Mutual solubility of the dibutylamine-water system between 10° and 60° was measured and this solubility curve was found to belong to the upper decreasing miscibility curves. Gelation discovered in the case of heptylamine, was not observed in amylamine or dibutylamine. Mutual solubility of the amylamine- and dibutylamine-water-sodium hydroxide system was measured. From these results, it was found that a linear relationship existed, within a certain concentration range, between the amine concentration (molar fraction) in the upper layer and that of sodium hydroxide (molar fraction) in the lower layer. Empirical formula of these straight lines was obtained and the slope of the straight line in the case of amylamine showed approximately the same value as those of butylamine and heptylamine.
Mutual solubility at 20° of the three systems of propylamine-water-sodium hydroxide, dipropylamine-water-sodium hydroxide, and dipropylamine-water-sodium chloride was measured and it was found that in these systems also, a linear relationship existed, within a certain concentration range, between the amine concentration (molar fraction) in the upper layer and the sodium hydroxide concentration (molar fraction) in the lower layer. Approximate straight line of these solubility curves was obtained and its slope was found to be about the same in these systems, and very similar to that of the aliphatic primary amines.
Total vapor pressure of the dipropylamine-water and butylamine-water systems was measured by a differential tensimeter and the partial pressure of each component of these systems was calculated from the values so obtained by the Van Laar equation. Mutual solubility of the amine-water-sodium hydroxide system indicated that the upper layer could experimentally be taken as a mixture of amine and water and lower layer, an aqueous solution of sodium hydroxide. Assuming that such two liquid phases exist in equilibrium, isopressure point of the partial vapor pressure of the amine water system and vapor pressure of the aqueous sodium hydroxide solution was obtained. The amine concentration of the amine layer at this isopressure point and sodium hydroxide concentration in the lower aqueous layer at this point were calculated and the values were found to agree well with the conjugation curve obtained from the measured values of butylamine- and dipropylamine-water-sodium hydroxide systems. These facts supply theoretical explanation of the approximate lineality of the conjugation curve of the mutual solubility of amine-water-sodium hydroxide system within a certain concentration range.
3-Acylhydrazino-6-chloropyridazines undergo cyclization in alkali hydroxide solution into s-triazolo [4, 3-b] pyridazines but not in neutral or sulfuric acid solution. 6-Chloro-s-triazolo [4, 3-b] pyridazine and 6-chlorotetrazolo [4, 5-b]-pyridazine formed their respective parent structure, s-triazolo [4, 3-b] pyridazine, m. p. 134°, and tetrazolo [4, 5-b] pyridazine, m. p. 104°, by catalytic reduction. These compounds reacted with 10-20% sodium hydroxide solution, alcoholic ammonia solution, and 80% hydrazine hydrate to form respectively 6-hydroxy-, 6-amino-, and 6-hydrazino-s-triazolo [4, 3-b] pyridazines.
Pyrogen is now said to be a complex polysaccharide. Some success was gained by an attempt to oxidize pyrogen in serum with periodic acid at room temperature. The consumption of periodic acid was found to increase with passage of time. There was some destruction of antibodies but there was no decrease after 10 minutes. Diphtheria and Weil sera, positive for pyrogen, were adjusted to pH 7.0, periodic acid was added to 0.02 molar concentration, and oxidized by allowing to stand for 10 minutes at room temperature (20°). Excess periodic acid was decomposed with glycerol, the solution was placed in a cellophane bag, and dialyzed against running water for 48 hours. The dialyzate was filtered through a Seitz filter and the filtrate was submitted to the measurement of antibody value and pyrogen test with a rabbit. It was found that pyrogen had been decomposed and that the decrease of the antibody value was around 20%.
Paper chromatography of steroids, possessing a δ-enol lactone ring which is a component of toad venom, and their derivatives, using a mixture of pyridine-water-acetone (4:1:10) for a stationary phase and heptane for a mobile phase, was carried out. Coloration with a chloroform solution of trichloride gave suitable Rf values.
