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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.

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.

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.

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.

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.

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.

Three products formed during the paper chromatography of glycine with water-saturated phenol as a solvent have been investigaetd. These unknowns were stable condensation products whose formation was dependent upon the presence of glycine ad formaldehyde. They were ninhydrin sensitive. Alkaline hydrolysis yielded glycine. For chromatographic practices, the amount of glycine or formaldehyde which may be converted into the condensation products will depend upon the amount of the other component in the mixture. Addition of 1% formaldehyde normally complexes all of the glycine. In usual chromatographic experiments part of the glycine may be complexed by residual formaldehyde present in chromatographic phenol. Glycine condensation may be minimized by redistilling the phenol solvent with aluminum turnings and sodium bicarbonate and by washing the filter paper with 1% oxalic acid.

A paper chromatographic system was developed for the resolution of mixtures of sulfated mucopolysaccharides. The effect of varying the ammonium formate buffer/isopropanol ratio in the developing solution on the RF values of heparin and chondroitin sulfate is presented. A complete separation of heparin from chondroitin sulfate was obtained with three solvent mixtures containing different proportions of buffer and isopropanol. One of these solvent mixtures, ammonium formate buffer-isopropanol (65:35) v/v, resolved commercial beef heparin into two principal components, on “fresh” paper. This system was used to compare beef heparin with mouse mast cell tumor heparin, rat heparin, sheep lung heparin, chondroitin sulfate, and β-heparin. A common metachromatic component with an RF of approximately 0.5 was0found in all of the heparin preparations chromatographed with this solvent mixture. “Aging” of the filter paper resulted in the loss of the ability of the buffer-isopropanol mixture (65:35) v/v to resolve the beef heparin. Increase of the buffer-isopropanol ratio to (70:30) v/v restored the resolution of the beef heparin on “aged” paper. Heparitin sulfate and α-heparin monosulfate were compared to beef heparin using the latter system.

A wide range of Δ 4 - and Δ 1 , 4 -3-ketosteroids, from weakly polar to strongly polar, may be chromatographed and detected on cellulose triaacetate paper, with benzene—methanol—water mixtures as developing solvents. Three systems have been worked out. In the first a one-phase mixture of benzene, methanol and water is used as the mobile phase and the paper is not accommodated. A “reversed phase” type of chromatogram is obtained, i.e. the more polar compounds have higher R F values than the less polar ones, as is usual for chromatography on acetylated paper. If, howerver, two-phase mixtures of benzene, methanol and water are used, as was the case in the other two methods, “reversed phases”, as well as “convetional” type chomatograms may be obtained. This depeds upon whether the lower or the upper layer is used as the mobile phase, the other layer being used for accommodation of the paper. Perhaps the first-mentioned method should, in contrast to the other two, be regarded as adsorption chromatography, rather than as partition chromatography. Well-reproducible R F values are obtained and, especially with the two-phase mixtures, compact spots are found. Detection methods and R F dta are given for various Δ 4 - and Δ 1 , 4 -3- ketosteroids, ranging from C 18 to C 27 compounds and including sex hormones and adrenoccortical hormones.

A silicic acid paper chromatographic method for the separation of four major classes of cholesterol esters, namely saturated cholesterol esters, cholesterol oleate, cholesterol linoleate and cholesterol linolenate plus cholesterol arachidonate is described. This method uses a benzene-hexane solvent. The use of combined column and paper chromatography as well as gas chromatographic analysis is presented for definitive identification of the various spots. The predominance of cholesterol linoleate in human plasma, and cholesterol arachidonate in rat plasma is confirmed.

A paper Chromatographic method for the analysis of phenylalanine in serum is described. The amino acids, including phenylalanine, are isolated from the deproteinised serum by means of a cation exchanger. The phenylalanine is then separated by paper chromatography at elevated temperature and estimated colorimetrically. Normal values are given. The excretion of phenylalanine and phenylpyruvic acid is correlated with the serum phenylalanine level. The method is specific and the reproducibility is good. Its application in a case of phenylketonuria is reported.

