Indirect quantitative determination of magnesium was carried out by dissolving Mg NH4PO4⋅6H2O, precipitated from a magnesium salt solution, in acid and the phosphoric acid thereby formed was led to oxine phosphomolybdate which was again dissolved in an acid and the oxine in the solution submitted to bromatometry. A good result was obtained by this method. Another method was the preparation of magnesium oxinate which was allowed to dissolve in a definite excess of an acid and the excess of the acid was titrated with the standard alkali solution. The error in this method was found to be comparatively large.
Histamine and tyramine are assumed to be one of the substances responsible for sideeffects attending the intravenous administration of protein hydrolysate preparations. Detection of histamine and tyramine and approximate determination of their contens in various protein hydrolysates and marketed parenteral preparations were carried out by paper chromatography in reference to the method of Urbach. The results obtained are shown in Figs. 1 and 2, and in Table I. These results indicated that histamine and tyramine were present in casein hydrolysate prepared with toluene as the antiseptic and the marketed parenteral preparations in larger amounts than was expected. It was also found that there were a difference in the amount of histamine formed by the manner of hydrolysis and that in protein hydrolysate preparations obtained by alkaline hydrolysis, care must be taken to remove these harmful amines.
In order to determine the effect of the difference in the method and extent of hydrolysis of proteins on the biological activity of protein hydrolysate preparations, Lactobacillus arabinosus 17-5 was cultured in basal media (Table II) containing various casein hydrolysates (Table I) as the source of amino acid, and multiplication of the bacillus against the same amount of total nitrogen of various hydrolysates was comparatively determined. As the results shown in Figs. 1 and 2 indicate, the multiplication on the hydrolysate by alkali was inferior compared to those on enzymatic and acid hydrolysates, even with the addition of chief amino acids which were assumed to disappear or decrease by secondary destruction during hydrolysis. These results were thought to indicate the effect of racemization by alkaline hydrolysis.
As one of the attempts on the simplified examination of the biological value of parenteral preparations of amino acid mixtures by the multiplication of Lactobacillus, Lactobacillus arabinosus 17-5 was cultured in a medium containing various marketed preparations as the source of amino acid and the multiplication against a definite amount of total nitrogen of each preparation was comparatively determined. As the results shown in Fig. 1 indicate, the multiplication was inferior in the hydrolyzate (C) of bovine blood by acid and alkali than the enzymatic hydrolyzate (A) and (D) of casein. There is, naturally, a difference between the nutrition demand in man and Lactobacillus but by making further examinations on corrections for such a difference, this Lactobacillus culture test may be used instead of animal test to estimate by simple means the biological value of complex amino acid preparations.
Relationship between the variation in the diuretic principle and the presence or absence of cinobufotoxin in Senso, a Chinese drug of toad secretion, was examined. It was confirmed that the disappearance of cinobufotoxin was not due to decomposition by Penicillium molds but to other reasons and that cinobufotoxin was a crystalline component that had no connection with the diuretic action.
Tomato was grafted onto the root of datura and N15H4NO3 with 3.46 atomic% excess of N15 was given as a fertilizer. Forty days after giving the fertilizer, over-ground portion and root were harvested to determine the amount of N15 in the alkaloids and proteins. It was found by this tracer method that the alkaloidal nitrogen in the root was 0.38% of the total nitrogen absorbed, and that in the overground portion, 0.08% The content of N15 in the protein-nitrogen was found to be the reverse, being higher in the over-ground portion than in the root. The content ratio of hyoscyamine and scopolamine in the total alkaloid from the tomatografted datura was found to be in reverse proportion in the root and leaves as the plant grew, which seemed to indicate that the change from scopolamine to hyoscyamine was occurring in the tomato leaf. When datura was grafted onto the root of tomato, a slight increase in the alkaloid content in the scion was detected as the plant grew. These results suggest that the alkaloid in datura is chiefly formed in the root but its formation in the over-ground portion cannot entirely be denied.
1) By the application of mono-, di-, and trichloroacetyl- and O, N-diacetyl-p-aminosalicylyl chlorides on 2-allylmercapto-6-aminobenzothiazole, corresponding 6-acylamino-benzothiazoles were obtained. One of the latter compounds, 6-p-acetamino-o-acetoxy-benzamino derivative was saponified to the 6-p-amino derivative. Dimethyl- and diethylamine were respectively applied to 6-monochloroacetamino derivative and 6-dimethyl-and 6-diethylaminoacetaminobenzothiazoles were obtained. 2) By the application of sodium and ethyl monochloroacetate to 2-mercapto-6-nitrobenzothiazole, 6-nitrobenzothiazolyl-(2)-thioglycollic acid and its ethyl ester were prepared. Condensation of ammonia, dimethylamine, or hydrazine hydrate to this 6-nitro ethyl ester respectively yielded the amide, dimethyl amide, and hydrazide. Reduction of the foregoing 6-nitro ester to the 6-amino ester and derivation to the 6-acetamino ester, with subsequent application of hydrazine hydrate to the 6-amino and 6-acetamino esters yielded the respective hydrazides.