Condensation-cyclization of 6-guanidinohexanoic acid (I) and chloromalonaldehyde in equimolar amounts in conc. sulfuric acid, by the application of chlorosulfonic acid under ice cooling, yielded 6-(5-chloro-2-pyrimidinyl) aminohexanoic acid (IV). Similar reaction with application of bromomalonaldehyde afforded the corresponding 5-bromo compound. Reaction of the hydrochloride of (I) and sodium nitromalonaldehyde in aqueous solution, with piperidine as a condensation agent, at room temperature yielded 6-(5-nitro-2-pyrimidinyl) aminohexanoic acid (VI), whose reduction with stannous chloride and hydrochloric acid gave the corresponding 5-amino compound (VII). Whereas (I) was resistant to esterification by the ordinary method, (VI) easily submitted to esterification. Fusion of a mixture of 2-methylthio-4, 6-dimethylpyrimidine (XI) and 6-aminohexanoic acid by heating to 200° in an oil bath afforded 6-(4, 6-dimethyl-2-pyrimidinyl) aminohexanoic acid. (XI) was prepared from acetylacetone and S-methylisothiourea according to the method of Hale and others. At the same time, some white needle crystals (X), m. p. 155.5-156°, were obtained as a by-product, whose analytical values corresponded to the formula of C7H10N4 and the substance was determined as 4, 6-dimethylpyrimidine-5-carbamidine by the synthesis of its derivatives.
By heating 6-(5-bromo-2-pyrimidinyl) aminohexanoic acid and thiourea in aqueous solution, it was derived to 6-(5-isothiouronyl-2-pyrimidinyl) aminohexanoic acid and application of sodium malonaldehyde to its hydrochloride in aqueous solution, with piperidine as the condensation agent, afforded 6-[5-(5-nitro-2-pyrimidinyl)thio-2-pyrimidinyl] aminohexanoic acid. This compound submitted to permanganate oxidation and the corresponding 5-(5-nitro-2-pyrimidinyl) sulfonyl compound was obtained. Its reduction with stannous chloride and conc. hydrochloric acid yielded the 5-amino compound. Diazotization of 6-(5-amino-2-pyrimidinyl) aminohexanoic acid by the usual method afforded 6-(5-hydroxy-2-pyrimidinyl) aminohexanoic acid. In a similar manner, 5-bromo and 5-chloro compounds were obtained, besides two kinds of crystals; one of white needles whose molecular formula agreed with C10H15O2N4Cl⋅HCl and the other of white rhomboprisms agreeing with C10H15O2N4Br⋅HBr. Both underwent condensation with anisaldehyde to form a hydrazone compound, so that they were determined as 6-[1-(5-chloro-2-pyrimidinyl) -hydrazino] hexanoic acid and its 5-bromo compound.
As one means of making biochemical and pharmacological studies on bacterial metabolites, effect of the metabolites on the activity of serum cholinesterase was examind in vitro. The coli bacilli group, putrefaction bacteria group, and intestinal pathogenic bacteria were used as the test organisms, cultured on bouillon, peptone, and synthetic media, and the effect of their culture filtrate on the activity of human serum cholinestrase was examined by the method of Hesterin. As a result, it was found that some of Escherichia coli, Alkalescens, Shigella; and Proteus had a strong inhibitive action. The bouillon medium showed the strongest inhibitive action while the synthetic medium showed no such inhibition. The inhibition increased with passage of time but reached the maximum after 48 hours and decreased by 72 hours later. These results indicated that a metabolic product of organic nitrogenous source effected inhibition of cholinesterase, that there is a difference in the degree of inhibition by each strain, and a few observations were gained on the periodic progress of such inhibition.
