mutagenicity in the Ames TA98 and TA100 strains with and without S9 mix. Mutagenic activity was found often in TA100 without S9 mix, though in moderate levels, i.e., CCE (36±39), CEE (46±54) and CAE (19±13 net revertants/1). Only one CCE, which was taken from Sept. to Nov., exhibited mutagenicity in TA98 without S9 mix. The highest mutagenic activity in the TA100 strain without S9 mix was observed for the water concentrates collected during the fall season, while the lowest activity was detected for the early spring samples. Organic residue and organic halogen contents in drinking water were determined, and the relationship between these parameters and mutagenicity of the water concentrates is discussed.

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Water samples collected from the Tama River were treated with chlorine under the conditions utilized for water renovation, in order to characterize and determine the halogenated organics formed in this reaction. Non-chlorinated water samples showed total trihalomethanes (THMs) and total organic halogen (TOX) levels ranging from 0.4 to 12.2μg/l (mean value, 4.1μg/l) and from 7 to 51.5μg/l (mean value, 25.9μg/l), respectively. It was found that the concentrations and compositions of organic halogen in chlorinated river water are strongly dependent on the sampling points and periods. The ranges of total THMs and TOX levels in chlorinated river water were from 2 to 158μg/l (mean value, 33.1μg/l) and 12 to 400μg/l (mean value, 151μg/l), respectively. Higher concentrations of both THMs and TOX and a high ratio of brominated THMs to total THMs were found on chlorination of water samples taken from the midstream of the Tama River, as compared with those observed from the upstream region. The concentrations of both THMs and TOX in chlorinated river water increased during spring and summer and decreased during fall and winter. In addition, the amounts of THMs and TOX in chlorine-treated river water were found to be dependent on the pH of the solutions and the time of contact with chlorine.

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The binding of ⁶³Ni to low-molecular-weight constituents of rat plasma, kidney cytosol and urine has been investigated in vivo and in vitro. Comparative tissue distribution and excretion of ⁶³Ni in rats after administration of ⁶³NiCl₂ through various routes revealed that among rat organs, the kidney showed the highest affinity for ⁶³Ni and that radio-nickel was excreted easily into the urine by way of the kidney. Sephadex G-25 and G-200 gel chromatographies of plasma, kidney cytosol and urine after iv or po administration of ⁶³NiCl₂ demonstrated that a major component of nickel metabolite in the rat urine may originate from one of three Ni-complexes with low molecular weight in the plasma and kidney cytosol. The behaviors of Ni-complex on thin layer chromatography and Sephadex G-25 gel chromatography suggested that one of the metabolites in the rat urine after iv or po administration of ⁶³NiCl₂ existed as a complex of nickel with a picrate-forming substance resembling creatine-related substances.

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Properties of hepatic aldehyde dehydrogenase (ALDH), a key enzyme for controlling the detoxication of alcohol, were compared in two strains of mice with different sensitivities to alcohol. The ALDH activities were also compared in four experimental animals including mice, rats, guinea pigs and rabbits, and humans under the same conditions. The activities were different among the animal species and strains. The enzyme activities were mainly localized in the cytosolic fraction in all animals tested. However, the subcellular distribution pattern of ALDH was different among the animals. Molecular properties of ALDH in hepatic cytosolic fraction of the animals were analysed by isoelectric focusing. The results indicated that each animal had a characteristic pattern. Under successive treatments of ethanol to two strains of mice, genetically different in the sensitivity to alcohol, ALDH activities and isozyme patterns were compared. In the alcohol sensitive strain, DBA/2J, ALDH activities in subcellular fractions did not change by giving 5% ethanol to this strain for 20 d. On the other hand, in the ethanol-resistent strain, C57BL/6J, given 10% ethanol as drinking water for 20 d, ALDH activities were elevated in microsomal and cytosolic fractions and the isozyme pattern of the cytosolic fraction underwent marked changes related to the treatment.

