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A procedure involving reaction paper chromatography for the determination of the number of nitrile groups in molecules from the shift in the RM values of the substance before and after reaction has been developed. A technique for the detection of substances with a nitrile group, which can be used with advantage even for other substances, has been devised.

High rates of acetylene (C 2 H 2 ) reduction (nitrogenase activity) were observed in woodroom effluent from a neutral sulfite semi-chemical mill under aerobic (up to 644 nmol of C 2 H 4 produced per ml per h) and under anaerobic (up to 135 nmol of C 2 H 4 produced per ml per h) conditions. Pasteurized effluent developed C 2 H 2 reduction activity when incubated under anaerobic but not under aerobic conditions. Activities were increased by addition of 0.5 to 3.0% glucose or xylose. Enrichment and enumeration studies showed that N 2 -fixing Azotobacter and Klebsiella were abundant, and N 2 -fixing Bacillus was present. Of 129 isolates of Klebsiella from pulp mills, lakes, rivers, and drainage and sewage systems, 32% possessed nitrogen-fixing ability.

The action of HClO 4 on rhodium hydroxide was studied with paper electrophoresis and ion exchange chromatography. The existence of chloro-complexes due to precipitation of co-ordinated Cl in the preparation of Rh(OH) 3 , was demonstrated. These can be decomposed by repeated precipitation or heating with AgNO 3 .

The RF′ values of a number of isomeric monosubstituted phenols have been determined for three solvent systems. This is a continuation of our previous data. The results confirm the earlier work showing that most of the phenols have significant variations in RF′ value between these three solvents. Some limited correlations between RF′ values and the molecular weight of the side chain were found for members of homologous series. The use of ceric ammonium nitrate as a spot locating agent for phenols was found to have certain limitations, particularly for ortho substituted phenols. In general, however, this reagent is quite suitable for most phenols.

A chromatographic separation on glass fiber paper of mixed glycerides of acetic, butyric, stearic, palmitic, and oleic acids in solvents consisting of 2% and 5% ether in isooctane has been described. In general, glycerides containing fatty acids of increasing chain length tend to move further up the chromatograms than those of shorter chain length.

A method was developed, combining reaction paper chromatography with electrolytic reduction, for the determination of the presence of -COOH groups and possibly their number by comparing the RF values before and after reaction. Moreover, in the solvent system used (n-propanol-ammonia), it is possible to assess the presence of other functional groups from the shift in the RF values.

Summary Some of the compounds of Sn(II) with platinum metals may be separated by adsorption chromatography on paper. Their behaviour in SnCl 2 -HCl and SnBr 2 HBr was noted.

An investigation of the electrophoretic data from patients with various malignant diseases shows that the possibilities of paper electrophoresis as a diagnostic tool are limited. The relative inaccuracy of the method may cover up important details; but more physiological information can scarcely be expected from data concerning only one physical property of a protein molecule. Quantitative evaluation of the data still gives rather qualitative results. With more material it may be possible to differentiate between various groups of diseases. The use of the “entropy” of a serum protein mixture does not give greater differentiation. Only in a limited number of cases can it be used to distinguish pathological and normal sera. The high correlation in the normal group between the albumin contribution to the total “entropy” and the total “entropy” provides a criterion for the detection of pathological serum protein mixtures in most cases.

The electrophoresis and immunodiffusion steps for an open, atmospheric-un-equilibrated system in dilute agar gel on vegetable parchment paper has been described. Studies were carried out on resolution and precipitin identification. Figures showing the attempt to identify individual components of serum were presented.

A simple technique for measuring diffusion rates on glass fibre paper is described. This technique was shown to be suitable for the study of polyacid formation and complexing in solution.

The adsorption paper chromatography of inorganic anions in acetate buffers was studied and the possible adsorption mechanism discussed. Some analytical separations were recorded.

Rapid reliable methods have been developed for the separation of mixtures of organic acids using one-dimensional and two-dimensional techniques of paper chromatography. New solvent mixtures are proposed which include acidic, basic and buffered types. These methods, which have been proved by application to some eighty well-known organic acids, offer considerable advantages over existing techniques in their ease of operation and ready reproducibility of results.

Operating variables in the use of recording ratemeters for scanning radioactive paper chromatographic strips are discussed. Bes sensitivity, and least statistical error in estimation of peak area, is obtained with a wide slit. However, wide slits give poor fidelity, that is, the shape of the record is not an accurate representation of the shape of the actual peak. Short time constnts give a record showing undesirable fluctuations due to statistical count rate variations, while long time constants give poor fidelity. Slow strip speed is required for strips of low activity. Optimal selection of operating conditions is considered from a quantitative standpoint.