Escherichia coli was cultured at 37° for 48 hours in (1) peptone water added with 1% of one of 19 kinds of carbohydrate, (2) bouillon and (3) synthetic media each added with 1% of one of 16 kinds of amino acids. Turbidity of such culture media and the inhibition of serum cholinesterase activity by their filtrate were examined. It was thereby found that, in the case of (1), addition of salicin, dulcitol, sorbitol, inositol, and sucrose was found to effect inhibition, and in the case of (2), the inhibition was the strongest when tryptophan was added, followed by alanine, isoleucine, and phenylalanine in that order, while in (3), the inhibition was in the order of l-cystine, tryptophan, and isoleucine. Inhibition of cholinesterase activity by tryptophan was assumed to be due to tryptamine and indole formed by its decomposition by Escherichia coli but in the case of other substances, further detailed examinations are warranted.
Homogenate, extract, and proteins of a squid were used as a medium for the culture of various bacteria and the changes in pH, amount of volatile basic nitrogen, trimethylamine-nitrogen, and degree of cholinesterase inhibition of each culture filtrate with passage of time were measured. It was thereby found that both the pH and basic nitrogen increased with passage of time, while trimethylamine-nitrogen and cholinesterase inhibition reached the maximum after 48 hours and decreased by 72 hours later. There were no cholinesterase inhibition or the presence of trimethylamine-nitrogen when squid protein was used as the medium, while these were extremely high when the squid homogenate and extract were used as the culture medium. Further, there was a significant relativity between these. The inhibition of cholinesterase activity by the culture filtrate using squids must be due to the trimethylamine formed by the bacterial reduction of trimethylamine oxide contained in the squid. Liberation of histamine was also measured in some of these culture media but the amount was extremely minute.
Spectroscopic studies were made on the formation of a complex salt between bivalent copper and compounds of p-hydroxyphenylazobenzene, Phenylazo-p-cresol, phenylazoresorcinol, and o, o′-dihydroxyazo series. These salts showed a fairly distinct difference in the ability to form a complex salt under the conditions of 1/6×10-4M concentration, 50% ethanol as a solvent, and boiling on a water bath for 30 minutes. The formation of a complex became difficult in the order of o, o′-dihydroxyazo compounds, phenylazoresorcinol, and phenylazo p-cresol. Phenyl rings having methoxyl, vitro, and other substituents were also found to have some difference in the formation of complex salts. Some discussions are made on the ease or difficulty in the complex formation by the kind of substituents present in compounds of azophenol series.
Examinations were made on the complex formation of o, o′-dihydroxyazobenzene, o-hydroxyphenylazo-β-naphthol, and o-hydroxyphenylazoresorcinol with copper, cobalt, nickel, magnesium, zinc, cadmium, and manganese under the conditions described in the previous paper. These dyes formed complex salt extremely easily. Copper was the most reactive, followed by cobalt and nickel, while magnesium and zinc formed a complex only with o-hydroxyphenylazoresorcinol. Cadmium and manganese did not form such complex salts under these conditions. The composition of some of these complex salts was determined by the Jop method and that of the copper complex of some compounds of phenylazoresorcinol series was also determined.
Formation of complex salts in 0.5N sodium hydroxide solution was examined spectrometrically in about 10 kinds of azophenol series compounds. Of these dyes, o, o′-dihydroxyazobenzene and o-hydroxyphenylazoresorcinol form complex salts with copper, cobalt, and nickel but not with zinc, cadmium, or manganese. The concentration of the dye and inorganic ions was 1/6×10-4M. Some of the dyes underwent decomposition in alkaline solution and Cu2+ was found to accelerate such decomposition.
A kind of methylpyridocarbazole was obtained by the Conrad-Limpach reaction of 3-aminocarbazole or its 9-acetyl compound. In order to determine the direction of this cyclization, methylpyridocarbazole was prepared from 6-hydrazino-quinaldine and cyclohexanone by the Fischer's indole synthesis and dehydrogenation. By their mixed fusion, the two substances were found to be identical, since the cyclization of 6-hydrazinoquinoline by the Fischer reaction always occurs in the 5-position.
