The review on our works mainly consists of metabolic activation of morphine related to in vivo and in vitro hepatotoxicity. Especially, the biotransformation of morphinone-glutathione conjugate from morphine, and the role of the reactive metabolite of morphine (morphinone) has been discussed in detail.

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Furazolidone (FZD) and sodium nifurstyrenate (NSA-Na), antibacterial nitrofurans, have been used as veterinary medicines for the prevention and treatment of bacterial infections in swine or fish. Information about the metabolism of FZD or NSA-Na is very important for the safety evaluation of these compounds. The present review describes isolation and identification of metabolites of the nitrofurans in mammalian species (rats, rabbits and swine) or fish (gold fish, eel and sea bream), nitrofuran-metabolizing activities of liver preparations and results of mutation test for the nitrofurans and their metabolites.

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Secondary ionization mass spectrometry (SIMS)-linked scanning analysis was used as a method for identifying a specific Aloe arborescens MILLER (Kidachi-aroe) component, aloenin, in health foods containing Kidachi-aroe. The compound was extracted with methanol and identified in positive mode and constant B/E using SIMS-linked scanning analysis without any further purification such as a chromatography. Kidachi-aroe was detected in the sample using aloenin as an indicator. In the SIMS-linked scanning spectra of health foods containing Kidachi-aroe, peaks were observed of a quasi-molecular ion (m/z 411[M+H]⁺) and its daughter ion at m/z 249, indicating the structure of aloenin. Kidachi-aroe was thus detected when present in health foods. The proposed method is easy, rapid and accurate for both characterization and estimation of aloe materials in health foods containing Kidachi-aroe.

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The characteristics of CHG (Chlorhexidene digluconate)-resistant activated sludge acclimated to the wastewater containing 0.5 ppm CHG were investigated. Six strains of 100 ppm CHG-resistant bacteria, Pseudomonas group and Flavobacterium group, were isolated from the effluent after treatment of the activated sludge with the wastewater containing 0.5 ppm CHG. The BOD₅ value of the aqueous solution containing 100 ppm CHG, which was measured by inoculating the effluent after treating the wastewater containing 0.5 ppm CHG with 0.5 ppm-CHG-acclimated activated sludge, was nearly equal to that of digluconate in the test solution and CH (Chlorhexidene) almost remained in the culture without biological degradation. The results showed that the CHG-acclimated bacteria could degrate digluconate but not CH or CHG and CHG-unacclimated activated sludge could not degrate even digluconate. Characteristic changes were observed in a number of 100 ppm CHG-resistant bacteria in the effluent during the batch activated sludge treatment of wastewater containing CHG at three concentrations. The results suggested that when the wastewater containing CHG is treated with batch activated sludge process, longer time aeration would be efficient to obtain high treatment efficiency, because the growth of CHG-resistant bacteria might be very slow.

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The POPHA (p-hydroxyphenylacetic acid) fluorometric method for the determination of hydrogen peroxide (H₂O₂) in rain water was investigated. The optimum conditions for peroxidase on the formation of fluorescent POPHA dimer from H₂O₂ and POPHA are over 2 units/ml of peroxidase concentration and above pH 5.0. The best conditions for catalase on the decomposition of H₂O₂ in the testing solutions as analytical blank are over 500 units/ml, over 2 min of reaction time and above pH 5.5. By this analytical method, the detectable range of H₂O₂ concentration is from 1 to 150 ng/ml and the coefficients of variation are below 5%. This method is hardly influenced by the coexistence of organic hydroperoxides, organic acids and aldehydes. However, the Fe²⁺ concentration of more than 1000 ng/ml showed negative interference for this fluorometric method. Recoveries of H₂O₂ added to rain waters are satisfactory, ranging from 92 to 94%. Therefore, it is considered that this method is applicable to the determination of H₂O₂ in the rain water.

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o-Dinitrobenzene (DNBz) and each nucleic base were made to react with NaOH in dimethylsulfoxide at room temperature for 3-60 h. All of the synthesized compounds were the 2-nitrophenyl adducts of nucleic bases. The mutagenicity test of the synthesized compounds and of o-DNBz were then carried out against Salmonella typhimurium TA98, TA98NR and TA100 without a mammalian metabolic activation system. o-DNBz has no mutagenic potency against all the strains at a dose up to 100 μg. In the compounds with the adducts of nucleic bases, the mutagenic potencies of the adducts of adenine and guanine in TA98 were 50 revertants/100 μg and 255 revertants/10 μg, respectively and the most potent mutagen in TA100 was an adduct of guanine (382 revertants/100μg). Finally, the structures of the adducts of adenine and guanine could be elucidated as N¹-(2-nitrophenyl)-adenine and N¹-(2-nitrophenyl)-guanine, respectively, by the acid and alkaline degradation and by the isopentyl nitrite treatment of these adducts.

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A mouse hepatic microsomal enzyme that catalyzes the oxidation of cannabinol (CBN) to 11-hydroxycannabinol (11-OH-CBN) was characterized. 11-OH-CBN formed was determined by HPLC. The reaction required NADPH as a cofactor and the optimal pH for the reaction was at 7.8. The Km and Vmax values for the reaction were 122 μM and 8.82 nmol/min/mg protein, respectively. The reaction was inhibited by inhibitors of cytochrome P450 such as SKF 525-A and by cannabidiol. The pretreatment of mice with phenobarbital significantly increased the enzyme activity but not with 3-methylcholanthrene, whereas that with cobaltous chloride significantly decreased the activity. These results indicate that cytochrome P450 plays a major role in the formation of 11-OH-CBN in the mouse hepatic microsomes.

