Manner of the formation of molecular compounds between p-aminomethylbenzene-sulfonamide (Homosulfamine) and sulfa drugs, and between sulfa drugs was examined by thermal analysis. The formation of molecular compounds was detected between Homosulfamine and sulfacetamide, sulfathiazole, phthalylsulfathiazole, sulfadiazine, Irgafen, and sulfisoxazole, and between diaminodiphenyl sulfone and sulfanilylthiourea. The molecular compounds are obtained as crystals from suitable solvents and their solubilities were found to be much greater than those of original sulfa drugs.
1) Nitration of isoquinoline N-oxide gives 5-nitro and 8-nitro compounds from which it was seen that the effect of N-oxide group does not extend to the benzene ring. 2) Catalytic reduction of 5-nitroisoquinoline N-oxide with palladium-carbon in acid medium gives 5-aminoisoquinoline and 5-amino-Py-tetrahydroisoquinoline while the same treatment in neutral medium gives 5-aminoisoquinoline. In the latter case, stopping the reduction after absorption of 3 moles of hydrogen gives 5-aminoisoquinoline and 5-aminoisoquinoline N-oxide from which it has been clarified that the N-oxide group in isoquinoline N-oxide possesses comparatively weak resistance against reduction but it still possesses the characteristics of the tertiary amine N-oxides. 3) Bucherer reaction of 5-aminoisoquinoline gives 5-hydroxyisoquinoline in a good yield.
Coumarin derivatives were prepared with a monosubstituent of hydroxyl, methoxyl, methyl, or acetoxyl group in one of the ortho (5), meta (6), or para (7) position of α, β-unsaturated carbonyl group and disubstituents of two hydroxyl, methoxyl, or acetoxyl groups in 5, 7-, 6, 7-, or 7, 8-position. The ultraviolet absorption spectra were determined in 95% ethanol solution. Methyl and acetoxyl derivatives showed spectra similar to that of coumarin, irrespective of the position or number of the substituents. Hydroxyl and methoxyl derivatives, on the other hand, showed extremely complicated spectra according to the position and number of substituents present. Assuming that the two absorption bands of coumarin at 270mμ and 312mμ were those of benzene around 200mμ and 240-260mμ that had shifted to these regions, the ultraviolet absorption spectra of these coumarin derivatives were qualitatively explained according to the latest theory regarding light absorption.
It was found that colorimetric determination of α-amino acids was possible by reacting them with ninhydrin in pyridine, in the pressence of ascorbic acid. The experimental error was less than 5%, and the minimum amount of α-amino acids possible to be determined was ca. 0.5 γ converting into amino-nitrogen. The hydrindantin produced from ninhydrin and ascorbic acid, pyridine, and an excess of ninhydrin all had an effect of preventing the decomposition of the pigment, diketohydrindylidenediketohydrindamine.
It is well-known that the isoquinoline cyclization of acyl β-methoxy (or -hydroxy)-β-phenethylamide results in the formation of isoquinoline compounds whereas the same reaction on acyl β-phenethylamide gives 3, 4-dihydroisoquinoline compound. There has been no report to date regarding the degree of ease or difficulty of this reaction. In order to find some basic principle of this reaction, isoquinoline cyclization was carried out on α-veratryl-γ-phenyl- and α-phenyl-γ-veratryl-α-hydroxy-β-benzamidopropane in an oridinary manner by which 1-phenyl-3-benzyl-6, 7-dimethoxyisoquinoline was obtained from the former and 1-phenyl-3-benzal-6, 7-dimethoxy-3, 4-dihydroisoquinoline from the latter. It was therefore concluded that no transition of the double bond occurred in the two compounds.
In order to examine whether rearrangement of the double bond occurred in isoquinoline derivatives, α-veratryl-γ-piperonyl-and α-piperonyl-γ-veratryl-β-benzamidopropane were prepared and isoquinoline cyclization with phosphoryl chloride carried out on them. The latter substance yielded two kinds of products one of which was found to be identical with the cyclization product of the former, there being no depression of the melting point on admixture and showing identical infrared absorption spectra. The other substance obtained from the latter was crystals of m.p. 180-183° which showed notable difference in the absorption range of double bonds and in a longer wave length range. It follows, therefore, that an identical substance had been obtained by the isoquinoline cyclization of the foregoing two kinds of acid amides as a consequence of the migration of the double bond. However, the cyclizable two aromatic rings in the acid amides both possess alkoxyl groups that the formation of two kinds of compounds can be expected and no definite confirmations can be made as to their structures. Further studies are being made on this point.