Hydrazine is a strong reducing agent but is comparatively rarely used for the reduction of nitro compounds. Reduction of nitrobenzene with hydrazine hydrate, in the presence of a metal, such as copper, iron, zinc, or aluminum, was carried out. It was found that zinc and aluminum do not show almost any catalytic activity while nitrobenzene was reduced in good yield to aniline in the presence of copper or iron powder. Application of this hydrazine reduction to o-, m-, and p-nitrotoluene, o-, m-, and p-nitrophenol, and m- nitrobenzoic acid showed these were all reduced to the corresponding amines. Polynitro compounds, such as dinitrobenzene, 2, 4-dinitrotoluene, 2, 4-dinitrophenol, and picric acid, were all partially reduced with hydrazine hydrate in ethanol solvent and respectively formed nitraniline, 2-nitro-4-aminotoluene, 4-nitro-2-aminophenol, and picramic acid.
2, 5-Dimethyl-4-methoxy-6-chloropyrimidine, b. p4 70°, was prepared from 2, 5-dimethyl-4, 6-dichloropyrimidine and was derived by hydrazine hydrate to 2, 5-dimethyl-4-methoxy-6-hydrazinopyrimidine, m. p. 142°. The reaction of the latter with R-COOH, ROCl, and pyridine, or with R-COOR' afforded monoacyl compounds whose heating with phosphoryl chloride or the treatment of the hydrazino compound directly with (RCO)2O and a minute amount of conc. sulfuric acid afforded 5, 8-dimethyl-7-methoxy-6-alkyl-s-triazolo [4, 3-c] pyrimidines, where R is hydrogen, methyl, ethyl, propyl, isobutyl, or phenyl group. Application of cold nitrous acid to the hydrazino compound yielded 5, 8-dimethyl-7-methoxy-1, 2, 3-tetrazolo-[4, 3-c] pyrimidine, m. p. 68-68.5°.
In order to find the effect of substituents in the 1-, 2-, and 3-positions of tetrahydroindole skeleton in adrenochrome monosemicarbazone on its hemostatic action, 1-methyl-5-semicarbazono-6-oxo-2, 3, 5, 6-tetrahydroindole (I), 1-methyl-2-ethoxycarbonyl-5-semicarbazono-6-oxo-2, 3, 5, 6-tetrahydroindole (III), and 1-β-hydroxyethyl-3-hydroxy-5-semicarbazono-6-oxo-2, 3, 5, 6-tetrahydroindole (VI) were prepared. However, all were found to be sparingly soluble in water and (VI) showed no increase of solubility in spite of the introduction of a CH2CH2OH group into adrenochrome monosemicarbazone.
3, 4-Dibenzyloxyaniline was methylated with dimethyl sulfate and sodium hydroxide to N-trimethyl-3, 4-dibenzyloxyanilinium hydrogen carbonate (II), whose pyrolysis afforded N-dimethyl-3, 4-dibenzyloxyaniline (III), and the debenzylation of (III) with subsequent oxidation finally gave o-benzoquinone type compounds (VI) and (VII). (VI) was extremely sparingly soluble in water that its hemostatic activity was not examined. On the other hand, methylation of 3, 4-dibenzyloxyaniline with formaldehyde and formic acid afforded needle crystals (XI) of m.p. 120-122°, which formed a monomethiodide and evolved formaldehyde on heating with acetic anhydride that (XI) was considered to be a kind of Tröger's base, discovered by Tröger. Its analytical values agreed well with those of the Tröger's base.
If the hemostatic action of adrenochrome and its monosemicarbazone were due to the o-quinonoid or o-quinonimide form, 6, 7-dioxo-1, 2, 3, 4, 6, 7-hexahydroquinoline type compounds should also show hemostatic activity. Under such assumption, 1-methyl- (XII) and 1-methyl-4-hydroxy-6-semicarbazono-7-oxo-1, 2, 3, 4, 6, 7-hexahydroquinoline (V) were prepared. However, both these compounds were found to be as sparingly soluble in water as other allied compounds of adrenochrome prepared to date, contrary to expectations, so that their pharmacological effect was not examined.
