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The paper chromatography of phenols (expecially phenolic acids), aglycones, glycosides and certain sugars is carried out with two new solvents. The authors have also examined these substances with the aid of paper electrophoresis and a borate buffer. A detection reagent for o -diphenols on paper is described.

Minimization of samples for infrared spectral measurement was attempted by the use of a Japanese folkcraft, hand-made paper, impregnated with liquid paraffin. By this method, it is possible to measure 0.1 mg. of a sample. This hand-made paper was also used for paper chromatography and the developed paper was submitted directly to infrared spectral measurement. Component of Penicillium islandicun and inorganic sulfate ion were determined by spectral measurement of their chromatogram.

An aqueous solution of sodium molybdate has been used as a specific reagent for the detection of phenolic compounds with o-dihydroxy groupings. The paper electrophoretic behaviour of phenolic compounds has been studied using a series of different buffers with pH values ranging from 5.2 to 10.0. o-Dihydroxy compounds can readily be detected by the use of ammonium molybdate buffer, and other structural features can be revealed by the choice of suitable electrolyte solutions. Paper partition chromatography of phenolic compounds using molybdate-treated paper in described.

The paper chromatography of vat and sulphur dyes, using reducing solvents at 80°, has been carried out, the dyes being separated as their leuco compounds. In both cases ascending paper chromatography was applied. The relation between dye constitution and chromatographic behaviour has been examined.

A new solvent system was given for the chromatography of food preservatives; benzoic acid, salicylic acid, p-hydroxybenzoic acid and it's esters, p-chlorobenzoic acid, sorbic acid, resorcinol and β-naphtol with the ascending method on Toyo Roshi No. 50 paper at 20-25°. Cyclohexanol-ammonium carbonate buffer gave results in 30 hours. Spots were detected by absorption when the paper was examined in ultraviolet light and with color reagcnts of the methyl red or diazo spray reagent. Rf values for this solvent were listed for these substances. Each substance was separated except in the case of mixture of benzoic acid and sorbic acid. However in this case, these two substances were identified by measuring spectrophotometrically the alcohol solution extracted from the part containing the substances on paper chromatogram.Horizontal paper electrophoresis on Toyo Roshi No. 51 paper for 1.5-2 hours at 20V per cm with buffer consisted of dimethylformamide, pyridine, acetic acid and water (pH 5.6) or of butanol, ammonia, acetic acid and water (pH 5.0) was useful. MNC (the ratio of the migration of the substances of New Coccine) and colors given with the methyl red, FeCl3 and diazo spray reagents were tabulated adding dehydroacetic acid to above substances. Data was also given for citric acid and tartaric acid.Sake (Japanese wine), soy etc., was acidified with HCl, extracted with ether, purified as ammonium-salts and submitted to paper chromatography and paper electrophoresis with similar technique.

A number of simple chromatographic applications are described in which sheet paraffin or other plastic film is used as an adhesive vapor barrier: 1. A funnel attached to a paraffin-enclosed narrow paper wick for spotting large volumes of sample solution as small spots of controlled size. 2. A funnel for the elution of a spot and its reapplication to another paper in a single operation. 3. A method for elution or chromatography which requires no jars or other rigid saturation chambers.

Modified paper chromatography procedures for the analysis of fatty acids produced by anaerobic bacteria are described. Both ethylamine and hydroxylamine derivatives of fatty acids were prepared from inoculated anaerobic culture broth. The derivatives were spotted on chromatography paper and developed with appropriate solvents. Paper chromatography is a valuable alternative to gas liquid chromatography as an ancillary procedure in the identification of anaerobic bacteria in the clinical bacteriology laboratory.

The automated dithionite test for the detection of hemoglobin S has been modified for use with AutoAnalyzers having a filter paper read-out. This method was compared with results of hemoglobin electrophoresis in 425 individuals, of whom 58 were positive for hemoglobin S. The two methods were in complete agreement. The automated filter paper dithionite test is a rapid, economical, accurate method for detection of hemoglobin S and could be easily incorporated into currently available mass screening equipment.

