The reaction of N-acetyl-p-toluidine, chloral hydrate, and conc. sulfuric acid for 20 hours at 70° yielded (5-acetamido-2-methylphenyl) glycolic acid (I) as colorless needles (from water), m.p. 229° (decomp.), as the chief product. (I) forms an acetate as colorless plates (from acetone), melting once at 115-125°, solidifying at 130-140°, and finally melting at 192°; ethyl ester as colorless prisms (from acetone), m.p. 153-154°. The corresponding amino-carboxylic acid, colorless plates (from 50% ethanol), m.p. 216° (decomp.), on diazotization, followed by treatment with hypophosphorous acid, and oxidation with permanganate yielded o-toluic acid. Cold oxidation of (I) with potassium permanganate yielded (5-acetamido-2-methylphenyl) glyoxalic acid as pale yellowish green prisms (from 50% EtOH), m.p. 198° (decomp.), while the same oxidation in acetic acid at 60° afforded 5-acetamido-2-methylbenzoic acid as colorless prisms (from 50% EtOH), m.p. 230° (reported m.p. 177°). The acid was confirmed by its synthesis from o-toluic acid, through 5-acetamido-2-methylbenzonitrile, m.p. 166°, and -benzamide, m.p. 247°, and the error of the reported m.p. was confirmed.
By the acylation of benzoylacetic acid m-aminoanilide hydrochloride with aryl or heterocylic sulfonyl chlorides, nine kinds of m-sulfonamido derivatives were prepared, including m-phenylsulfonamido, m-(4-methoxyphenyl) sulfonamido, m-(3-nitrophenylsulfonamido), and m-(8-quinolyl) sulfonamido derivatives.
The N-oxides of pyridine, quinoline, and their homologs are deoxygenated on heating with sulfur and liquid ammonia. When a nitro group is present, it is reduced to the primary amine by this reaction. Aliphatic N-oxides, such as dimethylaniline N-oxide, are also deoxygenated by this reaction to form the tertiary amines.
Ethyl 6-hydroxy-4-decaenoate, obtained by the Grignard condensation of ethyl 5-formyl-4-pentenoate and butyl iodide, was converted to the methyl ester, dehydrated to methyl 4, 6-decadienoate, and reacted with isobutylamine to 4, 6-decadienoic acid isobutylamide as colorless needles, m.p. 53°. This amide is a solid and only gives a mild but persistent paralytic action on the tongue, without any acrid taste, while the natural spilanthol is a liquid of b. p0·015 138°, possessing an acrid taste and a persistent paralytic action. From these facts, it is assumed that the two are geometric isomers.
1) Diacyldiaminoacetones, RCONHCH2COCH2NHCOR (R=C6H5-, C6H5CH2-, C6H5CH2CH-, C6H5CH: CH-, CH3-, -OC2H5) undergo condensation with benzaldehyde in ethanol-water mixture, in the presence of sodium hydroxide, to form 2, 6-diphenyl-3, 5-diacylaminotetrahydropyrones, though the yield never goes above 38%. 2) The concentration of ethanol affects the yield of the condensate and the condensation occurs with alcohols which are miscible with water, such as glycerol, propyleneglycol, dioxane, and triethanolamine. The condensate is formed in a very small amount even in water alone that these alcohols are found to act as a solvent in this condensation. 3) The concentration of sodium hydroxide has not as much effect as that of alcohol and other alkalis, such as potassium carbonate, sodium amide, and tertiary amines, or comparatively stronger alkalis such as triethylamine and N-ethylpiperidine, also work as alkaline condensation agent, although the yield of the product is smaller compared to the use of sodium hydroxide.
2-(5-Nitro) furanacrylanilide and 24 of its allied compounds were prepared and the effect of the kind and position of the substituent in the benzene ring on antibacterial action was examined with Staphylococcus aureus P 209, Streptococcus hemolyticus, and Shigella dysenteriae Komagome B III. It was thereby found that the presence of a hydroxyl in the benzene ring effected the strongest action and that the action was stronger in para- and meta-substituted compounds. Compounds with two or more substituents generally showed lower antibacterial action but an aminothymol derivative showed the strongest action against staphylococci and streptococci.
Eight kinds of naphthylamides, two kinds of naphthyl esters, and two kinds of phenyl esters, of 2-(5-nitro) furanacrylic acid derivatives were prepared and their antibacterial action were compared using Staphylococcus aureus, Streptococcus hemolyticus, and Shigella dysenteriae Komagome B III. 2-(5-Nitro) furanacryl-naphthylamide-(2), -4-chloronaphthylamide-(1), and -4-bromonaphthylamide-(1) showed powerful antibatcerial action against Strept. hemolyticus, but none of the substances examined were effective against Shigella dysenteriae.