Reaction of 1 part of 2-bromo-4-nitrophenol (III) with 1.5 parts of glacial acetic acid, 2.5 parts of glycerol, and 3 parts of conc. sulfuric acid by heating afforded a mixture of 5-bromo-6-hydroxyquinoline (II) and 6-hydroxy-7-bromoquinoline (I) in approx. 3:1 ratio. The use of conc. phosphoric acid in place of glacial acetic acid afforded a small amount of (I) alone. The same reaction with 1 part of 2, 6-dibromo-4-nitrophenol (VI) afforded 5, 7-dibromo-6-hydroxyquinoline (VII) and the use of 80% formic acid in place of the glacial acetic acid in this reaction chiefly gave (VII), accompanied with a small amount of (I). The use of conc. phosphoric acid in place of glacial acetic acid afforded (I) alone in a fairly high yield. These experimental results indicated that the synthesis of 6-hydroxy-7-bromoquinoline (I) alone and in a good yield can be effected by the modified Skraup reaction of 2, 6-dibromo-4-nitrophenol with conc. phosphoric acid, glycerol, and conc, sulfuric acid.
In order to examine their pharmacological actions, 3-amino-, 3-methylaminomethyl-, and 3-dimethylaminomethyl-1-phenyl-2-methyl-5-pyrazolone and 4-dimethylaminomethylantipyrine were prepared. However, none of these compounds showed antipyretic or analgesic actions.
3-Formyl- and 3, 4-diformyl-1-phenyl-2-methyl-5-pyrazolone were synthesized. The 3-formyl compound was obtained by the McFadyen-Stevens method. from 1-phenyl-2-methyl-5-pyrazolone-3-carboxylic acid or by the Ball reaction of the 3-hydroxymethyl compound, the latter being the more preferred due to the easy availability of the starting material and good yield of the product. The 3, 4-diformyl compound was obtained by the Ball reaction of 3, 4-dihydroxymethyl-1-phenyl-2-methyl-5-pyrazolone as an unstable oil, which was identified as the crystalline diphenylhydrazone and a pyridazine derivative.
The growth inhibiting concentration of p-aminosalicylic acid (PAS), isonicotinic acid hydrazide (INAH), thioacetazone (TB-1), streptomycin (SM), and dihydrostreptomycin (DHSM) in a Kirchner medium containing 10% horse serum against human-type tubercle bacilli H37Rv strain and its resistant strains to INAH, TB-1, SM, and DHSM, and PAS, is sometimes affected by the addition of excess of Ca, Cu, Ni, Fe (III), Al, Hg, Mg, Zn, Mn, Cd, and Co (II). The effect is either synergism or antagonism. The difference between synergism, antagonism, and no-effect occurs by the difference in the combination of antitubercular agents, the kind of metal used, and sensitivity or resistance of the bacillus used. The combination effective to sensitive strains may not affect resistant strains or simply remain effective. Sometimes, the combination may affect only the resistant strains. Detailed examination of the synergism-antagonism caused by different concentrations of the metal indicated that higher the concentration of the metal, the stronger was synergism while antagonism was found within a narrow concentration range of a combination of SM or DHSM and aluminum. The molar ratio of the antitubercular agent and metal combination which showed synergism was 5-0.005:1 in sensitive strains and 518-5:1 in resistant strains. In a combination of SM and mercury, this ratio was 0.5-0.15:1 in sensitive strains and 518-168:1 in resistant strains. The minimum effective concentration of mercury in such cases was 3.4×10-6M and 1.1×10-6M. The present experimental results would not support the hypothesis that the direct action of a metal on bacillus or a metal complex salt is effective.