Reaction of various aldoximes on benzoyl- or acetyl-eugenol by the utilization of the new isoquinoline synthesis (Kmetani method) gave the objective 1-substituted 3-methyl-6-methoxy-7-benzoyloxy (or 7-acetoxy)-3, 4-dihydroisoquinoline but the yield was inferior than that obtained by the use of safrole or methyleugenol.
Soy sauce, without any addition of antiseptics, usually showed growth of molds after 3.5-4 days. The addition of propyl p-hydroxybenzoate, a compound used as the control, was found to inhibit this mold-growth to some extent, 1mg. of the compound in 110cc. of the sauce, i.e. in 0.001% concentration, the mold did not grow until 11th day, with 3mg., until 37th day, and with 5mg. until 46th day. Some compounds found to prevent the growth of mold in a 61-day test period with 1mg., i.e. in 0.01% concentration, were p-chlorothymol, p-tert-amylphenol, 2-hydroxynaphthaldehyde-(1), 3, 7-dihydroxy-1, 9-dimethyldibenzofuran, thiodiphenylamine, 2-methyl-1, 4-naphthoquinone, and 2-ethyl-1, 4-naphthoquinone.
Effects of various essential oil components such as borneol, cinnamic alcohol and aldehyde, citral, citronellal, citronellol, geraniol, limonen, linalool, menthol, and terpineol, against pathogenic fungi and various bacteria were tested. In general, they were effective against fungi, fairly good against tubercle bacilli, but ineffective against Escherichia coli, Staphylococcus aureus, and Bacillus dysenteriae. As to the structural relationship, aldehydes were generally more powerful than corresponding alcohols, especially in α, β-unsaturated compounds. Terpenes were the same either in chain or cyclic forms. Hydrocarbons had the weakest action, and the most powerful was cinnamic aldehyde which was effective against Epidermophyton in 1:128, 000 dilution.
In order to obtain antifungal agents for medicinal drug preparations, 8 kinds of α, β-unsaturated ketones, 11 kinds of nitrofuran derivatives, 6 kinds of organic mercurials, and 37 kinds of essential oils and their components were tested in Czapek medium, using Penicillium chrysogenum, Aspergillum oryzae, and Mucor racemosus. It was found that Merthiolate in 1:512, 000 to 1:1, 024, 000 dilution and phenylmercuric acetate in 1:128, 000 to 1:256, 000 dilution show excellent results.
Using rats pretreated with a preparation of adrenal cortical extract (Adrex “Sankyo”) so as to suppress the secretion of ACTH from cerebral hypophysis, it was examined whether this could be used for the assay of ACTH. Sixteen hours after subcutaneous injection of Adrex, corresponding to 80g. of adrenal, with 2mg. of ascorbic acid, the rat was anesthetized with pentobarbital, its left adrenal taken out, and the test solution was injected intravenously. After 1 hour, its right adrenal was taken out and the ascorbic acid concentration in the two adrenals was compared. The difference value itself and the decreased rate were taken as the respective responses and analyzed statistically. In 0.625-5.0γ/100g. dose, lineality of the log dose-response curve was observed but the value λ was found to be too large and varied.
The thioformyl compounds, obtained by the application of potassium dithioformate to the hydrazides of benzoic, phenylacetic, and o- and p-hydroxybenzoic acids, were cyclized with conc. sulfuric acid to 2-phenyl-, 2-benzyl-, 2-o-hydroxyphenyl-, and 2-p-hydroxyphenyl-1, 3, 4-thiadiazole, respectively. Nitration of 2-phenylthiadiazole gave three kinds of nitro derivatives and that of m.p. 200° was confirmed as the p-substituent, that of m.p. 132° as the m-, and that of m.p. 113° as the o-substituent. Relationship between the ultraviolet absorption curve and structure of nitro and amino derivatives of 2-phenylthiadiazole was observed. Nitration of 2-amino-5-phenylthiadiazole first yielded a nitramino compound which underwent transition by sulfuric acid to 2-amino-5-p-nitro-phenylthiadiazole.