Aconite spp. were collected at Nenokuchi and Namariyama, by the Lake Towada in the northeastern regions of Japan. The plants collected in these locations were hard to distinguish from external appearances but some differences were observed in their respective alkaloids. The plants collected at Nenokuchi (sample designated as Ochiai No. 11) yielded mesaconitine and ignavine from the bases precipitated by ammonia and the presence of jesaconitine-like base was detected in the ultraviolet spectrum. The plant from Namariyama (Ochiai No. 12) yielded mesaconitine and Shiriya-base I, and the presence of jesaconitine-like base was detected as in the foregoing. A small amount of a crystalline base, m.p. 316-318° (decomp.), was obtained from the water-soluble portion. The aconites collected in Shiriyazaki and Iwaya, the northernmost tip of the Aomori Prefecture, were entirely the same both from external appearances and alkaloidal components. The plant from Iwaya (Ochiai No. 13) and that from Shiriyazaki (Ochiai No. 14) were found to contain aconitine, mesaconitine, and Shiriya-base I. Shiriya-base I recrystallizes from acetone in hexagonal plates, m.p. 158-160°, [α]D14: +43.5° (CHCl3), gives a nitrate of m.p. 214-215° (decomp.); the formula of C23H37-39O6N or C24H39O6N, with three methoxyl groups, is assumed for this base. The nitrate of mesaconitine was newly obtained as prisms (from methanol-ether), m.p. 177-180° (decomp.).
The alkaloidal components of the rhizome of aconites collected at Shimoburo and Sakuma in Kazamura Village, Shimokita Peninsula in northeastern Honshu (Aomori Prefecture), were isolated and purified. Until the specie names can be determined, the plants are designated as Specimen Ochiai No. 15 and No. 16. Ochiai No. 15 yielded 0.33% of crude bases (against the fresh rhizome) from which Shimoburo-base I, Shimoburo-base II, isohypognavine, and a crystalline base, m.p. 264-266°, were isolated. This component also contained a large amount of amorphous base. Ochiai No. 16 yielded 1.3% of crude base (against the dried rhizome) from which Shimoburo-base I, Shimoburo-base II, crystalline base of m.p. 267-267.5°, and small amounts of crystalline bases of m.p. 170°, m.p. 290° (decomp.), and m.p. 302° (decomp.), were isolated. This plant failed to yield isohypognavine. The tentatively named Shimoburo-base I comes as prisms melting at 198-202°, [α]D24: -141.2° (MeOH), and can easily be isolated as a sparingly soluble hydrochloride of m.p. 278-279° (decomp.). It also gives a hydrobromide of m.p. 275° (decomp.), picrate of m.p. 225-229° (decomp.), and methiodide of m.p. 258-260° (decomp.), and its analytical values correspond to the formula C21H29O3N or C22H31O3N, its nitrogen is tertiary, it possesses one N-CH3 or N-C2H5, but does not have a methoxyl or ester group. It forms a semicarbazone of m.p. 245-246° (decomp.). The tentatively named Shimoburo-base II comes as crystals of m.p. 208-213°, [α]19D: 25.52° (CHCl3), and formes a perchlorate of m.p. 223-226° (decomp.). Its composition agrees with C23H35O7N or C24H37O7N, possessing three methoxyls and one N-CH3 or N-C2H5. It is resistant to hydrolysis and the presence of a carbonyl can be assumed from its absorption maximum at 295mμ (logε=1.63). The base named isohypognavine was isolated as a hydrochloride of m.p. 195-198° (decomp.), [α]D22: +91.07 (MeOH). The free base is hard to crystallize but comes as prisms of m.p. 90-92°, which changes to m.p. 135° on drying. It forms a hydrobromide of m.p. 235-236° (decomp.), and a methiodide of m.p. 253-256° (decomp.), and its analytical values correspond to the formula C27H31O5N, possessing a benzoyl group, but not methoxyl or N-methyl group. The crystals of m.p. 267-267.5° from Ochiai No. 16 and m.p. 264-266° from Ochiai No. 15 are the same substances, giving [α]D12.4: +104.4° (MeOH), gives a picrate of m.p. 282-284°, methiodide of m.p. 286-287° (efferv.), and a perchlorate of m.p 213-214°. Its analytical values correspond to C20H27O2N, and its properties resemble those of kobusine but no confirmatory evidences have been obtained as yet.