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Identification of respective components of women's hair care products (hair tonics, hair treatments, hair creams and waveset lotions) remaining on human hair was investigated. Women's hair care products (hair tonics of 15 brands of 6 manufacturers, hair treatments of 23 brands of 9 manufacturers, hair creams of 24 brands of 14 manufacturers and waveset lotions of 40 brands of 12 manufacturers) were identified by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) after extraction with ether. Among 102 brands of women's hair care products from 16 manufacturers, components of 66 brands remained on hair (hair tonics of 7 brands, hair treatments of 19 brands, hair creams of 24 brands and waveset lotions of 16 brands) were identified. It was notable that remaining components were detected in all of hair creams. Those of 66 brands were grouped into 27 classes on the basis of components, and among them 12 brands were identified by their characteristic peak (s) detected by GC and GC-MS. Only diethoxyethyl adipate in two brands and docosamethyldecasiloxane in one brand were common components of men's hair care products (hair liquid, hair tonic and hair rinse), and the rest of 63 brands were identified without interference in men's hair care products peaks on gas chromatograms and mass spectra. Major components of hair tonics were detected even 5 d after treatment, hair treatments were 20 d, hair creams were 10 d, and waveset lotions were detected even 5 d after treatments. From these results, if major components were detected from treated hair, the brands of women's hair care products (hair tonics, hair treatments, waveset lotions and hair creams) can be identified.

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Determination of chlorhexidine in tooth paste was examined by using high performance liquid chromatography (HPLC). Chlorhexidine was extracted with methanol, isopropanol and acetonitrile. The best recovery was obtained with methanol containing acetic acid and sodium dodecyl sulfonate (SDS). Two g of sample was extracted with methanol-water-acetic acid (80 : 19.5 : 0.5) containing SDS (1%) and made up to 50 ml, and 20μl of this solution was injected to HPLC. A chlorhexidine peak was separated from other component peaks in commercial tooth paste by using an ODS column, 1-dodecansulfonate (DSA) as an ion pairing agent and methanol-water-THF (830 : 160 : 10) containing 5% acetic acid as an elution solution. The amounts of chlorhexidine in each 100 g of commercial tooth paste were ranged from 5.1 mg to 19.6 mg by this method.

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Laboratory wastewater contains many kinds and various forms of chemicals, and is discharged discontinuously. Especially, heavy metal wastewater which is released from research, educational and medical facilities of universities often contains organic acids as ligands. The aluminum coprecipitation process is one of the usual methods to remove heavy metals from inorganic wastewater. However, it is interfered with the organic acids contained in the wastewater. The wastewater was examined by a semibatch treatment system ; the aluminum solution was constantly fed into the vessel filled with wastewater adjusted to pH 7. A zinc solution containing oxalic acid was used as a model wastewater. In this paper, the efficiency of zinc removal was measured at various initial concentrations of zinc and oxalic acid. As a result, it was found that the zinc removal is interfered with such two reasons as forming complexes between aluminum and oxalic acid, and between zinc and oxalic acid. In order to improve efficiency of heavy metal removal, the following methods were discussed ; addition of heavy metal to compete with aluminum in the complex reaction of oxalic acid, or adjustment of the heavy metal concentration to its better removal coefficient. It is found that the laboratory wastewater can be reasonably adapted to such operations.

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A simple and rapid determination of virginiamycin (VGM) in chicken and swine tissues by quantitation of virginiamycin M₁ (M₁), which is a main component of VGM, was developed. M₁ was extracted from the tissues with a mixture of methanol-3% phosphotungstic acid (7 : 3), and the extract was cleaned up by liquid-liquid partitioning with chloroform. The chloroform extract was further cleaned up on a Sep-Pak silica cartridge using chloroform-methanol (97 : 3) as an eluent. M₁ was determined by reversed phase HPLC which was performed on a Kaseisorb LC-ODS-300-5 column with acetonitrile-water (35 : 65) solution as a mobile phase, and M₁ was monitored with UV detection at 235 nm. The caliblation curve was rectilinear in the range from 0.5 to 10.0μg/ml. The average recoveries were 93.6% and 95.2% for VGM added to chicken and swine tissues at the level of 1.0μg/g, respectively. A good correlation obtained between the HPLC method and the bioassay method shows that the present method will be applicable to determine VGM with the detection limit of 0.05μg/g.