Application of acylation agent, such as benzoyl chloride, acetic anhydride, and p-nitrobenzoyl chloride, to the thiol-type vitamin B1 respectively yielded crystals of m. p. 173-174° (decomp.) (VII), m. p. 122-123° (decomp.) (VIII), and m. p. 135° (decomp.) (IX). These compounds gave negative thiochrome reaction but the reaction became positive after treatment with acid or alkali. From such facts and from their elemental analytical values, they were respectively assumed as 2-methyl-4-amino-5-[N-(3′-benzoylmer-capto-5′- benzoy-Δ2′-pentenyl-2′)]-formaminomethylpyrimidine (VII), (O, S-dibenzoyl B1) 2-methyl-4-amino-5-[N-(3′-acetylmercapto-5′-acetoxy-Δ2′-pentenyl-2′)]-formaminomethyl-pyrimidine (VIII) (O, S-diacetyl B1), and 2-methyl-4-amino-5-[N-(3′-p-nitrobenzoylmercapto-5′-hydroxy-Δ2′-pentenyl-2′)]-formaminomethylpyrimidine (IX) (S-p-nitrobenzoyl B1). These compounds were tested for the prevention of B1-deficiency in Uroloncha domestica and were found to have almost equal efficacy as vitamin B1. It was also found that the absorption of (VII) and (VIII) through the alimentary canal was better compared to vitamin B1. Since (VIII) is easily formed from vitamin B1 and vice versa, it may be assumed that, as a part of biochemical actions of vitamin B1, the pyrophosphate of S-acetyl B1 undergoes transacetylation and acetylates coenzyme A to form acetylcoenzyme A.
The crystals (A) of m. p. 173-174° (decomp.), obtained by the application of benzoyl chloride to the thiol-type vitamin B1, when heated with hydrochloric acid, partially decompose into three components, 2-methyl-4-amino-5-aminomethylpyrimidine (II), formic acid, and γ-aceto-γ-benzoylmercaptopropyl benzoate (III). A part also decomposes into O-benzoyl B1 (IV) and vitamin B1 (V). Heating of (A) with alkali changes it into (IV) and (V). From these results, the structure of 2-methyl-4-amino-5-[N-(3′-benzoyl-mercapto-5′-benzoxy-Δ2′-pentenyl-2′)]-formaminomethylpyrimidine (I) (O, S-dibenzoyl B1) is assigned to (A). The crystals (C) of m. p. 122-123° (decomp), obtained by the application of acetyl anhydride to the thiol-type vitamin B1, when heated with hydrochloric acid, almost quantitatively form vitamin B1 (V) but heating with alkali change them into O-acetyl B1, (IX) and vitamin B1 (V). From these results, the structure of 2-methyl-4-amino-5-[N-(3′-acetylmercapto-5′-acetoxy-Δ2′)]-formaminomethylpyrimidine (VIII) (O, S-diacetyl B1) is proposed for (C).
By the application of alkyl alkylthiosulfinate or sodium alkylthiosulfate to the acetyl, propionyl, and benzoyl esters of the thiol-type vitamin B1, 24 kinds of 2-[2′-methyl-4′-aminopyrimidyl-(5′)]-methylformamino-5-acyloxy-Δ2-pentenyl-(3) alkyl disulfide (II) (O-acylthiamine alkyl disulfide) were prepared. (II) can also be obtained by the acylation of thiamine alkyl disulfide (I) in pyridine. All the compounds of (II) series were found to be approximately equal to vitamin B1 in preventing vitamin B1 deficiency in Uroloncha domestica.
The catalyst system used in the condensation of 5-nitrofurfural and acetaldehyde was considered as a basic and an acidic components. Piperidine, diethylaminoethanol, diethanolamine, and pyridine were selected for the former and acetic, tartaric, citric, succinic, maleic, valeric, and 5-nitrofuroic acids for the latter. A catalyst system was made by the combination of the two components and the condensation was carried out in benzene solution under a mild temperature conditions. 2-(5-Nitro)-furylacrolein, m. p. 118°, was obtained in 49% yield by the use of piperidine-acetic acid system and in 50% yield by the use of piperidine-5-nitrofuroic acid system. It was assumed that the electrophilic piperidinium ion, rather than the nucleophilic piperidine, took part in the catalytic action in this condensation.
Antifungal actions of 45 kinds of haloacyl compounds of aromatic primary amines against Trichophyton interdigitale in vitro were tested and by the examination of the relationship between chemical structure and efficacy, a definite knowledge was gained. The most strong antifungal action was shown by p-chloro-ω-chloroacetoanilide (1:300, 000) and o, m-dichloro-ω-chloroacetoanilide (1:70, 000) but these were found to be too irritating to the skin to permit their practical use.