Alkaloids were found to be more easily and rapidly separable by paper electromigration than by paper chromatography. The distance of migration to the cathode is always constant, whether in the free state or in a salt form. The alkaloids can, therefore, be identified directly from the extracts of crude drugs by using paper electromigration. Good results were also obtained by the combined use of chromatography and electromigration.
Acylation with phenyl benzoates possessing a substituent in the aromatic ring was examined. In the case of aromatic amines, substituent in either the benzoic acid nucleus or the phenyl nucleus, showed a striking influence while in aromatic sulfonamides, only the influence of the substituent in the phenyl nucleus was apparent. From these findings, the mechanism of acylation with a phenyl ester was examined.
Combined effect of Oxine with iron, zinc, magnesium, or copper sulfate was tested with Tricophyton and Achorion in the range of pH 5-9. Addition of iron and zinc salts was found to cancel the effect, that of magnesium, to decrease the effect, while the addition of copper was found to be about equal or in some instances increase the effect. The course of such a behavior was assumed to be due to the fact that iron and zinc bond completely with Oxine in pH 5-9 range thereby nullifying its effect while such an effect was assumed to remain in the case of magnesium due to incomplete salt formation. However, copper is known to undergo complete bonding in this pH range and this is somewhat inconsistent. The copper salt of Oxine was found to substitute with iron from which it may be assumed that the copper salt acts in the same way as Oxine itself. It was also found that the copper salts of isonictonic acid hydrazide, p-aminosalicylic acid, and hinokitiol, which are effective as the free compounds, also undergo substitution with iron. These experimental results support the theory of Zentmeyer regarding Oxine and confirmed that the inconsistent results of Sexton was due to the use of a specific material such as the copper salt.
Mercuration of azoxybenzene with mercuric acetate followed by treatment with sodium chloride yielded 2-chloromercuri-, m.p. 188°, and 4-chloromercuri-azoxybenzene, m.p. 230°. The former is more soluble in benzene than the latter. Mercuration of azoxybenzene with mercuric perchlorate yielded the same products.
Mercuration of pyridine 1-oxide with mercuric acetate, followed by treatment with sodium chloride yielded 4-chloromercuripyridine 1-oxide, m.p. 259-260°, which was confirmed by deriving it to 4-methoxypyridine 1-oxide. Mercuration of quinoline 1-oxide with mercuric acetate or mercuric perchlorate, followed by treatment with sodium chloride, yielded 8-chloromercuriquinoline 1-oxide, m.p. 235°, which was confirmed by deriving it to 8-bromoquinoline through its 1-oxide, m.p. 102-104°. 8-Bromoquinoline 1-oxide could not previously been obtained by any other method.
In order to study the oxide ring of desoxynupharidine, dihydrodesoxynupharidine was reduced by hydriodic acid and red phosphorus from which two optical isomers (V) and (VI) were obtained, whose reduction with palladium-carbon gave an identical tertiary amine (VIII) or (XIV). The latter was designated N-methyl-apotetrahyd-rodesoxynupharidine.
Karrer's method of C-methyl determination was applied to hydroxyapodihydrodesoxynupharidine, one of the products obtained by the cleavage of the oxide ring in desoxynupharidine. From the results obtained it was assumed that the oxygen atom in the oxide ring of the desoxynupharidine molecule is bonded to one of the carbon atom in another ring.
Paper chromatography was carried out on natural coumarin derivatives, umbelliferone, hernianin, daphnetin, citropten, esculetin, fraxetin, umbelliferone glycoside, daphnine, aesculin, and fraxin, using the simple descending method, with water-saturated isoamyl acetate, isoamyl alcohol, phenol, chloroform, and 4:1:2-mixture of isoamyl alcohol, acetic acid, and water, as the developing agents, to determine their Rf values. Applying the present method, components of the tree bark of Aesculus turbinata Bl. were examined and it was pointed out that esculin or esculetin could be present besides fraxin or fraxetin.