An ascending chromatographic method was used to identify acidic mucopolysaccharides. On silica-gel-treated glass papers, a displacement type of chromatography with ethanol-salt solutions separated seven mucopolysaccharides. The compounds were located by charring with sulfuric acid or different staining technics. The method is convenient, reproducible, and rapid and can be used to identify small quantities of mucopolysaccharides in mixtures. Certain difficulties as and identification procedure were observed, however, and are discussed. Observations on glass-fiber papers are useful in studying different reactions of mucopolysaccharides with various salts and stains.

The design and manipulation of an apparatus for centrifugal acceleration of paper chromatography has been described. The main novel features consists of using filter paper strips instead of circular sheets and a constant flow solvent reservoir enclosed in the chromatography tank. Our arrangement is particularly suited for the quantitative analysis of materials labelled with radioactive isotopes of short half-lives and examples are given of the separation of some 82 Br- and 131 I-labelled substances. Advantages of the above method over circular paper chromatography are discussed.

The separations of steroid oestrogens achieved by paper chromatography have been reviewed with special reference to studied involving epimeric oestrogens and to oestrogens which have been isolated recently. Methods of detection of these compounds on paper chromatograms have also been discussed. The mobilities of thirty-four different oestrogens and their derivatives in many solvent systems have been tabulated. It is hoped that this review will be found useful in providing a collection of data from which separations of mixtures of oestrogens can be planned.

1. (1) The binding of acid dyes by denatured protein on filter paper has been investigated with special reference to Light Green. 2. (2) The uptake of dye, measured by elution, has been shown to vary according to the method of denaturation, the nature of the dye solvent (particularly the salt and and ethanol concentration) and finally with the duration and temperature of the subsequent washing procedure. 3. (3) Albumin and globulin do not behave in the same manner with different denaturing agents so that different methods give different albumin/globulin dye uptake ratios. 4. (4) The stoichiometric relationship between dye and protein is consistently nonlinear even at lower concentrations of protein on the paper and the same pattern can be demonstrated with protein on cellulose acetate membrane. It is suggested that this non-linear relationship at low protein densities is due to interference by the cellulose supporting medium and other anions. 5. (5) Intrinsic constants and the number of binding sites at infinite dye concentration have been measured under standardized conditions for Light Green, Brom-Cresol Green and Lissamine Green. The results have been compared and differences discussed. 6. (6) In the quantitative measurement of protein based on dye uptake, the importance of restricting the protein density on the paper is stressed.

A technique is described by which samples of tissue weighing from 1 to 10 mg can be frozen and ground and then stuck to paper chromatograms and the soluble compounds separated by normal solvent development. The advantages and special applications of this technique are discussed.

1. 1. A method for the detection of Jaffe-positive substances in human blood serum using the technique of paper partition chromatography is described. 2. 2. A partial solution of the problem of apparent creatinine has been furnished by identification of pyruvate as a second Jaffe-positive compound beside creatinine in a water-soluble fraction of acetone-filtrate of sera; this finding was confirmed polarographically as well as spectrophotometrically . Up till now the quantitative significance of this substance has not been considered in connection with this problem. 3. 3. After a few preliminary experiments, paper chromatography of the insoluble residue of acetone-filtrate of serum has been left open for further investigation. It seems likely that the final solution of the problem of apparent creatinine will cover both possibilities which were discussed in the introduction, irrespectively of whether the chromatographic investigation of the insoluble fraction might show the presence of some new unknown Jaffe-positive component.

A method is described for rapid analysis of phenylalanine and tyrosine using eluates of whole blood that has been impregnated into filter paper. This method employs semi-automated ion-exchange column chromatography (amino acid analyzer) and will yield reasonably accurate and consistent quantitative values for phenylalanine and tyrosine concentrations within two hours. The technique has significant value in rapidly confirming the presence of increased phenylalanine with or without a concomitant increase in tyrosine concentration in autoclaved or unautoclaved blood filter paper specimens. Thus phenylketonuria can be differentiated from hyperphenylalaninemia due to tyrosinemia within hours after the initial identification of an elevated phenylalanine concentration by a screening method such as the Guthrie test. In addition, further evaluations of phenylalanine and tyrosine concentrations can be facilitated in infants on whom only blood filter paper specimens are obtainable.