2-Substituted and 2, 3-disubstituted arylthio-1, 4-naphthoquinones listed in Table I were prepared and their antibacterial action was examined (cf. Table II). The antibacterial mechanism of the naphthoquinones has been explained as their bonding with SH compounds, necessary for bacterial growth, but some of their derivatives containing a substituted SH group also showed bacterial action. It was therefore pointed out that the bacterial growth may possibly be inhibited by the oxidative mechanism in oxidoreduction potential of the quinoid structure which changes with the substituent present, as a more important factor of the antibacterial action of the naphthoquinones.
Amino derivatives of 1, 4-naphthoquinones shown in Table I were prepared and their antibacterial tests were carried out (cf. Table II). Amino derivatives showed antibacterial action against gram-positive bacteria, clearly indicating the effect of a substituent. Introduction of a halogen atom seems to give favorable effect on the antibacterial property. Aliphatic amines show antibacterial action against the gram-negative bacteria. Some of the 2-amino-3-thio-1, 4-naphthoquinones show considerable antibacterial action (Nos. 29 and 30), and it was pointed out that the antibacterial activity of naphthoquinones was due in part to some factors other than that caused by the bonding of such compounds with the -SH group, necessary for bacterial growth.
Sulfur in organic compounds was determined by the following procedures. The sample was heated with fuming nitric acid in a sealed tube, the cooled content was transferred to an evaporating dish, evaporated on a water bath to remove excess nitric acid, and barium chromate added to the acid solution to precipitate barium sulfate. Neutralization followed by filtration resulted in the presence of chromic acid ion corresponding to the sulfate ion and the filtrate was titrated with 0.05N ferrous sulfate, using barium diphenylaminosulfonate as the indicator. The endpoint of this titration was somewhat more clear than by other existing method. The semimicro- and micro-analyses gave good results, the maximum error being 0.30% and 0.29%, respectively.
Application of hydrogen peroxide to 2-mercapto-4-methyl-5-hydroxyethylthiazole and 2-mercapto-4-methyl-5-acetoxyethylthiazole, in the presence of hydrochloric acid, yields 4-methyl-5-hydroxyethylthiazole, while the use of conc. hydrochloric acid gives 2-chloro-4-methyl-5-hydroxyethylthiazole as a by-product. In case the amount of hydrogen peroxide is not enough, bis (4-methyl-5-hydroxyethylthiazolyl-2) sulfide is formed besides the foregoing. The last-named was confirmed by its synthesis from 2-chloro-4-methyl-5-hydroxyethylthiazole and 2-mercapto-4-methyl-5-hydroxyethylthiazole.
There are several compounds having powerful analgesic action in the series of 1-methyltetrahydropyridines possessing alicyclic radical in 2- or 4-position. These were obtained by the dehydration of hydroxypiperidines, which were prepared by the reaction with Grignard or lithium compounds of corresponding alicyclic compounds on 1-methyl-4-piperidones. The dehydration of 1-methyl-2-hydroxy-2-alicyclic piperidine was also tried but was not successful so far as the conditions employed were concerned.
Effect of various putrefaction amines on bacterial histaminase was examined by the Warburg manometric method, using Achromobacter sp. It was found that cadaverine and agmatine effected over 20% cross inhibition, while tyramine retarded adaptation and effected 46% inhibition (cf. Figs. 1-10 and Table I).
Catalytic reduction of α-isonitroso-β-hydroxypropiophenone, prepared from β-bromopropionyl chloride in four steps, was carried out under various conditions. Reduction in hydrochloric acid solution with palladium-carbon catalyst, or in dehydrated ethanol with platinum dioxide catalyst, afforded d, l-norephedrine derivative, formed by the reduction of the primary alcohol (-CH2OH) group. In this case, a small amount of the pseudo isomer formed as a by-product and the mixture was derived to the pure pseudo derivative by N→O1 acyl migration. Reduction in glacial acetic acid with palladium-carbon catalyst afforded d, l-1-phenyl-2-amino-1, 3-propanediol derivative which is a mixture of threo and erythro diasteroisomers. N→O1 Acyl migration of this mixture yielded the threo-type derivative which gave d, l-threo-1-phenyl-2-benzoyl-amino-1, 3-propanediol as the final product. It has been observed that the reduction in glacial acetic acid, if carried out at a higher temperature over a long period, resulted in the formation of benzoic acid as a by-product.
