Bacteria of the genus Vibrio are normal habitants in an aquatic environment such as river, estuarine, coastal and sea water, but include some species pathogenic for human. Currently 37 species are reported in the genus Vibrio, and 11 species in the genus Vibrio are recognized to be pathogenic for human. Of these pathogenic species, V. cholerae is well known as pathogen of epidemic cholera, and V. parahaemolyticus is the most important food poisoning bacterium in Japan. V. cholerae is serologically classified into two groups, V. cholerae Ol and non-Ol, namely NAG vibrio. V. cholerae Ol is an actual cholera vibrio causing epidemic cholera, characterized by severe watery diarrhea caused by cholera toxin. In the humen history, there were 7 worldwide cholera pandemics. On the other hand, some strains of non-Ol also cause diarrhea, but the symptoms are not so severe and not contagious. Therefore, non-Ol is defined as a food poisoning bacterium. In recent years, 40 to 60% of the food poisoning outbreaks are due to V. parahaemolyticus. V. mimicus and V. fluvialis are also recognized as food poisoning bacteria. In addition to these diarrhogenic vibrios, significance of V. vulnificus as a pathogen of opportunistic infection is suggested recently, because the vibrio occasionally causes fatal septicemia to aged patients having underlying disease, such as hepatic dysfunction. Sea foods in markets are frequently contaminated with V. parahaemolyticus or V. vulnificus in summer season, and V. cholerae non-Ol and V. mimicus are universal inhabitants in fresh or brackish water regions. Thus, the pathogenic vibrios are important bacteria in food sanitation control.

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The mechanism for the international regulatory control of drugs of abuse under the Single Convention on Narcotic Drugs and the Convention on Psychotropic Substances is outlined. A historical overview of the substances which have been added to the list of controlled drugs is presented, focusing on the review of dependence-producing substances by the World Health Organization (WHO) for international control decisions.

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An analytical procedure for morphine-3-glucuronide (M3G) and morphine (M) in the human urine was investigated by using high-performance liquid chromatography coupled with atomospheric pressure ionization mass spectrometry (LC/APCI-MS). The samples were purified with Sep-Pak C₁₈ cartridges. The LC separation was carried out using a L-column ODS in 50 mM ammonium acetate-methanol (86 : 14 v/v). The calibration graphs constructed by the absolute calibration curve method showed linearity over the concentration range of 30 to 1000 ng/ml (γ=0.9965) for M3G and 30 to 2000 ng/ml (γ=0.9970) for M. The detection limits by selected ion monitoring (SIM) were 3 ng/ml for M3G and 1 ng/ml for M, and by scan mode were 350 ng/ml for M3G and 80 ng/ml for M. The coefficients of variations for M3G and M were 4.79 and 3.15% at 1 μg/ml, respectively.

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Oxidative stress in the rat erythrocytes induced by interaction between organic selenium compound and reduced glutathione (GSH) was investigated in vitro. Erythrocytes simultaneously treated with selenocystamine or selenocystine and GSH revealed the increase of hemolysis and TAB-RS formation and the depressed GSH content. The elevation of TBA-RS formation induced by the interaction was depressed by the addition of catalase (Cat). The chemical interaction between organic selenium compound and GSH was confirmed by reduction of nitro blue tetrazolium. This reduction was depressed by the addition of SOD or Cat. Hydrogen peroxide was generated in the extracellular solution of rat erythrocytes which were treated with organic selenium compound and GSH. Hydrogen peroxide added in the erythrocytes, which were pretreated with GSH-depressor diethylmalate, increased TBA-RS formation. These results demonstrated that hydrogen peroxide which is formed by the interaction of organic selenium compound and GSH may permeate into erythrocyte membranes and induce lipid peroxidation.

