The enzyme solution was prepared from the heparynized serum of Wister rat diluted into 25-fold with Sorensen buffer (pH 7.0). p-Nitrophenylacetate was used as substrate, and the effect of phenylmercury acetate (PMA) and ethylmercury phosphate (EMP) on this enzyme was studied. The enzymic activities were examined by the colorimetrical determination of the amount of p-nitrophenol liberated from substrate. It was proved that both organic mercuric compounds inhibit this enzyme competitively.

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For the purpose of studying the effect of food additives on the activity of trypsin, the cup and disc methods were applied. Eight kinds of food additives, i.e., sodium benzoate, sodium dehydroacetate, dulcin, sodium cyclamate, sodium glutamate, glycine, Phloxine and Rose Bengale were tested. In each experimental run, one of these additives was dissolved in 20 ml of 1.5% agar solution (pH 7.6) which contained 0.5% of casein. The casein-agar medium which contained no food additives, was used as the control. These casein-agar solutions were poured into petri-dishes and solidified. Then, diluted trypsinsolutions were added through stainless cup or paper disc to the casein-agar plates in petri-dishes. The dishes were incubated at 37°for 24 hr. After incubation, transparent circles were formed in the plates by the action of trypsin on casein. The outlines of the digested casein circles became distinct by adding 9% trichloroacetic acid. After removal of trichloroacetic acid, the diameters of the circles were measured. The plot of the mean diameters of circles versus logarithm of enzyme concentration became linear. Thus the ratio of the enzyme activity in the presence of each food additive to that of control was calculated. The ratios were very small when dulcin, Phloxine and Rose Bengale were added to the reaction medium. These results showed that these three additives were inhibitory for the tryptic digestion of casein. It was also shown that other five additives had no effect on the activity of trypsin.

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In the first paper of this series, a new detection method of marihuana constituents by the thinlayer chromatography with the solvent system of benzene-n-hexane-diethylamine (25 : 10 : 1, v/v) was described. In the present investigation it was studied whether any constituent of the plant allied with marihuana or of tobacco mixed usually in the actual samples interrupt the identification of marihuana or not. It was also investigated if this method could be applied to examination of the smoke of marihuana. As the results, any interruption by the plant extract described above was not recognized in the detection of marihuana. It was also found that cannabidiol, tetrahydrocannabinol and cannabinol were effectively detected in the smoke condensate of marihuana as well as in marihuana resin. Using this new detection method of marihuana, the authors succeeded in identifying the illegally used marihuana samples.

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The previous study on marihuana smoking indicated that the addicts would inhale about half amount of three main constituents, cannabidiol (CBD), tetrahydrocannabinol (THC) and cannabinol (CBN) in marihuana cigarettes, and suggested that some of the constituents might be excreted as unchanged or conjugated forms into the urine of these addicts. The present study was undertaken to confirm this possibility using rabbits. It was found in this experiment that free marihuana constituents in urine was extracted effectively with CHCl₃ during 10 hr under the pH 3. It was also shown that treatment of the urine with either 5% HCl or β-glucuronidase was necessary for detection of marihuana constituents in the urine before the extraction. Applying these procedures, CBN was detected in the urine of rabbits injected either CBN or ganja extract which contained a considerable amount of CBN. However, the excretion of any constituent could not be confirmed in the urine of either CBD or THC treated rabbits.

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Thiamine related compounds (five samples) and thiamine decomposed products (seven samples) could be identified by the paper electrophoresis with 5 N acetic acid as the electrolyte. From further investigation of electrolyte, it was found that borax (1%) was effective for identification between allithiamine and DBT, and that formic acid (5 N) was effective for identification beetween TDS and thiazole·pyrimidine compound, and also between thiochrome and thiazolone compound. Applying this analytical methods, it was observed that alkali degradation process of thiamine to thiazolone compound was influenced by various pH, and that hydrolysis was occured exclusively in the condition of heating aqueous thiamine solution at 130°.

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The present investigation was undertaken to apply theoretical methods to the ventilation of the polluted air in room where carbondioxide was not completely diffused. C_m+1, C_m, and C_m-1 are concentrations of carbondioxide at places (m+1) δX, mδX and (m-1) δX at time T (=nδT), respectively, C⁺_m is the concentration of cabondioxide at a place mδX at (n+1) δT, where superscript+denotes time (n+1) δT, subscripts m, (m+1) and (m-1) denote spaces mδX, (m+1) δX and (m-1) δX, respectively. Assuming that concentracion of carbondioxide depends on time and space, the following equation can be obtained, ∂C/∂T=D (∂²C/∂X²) where X is a distance from a source of the evolution of carbondioxide, and D is a constant (not equal to 0). Implicit difference analogue for the above equation can be obtained as follows, (∂²C/∂X²)_m=(C_m+1-2C_m+C_m-1)/(δX)² According to the Taylor's expansion theorem, the Schmidt's and Dusinberre's methods, following equations can be obtained, C⁺_m=(C_m+1+C_m-1)/2 C⁺_m=(C_m+1+C_m+C_m-1/3 respectively. In two dimensional diffusion, C_{j, k+1} and C_{j, k-1} are concentrations of carbondioxide at places x=jh, y=(k±1) h at time T (=nδT), and C_{j+1, k} and C_{j-1, k} are at places x=(j±1) h, y=kh, respectively, and C⁺_{j, k} is at a place x=jh, y=kh at time (n+1) δT. The following equation can be obtained as one dimensional diffusion, C⁺_{j, k}=(C_{j+1, k}+C_{j-1, k}+C_{j, k+1}+C_{j, k-1})/4 It was found experimentally that above equations were appropriate from the concentrations of carbondioxide in room.

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The method for the determination of micro (0.5-5 ppm of NH₃) and macro (500-5000 ppm of NH₃) amounts of ammonia in air were studied. For the purpose of micro analysis of ammonia in air, the indophenol method, in which sodium nitroprusside was added as a catalyzer of indophenol formation, was more excellent in the sensitivity, reproducibility and stability of color than the other methods for micro analysis. For the purpose of macro analysis of ammonia in air, the cupric carbonate method was relatively simple and reliable. Furthermore, 0.5% boric acid solution was adopted as an absorption medium of ammonia in air.

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A quick and convenient procedure for microdetermination of mercury contained in various materials was established. The method involved a combustion of the materials in a steel oxygen bomb. The samples were burned in the oxygen-filled bomb and the combustion products were absorbed in 1.0 N nitric acid. Mercury was extracted with dithizon solution. The mercury dithizonate thus formed was chromatographically separated using an aluminum oxide column, and was determined colorimetrically. The method is recommendable for quick analysis of voluminous samples having low mercury content. By this method, mercury recoveries from rice bran, which contained mercury at levels between 0.5 and 1.5 ppm, averaged 93%. Unpolished rice, vegetable oil, hair, brain and internal organ of lives as well as rice bran were analyzed.

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