1) By the nuclear methylation of resacetophenone, 3-methyl-2, 4-dihydroxyacetophenone was obtained from which it has become clear that 3-methyl-2-hydroxy-4-methoxyacetophenone, obtained to date by the same reaction, had been formed by the etherification of the dihydroxy derivative. 2) In the nuclear methylation of resacetophenone, the chief product is paeonol when the molar ratio of resacetophenone, potassium hydroxide, and methyl iodide is equimolar or thereabouts, but the yield of 3-methyl-2-hydroxy-4-methoxyacetophenone increases as the molar ratio of potassium hyroxide and methyl iodide to resacetophenone increases, becoming the maximum at 2.5 moles of potassium hydroxide and 4 moles of methyl iodide to 1 mole of resacetophenone. There is no necessity for the great excess of methyl iodide. 3) In order to examine the reactivity of the 3-position in nuclear alkylation of resacetophenone, Legal and Ehrlich reactions of resacetophenone and its allied compounds were carried out.
The vapor-phase reaction of N-alkyldiethanolamine and ammonia, in the presence of various dehydration catalysts resulted in the formation of the corresponding N-methyl-, -ethyl-, -propyl-, and-butyl-piperazines in 2-20% yield. Catalytic vapor-phase reaction of N-alkyldiethanolamine and aliphatic and aromatic amines gave the corresponding N, N′-dimethyl-, N-methyl-N′-phenyl-, and N-ethyl-N′-phenyl-piperazines in 7-32% yield. Examination of various reaction conditions revealed that silica-alumina coprecipitate was the best as the catalyst and that the optimal reaction temperature was 325° for ammonia and aliphatic amines, and 450° for aromatic amines.
Reaction of equimolar amounts of 2-methyl-4-amino-5-aminomethylpyrimidine (IV), γ-aceto-γ-mercaptopropyl alcohol (V), and formaldehyde, in dil. ethanol, results in the formation of dihydrothiamine (II), m.p. 150°, in a good yield, and (II) is identical with the sample obtained by reduction of thiamine (I) with lithium aluminum hydride. When the foregoing reaction is carried out in the presence of sodium hydroxide or on the application of sodium hydroxide on (II), isodihydrothiamine, m.p. 160°, is formed. Solvation of dihydro- or isodihydro-thiamine in water with application of heat converts them into pseudodihydrothiamine. Both iso- and pseudo-dihydrothiamines possess the same composition and molecular weight as those of dihydrothiamine, show the same Rf values, and give identical reactions. Application of (V) to the condensate (VI) from equimolar reaction of (IV) and formaldehyde also yields (II) or its isomer. Thiamine is formed from (II) by the application of ferric chloride, sodium nitroprusside, or potassium ferricyanide. Warming the solution of (II) in hydrochloric acid effects its complete decomposition and formation of (VI). The same treatment psudo- or iso-dihydrothiamine gives the same results, forming thiamine or (VI).
Decomposition of procaine penicillin in buffer solutions was examined. From the fact that this decomposition is a primary reaction, a specific functional relationship was found to exist between the concentration of the buffer solution and the velocity constant, and beween pH of each buffer and the velocity constant, using phosphoric, citric, and acetic acids for the buffer solution.
The ketone group in C3-positon of tetrahydrosantonin (II), m.p. 153-156°, was protected as the ethyleneglycol-ketal, the lactone was ruduced by LiAlH4 to the diol-3-cyloketal (IV), m.p. 149-151°, and further hydrolyzed with an acid to 3-ketodiol (VIII), m.p. 117-118°. The hydroxyl in C5 of the diol is not benzoylated in the case of 3-cycloketal (IV) while it is benzoylated in the case of the 3-ketone (VIII).
The lactone in the desoxytetrahydrosantonin (II) was reduced with lithium aluminum hydride and the diol (III), m.p. 152-153°, was obtained, the hydroxyl in C5-position of this diol not undergoing benzoylation. Similar reduction of tetrahydroalantolactone (X) yielded the diol (XI), m.p. 113-115°, whose hydroxyl in C5-position submitted to benzoylation. The properties of the hydroxyl group in various lactones was examined and compared with the experimental results given in the 4th paper of this series.