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Enantiomeric compositions of methamphetamine (MA) and its major metabolite, amphetamine (AP), in the urine of mice administered with deprenyl (DPN) and MA were analysed by high-performance liquid chromatography using the tetra-O-acetyl-β-D-glucopyranosyl isothiocyanate (GITC) derivatization method for the discrimination between the use of l-DPN and MA enantiomers. l-DPN, d-MA, l-MA and dl-MA were intraperitoneally administered to mice at 1, 5 and 10 mg/kg. Urine samples were collected at 0-12, 12-24, 24-48 and 48-72 h after administration, Urine added with N-methylphenethylamine (internal standard) was applied to BOND ELUT CERTIFY (1 ml), washed with 1 ml of 0.1 M acetic acid and 6 ml of methanol, and eluted with 2 ml of conc. HCl-methanol (9 : 91). The eluent was evaporated, redissolved in 1 ml of water, basified with 1 N NaOH and extracted with chloroform. The amphetamines in chloroform layer was derivatized with GITC at room temperature for 30 min. The GITC-derivatives were analysed by HPLC with a UV detector (254 nm). As a result, administration of l-DPN could be clearly discriminated from that of d-MA and dl-MA by comparisons of the enantiomeric compositions of MA excreted in the urine. Discrimination between administration of l-DPN and l-MA could be carried out by the apparent difference of ratio of AP/MA in the urine. The AP/MA ratio of the former was 10 times that of the latter.

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A paraquat-sensitive electrode was constructed by use of a poly (vinyl chloride)-based membrane containing sodium tetrakis [3, 5-bis (trifluoromethyl) phenyl] borate as an ion exchanger and 2-fluoro-2'-nitrodiphenyl ether as a membrane solvent. The minimum detection limit of paraquat was 5×10^-7M. The calibration slope of the electrode in linear region was 28 mV/decade. The selectivity coefficients against Na⁺, K⁺, Mg²⁺ and Ca²⁺ were less than 10^-4, and substantially, these inorganic ions did not interfere with the response to paraquat. This electrode was utilized for a criminal investigation. Paraquat added intentionally to drinks such as juice, cola and milk in cases of murder can speedily be measured by the paraquat-sensitive electrode without separating the herbicide from assay mixtures. By using the present electrode, we could estimate paraquat down to 5×10^-5M contained in juice. This sensitivity was sufficient enough for the detection of paraquat used in a crime : when the lethal dose of paraquat (2.4 g/a man of 60 kg) was mixed with juice (250 ml), the concentration became about 40 mM. The electrode was also applied to a clinical analysis, and paraquat in the serum could successfully be detected down to 2μM.

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Discriminatory uptake of calcium (Ca) and strontium (Sr) by the intestine that had been shown to become more strict with age was examined to get a more insight into the regulatory mechanisms in rats. Active transports of Ca and Sr at the small intestine (duodenum, proximal and distal jejunum, and ileum) were compared using the everted gut-sac procedure between young and adult rats aged 5 and 25 weeks old, respectively. The ratio of S/M (the ratio of ⁴⁵Ca and ⁸⁵Sr transported against the concentration gradient from mucosal to serosal sides) indicated that Ca is transported actively only in young rats, while Sr is transported non-actively at any sites of the intestine examined in both young and adult rats. The discriminatory ability for Ca and Sr at the intestine was compared between the two age groups with the Sr/Ca ratio obtained by the ligated-loop procedure. The ratio was about 1.0 at the duodenum in young rats, while it was shown to be 0.4-0.6 at the other sites in young rats and also at all sites of the intestine in adult rats, indicating that the discriminatory ability is lowered to half at the duodenum in young rats. Phospholipid composition of the duodenum brushborder membrane was compared between young and adult rats to examine the effect of its change on the discriminatory uptake of Ca and Sr. In young rats, phosphatidylcholine increased and phosphatidylethanolamine decreased in composition. These results suggest that changes in phospholipid composition in the duodenum brushborder membrane may explain the change with age in the discriminatory uptake of Ca and Sr at the intestine.

