To study the effect of heavy metals on a living body, chlorella was used as the intermediate concentrator, and it was given as food to animals. For this purpose, accumulation of cadmium, mercury, and copper by Chlorella ellipsoidea was studied. Metalic ions of various concentrations (0-25 ppm) as chloride were added to the culture media, and chlorella was cultured by heterotrophic method. Chlorella growth was impeded by 1.0-2.5 ppm of cadmium, 1.0-5.0 ppm of mercury, and 0.5-1.0 ppm of copper. Chlorella died by the addition of morethan 25 ppm of mercury or 5.0 ppm of copper. Cadmium concentration coefficient in chlorella was 166.0 when using cadmium chloride.

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Mice divided into groups were maintained on an experimental diet containing 20 to 200 ppm of polychlorinated terphenyls (PCT) for 3 or 6 months. The concentration of PCT in the liver, kidney, brain, skin, and adipose tissue was analyzed by means of gas chromatography with ECD to clarify the mode of distribution and the ratio of accumulation of PCT from the diet. Contrary to the case of polychlorinated biphenyls and some organochlorinated pesticides, PCT showed a tendency to accumulate much more in the liver than in adipose tissue when compared on wet tissue basis. From the statistical anlyses of the results obtained from all individual mice, it became obvious that the accumulation of PCT in adipose tissue was significantly correlated with that in each of other tissues. It is notable that the concentration of PCT in the liver became 4 to 10 times higher than that of PCT in the diet fed to mice in each group, and the accumulated level of PCT even in the brain was about 10% of the concentration in the diet.

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Spectrophotometric Determination of acetaminophen in pharmaceutical preparations is described. Acetaminophen was hydrolysed by refluxing with perchloric acid to produce p-aminophenol which was reacted with p-dimethyl aminocinnamaldehyde in dilute trichloracetic acid solution to give a red color, which showed absorption maximum at 520 nm. Interference by the presence of phenacetin and sulpyrine can be eliminated by their extraction with chloroform in basic medium or extraction with ethyl acetate in acidic medium.

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The herbicides, 2, 4-D, 2, 4, 5-T, and PCP, which are drived from polychlorinated phenols, are usually contaminated with polychlorinated dibenzo-p-dioxins, and tetrachlorodibenzo-p-dioxin (TCDD) has been given a special attention for its high toxicity to mammals. An improved analytical method for TCDD in sea foods by gas chromatography with ECD and subsequent utilization of mass spectrometer was established. Samples were first hydrolyzed directly with alkali to remove organochlorine pesticides as well as esterified lipid materials. After the extraction of TCDD with hexane, nonsaponifiable materials contained in the extract was removed by treatment with concentrated sulfuric acid. PCB, which still overlap with TCDD in gas chromatographic analysis, can be removed by alumina column chromatography. Samples for which the cleanup by alumina column chromatography was insufficient were further subjected to Florisil column chromatography. Recovery of TCDD added to chopped fish samples by the present method was in the range of 85.4 to 92.5% and the detection limit was about 1.5 ppb by the above mentioned gas chromatography. Since the detection limit is not necessarily sufficient for the environmental monitoring of TCDD, mass fragmentography was employed for further elevation of sensitivity. Thus, 10 ppt of TCDD was become detectable from sea foods.

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In addition to the previous finding that the aliphatic amines react with the nitrite ion to form cyanide ion on distillation of a mixture in acidic condition, an interesting fact was found that glycine reacted with the nitrite ion even in a neutral phosphate buffer solution without heating to form a cyanide ion. This fact suggests the possibility that cyanide ion might be naturally produced in a polluted river water even in the absence of the source of a cyanide ion.

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Seven kinds of metabolites of o-sec-butylphenyl N-methylcarbamate (BPMC) were examined for their anticholinesterase activity to see whether metabolites more potent than BPMC itself were present or not. The products with N-methylhydroxylated or demethylated and alkyl side-chains hydroxylated or carboxylated showed no activity corresponding to BPMC, and it was recognized that metabolism was progressed towards detoxication. Relative intensity of anticholinesterase activity of each of these metabolites for pseudo- and true cholinesterase was similar.

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Bacteriostatic activity of Acridine Orange against E. coli was inhibited by the addition of anionic surfactants. The inhibitory activity of surfactants decreased with a decrease in the carbon number of alkyl chain of the surfactants. The apparent concentration of Acridine Orange to inhibit the growth of E. coli was increased by the addition of surfactants. It was concluded that the complex formation between Acridine Orange and surfactants resulted in a decrease of free Acridine Orange in the culture media.

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Ricin, a highly toxic protein, showed not only a pyrogenic activity but also a strong hemorrhagic activity in rabbit skin. Its activity was due to increased permeability of the blood vessel and therefore, it was more clearly demonstrated with 0.5% solution of Trypan blue, and this permeability was characterized by a long latent period and duration. Ricin had no caseinolytic activity or esterase activity using N-benzoyl-L-arginine ethyl ester and N-acetyl-L-tyrosin ethyl ester as substrates. These results suggest that ricin has a powerful inflammatory effect, but it is not an "inflammatory protease".