Dehydrogenation of dimethyl kainate in p-cymene, with palladium-carbon affords a dicarboxylic ester of 3-methyl-4-isopropylpyrrole whose saponification yields the dicarboxylic acid (III), m.p. 170° (decomp.) C10H13O4N. Heating of (III) with water gives a monocarboxylic acid (V), m.p. 119°, C9H13O2N, while decarboxylation of (III) and (V), followed by distillation yields 3-methyl-4-isopropylpyrrole. One of the two carboxyls in (III) is liberated easily. (V) is sublimable. From these evidences the structure of 2-carboxy-3-carboxymethyl-4-isopropylpyrrole or 3-carboxymethyl-4-isopropyl-5-carboxypyrrole was proposed for (III). From the foregoing results, from the facts that one of the pk′ of kainic acid is 2.1, and that kainic acid very easily forms an anhydride, the structural formula (A) was assigned to kainic acid, and (A'), i.e. 2-carboxy-3-carboxymethyl-4-isopropylpyrrolidine, for dihydrokainic acid.
Ozonolysis of dimethyl kainate in hydrochloric acid solution yielded formaldehyde as a volatile substance and a methylcarbonyl compound (I) of m.p. 181° (decomp.), C10H15O5N, pk' 2.1, 8.75, as a non-volatile substance. (I) Colors yellow with ninhydrin and its dry distillation yielded a substance giving a pyrrole reaction. From its pk' values, this substance was assumd to be a monoester, with a free α-carboxyl. The same portion also yielded the diester of (I), b.p3 128-132°, C11H17O5N. Kainic acid and its derivatives show intense absorption at 6.05 and 11.2μ, which disappear in the corresponding dihydro compounds. The foregoing results suggest that the double bond in kainic acid is present as an isopropenyl -CCH3CH2 group. From the fact that kainic acid easily forms are acid anhydride and that the infrared absorption of its N-acetylated anhydride is at 5.51 and at 5.66μ, kainic acid is assumed to form a six-membered anhydride. The proposed structure of kainic acid, from the experimental data obtained to date, is not the 2-methyl-3-isopropyl-4, 5-dicarboxypyrrolidine (B) proposed before, but 2-carboxy-3-carboxymethyl-4-isopropenylpyrrolidine (C).
1) The root of Coix Lachryma-Jobi L. was extracted and palmitic and stearic acids were isolated from the acid portion. 2) A new compound, coixol, m, p. 151-152°, C8H7O3N, was isolated from a portion soluble in alkali hydroxide but insoluble in petroleum ether. 3) The sterol fraction obtained from the neutral portion yielded stigmasterol and β- and γ-sitosterols. α-Sitosterol fraction was not examined in detail due to the small amount available. 4) From the water-soluble portion, after removal of the above, crystals of potassium chloride was isolated and the presence of glucose was detected.
Coixol, C8H7O3N, was isolated as colorless needles, m.p. 151-152°, from the portion soluble in alkali hydroxides, of the root of Coix Lachryma-Jobi L. var. frumentaced Makino. Decomposition of coixol with conc. hydrochloric acid, in the presence of stannous chloride, yielded 5-methoxy-2-aminophenol hydrochloride, m.p. 214°. Therefore, coixol was assumed to be 6-methoxybenzoxazolone and was confirmed by synthesis. Following coixol derivatives were prepared: N-Acetylcoixol, colorless needles, m.p. 147.5°. N-Benzoylcoixol, colorless needles, m.p. 162-162.5°. N-Ethoxycarbonylcoixol, colorless plates, m.p. 107°.
In the isoquinoline cyclization of N-acylethylamine, the cyclization is difficult when the substituents in the α- and β- positions are identical but progresses easily if a hydroxyl or a methoxyl is introduced into the β-position. As the most simple of this type, β-hydroxy and β-methoxy derivatives of N-acetyl-α, β-diphenylamine were taken up and their cyclization reactions were examined under various reaction conditions. It was thereby found that, when phosphoryl chloride was used, they were derived through oxazoline to ψ-ephedrine-type amide, the reverse of the original ephedrine-type amide, and N-acetyl-β-chloro-α, β-diphenylethylamine was obtained as a by-product. The use of a strong cyclization agent, such as phosphoryl chloride and phosphorus pentachloride, or phosphoryl chloride and polyphosphoric acid, finally yielded 1-methyl-3-phenylisoquinoline. The isoquinline compound was also obtained on submitting the ψ-ephedrine-type amide to drastic cyclization. It is therefore concluded that drastic isoquinoline cyclization, under the conditions which would inhibit the formation of a chloro compound, will yield the isoquinoline compound via the oxazoline compound. It is assumed also, that the facility of the formation of the chloro compound constitutes one of the factors for the ease or difficulty of the cyclization.