A study has been made of some of the fundamental factors involved in obtaining good separations and reproducible chromatograms, with the technique of contrifugally accelerated paper chromatography. A study was also made of variables which might influence R F values. Mixtures of bromphenol blue, methyl orange and methyl red were fractionated as well as solutions of lencine, methionine and glycine. For the separation of the dyes, Whatman No. 1 filter paper was used with a veronal buffer solution of ionic strength 0.05 and having a pH of 8.7. For this system, it was found that the R F values of the dyes were essentially unaffected by changes in rotational speed of the rotor from 300 to 925 r.p.m. It was found that the rate of addition of solvent was important in developing satisfactory chromatograms. A flow rate of 1.15 ml/min was found to be satisfactory at all rotational speeds above 250 r.p.m. The shape of the developed chromatogram was elliptical rather than circular, for all four types of paper which were studied. For Whatman No.1, the major axis of the ellipse was parallel to the machine direction of the paper. The R F values for the dye components were essentially the same for the following papers: Whatman No. 1, Eaton-Dikeman No. 613, and Cremer-Tiselius-Munktells. The amino-acid mixture was fractionated using Whatman-3MM paper and a solvent system of butanol-glacial acetic acid and water, in the volume ratio 40:10:10, respectively. The disk was rotated at 975 r.p.m. With a solvent flow rate of 1.2 to 2.1 ml/min, sharply defined zones were obtained. The time of development was 15 min for a flow rate of 1.2 ml/min and 7 min for a flow rate of 2.1 ml/min. The R F values obtained were 0.85 for lencine, 0.56 for methionine and 0.20 for glycine.

On account of differences in lipophilic character it is possible to separate p -nitrophenyl methylphosphonates and their sulfur analogs by reversed phase paper chromatography. Whatman No. 1 filter paper was impregnated with silicone oil 550; the upper layer of a chloroform-ethanol-water (5:5:3, v/v) system served as mobile phase. The relation R M = log (1/ R F —1) holds for homologous series of O-alkyl p -nitrophenyl methylphosphonates (I) and S-alkyl p -nitrophenyl methylphosphonothiolates (II) up to the n -pentyl homologues. Using a combination of Gibbs' and Dragendorff reagents it was possible to distinguish the phosphonates, phosphonothiolates and phosphonothionates.

After conversion to 2,4-dinitrophenylhydrazones, keto acids in biological fluids were extracted into isopropyl ether followed by NaHCO 3 and finally into ethyl acetate. The derivatives were chromatographed two-dimensionally on Whatman No. SG 81 paper. Limitations of the extraction and chromatographic procedures are discussed but it is considered that the method is superior to orthodox one-dimensional paper chromatography.

Following previous supplements in this series, a further 180 compounds were analyzed by paper chromatography in six solvent systems. The data originating from 15 standard colour reagents are also recorded and presented. The following types of compounds are described: flavonoid derivatives, aliphatic and aromatic aldoximes and ketoximes, substituted benzamide derivatives, aminobenzene derivatives and heterocyclic amines, N-nitroso compounds, N-oxides of tertiary bases, pyrimidine and indole derivatives and sulphonamide drugs. The paper chromatographic mobility patterns were characteristic, especially for 1,3- and 1,4-hydroxybenzaldoximes and ketoximes of the hydroxyacetophenone type. All the heterocyclic bases investigated showed mobility patterns similar to those of the alkaloids described in earlier supplements. Mobility patterns similar to the alkaloids were produced by some of the mercaptopyrimidines. In general amino-, diamino-, hydroxy and dihydroxy derivatives of pyrimidines showed low mobility in these solvents. Several flavonoid compounds were found to be Erhlich positive and a magenta colour was produced by l -epicatechin. Most of the N-unsubstituted benzamides were detected with p -dimethylaminocinnamaldehyde, and most of the N-nitroso derivatives of secondary amines and N-oxides of heterocyclic tertiary amines were revealed by this reagent.

