Mycotoxin production was investigated in 27 kinds of commercial foods artificially inoculated with following representative toxigenic fungi, i. e., afiatoxin-producing strain of Aspergillus parasiticus NRRL-3000, sterigmatocystin-producing strain of A. versicolor I-20-b, ochratoxin A-producing strain of A. ochraceus 10-21-N, and Fusarium toxinproducing strain of Fusarium solani F-135. It was observed that among vegetables and food products tested, dried sweet potato and salami sausage were favorable substrates for the production of several kinds of mycotoxins and dried persimon was favorable for sterigmatocystin production. Pepper, roasted coffee bean and cinnamon bark were not susceptible to mycotoxin contamination under the moistend condition. Substances contained in a chloroform extract of cinnamon and pepper inhibited the growth of the four toxigenic fungi. These results indicate that cinnamon and pepper might contain inhibitory materials.
To evaluate the safety of biological and chemical contents in “Tea fungus” as a kind of soft drink, microbial flora and organic acid content in this drink were investigated. As the result of microbial investigation, Acetobacter xylinum was isolated from the drink as sole source of bacteria and other yeasts such as Saccharomyces cerevisiae, S. inconspicus, Candida tropicalis, Debaryomyces hansenii, were also found. Among those isolates tested, A. xylinum was found to be a zoogloeal mats forming organism, and the product was increased in cooperation with S. cerevisiae. As the result of organic acid analysis by gas liquid chromatography, acetic acid, formic acid and lactic acid were found, especially acetic acid content occupied more than 95% of the total of them in this drink. Several kinds of pathogenic bacteria such as Salmonella typhi, Shigella sonnei, pathogenic Escherichia coli, Staphylococcus aureus, were all died within 24 hours, when experimentally inoculated into the drink and incubation at both of 25°C and 37°C, though several kinds of filamentous fungi were found grow well in it.
In order to clarify the chemical form and role of cadmium in tissues which contain high levels of the metal, the present study was carried out on the subcellular distribution of cadmium and characterization of cadmium-containing protein fraction from watersoluble fraction in the liver of whelk, Buccinum tenuissimum. The concentration of cadmium accumulated by whelk liver was found often more than 1mM and about 30% of which distributed to the 105, 000×g supernatant, about 60% to the nuclear and cellular debris fractions, and residual small amount of the element to the mitochondrial and microsomal fractions. When the liver homogenate was treated with high ionic strength such as 2.0M NaCl and 10mM Na2-EDTA, cadmium content of the soluble fraction increased in both cases by 35% compared with control, which appeared to be explained by ionic adsorption. Cadmium in the soluble fraction did not markedly increase by boiling the liver homogenate for 25min, and also by digestion with proteases; pepsin and trypsin. On the other hand, cadmium in the soluble fraction under the acidic pH, whereat concentration of the metal increased 4-fold compared with that extracted with 0.25M sucrose, was found to be inorganic free state by gel filtration, but under the alkaline pH, cadmium in the soluble fraction almost was found to be bound to proteins. By means of gel filtration over Sephadex G-75, cadmium in the soluble fraction was found to be principally bound to fraction F-I of >75, 000 molecular weight and fraction F-III of <1, 500 molecular weight and a significant amount of cadmium was also associated with fraction F-II of 9, 000 to 13, 000 molecular weight. Under the presence of 2-mercaptoethanol, cadmium in the soluble fraction was almost associated with fraction F-II. Cadmium-binding protein F-I and F-II equilibrated with free cadmium ion, and the equilibrium was influenced with pH and 2-mercaptoethanol. At the acidic pH under pH 5.0, protein bound cadmium decreased sharply, but over pH 7.0, cadmium-protein complex was very stable, and therefore it was difficult to remove completely cadmium from F-I or F-II by dialysis with 1mM Na2-EDTA. These evidences show that cadmium in the liver of whelk are bound firmly to be the nucleus and protein in the soluble fraction.
From the standpoint of sero-epidemiology, a technique measuring antibodies against the enterotoxin produced by Cl. perfringens was devised using Vero cells, and the system, thus established, applied to the determination of the antibody levels in the sera of humans and animals. 1. Antibodies neutralizing the quantity of enterotoxin as small as 1.2μg/ml (corresponds to 8 minimum cytotoxic dose) could be detected by the micro cell culture method, where the specific neutralization of Vero cell toxicity by the enterotoxin was utilized as an index. 2. The positive ratio for the antibodies (neutralization of 8 MCD or more enterotoxin) in humans was 19.9% (29/146), whereas, in other animals, 0% (0/35) for tattles, 0% (0/60) for pigs, 0% (0/55) for chickens, 0% (0/41) for brown rats. 3. The maximum antibody level detected in a healthy human serum (1ml) was capable of neutralizing enterotoxin of 4.8μg, and this antibody seemed to belong to the IgG class.