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A simple method for the collection and determination of volatile hydrocarbons in air was examined. The investigated thirteen volatile hydrocarbons were benzene, toluene, ethylbenzene, three kinds of xylene, cumene, three kinds of ethyltoluene, mesitylene, octane and nonane. Air dehumidified by magnesium perchlorate was sampled at 0.5 l/min for 24 h through a glass tube (2.6 mm i.d.) packed with 400 mg of Carbosieve S-III (60/80 mesh) which was divided into a 200 mg front bed and a 200 mg rear bed. Each sampled Carbosieve S-III was extracted ultrasonically with 1 ml of carbon disulfide for 10 min. An aliquot of the extract was analyzed by GC equipped with a FID on a glass column (3 mm i.d. by 2 m long) packed with 5% SP-1200 and 1.75% Bentone 34 on Chromosorb W (AW, DMCS, 60/80 mesh). The efficiencies and recoveries of the collection of the volatile hydrocarbons were quantitative ; the minimum range of detectable concentrations were from 0.03 ppb for benzene to 0.16 ppb for cumene, m-ethyltoluene and mesitylene. This method could be applied to analyze the volatile hydrocarbons in indoor air from laboratories and houses, and in the atmosphere.

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To investigate whether any toxic lipophilic substances were contained in the anchovy (Engraulis japonicus) infected with Anisakis simplex or not, correlations among mouse lethal toxicity in the lipophilic fraction, the crude lipid content from the whole body or viscera of the anchovy and the prevalence of Anisakis infection were examined, but no significant correlation was found in the present survey data. However, the anchovy having Anisakis simplex showed a significantly higher level (p<0.01) of free fatty acids in viscera, in particular, C16 : 0, C16 : 1 (n-7), C18 : 0, C18 : 3 (n-3), C20 : 3 (n-6), C20 : 4 (n-6), C20 : 5 (n-3), compared with that free of Anisakis. Total polyunsaturated fatty acid in phospholipid and triglyceride tended to decrease in the anchovy viscera having Anisakis simplex compared with those without parasites. It is possible to postulate that an increase of free fatty acids in the anchovy with parasites may be related to the outbreak of human allergic symptoms accompanied by eating raw anchovy.

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We have reported previously the presence of mutagenicity in the extract from the sediment collected from some points of the Nakatsu river. In order to characterize mutagens in the river sediment, the extract from the Nakatsu river sediment was fractionated into 9 fractions by HPLC and tested using Salmonella typhimurium TA98, YG1021 and YG1024. The middle-high polar fractions in the extract from the sediment showed mutagenicity to YG1024 by the addition of S9 mix. Then, some aminoarenes were investigated in these fractions by NPD-GC. More polar two fractions revealed mutagenicity to TA98 without S9 mix. In these fractions some mutagenic substances were surveyed and identified by GC-MS, but unknown peaks were present. Mutagenicity, PAH concentration, amount of extract and ignition loss of the sediments from the Nakatsu river were compared with those of the Sagami river. As the result it is comfirmed that the Nakatsu river is specific on mutagenic contamination.

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HPLC using a large-volume injection technique has been developed to determine trace levels of pesticides in water. To prevent disturbance of the base line from excess water injected, a reversed-phase guard cartridge is substituted for an injection loop as an enrichment column. This procedure is applied to Asulam, N'-methoxycarbonyl sulfanylamide, one of the herbicides used in golf links. One milliliter of sample made up in a solution of 0.5 mM in cetyltrimethylammonium chloride is passed through the cartridge, where Asulam is trapped quantitatively. Asulam is then back flushed from the cartridge onto an analytical column, where the separation is achieved. Asulam below ppb level can be readily detected. The precision of the measurements, evaluated by the analysis of spiked samples, is 3.3% at 1 ppb level.

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To assess the components and the contents of several health foods, especially processed by condensing, the contents of amygdalin (AM), benzoic acid (BA), levulinic acid (LVA), 5-hydroxymethylfurfural (HMF), 5 kinds of organic acids (citric acid, malic acid, succinic acid, tartaric acid and fumaric acid), potassium and sodium were determined for 15 Japanese apricot extracts (10 paste types, 5 pill types) and 7 prune extracts. AM was detected in all the samples of Japanese apricot extracts (0.067-4.64 mg/g), but was not detected in the prune extracts. The average contents of BA, LVA and HMF were 0.17 mg/g, 0.61 mg/g and 5.32 mg/g for the paste type of Japanese apricot extracts, 0.086 mg/g, 0.24 mg/g and 0.99 mg/g for the pill type, and 0.011 mg/g, 0.005 mg/g and 3.47 mg/g for the prune extracts. By the dry concentration of Japanese apricot juice at 95°C, HMF was produced in large quantities accompanied with a change in the state from liquid to semisolid, and then LVA was produced. Main organic acids in the components of Japanese apricot extracts were citric acid and malic acid. The average contents of citric acid and malic acid were 319 mg/g and 106 mg/g for the paste type of Japanese apricot extracts, and 94 mg/g and 29 mg/g for the pill type. In the prune extracts, the content of malic acid was detected to be 8 mg/g on the average. The contents of potassium were 24 mg/g for the paste type of Japanese apricot extracts, 16 mg/g for the pill type, and 9 mg/g for the prune extracts on the average. All these values were larger than those of respective contents of sodium in the three extracts.

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