Isoquinoline cyclization of (p-nitrobenzoyl)-β-alanyl β-veratrylethylamide, by the ordinary process, yields two substances of different decomposition points; yellowish substance of m.p. 242° (decomp.), and yellow substance of m.p. 193° (decomp.). Catalytic reduction of the substance of a higher melting point in the presence of platinum catalyst, yielded by absorption of 5 moles of hydrogen, 2′-(p-aminophenyl)-1′, 2′, 3′, 4′, -5′, 6′-hexahydropyrimidino-1′ 6′: 1, 2-(6, 7-dimethoxy-3, 4-dihydroisoquinoline).
The Grignard reagents prepared from methyl, ethyl, propyl, isopropyl, allyl, butyl, phenyl, benzyl, and cyclohexyl halides were reacted respectively with 2-dimethylamino-methylcyclopentanone and the corresponding 1-alkyl (allyl, phenyl, benzyl, or cyclohexyl)-2-dimethylaminomethylcyclopentanols were prepared. Application of benzoyl, p-nitrobenzoyl, and cinnamoyl chloride to the 1-ethyl, -propyl, -phenyl, -benzyl, and -cyclohexyl derivatlves yielded the corresponding esters. 1-Ethyl-1-p-nitrobenzoyloxy-2-dimethylaminomethylcyclopentanol was reduced to the amino compound and condensed with ethyl chlorocarbonate to 1-ethyl-1-p-ethoxycarbonylamino-benzoyloxy derivative.
6-Acetylated compound (VII), obtained by the reaction of 5-hydroxy-7, 4′-dimethoxyflavone (VI) and glacial acetic acid or acetic anhydride, in the presence of polyphosphoric acid was heated with morpholine and sulfur, and saponified by 100% phosphoric acid to 6-carboxylmethyl compound (XI). Its methylation and saponification yielded 6-carboxymethyl-5, 7, 4′-trimethoxyflavone (XIII). Condensation of (XIII) with anisole, in the presence of polyphosphoric acid yielded the 6-(p-methoxyphenacyl) compound (XV) which underwent demethylation by aluminum chloride to 5-demethylated compound (XIV). These compounds showed obvious depression of the melting points in admixture with the corresponding methylated compounds of the flavones obtained by the decomposition of ginkgetin. These facts indicate that further studies are required for the elucidation of the structure of ginkgetin.
The magnesium salt present in the aqueous extract of Digenea simplex suppresses oxygen uptake of the homogenized muscle of Ascaris suilla but such action is not observed with kainic acid. Anthelmintic action of the magnesium salts of lactic acid and kainic acid, the components of Digenea, was examined in vitro from which it was found that these salts possessed strong effect on the nonoxygen respiration of the nerve muscles of Allolobophora eoetida, earthworm, nerve muscle preparation of the upper tail of hog ascaris, and homogenized muscles of hog ascaris. Oral administration of these magnesium salts in a puppy showed that magnesium kainate was extremely effective in a small amount and that magnesium lactate was effective, though the efficacy was somewhat weaker. The anthelmintic mechanism of kainic acid is assuumed to be the excitement and paralysis of the muscle movement and inhibition of respiration in ascaris. Suppression of tissue respiration by kainic acid was found to be due to the decomposition of iron-containing respiration enzyme contained in the epithelial cells of the digestive tract of hog ascaris, as a result of histological examination of the hog ascaris immersed in a solution of kainic acid and of chemical examination of enzymes in the tissues of digestive tract of the ascaris.
Bromatometry of Oxine, the Berg's method whereby the metal which combines quantitatively with Oxine is indirectly determined, is rapid and reliable and is widely used, but it has not been adapted for micro-determination. The iodometric titration in the final stage of the Berg method was changed to azotometry, the whole procedure was scaled down, and 17 kinds of metals, including calcium, magnesium, zinc, aluminum, lead, iron, cobalt, and nickel were found to be determinable in micro-quantities. The determinable range is 10-200μg./cc., and the error is within ±1%. Cr+++, if derived to the bivalent form by reduction with hydroxylamine, forms an Oxinate of definite composition (Cr(C9H6ON)2) and can be quantitatively determined.
In the Friedel-Crafts reaction of six kinds of 4-substituted diphenyl ethers and acetyl chloride, their relative rate constants were measured with toluene as the standard substance. The values obtained were as follows: CH3O 1166, CH3 687, Cl 121, H 90.5, CH3CO 35, NO2 1.3.