Mean R F and R M values have been obtained for a number of steroids and dyes in Bush aqueous methanol systems. Scrutiny of the Δ R M values verifies the stability of the condition achieved.

Summary The chromatographic behaviour of metal ions on acetylated paper with butanol-HCl was investigated. The lower valencies of polonium may be differentiated in this system.

Single and two-dimensional paper chromatograms were prepared of normal and de-coated embryonated eggs of Ascaris suum, vitelline membranes, egg fluid, and larvae. Eighteen amino acids (cystine, lysine, arginine, histidine, aspartic acid, glycine, serine, alanine, proline, glutamic acid, threonine, tyrosine, methionine, valine, tryptophan, phenylalanine, leucine, isoleucine) were identified in larvae and in normal and decoated embryonated eggs. Nine amino acids (leucine, tryptophan, cystine, isoleucine, phenylalanine, tyrosine, proline, aspartic acid, and arginine) were identified from the vitelline membrane. Seven amino acids (cystine, arginine, aspartic acid, glycine, serine, proline, tyrosine) were identified from a fraction in the middle coat. Using the DNFB technic, the N-terminal amino acid in the vitelline coat was determined to be cystine. Sixteen amino acids (cystine, lysine, histidine, aspartic acid, glycine, serine, glutamic acid, threonine, proline, tyrosine, methionine, valine, leucine, tryptophan, phenylalanine, isoleucine) were identified from the outer protein coat.

The separation of cis- and trans-dihalogenodiammineplatinum (II) complexes was effected by paper chromatography using neutral fast flowing solvents such as water at 5° or acetone—water (9:1). Practically all mixtures of cis- and trans- as well as of dichloro-, dibromo-, diiodo as well as chloroiodo- and chlorobromo-complexes could be separated.

The use of ethylene glycol as the stationary phase for the separation of corticosteroids and of 17-ketosteroids by paper chromatography is proposed. Some characteristics of systems with ethylene glycol in the stationary phase are described.

The rapid, quantitative and qualitative analysis of particulates on air filter samples is becoming increasingly important as more air pollution sampling stations are set up throughout the countiy. Although atomic absorption provides a sensitive technique for the analysis of many elements, the disadvantages of complex sample preparation, sample destruction and the necessity to analyze one element at a time make this technique unsuitable for a large volume of samples. X-ray energy spectroscopy when combined with automated sample handling and the latest dedicated data reduction systems provides a technique which enables the analyst to process large numbers of samples and obtain precise quantitative and qualitative data rapidly. This paper will describe the preparation and analysis of typical air filter type samples, and the steps taken to identify the elements in the samples and obtain computerized reduction of the data in μg/cm2, ppm or percent.

The authors studied the use of chloranilic acid as a reagent in the paper chromatography of some of the heavier metals. Sensitivity was found to vary from 0.01 μg for ferric iron, to 5.0 μg for mercury (II) and zinc. It was found that iron gave tailing spots with most of the solvent systems, which interfered with the separation of the other cations. This interference was avoided by using acetone—acetic acid, which displaces iron to the solvent front. Efficient two-dimensional separations were obtained by carrying out a second run with ethanol—acetic acid.

A RADIOIMMUNOASSAY FOR HUMAN PITUITARY LUTEINISING HORMONE USING PAPER CHROMATO-ELECTROPHORESIS

I investigated amino acids composition of the protein from the tuberculous pus plasma in 3 cases and free amino acids in the tungstic acid filtrates of tuberculous pus plasma by a paper-chromatographic procedure. 1) I found tuberculous pus plasma proteins do not essentially differ from usual proteins regarding the qualitative composition. 2) In 3 cases of closed tuberculous abscess never punctured free amino acids were not detected. 3) Free amino acids were present in the protein-free filtrates of tuber-culous pura of 5 closed abscesses often punctured before. 4) The number of the free amino acids found there, however, far less than that of the corresponding amino acids in hot true pura. There is a striking contrast between tubercuous and non-tuberculous true pura.