A paper chromatographic solvent system and an extraction and concentration microtechique were used to determine air-borne 1,2:3,4-dibenzopyrene, 3,4-benzofluoranthene, 3,4-benzopyrene, 1,12-benzoperylene and anthanthrene in an automobile tunnel, in frequented cross roads, ad in sites with little or no automobile traffic. Fluoranthene, pyrene, chrysene, 1,2-benzanthracene, 1,2:5,6-dibenzanthracene, 11,12-benzofluoranthene and coronene were identified on the same chromatogram, 1,2-benzopyrene was presumed. Using at least three filters in series, quantitative air-sampling of penta- and higher cyclic hydrocarbons is claimed. The results are tabulated.

A two-dimensional paper chromatographic method is described which allows good separations and spectrophotometric determinations of nanomole amounts of hypoxanthine, xanthine, uric acid and uric acid riboside. The method can be applied to biological material containing adenine nucleotides and considerable amounts of free amino acids.

The RF values of inorganic ions on paper pre-treated with 0.25–2.0 % cellulose acetate and impregnated with various concentrations of 2-thenoyltrifluoroacetone (TTA) was studied by using acetate buffer as a developing agent. 1% cellulose acetate pre-treated paper impregnated with o.2 M TTA was found suitable for reversed phase paper chromatography. The RF values of inorganic ions were examined on the impregnated paper by using various pH's of the developing agent (0.1 M acetate buffer). Separations of PbCu, CoCu, ZnCu and FeCoNi were performed in this system.

The mammalian respiratory tract produces a secretion in which the major macromolecular components are mucin-type glycoproteins. High-voltage paper electrophoresis has proved a valuable research tool in studies of the formation of these glycoproteins. Separations of hexoses, amino sugars, oligosaccharides, sugar nucleotides and mucin glycoproteins are rapidly and easily obtained by paper electrophoresis at voltages of 50 to 100 V/cm. The use of high-voltage paper electrophoresis to follow amino sugars interconversions and the transfer of glycosyl units to mucin acceptors is described.

Preparative paper chromatography (PPC) for the purification of synthetic compounds labelled with 14C : choline, chlormequat, glutamic acid, indoleacetic acid and urea is described. The influence of some parameters as time, amount of substances used and geometry factors (dimensions of paper sheets) on the repeatibility of PPC are given.

A machine is described for the reproducible treatment of paper chromatograms with liquid chemical reagents. Strips of filter paper 5 cm wide are passed through the machine by two pairs of belts which grip the edges of the strip leaving the central area exposed on both sides. A film of liquid reagent is fed on to the undersurface of the paper strip by means of a roller and trough which is fed from a simple constant level device. The reagent-soaked strips pass immediately into a straight tunnel into which a stream of hot air is introduced by ports opening from manifolds running along the sides of the tunnel. The air stream is passed out of the top of the tunnel by means of an exhaust tube and fan connected to its distal end. The strips are passed out of the machine by disengagement of the belts which are returned to the starting point of the machine after being washed, and dried on their return passage through the hot air tunnel. Operating temperatures up to 150° can be achieved and are adequate for producing quantitative and complete reactions with the majority of color reagents that are useful for the quantitative estimation of substances by direct scanning of paper chromatograms (e.g. Zimmerman reagent, ninhydrin, phosphomolybdic acid). The mechanization of the treatment of paper chromatograms with color reagents improves greatly the reproducibility of the results, the evenness and reproducibility of background, and enables the successful use of color reactions which cannot be controlled sufficiently by manual methods to allow their quantitative estimation. It also enables the treatment of very large numbers of chromatograms: the present apparatus is capable of treating 40 cm long chromatograms at a rate of up to 88 per h or over 500 per day. The development, important features of design, and method of operation of the machine are described.

Reversed-phase partition chromatography was used to separate the rare earths, thorium and uranium on papers treated with methylenebis(di-n-hexylphosphine oxide), MHDPO. The separation factors between adjacent rare earths were found to increase with the acid concentration in both nitric and hydrochloric media with the larger separation factors in nitric acid. Opposite trends were observed in hydrochloric and nitric acid. In the former case the partition coefficient increased with acid concentration, but in the latter case it decreased. This was explained on the basis of the solvation energies of the cations and anions, the mass action effect and the competitive extraction of nitric and hydrochloric acid. Solvation number studies indicated a 1:1 adduct between MHDPO and the rare earth nitrates. Corresponding studies on the rare earth chrlorides gave anomalous results. It was postulated this was caused by non-equilibrium conditions.