Formation of a complex salt was examined in 55 combinations of antitubercular agents, PAS, INAH, TB-1, SM, and DHSM with Ca, Co, Ni, Fe (III), Al, Hg, Mg, Zn, Cd, and Co (II). In order to find out the state in lower concentrations, effect of metal addition on the ultraviolet absorption curve of aqueous solutions of PAS, INAH, and TB-1 and that of addition of SM or DHSM on the ultraviolet absorption curve of aqueous solution of metals were observed. The breaking point in the high frequency titration curves obtained by the dropwise addition of an aqueous solution of antitubercular agent into metal solution was also examined. As for changes in the ultraviolet absorption curves, a shift of the maximum absorption was observed in a combination of PAS with Al or Hg, INAH and Hg, TB-1 with Cu and Hg, SM and Cu, and DHSM and Cu, a change in wave form in that of PAS and Fe (III), and a change in absorbancy at the maximum absorption in combinations of PAS with Cu, Al, and Hg, INAH with Cu, Ni, Hg, and Mn, TB-1 with Cu, Fe (III), Al, and Hg, SM with Co, Fe (III), Hg, Zn, and Mn, and DHSM with Ca and Cu. Appearance of a breaking point in high frequency titration curve was observed in combination of PAS with Co, Ni, Fe (III), Al, Hg, Mg, and Mn, INAH with Ca, Co, Ni, Fe (III), Hg, Zn, and Co (II), TB-1 with Ca, Co, Ni, Fe (III), Al, Hg, and Mn, SM with Al, Hg, and Mg, and DHSM with Al and Hg. The result from the second method agreed by 63.6%, complex formation was observed in eight combinations by the ultraviolet absorption and in 12 combinations by the high frequency titration alone.
It was found that the effect of Ca, Cu, Ni, Fe (III), Al, Hg, Mg, Zn, Mn, Cd, and Co (II) on the in vitro action of PAS, INAH, TBI, SM, and DHSM against sensitive and resistant strains of human-type tubercle bacilli and the presence or absence of tendency of these metallic ions to form a complex with these antitubercular agents were in parallel relation. This suggests the role of complex formation in the mechanism of these metals affecting the in vitro action of antitubercular agents. As was pointed out earlier, the effect of metals cannot be explained by the direct action of the metal on tubercle bacilli or the toxic action of the metal complex itself. In this connection, a hypothesis has been presented on the effect of metals on the in vitro action of these antitubercular agents. The antitubercular action of these agents seems to be displayed in connection with a requisite metal in an enzyme. When a metal forms a complex salt with a substance which competes with the antitubercular agent for this particular metal, a synergism between a metal and the agent occurs, but when the antitubercular agent and the said metal forms a complex salt, then antagonism occurs. Which complex is formed depends on the stability of the complex. This hypothesis can well explain the facts observed and reported in Parts I and II of this series.
In order to confirm the conclusions reached in the preceding paper, antibacterial activity of fatty acids and 4-acylaminosalicylic acids against tubercle bacilli was examined. It was thereby found that the relationship between such activity and number of carbon atoms in the acyl group existed as in the alkyl compounds and could be represented as four heaped curves, with C12 as the peak. Moreover, as was anticipated, 4-dodecanoylaminosalicyilc acid showed antibacterial activity equal to or stronger than that of its parental compound, PAS. The same relationship between antibacterial activity and the length of alkyl chain was observed to exist in the case of alkyl compounds against bacteria other than tubercle bacilli, and this phenomenon can also be explained by the same conclusion reached in connection with tubercle bacilli. However, permeability through the bacterial surface is limited by the nature of such surface layer and properties of the particular alkyl compound involved, so that the shape of the descending curve and position of the peak differs according to the kind of bacteria used when the compound is fixed, and according to the structure of the compound, if the bacterial strain is fixed.
A new synthetic method for estradiol 17-monoesters by heating estradiol with an acid is described. The acid used is limited but the corresponding estradiol 17-monoesters were obtained in a good yield by heating estradiol with acetic, propionic, butyric, isobutyric, valeric, and ethoxyacetic acids.
Esterification of steroidal alcohols by ester exchange reaction was examined. Hydrochloric, sulfuric, and p-toluenesulfonic acids were used as a catalyst and the last-named gave the best result, Testosterone afforded the corresponding esters by exchange reaction with methyl acetate, propionate, and valerate, Δ5-androstene-3β, 17β-diol formed a diacetate by reaction with methyl acetate, and estradiol formed the corresponding 17-monoesters by reaction with methyl acetate and propionate.