Summary The electrophoretic pattern of serum proteins has been studied in 30 infants suffering from toxicosis. During the stage of acute dehydration the main changes were a moderate or marked increase of alpha globulin, particularly of the α2 fraction and a decrease of albumin. In some cases there was also an increase of the gamma globulin. During recovery the alpha globulin decreases while albumin increases. However, the return to normal of the electrophoretic pattern is not always achieved when clinical recovery occurs. The absolute amounts of total serum proteins have also been estimated and their significance has been discussed. Electrophorese sur papier des proteines du serum au cours de la toxicose. L'aspect electrophoretique des proteines du serum a eteetudie chez 30 ehfants presentant une toxicose. Au cours de la periode de deshydratation aigue, on a pu noter une augmentation nette des alphaglobulines (en particulier la fraction alpha 2), et une diminution de l'albumine. Dans quelques cas, on a pu noter aussi une augmentation des gammaglobulines. La diminution des alphaglobulines, et l'augmentation de l'albumine est survenue avec la guerison. Neanmoins, le retour a la normale de la courbe d'electrophorese n'est pas toujours acheve au moment de la guerison elinique. Papierelektrophorese von Serum-Protein bei Sauglingstoxikose. Die Elektrophoresebefunde von Serumproteinen sind bei 30 an Toxikose leidenden Sauglingen untersucht worden. Wahrend des Stadiums der akuten Dehydration waren die Hauptveranderungen eine massige oder ausgepragte Zunahme des Alpha-Globulins, insbesondere der α2-Fraktion und eine Abnahme des Albumins. In einigen Fallen be-stand auch eine Zunahme des Gamma-Globulins. Wahrend der Genesung nimmt das Alpha-Globulin ab, wahrend das Albumin ansteigt. Jedoch ist der Ruckgang zum nor-malen elektrophoretischen Bild nicht immer vollendet wenn die klinische Genesung eintritt. Electroforesis en papel de las proteinas sericas en la toxicosis. Se estudiaron las imagenes electroforeticas de las proteinas sericas en 30 lactantes afectos de toxicosis. Durante el periodo de deshidratacion aguda, los cambios mas notables fueron un moderado o marcado aumento de la alfa globulina, especialmente de alfa 2. Durante la convalecencia, la fraecion alfa globulina disminuyo mientras au-mentaba la albumina. Sin embargo, la vuelta a la imagen electroforetica normal no siempre es completa al ocurrir, la mejoria clinica.

Phenol and butanol-acetic acid-water are suitable solvents for bidimensional paper chromatography of amino compounds in the routine clinical laboratory. A chart illustrating the positions of 56 amino compounds on bidimensional chromatograms with these solvents is presented. The behaviour of the compounds studied can be correlated with their chemical structure. Members of homologous series behave in a predictable manner. The effect of inorganic salts is discussed and the importance of maintaining the room temperature at a suitable level stressed. The former affects separation in phenol, the latter in butanol-acetic acid. Desalting of urine and packed red and white blood cells is unnecessary when runs are made at the proper temperatures. Illustrative chromatograms of normal and pathological untreated urines are presented.

The object of this work was to find a rapid and precise method for the separation of the principal monosaccharide constituents of the carbohydrate fraction of wood: glucose, galactose, mannose, arabinose and xylose. Discarding the imprecise methods of classical analysis, we have considered the possibility of the application of paper chromatographic methods. None of the eluents suggested in the literature gives a satisfactory solution to this problem. We have studied the use of a solvent composed of a mixture of ethyl acetate, pyridine and water in the proportions 3.6:1:1.15; we have found that this eluent gives a complete separation of the sugars and allows both qualitative and quantitative determinations.

1. The estimation of unsaturated fatty acids of three varieties of linseed oil is carried out by the paper chromatography technique using131I for saturating the acids. 2. The relationship between the total and individual unsaturated fatty acids content on one side, and the maximal spot activity on the other side, is linear and can be used as standard reference line. 3. Determination of the Rf values of the three occuring unsaturated fatty acids, depends mainly on the change of the specific activity of the iodinating mixture.

The effect of methylamine and methylamine hydrochloride has been systematically investigated in the paper chromatography of various metal ions. It has been found that methylamine hydrochloride like EDTA helps in preventing tailing. A new separation of Fe, Mn, Ni, and Co has been developed. The Rf-values are 0.97, 0.49, 0.08, 0.79 respectively and the solvent system used is ethylacetoacetate/methylamine hydrochloride/conc. hydrochloric acid (20∶2∶7). The values are equally spaced, the spots are compact and the separation is probably the best yet recorded.

The supernumerary (third) spots which separate when binary mixtures of copper(II) complexes of histidine and some other amino acids are subjected to paper electrophoresis were re-examined. It is concluded that, in the early stages of electrophoresis, the spots consist of mixed-ligand complexes composed of copper(II), histidine and other amino acid and that, as electrophoresis proceeds, the complexes undergo tow different reactions, simultaneously. A portion of them re-equilibrates to form homogeneous copper(II) complexes of each of their constituent amino acids, while the remainder decomposes to the relatively stable 1:1 histidine—copper(II) complex and the other amino acid which is liberated as the free, uncomplexed ion. This interpretation is based on electrophoresis in mildly alkaline electrolytes of mixtures containing copper(II) histidnate and the copper(II) complex of either an acidic or a basic amino acid.

