Recent works on the periodate oxidation analysis of carbohydrates are reviewed, placing emphasis on the simultaneous analysis of the products of oxidation. A number of examples are presented for the application of the product analysis to the elucidation of carbohydrate structures, to demonstrate the usefullness of this newly developed technique.
The effect of temperature on the compaction behavior and the adhesive properties of methyl and butyl p-hydroxybenzoate (abbreviated as Me-POB and Bu-POB, respectively) powders was studied. The adhesive force at a contact point between two particles (H) determined by tensile strength measurements showed maxima at the value of homologous temperature (ratio of temperature TK to melting point TmK, T/Tm) between 0.94 and 0.96. The value of the compression coefficient (k) in the Kawakita equation, a measure of the ease of powder compaction, remained nearly constant up to about 0.9 of T/Tm. Above 0.9 Tm it increased remarkably, which may be caused by the reduction of friction among particles and between particles and die wall at high temperature. Under given tapping conditions, the initial rate of compaction (v1) decreased and the final porosity (εf) increased with increasing temperature in the range of T/Tm below 0.9. It was thought that the probability of filling the vacant spaces existing in the lower layer of the powder bed with particles diminished as the adhesive force between particles increased. Then, an empirical equation relating εf to both H and impact accerelation α* was presented.
The minimal inhibitory concentrations of cercosporin, isocercosporin, and various quinones on bacteria and fungi were determined by the agar dilution method with Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, Escherichia coli, Candida japonica and so on. Gram-positive bacteria were more sensitive than gram-negative ones.
An inclusion complex of acetohexamide (AHA) with β-cyclodextrin (β-CyD) in the molar ratio of 1 : 2 was prepared, and its permeation behavior through a cellophane membrane, in situ absorption behavior in rabbit small intestinal tract, and in vivo extent of bioavailability in rabbit were examined in comparison with those of AHA alone. The membrane permeation constant and in situ absorption rate of AHA in solution were found to decrease by the addition of β-CyD. However, the lesser permeability and absorptivity of the complex were significantly overcome by an improved solubility of AHA. The inclusion complexation of AHA with β-CyD was also effective in releasing drug from a hydrophobic suppository base such as Witepsol W35. The complex increased highly the levels of plasma concentration and cumulative urinary excretion of AHA and hydroxyhexamide, one of the metabolites of AHA, after oral or rectal administration to rabbits. The increase of bioavailability of AHA by means of the inclusion complexation suggested the possibility of smaller doses and fewer side effects in sulfonylurea therapy.
From cut 2 fraction (bp 215-250°) of Colorado shale oil seven isoprenoid hydrocarbons were isolated and their structures determined by mass and 13C-NMR spectral analyses were as follows : 2, 6-dimethylundecane, 2, 6, 10-trimethylundecane, 2, 6, 10-trimethyldodecane, 2, 6, 10-trimethyltridecane, 2, 6, 10-trimethyl-1-undecene, 3, 7, 11-trimethyl-1-dodecene, and 4, 8, 12-trimethyl-1-tridecent. These isoprenoid components were isolated from the distillate by successive fractional distillation, chromatography on silica gel impregnated with silver nitrate and preparative gas chromatography.
An optically active sesquiterpenoid hydrocarbon was isolated from cut 3 fraction (bp 250-280°) of Colorado shale oil and its structure was determined to be (+)-drim-8-ene (1) on the basis of chemical and spectral properties and by comparison of its 13C-NMR spectrum with that of an authentic specimen derived from dehydroabietic acid.
The relationship between the growth of the fruit of Gardenia jasminoides f. grandiflora and the formation of its main secondary metabolites, crocin and geniposide, was investigated. The growth of the fruit appeared to be divided into two stages. In the first stage (1-6 weeks after flowering), the fruit weight as well as the geniposide content increases rapidly but no crocin was detected. In the second stage (8-23 weeks after flowering), the geniposide content per fresh weight of the fruit hardly changed, while the production of crocin began a few weeks after the beginning of this stage and its content increased almost linearly with time until full ripeness. TLC analyses of the fruit extracts failed to detect, at any growth stage, the presence of unoxygenated carotene that had been proposed as a possible precursor of crocin. However, the organ culture showed that crocin is produced de novo in the detached fruits, suggesting that crocin in Gardenia fruits might be synthesized through an unknown colorless precursor.