Periodate oxidation of the N-glucosides of o-, m-, and p-toluidine, m- and p-nitraniline, and o-, m-, and p-aminobenzoic acid was carried out by the usual procedure. After about 10 hours, m- and p-nitraniline N-glucosides consumed about 5 moles of the oxidant but the other N-glucosides consumed various amounts of the oxidant above 5 moles. The consumption curves of these latter glucosides became straight lines after 2-10 hours and were almost parallel to each other under the same conditions.
In order to find out the influence of the S-alkyl group on the effect, 7 kinds of N, N′-disubstituted benzamidines, 4 kinds of N, N-disubstituted benzamidines, and 14 kinds of naphthoamidines, all possessing a thioalkyl group, were synthesized. Preparation of N, N′-disubstituted compounds was carried out by the halogenation and amination of the corresponding thioalkylbenzoic acid alkylamides, and the N, N-compounds by the Grignard reaction of the corresponding benzonitriles. The hydrochlorides of the former were of low hydrophilic properties when the alkyl in N and N′ were of aromatic nucleus and were assumed to be unsuitable as local anesthetics. On the other hand, the physical properties of the N, N-disubstituents were found to be more suitable for such application. Their pharmacological actions are shown in Report V.
In order to obtain local anesthetics which are not antagonized by sulfanilamides, various substances possessing thioalkyl group in the nucleus as the anesthesis-strengthening group were synthesized. The pharmaceutical efficacy of some of the promising of these compounds was determined. Surface anesthetic power was tested on rabbit cornea and irritation on aural canal of a guinea pig, infiltrating anesthetic power by human subcutis and rabbit eye lid, and the lethal dose, LD50, by mouse. As a result, it was found that the introduction of S-alkyl group extremely heightened the rate in alkamine and dialkylaminoacetanilide types while such an increase was not observed in amidine type, although the infiltrating anesthetic power in the latter was strengthened. The compounds showing a large pharmaceutical efficacy rate were diethylaminoethyl p-thioethylcinnamate and diethylamino ethyl 3-thioethyl-4-hydroxybenzoate which were respectively 5 and 3 times that of procaine and 20 and 7 times that of cocaine. p-Thioisoamyl diethylamino acetanilide possessed a small minimum concentration, showing surface anesthetic power even in 0.1% concentration, and was slightly better than cocaine.
Solubility of 2-methyl-1, 4-naphthoquinone in the aqueous solution of nonionic surface-active agents possessing a polyoxyethylene group was determined and following characteristics were observed. The solubility of 2-methyl-1, 4-naphthoquinone was approximately proportional to the concentration of the surface-active agent but the solubilizing efficiency was better at a lower concentrations. At a very high concentration region of the active agent, the solubilizing efficiency was found to increase rapidly. Compared in the same weight/volume concentration, active agents with less oxyethylene group possessed better solubilizing efficiency. These results were compared with the solubility in an aqueous solution of a few ionic surface-active agents and simple observations were made.
Japanese official water-soluble food colors were divided chemically into four groups by the existing A.O.A.C. method. Each of the groups was developed by paper chromatography (ascending method) using a different solvent for each group and divided into each dye. The isolated dye on the filter paper was detected by its Rf value and colorations with conc. HCl, conc. H2SO4, 10% NaOH, or dil. ammoniac water. It was found through this study that the Rf values were dependent on the molecular weight and dissociation constant. Examples of isolation of marketed mixed dyes by this method are given.