2, 2′-Bipyridyl shows strong resistance to electrophilic reagents, similar to pyridines, and is hard to undergo substitution reactions. It is comparatively reactive to nucleophilic reagents but the yield is poor in amination by sodium amide. 2, 2′-Bipyridyl forms N, N′-dioxide and the N-oxide bond is severed by phosphorus chloride, thionyl chloride, and hydriodic acid. It is difficult to effect transition of the N-oxide to the pyridone-type compound by acetic anhydride but the N-oxide compound is nitrated comparatively easily, forming 4, 4′-dinitro compound.
The nitro groups in the dinitro-2, 2′-bipyridyl N, N′-dioxide (I) are extremely reactive and undergo various substitution reactions. For example, heating (I) with conc. hydrochloric acid at 100° for 4 hours yields a dichloro derivative, with 30% sulfuric acid at 100° for 4 hours a dihydroxy derivative, and reaction with benzyl alcohol, by standing with the alcohol in the presence of sodium for two days at room temperature, yields dibenzyloxy derivative, all in a good yield. Warming (I) with phosphorus trichloride for 3 hours effects deoxygenation of the N-oxide groups and halogen substitution of the nitro group, forming dichlorobipyridyl. Oxidation of this substance with neutral permanganate yields 4-chloropicolinic acid, showing that the nitro groups in (I) are in 4- and 4′-positions. The nitro groups in (I) can be used as the nitrous acid-supplying agent. For example, aminoazobenzene is obtained as crystals from aniline or aniline hydrochloride, containing a small amount of potassium iodide and water. Catalytic reduction of (I) is somewhat difficult, reduction with palladium-carbon yielding 4, 4′-diaminobipyridyl N-oxide and further reduction requires a long time. Catalytic reduction of (I) in conc. hydrochloric acid solution results in the substitution of one of the nitro groups with chlorine and the other nitro group is reduced to the hydrazo group, the reaction becoming extremely slack thereafter. The hydrazo compound is rapidly oxidized to the azo compound in air but its catalytic reduction in weakly acidic medium, with a fresh catalyst, affords the amino compound.
Ursolic acid, m.p. 280°, oleanolic acid, m.p. 297-300°, and acetyloleanolic acid, m.p. 262-263°, were isolated in pure state from the pericarp (including pulp) of Ligustrum japonicum Thunb. (Oleaceae).
In order to prepare intestinal and digestive preparations of living bacteria from the so-called spore-bearing lactic acid bacteria, different from the ordinary Lactobacillus in that they form heat-resistant spores, bacteriological and biological properties of six strains of this bacteria were examined. One of the strains, P 43, was found to be suitable for the present purpose. P 43 is a strain of spore-bearing Lactobacillus which grows in aerobic or in facultative anerobic conditions at 37° and the living cells themselves are also extremely resistant to heat. It has no hemolytic or toxic properties, does not generate indole or hydrogen sulfide, and does not require special vitamin B1 and B2 for growth. It coagulates milk and forms a large amount of lactic acid from glucose by homofermentation, just as in ordinary Lactobacillus, and a large number of sugars are fermented by this bacillus.
The formation of spores in the so-called spore-bearing Lactobacillus is observed only under specific conditions of cultivation. Conditions for the spore-formation in P 43 strain were examined and a process for abundant spore formation was found to be a shaking culture in a specific bouillon medium, practicable for industrial method. The spore-bearing cells of this P 43 were used to paepare powders in which the presence of living cells was found to be extremely good and its use for making marketable product seemed possible.
A strain, P 43, of the so-called spore-bearing Lactobacillus was examined in vitro and it was found to be strongly resistant to synthetic gastric and enteric juice. It was able to co-exist with the Lactobacillus commonly present in human intestines but its antibacterial action against aneurinase, coli, proteus, pyogenes, and subtilis bacilli was not very marked. In vitro test of this bacillus, by oral administration in man and rats, revealed somewhat different reaction. Enteric fixation of this bacillus was found to require spore-bearing cells and administration of such cells in thiaminase patients was found to have effected complete elimination of thiaminase bacillus shortly after administration and marked decrease of intestinal fermentation.
In the determination of isonicotinic acid hydrazide in hydrochloric acid solution with 0.1N potassium bromide and potassium bromate, the concentration of the acid at the time of reaction is the optimal at 0.70-0.86N of hydrochloric acid. The oxidation reaction can be quantiatively accelerated by shaking vigorously, especially in the initial stage of the reaction. The most suitable reaction period is 15-30 minutes and the evaporation of the free bromine can be prevented and addition of potassium iodide facilitated by cooling with ice. Prevention of the liberation of iodine from unreacted potassium iodide can be effected by lowering the acidity to around 0.38N hydrochloric acid. The foregoing precautions afford stabilized analytical values.