The mobilities in free solution of twenty organic ions have been calculated from their sizes, shapes and charges. From these mobilities the zone mobilities of the ions in paper electrophoresis, relative to the zone mobilities of butylammonium ion (for cations) or picrate ion (for anions), have been calculated by taking account of the extent to which the ions are retarded by reversible adsorption on the paper. Results in fair agreement with experiment were obtained for most ions having approximately ellipsoidal shapes; however, the experimental mobilities of cations and anions derived from l -amino acids are systematically lower than calculated. A comparison is made with relative mobilities calculated according to JOKL.

The mechanism of peak broadening in paper chromatography is investigated by comparing the peak widths obtained in chromatography with those caused only by diffusion, for a set of amino acids of widely differing RF values and with eight kinds of Whatman paper. The results show that longitudinal diffusion occurs both in the mobile and the stationary phase in paper chromatography. The mass transfer term is composed of a contribution of unevenness of flow throughout the paper and a contribution of the slowness of attainment of the partition equilibrium between the mobile and the stationary phase. The resistance to attainment of equilibrium resides predominantly in the mobile phase.

Two phase solvent mixtures are described in which di-substituted steroid ketones and esters have partition coefficients such that they may be readily separated by partition chromatography on paper. 80 steroids and derivatives have been characterised in these solvent systems and their RF values are presented. Notable features include the resolution of otherwise similar and ostane and pregnane derivatives, 3-hydroxy-17-ketone from 17-hydroxy-3-ketones in the androstane series, aetiocholanolone from its -9: ii - unsaturated analogue and substantial inversion of general order of separation as compared with the order observed in procedures for unesterified steroids. Up to 0.5 mg of steroid or derivative per cm2 may be chromatographed from each origin.

A simple quantitative procedure for sugar analysis of plant materials by paper chromatography with good reproducibility is described. The alcoholic extract was partially purified by a tertiary solvent system and ion exchange resins and then paper chromatography with the following solvent: water-saturated n -butanol, 95% ethanol and trichloroethylene (6:2:2, v/v). The sugars were selected by a guide strip technique, and eluted from the “cut-out squares” of paper with 3 ml distilled water by a micro-elution tube, here described. Theyb were determined by a micro-colorimetric method. The method permitted 72 determinations to be done at one time.

A method for the determination of several impurities in reactor grade uranium compounds by post-irradiation paper chromatography is described. The bulk of the uranium is removed before irradiation by spotting the uranyl nitrate solution on to a Schleicher & Schull 2043 A paper, then chromatographing with diethyl ether containing 5 vol. % nitric acid as solvent.

A method for the quantitative determination of gibberellic acid in fermentation media, using descending paper chromatography in butylacetate-water is described. A sample is adjusted to pH 2.5–3.0, extracted with n-butanol, and 0.05 ml. organic layer is spotted on Chromatographic paper. After equilibration of the atmosphere in the jar, the chromatogram is developed in butylacetate saturated with water, for 7 hours, and the solvent is allowed to drip off the bottom of the sheet. Detection is carried out by spraying with a 3% solution of sulphuric acid in methanol and after drying the paper, spots with blue u.v. fluorescence are observed. The area of the spots is evaluated with the aid of a calibration curve, plotted from values of standards corresponding to 20, 60 and 120 μg. gibberellic acid. The error of estimation is ±10–15% when estimations are carried out carefully. The lowest limit of sensitivity is 5 μg.

Ion exchangers with few reactive groups such as cellulose sulphonic acid paper do not distinguish between ions of different charges in the law of mass action equation. It is suggested that this is due to the large distance between the exchange groups. Similar results are obtained with papers impregnated with very dilute solutions of liquid ion exchangers.