Growth inhibitory action against three kinds of bacteria was tested in 61 kinds of compounds. The compounds that were effective in completely inhibiting the growth of Micrococcus pyogenes var. aureus in 80, 000 dilution were 5, 4′-dichloro-2, 4-dihydroxy-diphenylmethane, 2, 4-dihydroxy-5-heptylbenzaldehyde, 2, 4-dihydroxy-3-chloro-5-ethylbenzaldehyde thiosemicarbazone, 2, 4-dihydroxy-3-chloro-5-butylbenzaldehyde thiosemicarbazone, and α-collatolic acid dithiosemicarbazone, in 160, 000 dilution by 2, 4-dihydroxy-3-chloro-5-hexylbenzaldehyde and methyl monochloroisohaematommate thiosemicarbazone, in 320, 000 dilution by 2, 4-dihydroxy-5-isoamylbenzaldehyde thiosemicarbazone, and in 640, 000 dilution by 2, 4-dihydroxy-3-chloro-5-hexylbenzaldehyde thiosemicarbazone. The compounds effective in inhibiting growth of Bacillus subtilis in 80, 000 dilution were 5, 4′-dichloro-2, 4-dihydroxydiphenylmethane, 2, 4-dihydroxy-3-chloro-5-ethylbenzaldehyde thiosemicarbazone, 2, 4-dihydroxy-3-chloro-5-butylbenzaldehyde thiosemicarbazone, α-collatolic acid dithiosemicarbazone, and methyl monochloroisohaematommate thiosemicarbazone, in 160, 000 dilution by 2, 4-dihydroxy-5-isoamylbenzaldehyde thiosemicarbazone, and in 320, 000 dilution by 2, 4-dihydroxy-3-chloro-5-hexylbenzaldehyde thiosemicarbazone. Some compounds were able to inhibit the growth of Escherichia coli communior in 10, 000 dilution but none were effective in higher dilutions.
Course of oxidation of trans-π-oxocamphor (I) was followed by passing air through its aqueous solution and determining the amount of (I), peracid (II), and isoketopinic acid (III) formed. It was thereby found that the air oxidation of (I) was accelerated by the presence of a minute amount (1×10-6 mol./L.) of iron ion, and in its absence, irradiation of light resulted in the formation of peracid (II) and the oxidation proceeded.
Formation of peracid during the air oxidation of various aldehydes was found to vary greatly. The peracid formation was found to be extremely easy in trans-π-oxo-camphor where irradiation of light promoted formation of a free radical which in turn produced chain reaction (Fig. 1) and the oxidation proceeded. The peracid (II) formed in this case could not be isolated but it was found that (II) oxidized sodium chloride added during this oxidative reaction to liberate chlorine. By this indirect means, the formation of the peracid during this oxidation was confirmed.
The reduction product of m.p. 175°, obtained from the chloromethyl compound, m.p. 185° (decomp.), was found to be identical in various properties, melting point, and by mixed fusion with 6-methylacacetin-7-methyl ether (XX), m.p. 179°, formed by the methylation of acacetin with methyl iodide and sodium methoxide in methanol solution. This has clarified that chloromethyl group was present in the 6-position. The structure of (XX) was determined by the following routes. The permethylated compound was submitted to hydrolytic ring cleavage with 15% alcoholic potassium hydroxide to the diketone (XXII) which was decomposed with 30% potassium hydroxide. The phenolic compound, m.p. 148°, thereby formed, was found to be identical with the reduction product (VI), m.p. 148°, of a formyl compound (V), prepared from phloroglucinol dimethyl ether (IV), from which (XX) was determined as the 6-nuclear methylated derivative.
Antifungal action, in vitro of various surface active agents against Trichophyton was tested by which cationic agent, trimethylcetylammonium halide, and anionic agent, sodium p-laurylphenylsulfonate, were found to possess excellent properties. Nonionic surface active agents were found to be practically devoid of antifungal activities. The increase in the efficacy of surface active agents by the concurrent use of 2-dimethylamino 6-(β-diethylaminoethoxy)-benzothiazole dihydrochloride (Asterol), a chemotherapeutic for Trichophyton, was found to be 3-30 times, and the surface active agents that failed to show any efficacy by themselves were also found to have acquired such efficacy. This was assumed to be due to the increase of permeability of fungal membrane of Trichophyton by surface active agents.
Six kinds of compounds, of the following general formulae, were prepared in order to find a powerful analgesic. For their preparation, Grignard reagent or lithium compound was applied to N-methyl-2-phenacylpiperidine or ethyl N-methylpiperidyl-2-acetate, and the carbinols thereby formed were boiled with hydrochloric acid in glacial acetic acid solution to form the corresponding ethylene compounds.
In order to test the curare-like action, following three ammonium compounds possessing diphenyl ether as the functional group were prepared: 4, 4′-bis(Dimethylamino)-diphenyl ether dimethiodide (II), 4, 4′-bis(dimethylaminomethyl)diphenyl ether dimethiodide (III), and 2-methoxy-5, 4′-bis(dimethylaminomethyl)diphenyl ether dimethiodide (IV). For comparison, 4, 4′-bis(dimethylaminomethyl)biphenyl dimethiodide (V) was also prepared. For their preparation, diphenyl ether, 2-methoxydiphenyl ether, or biphenyl was derived to bis(chloromethyl) or a corresponding bromo derivative to which was applied liquid dimethylamine. Methyl iodide was then applied to the dimethylaminomethyl derivatives thereby formed to the corresponding objective compounds.