In the previous paper, it was reported that 3-(1, 2, 3, 4-tetrahydro-7-naphthyloxy)-1, 2-propanediol was a better compound than Myanesin from the point of its action in relaxing the muscles and in inhibiting convulsant poisons, toxicity, and action time. Thirteen kinds (including 9 new compounds) of allied compounds containining bromine or nitrogen, and the glycerol ethers were prepared and their pharmacological action examined. Of these compounds, 3-(o-cyclohexylphenoxy)-1, 2-propanediol and 3-(o-β-bromoethoxyphenoxy)-1, 2-propanediol were found to relax the muscles, the former showing stronger action than that of Myanesin, but these two compounds were found to possess a small safety margin. Examination of the head-drop dose of 2-hydroxy-3-(1, 2, 3, 4-tetrahydro-7-naphthyloxy)-propyltrimethylammonium iodide and β-(o-2, 3-dihydroxy-propoxyphenoxy)-ethyltrimethylammonium bromide with Waser's method showed that although these were extremely effective, they caused death by respiratory paralysis at the effective dose.
Cultivation tests were carried out (at Kasukabe, in 1950) on summer buckwheat (Fagopyrum esculentum M. var. aestivum Makino), autumn buckwheat (F. esculentum M. var. autumnale Makino), diploid and tetraploid buckwheat, and Tartarian buckwheat (F. tataricum Gaertner), for the purpose of examining their rutin content. Chief studies were made on the relationship between sowing time and yield of crop and rutincontent and following results were obtained: 1) Both the summer and autumn buckwheat were found possible to cultivate twice a year, in the spring and autumn. 2) Both the summer and autumn buckwheat cultivated in the spring were found to have larger content of rutin and greater yield of rutin per 10 ares (Tables I and II). 3) Cultivation of autumn buckwheat in the spring was found to give a larger rutin content and greater yield of rutin per 10 ares (max. 3.49% and 1.896kg., resp.) than the cultivation of summer buckwheat in the spring (max. 3.13% and 1.291kg., resp.) (Tables I and II). 4) No apparent difference was observed between the diploid and tetraploid buckwheat as to the yield of crop, rutin-content, and yield of rutin per 10 ares (Table III), although, naturally, tetraploid organs (cotyledons, first foliage leaves, pollen grains, and stomata) were larger than those of the diploid (Table IV). 5) Rutin-content and yield of rutin per 10 ares of the Tartarian buckwheat were not necessarily greater than those of summer and autumn buckwheat (Table V).
Extracts of several kinds of tonic crude drugs were added to synthetic medium not containing nicotinic acid and the growth of Lactobacillus arabinosus was examined. Growth was found in all the media and the content of total nicotinic acid calculated from the degree of growth was 10-135γ/g., with an average of 38γ/g. In order to see whether this growth was actually due to nicotinic acid or not, examination was made by paper chromatography using Nuphar japonicum as the sample and a growth of microörganism was found in the portion giving the same Rf value as that of the standard nicotinic acid. By the same method, presence of vitamin B12 was proved in Ligusticum acutilobum the fact of which is significant as this crude drug is used for anemia.
Application of four or two moles of sodium alkoxide to 2, 6-dichloroisonicotinic acid gives either 2, 6-dialkoxy- or 2-alkoxy-6-chloroisonicotinic acid. Catalytic reduction of the latter in methanol, in the presence of alkali and with palladium hydroxide as a catalyst, yields 2-alkoxyisonicotinic acid. Esterification of these acids by the usual method and derivation to respective hydrazide gave 2, 6-dimethoxy-, -diethoxy-, -dipropoxy-, -diisopropoxy-, -dibutoxy-, -diisoamyloxy-, and -diphenoxy-isonicotinic acid hydrazide, 6-chloro-2-methoxy-, -ethoxy-, -propoxy-, -isopropoxy-, -butoxy-, -isoamyloxy-, and -phenoxyisonicotinic acid hydrazide, and 2-methoxy-, and -ethoxy-isonicotinic acid hydrazide. Their antitubercular actions will be reported in due course.
In the synthesis of phenolic glycosides, the yield of the α- and β-types of tetraacetylphenyl-D-glucoside by the Helferich method, with zinc chloride as a catalyst, is very poor. In order to increase the yield of the α-compound, over ten kinds of compounds were used in place of zinc chloride and it was found that zinc bromide gave a far better yield of the α-compound. In the screening of these catalysts, it was found that the mixing ratio of the α- and β-types in the crude tetraacetylphenyl-D-glucoside obtained by the condensation could be calculated by the use of a graph showing the specific rotation and mixing ratio of various mixtures of pure tetraacetyl-α, β-phenyl-D-glucoside.