The determination of ketosteroids in biological fluids by a fluorescence highperformance liquid chromatographic procedure and the application of this method to clinical analysis of steroidal compounds in the human blood and urine are described. Corticoids are extracted with methylene chloride from 0.5 ml of plasma or 1 ml of urine, and 17-ketosteroids are extracted with methylene chloride after enzymatic hydrolysis or solvolysis (1 ml of urine or 0.1 ml of serum). Δ4-3-Ketosteroids and 17-ketosteroids are labelled with dansyl hydrazine in HCl-EtOH solution and in trichloroacetic acid-benzene solution, respectively, and chromatographed on a microparticulate silica gel column by use of CH2Cl2-EtOH-H2O mobile phase systems with a fluorophotometric detector at 350 nm (excitation) and 505 nm (emission). Comparisons with radioimmunoassay gave correlation coefficients of 0.978 (n=36) and 0.964 (n=81) for plasma cortisol and serum dehydroepiandrosterone sulfate, respectively. Metabolite of carbamazepine did not interfere with the analysis of urinary free cortisol. The serum concentrations of cortisol, 11-deoxycortisol and dehydroepiandrosterone sulfate after a single oral administration of 1.5 g metyrapone to human subjects were determined and the serum 11-deoxycortisol could be measured by this method as a rapid metyrapone test. The method is thus useful for the clinical analysis of ketosteroids in biological fluids.
The species differences in the metabolism of befunolol were investigated by using the cell fractions prepared from the liver or other tissues of several species. 2-Carbonyl-reducing activity forming 2-(1-hydroxyethyl)-7-(2-hydroxy-3-isopropylaminopropoxy) benzofuran (MI) was detected in the hepatic 103500×g supernatant from human and experimental animals such as rabbit, rat, hamster, guinea pig and mouse. 4-Hydroxylating activity forming 2-acetyl-4-hydroxy-7-(2-hydroxy-3-isopropylaminopropoxy) benzofuran (M II) and oxidizing activity at 7-side chain of befunolol were detected in the hepatic microsomal fraction from the animals, but not in that from human. The differences among the species were observed in the degree of each befunolol-metabolizing activity. The reducing activity in the animals was inhibited by the presence of microsome, but the activity in human did not vary under similar conditions. At low concentration of befunolol, the 9000×g supernatant of the human liver in which neither 4-hydroxylating nor 7-side chain oxidizing activity was detected apparently showed higher reducing activity than that of the animals. The reducing activity was detected in the cell fraction of several organs besides liver. The species differences in the pattern of tissue distribution of the activity were also observed among rabbit, guinea pig and human.
The relationship between the antioxidative action of sodium copper chlorophyllin (Cu-Chl-Na) and its chemical structure was examined. By the aerobic incubation of Cu-Chl-Na in aqueous solution, the spectral absorbances of the solution at Soret (406 nm) and red (632 nm) maxima decreased while the ratio of the absorbances (Soret/red) increased. Since the Cu-Chl-Na incubated aerobically lost its original antioxidative activity, the thiobarbituric acid values in the rat liver homogenates treated with Fe2+ and ascorbic acid increased, and a decrease in the reduction of 1, 1-diphenyl-2-picrylhydrazyl was observed. Further, Cu-Chl-Na reacted directly with 1, 1-diphenyl-2-picrylhydrazyl to decrease the spectral absorbances at both Soret and red maxima and to increase the ratio of the absorbances (Soret/red). Since the spectral absorption in the red-band region is attributable to the absorption of copper chlorin structure, the above results suggest that copper chlorins are responsible for the antioxidative activity of Cu-Chl-Na.
Some N, N-dialkylglycylbenzhydrylamides were synthesized. Phthaloylglycine was condensed with benzhydrylamine in the presence of DCC to give 4, which was treated with hydrazine hydrate to give 3a. The dialkylamides, 3b-3e, were obtained by the reaction of chloroacetylbenzhydrylamine with the corresponding secondary amines. In antihistaminic tests the compounds, 3b-3e, showed weak activities.