In view of the fact that a folic acid isomer in which the p-aminobenzoic portion of folic acid had been replaced with m-aminobenzoic acid was antagonistic to folic acid, the biological effect of still another folic acid isomer in which the same portion had been replaced with anthranilic acid seemed to be of interest and its synthesis was carried out by the method elaborated by Uyeo and Mizukami. The compound thereby obtained was a yellowish white amorphous substance, even after repeated purification by reprecipitation, and its biological tests showed it to possess neither the antagonistic nor synergistic action against folic acid.
Intermediates of food coloring matter which are difficult to be obtained in a pure state by synthesis and are impossible to purify are R acid (2-naphthol-3, 6-disulfonic acid) and G acid (2-naphthol-6, 8-disulfonic acid). Their complete isolation was effected by a quantitative spot test using a round filter paper, and developing in a low-pressure, closed vessel, using acetic acid-butanol solution (butanol: water: acetic acid=5:4:1). The separated zones were cut out, extracted, and color developed by coupling with naphthionic diazonium solution in alkaline medium as Amaranth and New Coccine. These red dyes were colorimetrically determined by an electrophotometer. Amaranth, New Coccine, R acid, and G acid were purified by the chromatopile from which pure products of these were obtained. Quantitative spot test could be utilized for the quality tests and quantitative determination, rapidly and simply, for tar dyes used for food coloring, such as the Amaranth, New Coccine, Indigo carmine, and mixed dyes.
Domestic Fagara genus plants contain Fagara ailanthoides Engl. and Fagara schinifolia Engl. The former yielded yellow needle crystals of m.p. 146-147°, corresponding to C13H10O5, and this was proved to be isopimpinellin. The latter yielded yellowish white needle crystals of m.p. 188°, corresponding to C12H8O3, and of m.p. 121°, corresponding to C7H6O2, which were respectively proved as bergapten and benzoic acid.
Aesculetin-glucoside (aesculin) was isolated and determined from the trunk bark of Fraxinus japonica Blume. According to the existing literature, fraxin (Iida), fraxinin (Okui), and aesculetin (Shimada) are reported as the component but fraxin is identical with aesculin obtained by the authors.
Antibacterial tests aginst H37Rv strain of tubercle bacilli were made with the hydrazides of nine kinds of normal dibasic acids between oxalic and sebacic acids. The growth inhibitory action was the weakest in glutaric acid hydrazide, the action strengthening with the increase in the number of carbon stoms above that, the action becoming the strongest with suberic acid hydrazide with eight carbon atoms. Above that number of carbon, the action was found to again decrease.
The leaves of Mahonia japonica DC. was found to contain all the alkaloids contained in the wood portion, with the exception of berbamine. The contents of the alkaloids, however, differed in different portion of the plant, and these ratios are shown in Table I.
Examination on the sensibility and the weights of E. Mettler's microbalance was made and it was found to have enough precision to allow the use for microanalysis under the same conditions as the existing Kuhlman's microbalance.
In order to examine the physiological action, arecoline hydrochloride, arecaidine hydrobromide, and arecaidine hydrochloride were prepared from arecoline hydrobromide. Esterification of arecaidine hydrochloride was carried out by the saturation of dry hydrochloric acid gas in its dehydrated alcohol solution to form ester salts. The free bases of the esters were so unstable that they were derived to stable picrates and again led to the free bases by the use of a liquid chromatography. The bases were then used after neutralization with various acids.
Ammi visnaga Lam., the source of khellin is an annual or biennial crop, which can be cultivated in Japan. The content of khellin in the fruit, as determined by the present authors by the modified method of Abd El-Aziz Abd El-Rahman, was 0.1%, but none was detected from undergrown fruits, and primary and secondary rays of compound umbels. It seems to be more profitable and desirable to cultivate this plant as a biennial crop by sowing in autumn, in Japan. The cultivation of this plant is practically possible by means of transplantation, too, which should preferably be made by the end of April at the latest. The autumn sowingshould be done by the middle of October and spring sowing by the beginning of April at the latest. The cultivation of this plant is suited in a rich and well-drained soil, containing a large amount of clay.