Antibacterial tests, in vitro, against human type tubercle bacilli, Staphylococcus aureus, and Escherichia coli were carried out with 9 kinds of benzylidene compounds and seven kinds of 2-phenyl-1, 3, 4-thiadiazole derivatives prepared by the condensation of 2-amino-1, 3, 4-thiadiazole derivatives and aromatic aldehydes. 2-Amino-5-(p-nitrobenzylideneamino)-, 2-amino-5-cinnamylideneamino-, and 2-o-hydroxyphenyl-1, 3, 4-thiadiazoles showed approximately the same efficacy as that of Tibione. None of the compounds tested seemed to be effective against staphylococci and coli bacilli.
One of the three structures proposed for neocyanine, the Brooker-type dye, was prepared in order to comparatively examine various properties of this dye and those of neocyanine. Corresponding amount of 2-phenylaminoethenylquinolinium ethiodide was applied to 1, 1′-diethyl-2, 2′-trimethylene-10-anilinoquinocyanine dichloride trihydrochloride in acetic anhydride, in the presence of anhydrous potassium acetate and a Brooker-type dye with quinaldine nucleus was prepared. Another Booker-type dye with lepidine nucleus was prepared by the similar manner from 1, 1′-diethyl-10-anilino-4, 4′-trimethyl-enequinocyanine diiodide dihydrochloride and 4-phenylaminoethenylquinolinium ethiodide. Brooker-type dye with benzothiazole nucleus was obtained similarly from 1, 1′-diethyl-2, 2′-trimethylene-10-anilinothiocyanine dichloride trihydrochloride and 2-phenylaminoethe-nylbenzothiazolium ethiodide. These dyes showed identical absorption maxima and absorption curves with the corresponding binuclear trimethine dyes but the analytical values of these dyes were found to agree with those of the Brooker-type, and the absorption maxima were totally different from those of neocyanine dyes.
Kendall prepared [2-(3-ethylbenzoxazole)] [2′-(3-ethylbenzothiazole)] [γ-2″-(3-ethylbenzoxazole)]-pentamethinecyanine diiodide in order to experimentally prove that the structure of neocyanine corresponded to the formula proposed by König. In order to prepare unsymmetric Brooker-type dye corresponding to the above, 1, 1′-diethyl-meso-anilino-2, 2′-trimethylenethiaoxacyanine dichloride dihydriodide and 2-phenylaminoethe-nylbenzoxazolium ethiodide were reacted in acetic anhydride, in the presence of anhydrous potassium acetate, and the objective dye was obtained. Similarly, the objective dye was obtained from 1, 1′-diethyl-mesoanilino-2, 2′-trimethylenethiocyanine dichloride trihydrochloride and 2-phenylaminoethenylbenzoxazolium ethiodide. It was thereby found that substitution of nuclei occurred during the preparation of unsymmetric Brooker-type dyes. The objective dyes here obtained showed absorption maximum at 5, 220-5, 240 Å which was different from that (λmax=6, 100 Å) of the neocyanine obtained by Kendall.
Titration of the alkaloids in the Japanese Pharmacopoeia and some weak bases were carried out with high frequency titration apparatus. Good results were obtained by the use of a mixed solvent of alcohol and water for substances which tend to give a curve by the strength of dissociation. In general, substances with dissociation constants over 10-9 can be determined well. The amount of samples needed is usually around 30 mg., and the error, approximately 0.5-1%.
When santonin is dissolved in dehydrated methanol, 10% sodium methoxide solution applied to it at a room temperature, and diluted with dehydrated methanol after a definite interval, a comparatively stable coloration is obtained and this can be utilized for colorimetric determination. The minimum amount determinable is less than 1mg. and the experimental error is not more than 3% in majority of cases.
Santonin was derived to 2, 4-dinitrophenylhydrazone and isolated by chromatographic adsorption on alumina. Colorimetric determination was carried out on the coloration of the eluates thereby obtained. Range of determinable amount was 10-100γ and the experimental error was around ±3% in the majority of cases.
1) 2-Amino-5-isopropylthiazole, obtained by the reaction of α-chloroisovaleraldehyde and thiourea, was diazotized and substituted with chlorine to prepare 2-chloro-5-isopropylthiazole. 2) Four kinds of 1-phenyl-1-pyrimidyl-3-dimethylaminopropane derivatives and thirteen kinds of 1-phenyl-1-thiazolyl-3-diethylaminopropane derivatives were prepared. All of these compounds were found to possess antihistaminic and antiacetylcholinergic activities but 1-phenyl-1-(5-methyl-2-thiazolyl)-3-dimethylaminopropane and its p-chloroderivative displayed especially powerful activities.