Proteolytic enzymes are contained in many digestive and gastrointestinal drugs. The proteolytic activities are commonly determined by casein-folin method. Since this method requires complicated procedure and long time, it was attempted to develop a simple method of determining proteolytic enzymes in gastrointestinal drugs for the routine quality control of the drugs. Azocoll used as substrate is a collagen combining with an azo dye to provide an insoluble substrate. Proteolytic enzymes digest the dispersed substrate and release colored hydrolyzed products to the solution. The reaction is simply terminated by filtering off the substrate. The activities can be given by determining the absorbance of the filtrate as an index. When this method was applied to the determination of the activity of proteolytic enzymes contained in the gastrointestinal drugs, the following several advantages were found compaired with the casein-folin method ; i) simple and quick procedure, ii) high sensitivity, iii) long time stability of the colored hydrolyzed products, iv) good correlation with the conventinal method.
Photostability of eight kinds of photosensitive solid drugs was evaluated from the point of coloration under various light sources such as mercury vapor lamp, daylight lamp, and two kinds of fluorescent Iamps. Sample drugs examined were sulpyrine, chlorpromazine hydrochloride, perphenazine, indomethacin, promethazine hydrochloride, imipramine hydrochloride, furosemide, and phenytoin. Coloration was determined by tristimulus colorimetry. In order to investigate the photosensitivity of these samples, their action spectra for coloration was obtained with a grating monochrometer in the range of irradiation wavelength 306-475 nm. Coloration process of each sample could be expressed as a straight line on double-log scale. These processes changed corresponding to the combination of drug and light source, indicating different coloration reaction orders. These apparent orders and the degree of coloration were discussed in relation to the action spectrum of each sample. In some of the samples the existence of photosensitive similarity in coloration between exaggerated and ordinary irradiation conditions was suggested.
A rapid, simple and accurate high-performance liquid chromatographic procedure was established for the quantitative determination of paeoniflorin in pharmaceutical preparations. Paeoniflorin, extracted from these preparations containing multiple components with water, was separated on Zorbax CN column (4.6 mm i.d. × 25 cm) with a mobile phase of 0.1 M HClO4 (pH 3.5)-CH3CN (9 : 1, v/v) at room temperature. The recoveries in all preparations were good. This method is considered to be useful for the evaluation of these crude drug preparations.
It has been ascertained that diisopropylamine was effective as a coexisting base when aliphatic and aromatic amines could be easily alkylated with primary alkyl halides. Pyrrolidine, piperidine, aniline, and 1, 2, 3, 4-tetrahydro-β-carboline derivatives as amine and ethyl, propyl, butyl, allyl, and cyclohexyl bromides or iodides as alkyl halide were used for this examination.
The biological fate of l-1, 4-dimethyl-10-hydroxy-2, 3, 4, 5, 6, 7-hexahydro-1, 6-methano-1H-4-benzazonine hydrobromide (l-ST-2121) was studied in mouse, rat, rabbit, dog, monkey and man after i.v. or s.c. administration. Plasma levels of unchanged l-ST-2121 showed typical two-phase elimination curve after i.v. adminstration to rat and rabbit, and conjugated l-ST-2121 reached the maximum in about 30 min after the injection to rabbit. After s.c. administration to mouse, rat, rabbit, dog, and man, plasma levels of the unchanged drug reached the maximum (for example about 2 μg/ml in mouse) within about 40 min and the peak was shifted to the later time in larger animals. The plot of the biological half-life vs. body weight of each animals in log-log scale gave a linear function curve except for rat, and the maximum plasma levels of the unchanged form were approximately proportional to each dose. Urinary excretion percentage of the unchanged form, the conjugated, 9, 10-dihydroxy compound and nor-compound were measured after s.c. administration to each animals and man. Total excretion percentages were 65-99% and the speeies differences were observed in the excretion pattern of the unchanged and nor-compound, that is, the largest excretion of the unchanged form is 36% in mouse and the least is 5% in dog, and the largest excretion of nor-compound is 13% in man and the least is 1% in dog.