Inhibition of succinic oxidase of Escherichia coli K12-W strain by p-hydroxybenzoic acid and its methyl, ethyl, propyl, butyl, amyl, hexyl, and heptyl esters was examined with the Warburg manometer. It was thereby found that the free acid did not show any inhibition even in higher concentrations but the esters showed a strong inhibition with increase of concentration and the degree of inhibition increased with the increasing number of carbon atoms in the alkyl group comprising the ester. It was also found that the molecules of alkyl p-hydroxybenzoate not in dissociated state show marked respiratory inhibition and this action decreased with the increase of the dissociated forms.
For the purpose of stabilizing thiamine in various dry preparations and in order to find out the relationship between properties and stability of various thiamine salts, scores of thiamine salt preparations, with different solubility in water and acidity, were manufactured. The solubility of such solutions, preparations and stability of simple aqueous solution of each amine salt were examined and it was found that in a solution, its pH was the chief factor in the hydrolysis and oxidative decomposition of the thiamine salt. Acid salts were stable and neutral salts were generally unstable.
Homostephanoline, m. p. 233°, one of the tertiary phenolic bases of Stephania japonica MIERS, had been given a molecular formula of C32H44O7N2. Reëxamination of this point was carried out and the formula now proposed is C20H25O5N=C16H12O(OH)-(N-CH3) (O-CH3)3. It was confirmed by the present series of experiments that O-methylhomostephanoline, obtained by the methylation of homostephanoline with diazomethane, is identical with hasubanonine, one of the tertiary non-phenolic bases from the same plant.
Examination of the presence of water-soluble quaternary base in Sinomenium acutum REHD. ET WILS. (Japanese name, ohtsuzurafuji), which had never been studied, revealed the presence of magnoflorine, an aporphine-type base, in a large amount.
Of 2, 2′-, 3, 3′-, and 4, 4′-bipyridyl 1, 1′-dioxides, 4, 4′-compound is the most easily reduced with sulfur dioxide, followed by the 2, 2′-compounds, and the 3, 3′-compound is the most resistant to this reduction. As would naturally be expected, reduction potential is the lowest in the 4, 4′-compound and the highest in the 3, 3′-compound, though all possess low potential than that of pyridine 1-oxide.
Circular paper chromatography was applied to the determination of ephedrine in Ephedra. A definite amount (2.3×10-3cc.) of methanolic solution of pure ephedrine in various concentrations was spotted on the center of a circular filter paper, developed with butanol saturated with water, and colored with iodine gas. The relative graph of the area of ringed bands on the circular chromatogram and a logarithm of the amount (in γ) of the sample ephedrine was prepared and a quantitative relationship was found to exist between the two values (Fig. 1). The same chromatography of the methanolic extract of Ephedra was carried out and the percentage content of ephedrine was calculated from the colored ring bands, using the foregoing graph. The values thereby obtained was found to compare well with the percentage content obtained by the assay method given in the Japanese Pharmacopoeia (Table II).
The potassium content in the rhizome of Imperata cylindrica BEAUV. VAR. Koenigii DURAND ET SCHINZ. is almost equal to that in diuretic crude drugs and amount to 0.75% of the dried specimen. The dried rhizome also contains 18.8% of sugars in which sucrose and glucose constitute the main portion, and a small amount of fructose and xylose were also detected. Determination of the reducive sugars in the sugar mixture as glucose gave values corresponding to 36.5%, which corresponds to about 6.8% content in the dried rhizome.
Daily componental variation in the leaves of Datura Tatula L. was examined by its flower pot cultivation under uniform conditions, picking the young and mature leaves at 5.00 a.m., and 12.30 and 8.00 p.m., when the plant growth was at its height. There were no variation in alkaloidal content on the amount of total and protein nitrogen in one day, while the content of all these components were higher in young leaves than the old. In other words, variation in the content of the alkaloid and amount of various forms of nitrogen showed the same tendency. There was no great variation in the ratio of hyoscyamine to scopolamine. On the contrary, daily variation of the sugars was extremely great and this phenomenon was extremely marked in oligosaccharides, the amount being small in the morning and increasing towards the night. However, no difference in the amount of sugars was observed between young and mature leaves.