Growth inhibition on human type tubercle bacilli, Aoyama B strain, was tested with 17 kinds of isonicotinic acid hydrazone series, 12 kinds of furfural derivatives, 6 kinds of thymol derivatives, 4 kinds of p-hydroxybenzaldehyde derivatives, 5 kinds of 3-chloro-2, 4-dihydroxy-5-alkylbenzaldehyde derivatives, and 6 other compounds. Growth of the tubercle bacilli was inhibited by the isonicotinylhydrazone of p-thymotinaldehyde in 5, 120, 000 dilution, of p-methoxybenzaldehyde in 2, 560, 000 dilution, and of 2, 4-dihydroxy-5-isoamylbenzaldehyde, p-nitro-o-methoxybenzaldehyde, p-hydroxy-m-methoxybenzaldehyde, p-dimethylaminobenzaldehyde, p-sulfanilamidobenzaldehyde, and 2-hydroxy-3-carboxybenzaldehyde in 1, 280, 000 dilution, and by furfural isonicotinylhydrazone in 1, 280, 000 to 2, 560, 000 dilution. Isonicotinic acid hydrazide, used as a control was able to inhibit bacillary growth in 12, 800, 000 to 25, 600, 000 dilution, sodium p-aminosalicylate in 160, 000-320, 000 dilution, and Tibione in 40, 000 dilution.
Examination of the alkaloids in Berberis amurensis Rupr. var. japonica (Regel) Rehd. forma Bretschneideri (Rehd.) Ohwi was carried out. As tertiary bases, a large amount of berbamine, a small amount of hydroxyberberine, and a new base of m.p. 190-191°, were isolated, and berberine, palmatine, and jatrorrhizine were obtained as the quaternary bases. Berberis spp. generally contain oxyacanthine, together with berbamine, but the present plant did not contain oxyacanthine.
Saponification of 2-nitro(amino)-6-alkoxybenzonitriles by the method of Radzisgewsky yielded the corresponding acid amides. However, 2, 6-dimethoxybenzonitrile did not submit to saponification by this method and warming 2-nitro-6-methoxybenzonitrile with conc. sulfuric acid at 60-65° effected its decomposition, forming m-nitro-phenol.
The blood staining with the Giemsa solution markedly varies by the pH of the liquid used for dilution. The use of a buffer in place of distilled water makes it possible to obtain a clear stain by eosine and azure staining. It is proposed that the quality of the Giemsa stain solution be judged by the state of staining of normal blood corpuscles at pH 5.8.
Reconstituted milk in natural milk can be detected, up to 5%, by the following method. Ten cc. of reconstituted milk is diluted with the same volume of water, adjusted to pH 4.5-4.6 with acetic acid, and the precipitated curd is washed several times by centrifuging. Two cc. of acetone is mixed with the washed curd, 2cc. of 10% potassium hydroxide solution is added, and the mixture is warmed at 50°. On standing for a few minutes, the fat will float as a white mass and the lower alkali layer becomes a clear, yellow liquid. This alkali solution is submitted to colorimetry under irradiation of ultraviolet rays in a dark room.
Penicillium islandicum Sopp (D-strain) was cultivated in a Czapek-Dox (solution) containing 5g. glucose, at 32-33° for 14 days. The culture filtrate was mixed with activated carbon and the substance which was adsorbed on the activated carbon was dissolved out with acetone. The extract obtaind by evaporation of acetone showed a toxicity (M. L. D.) of 10mg./10g. mouse. From the ether-soluble portion of this extract, malonic acid (LD50 1.4mg/10g. mouse) was obtained as colorless plates, m.p. 135-136°, and a substance which easily sublimates at 90° at 1mm. Hg, as colorless needles, m, p. 121-127°, was obtained. The latter did not show any toxicty and was assumed to have been formed secondarily from malonic acid and acetone. Extraction of the ether-insoluble portion with ethanol yielded d-mannitol as colorless leaflets, m.p. 165-166°.
Infrared absorption spectra of 18 kinds of diphenyl ether derivatives, and xanthone and diphenylene dioxide were examined from which the following facts were revealed. 1) The ether bonding between two benzene rings shows a very intense absorption at 8.0-8.25μ, and there are absorptions of medium intensity at about 8.6 and 7.7μ. 2) In 4-acetyldiphenyl ether derivatives, the carbonyl band of the acetyl group is approximately constant and appears at around 6.03μ, irrespective of the kind of other substituents.