CHANG (1955) reported the lowering of the blood hemoglobin of chicks due to free gossypol feeding and postulated that free gossypol in the diet leads to interference with the synthesis of the proteins of the blood in these chicks. In view of the useful information that has been obtained in recent years using the electrophoresis technique in the study of normal and pathological sera, an experiment was planned to study the electrophoretic patterns of serum samples obtained from birds fed a normal diet, as well as from birds fed free gossypol in the diet. An electrophoresis cell using filter paper strips, as originally designed by Durrum (1950) and marketed by Spinco, was employed. Later it was discovered that the resolution of the globulins of chick serum is very poor on filter paper. More serum samples were obtained from birds used in a study of the effect of protein level on …

Chromatographic separation of oleic acid-I131 and triolein-I131 from free iodide has been studied in a number of solvents on ordinary chromatographic paper as well as paper impregnated with various heavy metals. The latter help in firmly fixing the iodide at the point of spotting. Copper and lead oleates which can form on the impregnated papers move along with oleic acid. Bismuth, mercuric, palladium, and silver oleates are not formed on the impregnated papers.

This paper describes the method for the separation and identification of zinc dithiophosphates having alkyl or aryl substituents by partition paper chromatography.Of a variety of solvent systems examined, the solvent system of cresyldiphenyl phosphate (stationary phase)-90/10 isooctane•n-cetane (mobile phase) was most suitable for the separation and identification of zinc dialkyldithiophosphates. Their Rf values with this solvent system increase with the number of carbon atoms in the alkyl group. Among zinc dialkyldithiophosphates having isomeric alkyl groups respectively of straight primary, branched primary and secondary, the Rf values increase in this order. The solvent system of liquid paraffin-isopropyl alcohol was found to be satisfactory for partition paper chromatography of zinc dialkyldithiophosphates having long chain alkyl groups.A large difference in the Rf values between zinc dialkyldithiophosphates having isomeric alkyl groups respectively of primary and secondary, was observed when the solvent system of polyoxyethylene octylphenylether (E. O. 10mol)-n-heptane was used and similarly, a large difference in the Rf values between zinc dialkyldithiophosphates having isomeric alkyl groups respectively of straight primary and branched primary, was observed when the solvent system of chlorinated paraffin-isopropyl alcohol was used. It is effective especially for the analysis of mixtures of zinc dialkyldithiophosphates to use these solvent systems together with the solvent system of cresyldiphenyl phosphate-90/10 isooctane•n-cetane.It was found that zinc diaryldithiophosphates can be distinguished from zinc dialkyldithiophosphates by the solvent system of chlorinated paraffin-isopropyl alcohol and identified by the solvent system of cresyldiphenyl phosphate-90/10 isooctane•n-cetane or liquid paraffin-isopropyl alcohol.Zinc dithiophosphates in commercial antioxidants and EP hydraulic oils could be identified by the above mentioned solvent systems.

The application of cellulosic anion-exchange papers to the separation of tracer amounts of rare earths has been investigated, using citric acid solutions as eluents. It was found most satisfactory to use diethylaminoethyl-cellulose paper with 0.026 M citric acid (pH 2.4) as eluent. Under these conditions a practical separation factor of 2.6 between europium and cerium was obtained. Slightly higher separation factors could be obtained by adding ethanol to the citric acid solutions.

A quantitative study has been made of the separation of uroporphyrin octmethyls esters I and III by thin-layer chromatography using cellulose powder containing 10% gypsum (MN300G) and a kerosen—dioxan system. The separation by paper chromatography has been further studied. Thin-layer chromatography offered no apparent advantage over paper chromatography. Results by both methods indicated some interaction between the two porphyrin isomers leading to inability to separate quantitatively uroporphyrin ester mixtures when the III isomer comprised 65% or over. Paper chromatography is still of great value for separation of the I and III isomers when unoporphyrin III ester is less than 65% and also for the detection and determination of pseudouroporphyrin.

A method is described for the fractionation fo DNA preparations on DEAE- cellulose paper. This paper can be used as strips or as centrifuged paper pulp. In both cases, similar results are obtaiend. The reproducbility and the senstivity of the fractionation are very high. During the fractionation process, the desorption of the macromolecular species is complete. Some of the fractions obtained can be rechromatographed and eluted in the same position. The additivity of the fractions evidences their preexistence in the preparation. The use of cellophane strips enables the complate fractionation to be run in ten hours; with centrifuged paper pulp, a complete elution requires two hours. The physicochemical properties of the DEAE-cellulose paper allow a desorption of the highest molecular weight DNA molecules at neutral pH. These results are discussed and several examples of fractionation of polynucleotides derived from DNA are given.