The effect of hemicellulose contents in pulp on beating rate and strength of paper made from it was studied.In this study two methods were used, one the adding method (adding isolated hemicellulose to the standard pulp) and the other the extracting method (removing hemicellulose stepwisely).In the former when the hemicellulose is isolated with drastic agents, chemical nature of hemicellulose is liable to change, so that it is impossible to say that the isolated hemicellulose will act in the same manner as it does in the original state. Therefore extracting method was employed in the present research.For the preparation of chlorite holocellulose, thin pieces of beech and aspen wood were treated with hot acidified sodium chlorite. These chlorite holocelluloses were treated with KOH solution under nitrogen gas, in order to minimize the oxidation of cellulose. For hemicellulose extraction, KOH solutions of various concentration (from 0.3 to 20%) were used in order to get a series of pulp in which the hemicellulose content was stepwisely arranged.The beating rate and the sheet strength of this series of pulp were examined. From the result of this study, it was concluded that there appeared to be the optimum hemicellulose content in pulp as for as the paper strength was concerned. Other observations were also made with regard to the result observed.

Examination of urinary product of rabbits administered with MHB, by filter paper chromatography, indicated that ureide, 3-OH-MHB, 3-keto-MHB, 3-keto-nor-MHB, and unknown compounds (MHB-M (VI)) were excreted. An excellent method for the separation of these compounds is the use of a buffered filter paper and butanol saturated with borate-NaOH buffer (pH 11). For detection of the located metabolites, 1% solution of sodium periodate and 1% solution of potassium permanganate were used and ultraviolet spectrophotometry was adopted for the characterization of barbiturates.

A simplified method for the quantitative determination of keto-acids by means of paper chromatography of their 2,4-dinitrophenylhydrazones is reported. 1. 1. The identification of these compounds on the filter paper is discussed, especially of those compounds which give more than one spot. 2. 2. The method is described in detail, espicially with regard to the pyruvic and α-ketoglutaric acid content in biological material. The recovery of these two keto-acids, starting from known aqueous solutions, averages 94%; on the basis of these results a standard curve can be drawn. According to this curve, recovery from deproteinized extracts of liver and blood averages 91.6% for pyruvic acid and 96.7% for α-ketoglutaric acid. 3. 3. Using this method the pyruvic and ketoglutaric acid content of rat blood, muscle, liver and brain is estimated.

Food preservatives—sorbic, benzoic, p-hydroxybenzoic acid and p-hydroxybenzoates—have been extracted from foods directly on to chromatographic paper with ethyl ether. Paper chromatographic separation of the extracted compounds was effected on the same paper. Methyl, ethyl and propyl p-hydroxybenzoates have been separated from each other during ether extraction and from the critical pair, the butyl and benzyl p-hydroxybenzoates, on a paper impregnated with sodium hydroxide. An examination was made of the reactions which distinguish sorbic and benzoic acids when both are present.

A technique has been developed by which complex mixtures of amines can be sepated by column chromatography on the ion exchange resins Amberlite CG-50 and Amberlite CG-120. Identification of amines is made on the basis of the speed of their elution from the resin column, as well as by subsequent paper chromatography of effluent fractions. The method can be applied to amines derived from various physiological fluids, and offers the possibility of isolating unidentified compounds for chemical characterization. Using the combined technique of ion exchange column chromatography followed by paper chromatography, 26 amines were identified in human urine. these were: methylamine, dimethylamine, ethylamine, ethanolamine, β-hydroxypropylamine, pyrrolidine, piperidine, putrescine, cadaverine, 2,2'-dithiobis(ethylamine), histamine, N-acetylhistamine, i -methylhistamine, p -hydroxybenzylamine, 3-methoxy-4-hydroxybenzylamine, p -tyramine, m -tyramine, octopamine, synephrine, 3-methoxytyramine, normetanephrine, metanephrine, tryptamine, serotonin, bufotenin, and kynuramine. An additional 20 unidentified bases, presumably amines, were found to be regularly excreted in urine. Evidence is presented supporting the identification of kynuramine, β-hydroxy-propylamine, putrescine, cadaverine, and 2,2'-dithiobis-(ethylamine). These 5 amines have not previously been considered to be constituents of normal human urine. It is suggested that study of the urinary excretion of amines may provide useful clues as to the mechanisms involved in various metabolic disorders, including those characterized by mental dysfunction.