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Concentrations of trichloroethylene (TCE) in air, soil and groundwater were measured around a metalware factory, where TCE was used for degreasing and cleaning of metalware. The mean level of TCE exhausted from the cleaning system through the stack at the factory was 360 ppm at the operating system or 130 ppm at the stand-by system, and was estimated to 30% of TCE used at the factory ; most residual of the used TCE (70%) was introduced into air through ventilators. TCE exhausted from the factory to the air was diluted rapidly, and TCE concentration in air decreased as the distance from the stack lengthened on the leeward. Some of TCE in air was supplied to soil within at least 400 m from the stack. TCE concentration in soil decreased as the distance from the stack lengthened. TCE concentration in groundwater from a well of 10 m depth located at the factory was 15000 μg/l at the average. cis-1, 2-Dichloroethylene, a product from TCE by biodegradation, were also detected in groundwater at 1700 μg/l at the average. TCE exhausted to the air distributed in air and soil, but this TCE did not cause groundwater contamination at the level of more than several mg/l.

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A high performance liquid chromatography-mass spectrometric (LC-MS) method for the determination of oxazepam glucuronide in the human urine was investigated. Oxazepam glucuronide was extracted from the urine by using Sep-Pak C₁₈ cartridge with methanol-50 mM ammonium acetate (70 : 30 v/v) as eluent. The eluate was analyzed by LC-MS equipped with atmospheric pressure chemical ionization (APCI). High performance liquid chromatographic (HPLC) analyses were performed on a L-column (C₁₈ reversed phase column) in the solvent system of methanol-50 mM ammonium acetate (70 : 30 v/v) at a flow rate of 1.0 ml/min. The vaporizer temperature was set to 230°C, desolvation temperature to 390°C, drift voltage to 108V, focus voltage to 96V and multiplier voltage to 2400V. Peak areas were measured using selected ion monitoring (SIM) of a base ion peak at m/z 287. The calibration curve gave a good linearity over the range of 0.25-1 μl/ml. The detection limit was 50 ng/ml by recording the base ion peak at m/z 287.

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The analyses of dialkylphosphates, aqueous metabolites or degradation products of organophosphorus pesticides, with extractive pentafluorobenzylation in the presence of polymeric phase-transfer catalysts (tri-phase catalysts) were investigated. Dialkylthiophosphates and dialkyldithiophosphates (alkyl=methyl, ethyl) in aqueous samples were extracted and derivatized in more than 98% yield by pentafluorobenzyl bromide in 15 min at 40°C using tri-n-butyl methyl phosphonium bromide polymer bound as a tri-phase catalyst and MIBK as an organic solvent. Dimethylphosphate and diethylphosphate were also derivatized in 43% and 100% yields, respectively, in the presence of this tri-phase catalyst in 90 min at 90°C. The simultaneous analyses of these 6 phosphates in the spiked urine sample were carried out with excellent detection limits, 0.44-0.79 ppm by scan-mode GC-MS and 0.027-0.081 ppm by ECD-GC) using this tri-phase procedure.

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Ten samples of powder mixtures containing known quantities of methamphetamine·HCL and ephedrine·HCl were measured by X-ray diffraction. The respective I/I_O values of 19 peales were selected for calibration. The data was used for the multivariate analyses including a cluster analysis, and a cluster analysis derived from the deviation and a principal component analysis. Subsequently the approximate values of the methamphetamine·HCl contents of unknown samples were determined from the 12×19 matrix. The accurate methamphetamine·HCl contents of unknown samples were calculated from the principal component score. Differences between the theoretical and calculated values of 10 samples ranged from -1.4 to 1.7% (the respective methamphetamine·HCl contents are 3.9, 14.5, 20.2, 34.4, 44.0, 49.8, 60.0, 75.2, 96.1 and 99.1%).