1) Sulfuric acid decomposition of β-phenyl-β-(2-pyridyl)-β-cyanopropionaldehyde diethyl acetal, obtained by the condensation of phenylacetonitrile, 2-bromopyridine, and bromoacetal, yielded β-phenyl-β-(2-pyridyl)-propionaldehyde. p-Chloro derivative was also obtained in a similar manner. 2) The aldoxime obtained by the application of hydroxylamine and β-phenyl-β-(2-pyridyl)-propionaldehyde was reduced and methylated to give 1-phenyl-1-(2-pyridyl)-3-diethylaminopropane. 3) Reductive amination and Leuckart reaction of β-phenyl-β-(2-pyridyl)-propional-dehyde yielded 1-phenyl-1-(2-pyridyl)-3-dimethylaminopropane. 1-(p-Chlorophenyl)-1-(2-pyridyl)-3-aminopropanes, e.g. its N-dimethyl, N-diethyl, N-methyl, and N-ethyl derivatives, were prepared in a similar manner. 4) It was observed that during the formation of p-chlorophenyl-2-pyridylmethane by the sulfuric acid decomposition of p-chlorophenyl-2-pyridylacetonitrile, p-chlorophenyl 2-pyridyl ketone is formed as a by-product.
1) Comparative examination of seasonal variation in santonin content and yield of crop was carried out between July 2, 1952, and December 26, 1952, with the second year growth of Artemisia kurramensis Qaz. sown respectively in March and October, 1951, at Kasukabe, in order to establish cultural methods of autumn sowing. The results (Table I) showed that the highest yield of crop per 10 ares on airdry basis and the santonin content in the spring-sown plot were 169.3kg. (harvested on July 15) and 1.993% (harvested on July 2), respectively, and those in the autumn-sown plot were 174.9kg. (harvested on August 15) and 2.052% (harvested on August 1), respectively. Although there is one month's difference between the two, the yield and santonin content are similar showing that the cultivation by autumn sowing is possible. 2) In order to observe the influence of acclimatization, comparative examination of seasonal variation in santonin content and yield of crop was made between September 5, 1952, and December 26, 1952, with the first year growth of plants sown in March, 1952, at Kasukabe, grown from the seeds collected from the plants cultivated at Kasu-kabe during 1951 and the fresh seeds of Pakistan origin. The results (Table III) showed that the highest yield of crop per 10 ares on airdry basis and the santonin content in the acclimatized plot were 132.0kg. (harvested on September 16) and 1.754% (harvested on September 5), respectively, and those in the original Pakistani seed plot were 74.6kg. (harvested on October 30) and 1.253% (harvested on September 5), respectively, indicating the former to be better. The flowering in the acclimatized plot was about one month earlier than that in the original Pakistan seed plot, and seeds were found to have formed only in the acclimatized plot, where there was less damage from decay after harvest than the original seed plot. 3) The period at which the second year growth showed the highest yield of crop and santonin content was at the maximum of vegetative growth, irrespective of the appearance of flower-heads. 4) The size of plants and santonin content were not necessarily parallel in the second and third year growths, observed individually and by harvested periods (Tables II and IV). Some plants of the second and third year growths harvested in July and August showed a high content of santonin such as 2.701% and 2.895%. It is realized that a propagation system by clonal separation must be established.
The degree of the hydrolysis of m- and p-nitraniline N-glucosides under the usual conditions of periodic oxidation is below 30% and the volume of formic acid produced by such oxidation from the N-glucosides, which consume exactly five moles of the oxidant, is the same as that from glucose itself. Based on such result, following mechanism is suggested for the periodic acid oxidation of the N-glucosides: RNHCH(CHOH)nCH⋅CH2OH IO4′→RNH2+(n+2)HCOOH+HCHO RNHCH(CHOH)nCH2 IO4′→RNH2+(n+1)HCOOH+HCHO
It was found that the addition of copper powder in an amount 1/10 to 1/6 of that of zinc dust used in the Reformatsky reaction was found to give 10-30% increased yield compared to the absence of copper powder.
Glucogenkwanin, C22H22O11⋅2H2O, faint yellow needles, m.p. 273-274°, was isolated from the bark of Prunus verecunda (Koidz.) Koehne and two kinds of P. Jamasakura variety (Japanese hill cherry). From the bark of P. Lannesiana variety (double-petalled species), sakuranin was obtained. When hydrolyzed with 5% H2SO4, glucogenkwanin gave genkwanin and glucose. By the methylation of glucogenkwanin with dimethyl sulfate and 30% KOH solution and following hydrolysis of the methylated product with 5% H2SO4, apigenin-7, 4′-dimetyl ether, m.p. 174-174.5°, was obtained. Glucogenkwanin, therefore, is genkwanin-5-glucoside.