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Whether sandwich enzymeimmunoassay (EIA) could be used for the determination of x-carrageenan (x-CA) or not was studied. Furthermore, EIA was compared with double diffusion method and quantitative precipitation reaction. Antisera were obtained from rabbit and guinea pig immunized with x-CA, methylated bovine serum albumin and complete Freund's adjuvant. In EIA, the anti-x-CA rabbit serum as a capture antibody on the solid phase and anti-x-CA guinea pig serum as a detector antibody were used. Further, peroxidase-conjugated anti-guinea pig IgG rabbit antibody and a combination of hydrogen peroxide and orthophenylenediamine were used for color development. In EIA, most suitable dilutions for the determination of x-CA were 20000-fold in both rabbit and guinea pig antisera. The range of determination were from 64 to 512 ng/ml. Cross reaction with some additives with similar uses to x-CA was not observed except that agar slightly showed the reaction. EIA was superior in the sensitivity, the range of determination and the amount of antiserum required to two other methods, double diffusion method and quantitative precipitation reaction. Thus, it was suggested that EIA may be a useful method for the determination of x-CA.

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The simultaneous detection and confirmation of paraquat and diquat which are components of Preeglox L, a bipyridylium-type herbicide formulation, by fused silica capillary gas chromatography/mass spectrometry was investigated. After reduction of paraquat and diquat with a mixture of sodium borohydride-nickel (II) chloride, the reduction products were gas chromatographed on a non-polar (DB-1) and two polar (DB-17 and Supelcowax 10) fused silica capillary columns by using a mass spectrometer as a detector or an ion trap detector. Of the above three columns, a DB-17 column was the most suitable for the simultaneous detection and confirmation of paraquat and diquat as their perhydrogenated products. Further enhancement of specificity was achieved by mass chromatography at the molecular ions of the perhydrogenated products (m/z 196 and 194) and their base peak ions (m/z 96 and 83). The retention indices of the perhydrogenated product of paraquat, and the trans- and cis-isomer of perhydrogenated diquat were 1525, 1530 and 1588 for DB-1, 1740, 1800 and 1840 for DB-17, and 1895, 1920 and 2035 for Supelcowax 10, respectively. Paraquat and diquat as their perhydrogenated products could be also identified by GC with a Fourier transform infrared detector on a DB-17 column.

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We have previously proposed an atomic absorption spectrophotometric method for determination of total phosphorus in water. In this paper, the proposed method was applied to the rain water collected for a year (July 1984-June 1985) from three different locations : Kobe, Amagasaki and Miki. The concentrations of total nitrogen in the water samples were also determined by a conventional method as described in the Official Gazette. Kobe, Amagasaki and Miki, situated in the southern part of Hyogo prefecture in Japan, were chosen as representatives of residential, industrial and agricultural areas, respectively. The water samples from Miki contained significantly higher concentrations of total phosphorus than those from Kobe and Amagasaki, this might be due to fertilizer used in the agricultural area. In contrast, the concentrations of total nitrogen in the samples from Amagasaki were significantly higher than those from the other two locations, which suggested that the nitrogen in rain water might be derived from air pollution with nitrogen oxides owing to motor vehicles and factories in the industrial area.

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COD values (with permanganate) of dicarboxylic acids and unsaturated acids were investigated by the micro titration method previously reported. Dicarboxylic acids will be contained in various industrial effluents and produced during the COD treatment of unsaturated acids. Unsaturated carboxylic acids are constituents of oils extracted from plants or fishes and contained in the domestic effluents. In the case of dicarboxylic acids having longer carbon chain, marked promotion effects of AgNO₃ added into a reaction medium on the COD values were recognized. Average oxydation rate to the perfect oxydation was 12.4%. In alkaline method, oxydation rate was very low values of 0.636%. Oxydation rate by BOD determination was very high rate of 72.1%. In the case of unsaturated acids, the COD values were corresponded to the numbers of unsaturated bonds. Oxydation rate was 39.3% in the acidic and 31.8% in the alkaline method. Determination of reaction products will be reported in the latter.

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