Reaction of butyl hydroxy anisol (BHA) with sodium nitrite or potassium nitrate by irradiation of ultra violet ray were studied. BRA and sodium nitrite or potassium nitrate in ethanol-water (4:1) solution were irradiated by ultra violet ray for 24hr at room temperature. As the results, 4 species of reaction products were identified, and these structure were determined to be 1, 4-dimethoxy-2-nitrobenzene, 1-hydroxy-2-tert-butyl-4-methoxy-6-nitrobenzene, 1, 4-dimethoxy-2-tert-butyl-6-nitrobenzene, 1, 4-dihydroxy-2-tert-butyl-6-nitrobenzene. At the same time, the degradation products of BHA by ultra violet ray irradiation was studied, as a results, 7 species of degradation products were identified, and these structure determined to be 2-tert-butyl-quinone, 2-tert-butyl-hydroquinone, 2-tert-butyl-1, 4-dimethoxy-benzene, 2′, 3-di-tert-butyl-2-hydroxy-4′, 5-dimethoxy-biphenil ether, 3, 3′-di-tert-butyl-2, 2′-dihydroxy-5, 5′-dimethoxy biphenyl, 3, 3′-di-tert-butyl-2′-hydroxy-2, 5, 5′-trimethoxy biphenyl and 3, 3′-di-tert-butyl-2, 2′, 5, 5′-tetramethoxy biphenyl.
Reaction of buthyl p-hydroxybenzoate (PHBA-Bu) with potassium nitrate or sodium nitrite by irradiation of ultra violet ray and mutagenicity of its reaction products were studied. 1% aqueous solution of PHBA-Bu and potassium nitrate or sodium nitrite were mixed and irradiated with stirring for 5 days under the high voltage mercury lamp (230W). The reaction products were separated by thin-layer chromatography with benzenechloroform system (4:1, v/v), and colored spots of four were appeared on the plate. The yellow substance which was showed most high Rf-value was isolated, and was identified as butyl 3-nitro-4-hydroxybenzoate (BNH). BNH showed mutagenicity by using Bacillus subtilis and Yeast in comparison with original material.
Impurities in piperonyl butoxide were analysed by gas chromatography. Each of the samples contained 80.5-98.6% pure piperonyl butoxide and another 5 kinds of foreign matters. The sample was fractionated with a column and thin layer chromatography. Each of the cleaned up components showed a single peak by gas chromatography. For the identification of each component, the elementary analysis and spectrometry (FD-Mass, EI-Mass, IR and NMR) were carried out. The following substances were identified as the impurities; 6-hydroxymethyl dihydrosafrole, 6-propylpiperonyl n-butylethyleneglycol ether, 6-propylpiperonyl n-butyldiethyleneglycol ether, 2-propyl-4, 5-dimethoxybenzyl n-butyl-diethyleneglycol ether, bis (2-propyl-4, 5-methylenedioxyphenyl) methane and di (2-propyl-4, 5-methylenedioxbenzyl) ether.
Urinary excretion rate of nitrate in man was studied by determining the amount of nitrate ingested with foods and beverages and that excreted into urine for 24hr and 48hr. The mean nitrate concentration ±S. D. in the urine (n=33) which were collected from 11 subjects at random was 74.4±42.5ppm. The excretion amounts of nitrate in the 24hr urine obtained from 9 subjects who ingested conventional diet containing from 44.1 to 864.2mg of nitrate was from 45.6 to 236.9mg, indicating a remarkable and individual differences of urinary excretion rate of nitrate (27.4 to 133.8%). The excretion amount of urinary nitrate every 2hr in two subjects was less than 10mg when only the low-nitrate diet containing a small amount of nitrate was ingested, but increased more than 25mg every 2hr by the ingestion of 200mg of sodium nitrate or 100g of salted Chinese cabbage, containing 75.1mg of nitrate, with the low-nitrate diet. The range of the urinary excretion rates of nitrate for 48hr was from 78.2 to 86.2% (n=3). No nitrite was detected in any urine specimens, and neither nitrate nor nitrite was detected in any feces (n=6).
Extraction technique and determination method of 3, 5-dinitro-o-toluamide (zoalene) were described in this paper. The mixture of sample and anhydrous sodium sulfate was ground to fine powder by a homogenizer. The mixture was packed in a glass column of 38mm in diameter tightly. As eluting agent acetonitrile was used. Zoalene was eluted efficiently by the column chromatography, and after clean up it was treated by florisil column chromatography, condensate of eluent was made to react with heptafluoro-n-butyric anhydride. The determination was performed by ECD gas chromatography on the solution of reaction product. A linear calibration curve was obtained in range of 0.1μg/ml to 1.0μg/ml of zoalene. The lower limit of the sample size of zoalene for identification was 0.03ng.
Elimination, distribution and metabolism of morpholine salts in rats were investigated by means of chemical analysis and/or radioassay. Gas-liquid chromatography was used for chemical analysis of morpholine in the rat urine and faeces. The analytical results of the excreta accorded with those made by the tracer technique. When rats were given morpholine-HCl or -palmitate, it was found that about 90% of the dose was excreted in the urine over a period of 3 days and the remaining in the faeces. The faecal excretion of morpholine palmitate appeared to be a little more than that of morpholine-HCl after an oral dose of each labelled compound. Morpholine was largely excreted unchanged in the urine. The elimination of morpholine from organs, tissues and blood was generally rapid and specific organ-affinity was not observed in other organs except the intestine. The lowest affinity was found for adipose regardless of routes of administration.