Two-dimensional paper chromatography has been adapted to the study of fat-soluble materials, which are thereby displayed mapwise on one sheet. The procedure, applied to a variety of plant tissues, has led to the discovery of reducing substances, pigments (including a new chlorophyll), fluorescing substances, quinones and unidentified phyllolipids; has helped in the differentiation of viruses; has uncovered a new enzyme or a new activity of an old one; has amplified knowledge on 20 substances that accompany chlorophyll and that may form part of the photosynthesizing apparatus; has aided the study of carotenoids from flowers, and indicated how mapping may be used in studying the origin of hybrids; has shown that mono-esters of carotenoids occur naturally, and given a hint that they may be common; and has played a major part in investigating the distribution, fluctuation and biochemical role of tocopherols in plants.

After tretment of paper with a liquid anion exchanger (tri- n -octylamine) the differences in the stability of the metal complexes may be used to improve the analytical separation of some important fission products and actinides by paper chromatography. The R F values of Y, La, Ce, Zr, Ru and Th, Pa, U, Np, Pu, Am with nitric acid and nitrates as eluents are given. Quantitative determination by scanning the radioactive nuclides is possible and some methods are proposed. The oxidation states on Np and Pu are identified by their different R F values. The composition of mixtures with different valency states of the metals may be determined quantitatively.

Summary The properties of a commercial chromatography paper (Whatman No. 4) were studied with water and nine organic fluids. Imbibition was greater with the more polar fluids and ranged from 22.0 mm3/cm2 for water to 11.9 mm3/cm2 for benzene. Measurements of flow rates under the influence of gravity showed that, the more polar the fluid, the smaller the resistance to flow offered by the paper. The rate of water was 0.23 ml per hour per cm width at 20°.

A method for the determination of urinary normetadrenaline and metadrenaline is described whereby these 3-O-methoxy metabolites of noradrenaline and adrenaline are separated by paper chromatography before estimation by Pisano's method. Results in 80 normal subjects are given.

Simple paper chromatography of urine obtained in the Figlu test has been found satisfactory for determining excess excretion of the imidazole metabolites of histidine. It has been confirmed that examination for these in addition to formiminoglutamic acid is necessary for a full assessment of this test. When excretion of the latter is not abnormal in folic-acid and/or vitamin B(12) deficiency, that of urocanic acid, a glycine conjugate of urocanic acid and/or imidazole propionic acid, is usually raised. An abnormally high excretion of hydantoin-5-propionic acid after histidine has also been found in these deficiencies, and its estimation may be of additional use in the evaluation of the Figlu test.

The technique of paper chromatography has been adapted to the identification of gluconic, 2-ketogluconic, and α-ketoglutaric acids. Combinations of methyl and ethyl alcohol were found to be the most suitable solvents and ammoniacal silver nitrate was found to give the most satisfactory reaction. When grown under normal physiological conditions where glucose was metabolized to carbon dioxide and water, Pseudomonas aeruginosa 9027 was shown to have oxidized glucose by way of gluconic and 2-ketogluconic acids. Since a strong system for oxidizing both gluconic and 2-ketogluconic acids was demonstrated, the presence of these acids over at least an eight hour period of growth is taken as evidence of their importance as intermediates in the oxidation of glucose by this organism.

Both ascending thin-layer chromatography on Silica Gel G with developers containing ethers and ascending paper chromatography with dialkyl ethers on papers impregnated with formamide were successful in the separation of the monoximes and dioximes (including their possible stereoisomers) of furil, benzil, biacetyl and cyclohexane-1,2-dione. Selectivities of various separation and visualisation methods as well as the detection limits for some individual oximes were examined and evaluated. Spraying of the developed chromatograms with ammoniacal solutions of Ni(II) and Cu(II) salts followed by UV irradiation for the isomeric oximes (γ, β) and/or by oximation of the monoximes gave the best results in checking the purity and isomeric composition of the oxime reagents tested.