Methods are given for the separation and analysis of(i) the methylated sugars obtained from the methylated polysaccharide, and (ii) the glycosides obtained by a SMITH degradation of the mannan. Analysis of a mixture of isomeric methylated sugars was facilitated by successive borohydride reduction, periodate oxidation and borohydride reduction. In this way isomeric C6 sugars were selectively degraded into C6, C5, C4, C3 and C2 alditols which were readily separated by vapour-phase chromatography (v.p.c.). Details are given for the separation of tri-, di-, and mono-O-methyl-d-mannoses. SMITH degradation of the mannan gave a series of glycosides of which i-O- (α- and β-d-mannopyranosyl)-glycerol, i-O-(α- and β-d-lyxofuranosyl)-erythritol and 2-O-(d-lyxofuranosyl)-d-erythritol were separable by paper chromatography. Other glycosides separated by v.p.c. of their trimethylsilyl derivatives were 2-O-(α-d-mannopyranosyl)-d-erythritol, the β-anomer (p-nitrobenzoate, m.p. 235-238°); i-O-(β-d-mannopyranosyl)-erythritol (p-nitrobenzoate, m.p. 239–243°) and the αanomer. The mannan was shown to contain d-mannofuranose and d-mannopyranose residues, to be highly branched and to contain mainly (i → 4) and (i → 6) linkages.

A study has been made of the spreading of the stains produced by low vapor pressure liquids in various types of filter paper and in printflex card. Both the rate of spreading and the distribution of the liquid in the stains have been measured. Semiempirical equations describing the data have been developed. The spreading studies have led to a theoretical stain size drop size relation for low vapor pressure liquids, which may be used for estimating the drop size distribution of a liquid spray. A short time after collection, the stain diameter, D, will be proportional to the drop diameter, d. With such liquids there is no “final stain size.” The spreading factor, Dd, should be proportional to t16, where t is the time after collection. These conclusions have been verified in subsidiary experiments.

Four different chromatographic solvents containing borates of organic bases are described. Two of them permit the separation of sugar nucleotides differing only in the sugar moiety and give also a good resolution for some sugar 1-phosphates. The other two systems permit, in a single run, the separation of a group of sugars which is usually difficult to achieve.

Metabolism of the antioxidant, 2, 6-di-tert-butyl-p-cresol (BHT), was studied in a rabbit. After administration of 0.5g./kg. of BHT by mouth, 37.5% of it was excreted as glucuronides, 16.7% as ethereal sulfates, and 6.8% as free phenols. α-Hydroxy-2, 6-di-tert-butyl-p-cresol, 3, 5-di-tert-butyl-4-hydroxybenzoic acid and 4, 4'-ethylenebis (2, 6-di-tert-butylphenol) were identified by paper chromatography from the rabbit urine 24 hours after the administration of 0.5g./kg. of BHT. Unchanged BHT was isolated from the feces, but not from the urine.

Plasma and sera from normal and leukemic (erythroblastosis) chickens have been studied with the aid of paper electrophoresis. Differences are more easily demonstrable with pH 6.4 phosphate buffer than with barbiturate buffer. There is a relative depression in serum albumin and an apparent 'shift' in the positions normally occupied by the α- and β-globulins. The latter regularly appear to move as one band in electropherograms of sera from leukemic birds. It has been possible to induce this 'shift' in normal sera by storage or incubation.

Selected organic compounds commonly occurring in plant and animal tissues have been chromatographed on glass fiber, paraffin-coated and untreated cellulose paper in a number of different solvent systems. The RF values of these compounds represent new additions to the literature in the solvent systems studied.

Solubilized vat dyes wre separated by paper chromatography, using a methanolic solution of ammonia as solvent system. A system containing pyridine, isoamyl alcohol and ammonia, as well as a mixture of methanol and acetic acid were also used. Both descending and centrifugal chromatography were employed. The relation between dye constitution and chromatographic behaviour has been examined.

The low-voltage and high-voltage paper electrophoretic behaviour of iodide, iodate, periodate, tellurite and tellurate ions has been studied in sodium hydroxide and sodium chloride solutions used as electrolyte. Iodide, iodate and periodate can be very well separated in 0.01–1.0 N NaOH or in sodium chloride solutions of concentration less than 0.5 N. Simultaneous separation of iodide, iodate, periodate, tellurite and tellurate could not be realised. One could, however, obtain the following separations: tellurate, tellurite, (iodate + periodate), and iodide in NaCl by low-voltage electrophoresis, periodate, tellurite, iodate and iodide by high-voltage electrophoresis in sodium chloride (< 0.5 N).

Summary Six solvent systems, two of them of “reversed phase” type, are recommended for the paper chromatographic separation of narcotics. A chromatographic pattern obtained in these systems will facilitate the identification process.