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In addition to the previous findings that three componental groups, volatile fatty acids, aromatic hydrocarbons, and phenol, had been recognized as the substances responsible for offensive odor in odorous fish, high-resolution GC-mass measurement was made, in order to detect micro-quantity of other components, and the structures of almost all of the peaks were determined. The above three components appeared as large peaks in dichloromethane extract. DOP, DBP, and PCB were detected and they were assumed to have been the pollutants in water, but amines and sulfur compounds were not detected. Higher fatty acids, cholesterol, and aliphatic hydrocarbons (C₁₀ to C₁₈) were detected as normal components in fishes. As a new component, tetramethylpyrazine was detected and its role is now being examined. Structure of other peaks has not been determined. This high-resolution GC-mass measurement has indicated volatile fatty acids, aromatic hydrocarbons, and phenol as the main substances responsible for the offensive odor in fish.

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Effect of bile acid salts on lipolysis catalyzed by porcine pancreatic lipase was examined by kinetic treatment using pH-stat. 1) The shift of the wavelength of maximum absorption of a dye, Rhodamin 6G, was used to determine the critical micelle concentration of seven bile salts ; sodium cholate, taurocholate, deoxycholate, chenodeoxycholate, ursodeoxycholate, glycoursodeoxycholate and tauroursodeoxycholate. No data are available on the melting point, optical rotation, critical micelle concentration, and effect on lipase activity of the last two bile salts. 2) In this experiment, available surface area of oil droplets was estimated from turbidity, which was proportional to oil-water interface area. By using this method, reproducible data were obtained. This method may enable generalization of available surface area. 3) When tributyrin emulsion was used as a substrate, at the concentration higher than 1 mM, all of these bile salts inhibited lipase activity. In the presence of bile salts, the apparent Km increased, but Vm was not affected, except for chenodeoxycholate. Among the bile salts used, taurocholate was the most and cholate was the least inhibitory at 1 mM for lipase activity, judging from the values of Vm/Km. 4) When olive oil emulsified by polyvinyl alcohol was used as a substrate, effect of bile salts on lipase differed from the use of tributyrin. In the presence of bile salts, the apparent Km and Vm increased. In the presence of 1 mM taurocholate, Vm/Km increased 2-fold of that in the absence of bile salts. As a stabilizing emulsifier, arabic gum was not favorable for estimating the effect of bile salts.

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Distribution of ⁷⁵Se in whole blood, serum, and erythrocytes, after subcutaneous injection in the form of L-selenocystine and sodium selenite to rabbits was examined. During the first 4 hr after the injection of both selenium compounds, ⁷⁵Se appeared in greater concentration in the serum than in erythrocytes but after 10 days, concentration in the serum decreased to roughly epual level as that in erythrocytes. Biological half life of ⁷⁵Se, calculated from the result of successive measurement of radioactivity, 7 and 6 days in whole blood, and 6 and 3 days in serum, respectivly, for L-selenocystine and sodium selenite. Distibution of ⁷⁵Se in whole blood, serum, and erythrocytes was examined with sampled from a rat 30 min after intravenous injection of sodium ⁷⁵Se-selenite (in vivo), and with rat blood incubated with sodium ⁷⁵Se-selenite at 37°for 30 min (in vitro). A higher radioactive concentration was found in erythrocytes in in vivo experiment than that in in vitro experiment. Results of dialysis of sera obtained in vivo and in vitro experiments suggestad that there was some difference between the two sera in the manner of affinity of seleninm to serum protein. Concerning the distribution in erythrocytes, most of selenium was detected in hemoglobin, especially in globin. Globin tagged with ⁷⁵Se obtained by in vivo experiment was hydrolyzed and separated by two-dimentional paper chromatography. The autoradiogram of this paper indicated that the hydrolyzate contained at least 4 differnt componds bearing ⁷⁵Se, two of these seemed to be methionine and cystine or cysteine.

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Determination of organic mercury in various kinds of household necessities, such as clothes, belts, shoe polish, etc., was examined. The organic mercury was extracted with carbon tetrachloride in a hydrochloric acid medium, reextracted from the carbon tetrachloride layer with aqueous solution of cysteine acetate, and then determined by atomic absorption spectrophotometry combined with pyrolysis and the gold amalgamation technique. The recovery of phenylmercuric acetate from cotton and wool cloths was tested in hydrochloric acid of various concentrations. The most appropriate concentration of hydrochloric acid was 1 N for cotton cloth, and 2 N for wool cloth. Recovery was 97 % in the former and 73 % in the latter. All samples analysed contained organic mercury but not more than 0.03 μg/g. There were no articles sanitized with phenylmercuric compounds.

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The previously reported Acridine Orange method for the determination of alkylbenzene sulfonate was applied to the survey of pollution in rivers water and in kinokawa. The proposed method gave a little higher values than the official method using Methylen Blue, but the former was better because of its small coefficient of variation. It was concluded from the results that the proposed method would be available as a routine one for the survey of pollution of alkylbenzene sulfonate in river water.

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A rapid microanalytical method for the determination of chromium in urine utilizing flameless atomic absorptiontechniqe has been developed. By the use of a 2-channel type of atomic absorption spectrophotometer, urinary chromium concentration can be determined rapidly because the interference due to the smoke of other component which occure at the atomization of samples is corrected automatically. This method has been used for the determination of chromium in urine samples of individuals exposed to chromium and nonexposed. The chromium level, mean and standard deviation, found in urine of nonexposed group was 3.1±2.2 ppb, and that of exposed groups was 4.4±6.0 ppb inhabitants near the factory and 6.6±4.3 ppb for workers in the factory handling chromium.

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