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The concentrations of seven non-volatile amines, tyramine (Tym), histamine (Him), phenethylamine (Phm), putrescine (Put), cadaverine (Cad), spermidine (Spd) and spermine (Spm), during miso fermentation were studied. Tym of these amines which is generally known to be present in miso was detected during fermentation by the conventional production method. Him and Phm were not detected, and concentrations of Put, Cad, Spd and Spm, which were present in the raw starting material for miso, did not change significantly during fermentation. However, Tym and Him were produced when the miso starting materials, i. e., cooked soybeans, rice koji and mashed miso, were added to liquid medium and allowed to stand. Tym- and Him- producing bacterial strains were isolated from the liquid culture media containing the miso starting materials. The Tym-producing strain was a gram-positive coccus, and the Him-producing strain was a gram-positive rod. Thus, the results show that the Tym and Him observed in culture were produced by the two different species. During miso fermentation, Tym and Him are apparently produced within a short period of time when the respective optimum conditions for their growth are present. With the exception of Phm which did not exist in the miso starting materials, other amines including Put, Cad, Spd and Spm were not produced and microorganisms producing them are not believed to be present during miso fermentation.

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A simple and rapid method for the quantitative analysis of oxine-Cu (copper 8-quinolinolate) in the environmental waters was studied. The sample waters were passed through a polychlorotrifluoroethylene (PCTFE) resin powder-cartridge for preconcentration and the passing solution was subjected to reversed-phase high performance liquid chromatography on a column of C₁₈-bonded silica-gel (ODS-Cu) with spectrophotometric detection at 240 nm. The standard curve was linear in the range from 1.0 to 100 ng. The recoveries of oxine-Cu added to river waters (n=6) and tap-waters (n=2) were from 92.0 to 100.0%. The present method was successfully applied to the analysis of oxine-Cu in the environmental waters.

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Morphine and codeine in poppy seeds (four brands) available in Japanese market were measured by using gas chromatography-mass spectrometry. These alkaloids were detected in all poppy seeds and their contents were 7.3 to 60.1 μg/g of morphine and 6.1 to 29.8 μg/g of codeine. These two alkaloids were also analyzed in several kinds of seed foods (bread and dumpling), on which poppy seeds (0.39 to 4.85 g/piece) were decorated. The poppy seeds on these foods contained 1.8 to 93.0 μg/piece of morphine and 1.4 to 56.9 μg/piece of codeine. From these results, it was strongly proposed that the cut-off values of morphine and codeine in body fluid for the forensic detection of illicit use of morphine, heroin or opiates have to be established.

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Determination of Saccharin in foods by high performance liquid chromatography (HPLC) after extraction with dialysis was developed. Five to twenty grams of samples were cut into pieces and transferred into a cellophane tubing with 30 ml of 0.1 N HCl. The tubing was submerged in 0.1 N HCl in a messcylinder so that the total volume reached 200 ml. After 24 h of dialization with occasional mixing, the dialysate was analysed by HPLC on a column of NH₂ using methanol -1% phosphoric acid (4 : 6) as mobile phase, and the elution was monitored by the absorbance at 230 nm. The minimum measurable amount was 0.005 g/kg, and the recoveries when added to soysauce and grape juice at concentrations of 0.1 to 0.5 g/kg were 96-101.7%.

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A rapid and simple method for the simultaneous determination of trace amounts of asulam and 4 kinds of phenoxy acid herbicides (2, 4-PA, MCP, 2, 4, 5-T and MCPP) in water is described. A 100 ml volume of sample water was preconcentrated by passage at a flow-rate of 5 ml/min through Bond Elut SAX. These herbicides absorbed were eluted with 4 ml of 0.2 M NaCl-acetonitrile (1 : 4). The mixture was concentrated to 1.0 ml under vacuum at 35-40°C and analysed by high performance liquid chromatography with mobile phase of acetonitrile-0.05 M phosphate buffer (pH 4.5) (30 : 70) containing sodium 1-heptanesulphonate (5 mM). The limit of detection with UV detection at 280 nm was 1.0 μg/l in these herbicides. The recoveries for each herbicide from 5 kinds of sample water were 80-91% and the coefficients of variation were 2.4-4.1%.

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