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Filter paper moistened with solutions used in electro-chromatography was spotted with 0.5-1.0μl of solutions of mucopolysaccharides and allowed to air dry. Substances tested with respect to their staining reaction were as follows: (a) From commercial sources: hyaluronic acid, heparin, chondroitin sulfate, ovomucoid and gastric mucin, (b) From natural sources: blood serum, saliva, tears, vitreous filtrate and aqueous humor. Alcian blue was found to be a good general stain for mucopolysaccharides and for locating such material on filter paper, especially when more specific means were used subsequently for identifying the kind of mucopolysaccharide present. Staining by colloidal iron of materials on filter paper was similar to that by the periodic acid-Schiff reaction. However, heparin and chondroitin sulfate were not stained by iron when on filter paper but were stained when placed on glass slides.

Cernik, A. A., and Sayers, M. H. P. (1971). Brit. J. industr. Med., 28, 392-398. Determination of lead in capillary blood using a paper punched disc atomic absorption technique. Application to the supervision of lead workers. The presence of lead in blood is the most incontrovertible evidence of absorption but hitherto the need for venepuncture has limited its determination in the supervision of industrial workers. Micro-methods using atomic absorption spectrophotometry (AAS) have, however, made possible the development of a sufficiently reliable test using a drop of blood obtainable by ear prick for use in the field for screening purposes. A micro-sampling method by AAS is compared with a routine polarographic procedure (POL) using venous blood (corr. coeff. = 0·990). The pipetting of microlitres of blood can be eliminated by spotting the blood onto filter paper, allowing it to dry in air, and then using a punched-out standard disc of dried blood for analysis instead. Correlation of this method (PD) with the micro-sampling technique (AAS) is good (r=0·981). The PD method using capillary blood also correlates acceptably with the micromethod using venous blood (r = 0·913). A pilot field study using capillary blood estimated by the PD technique showed that with this method blood can be collected by ear prick in factories for monitoring workers in the lead industry, thus eliminating the need for routine venepuncture.

Paper chromatography has recently been applied to the separation and estimation of the individual tocopherols. The present communication draws attention to errors that are likely to arise if a solution of ‘Vaseline’ petroleum jelly in ethyl ether is used to impregnate the filter paper in order to achieve phase-reversal. The use of liquid paraffin B.P., dissolved in light petroleum, not only overcomes these objections, but enables the elution after chromatography to be simplified. Losses of tocopherols are minimized by chromatography in an inert atmosphere. The modified technique, which is described in detail, has consistently given recovery figures of 95%, or better, when tested with mixtures of synthetic tocopherols.

Methodology for obtaining reproducible measurements of the relative mobility of organic compounds over pH 2–12 using high-voltage paper electrophoresis is described. Use of the method for the elucidation of some structural features of simple molecules is illustrated by applying it to the characterization of metabolites of drugs excreted in the urine and bile of rats. The compounds studied included p -nitrophenol, 1-naphthol, salicylic acid, 4-nitrocatechol, probenecid, phenol red, and their metabolites.

Methods are given for determining individual tocopherols in natural oils and food-stuffs. These methods consist of either a single-dimensional separation on zinc carbonate-impregnated paper, or this separation in conjunction with a second-dimensional partition separation on paraffin-coated paper. Chromatographic runs are short and rapid and can be made without preliminary removal of carotenoids and moderate amounts of vitamin A or sterols. Over 98% recoveries are obtained with pure tocopherols. The presence of the recently described 5-methyltocol in wheat products has been confirmed, and, in addition, using two-dimensional methods, a substance has been isolated from some cereal grains, which appears to be a new tocopherol.

SummaryUsing paper electrophoresis, satisfactory separation of protein components in salivary secretions of the rat was most conveniently obtained with parotid saliva. This secretion was readily ob...

A simple method is described for determining the uric acid: creatinine ratio (UA/C ratio) in urine collected on filter paper. Uric acid and creatinine can be eluted quantitatively from Schleicher and Schuell 903 filter paper in alkaline buffer; the eluate is analyzed for the two compounds simultaneously by an automated method. The mean UA/C ratio was defined in 820 newborn infants in the postnatal period of life. After the third day, the average is 1.49 ± 0.60 (mean ± 1SD), falling from a high value of 2.25 ± 0.85 on the second day. There is no significant sex difference or diurnal variation at this age and neither gestational age nor birth weight apparently affects the ratio. The technique can be used to screen population, particularly the neonate after the 3rd day of life, for disorders of purine metabolism. An abnormal ratio (i.e., above 3.29 or > 3SD above the mean) would indicate such a disorder. Transient high UA/C ratios can be anticipated in at least 0.5% of samples often because of acquired perinatal complications.

A simple paper chromatographic method was found to be effective for the study of phenylketonuria and tyrosyluria. It proved to be much more reliable than conventional tests for the detection of abnormal amounts of ketoacids in urine and was suitable and convenient for following the effects of dietary variations on the excretion of these compounds. Information obtained about ketones other than those of primary importance to the conditions studied included confirmation of the excretion of p-hydroxyphenylpyruvic acid in phenylketonuria. A modification of the method applicable to histidinaemia was devised.

Results are reported on the analyses of milligram quantities of some pure and crude steroid preparations. Samples are chromatographed on paper, and the entire sheet is rapidly eluted by a previously described technique. Components, in concentrations of 0.2 per cent or less, can theoretically be separated and detected. This technique may be found of equal benefit in the study of other compounds which can be separated by paper chromatography.

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.

A paper Chromatographic technique on salt-buffered papers for separating the alkaloids of opium has been devised. The method has been used for determining the origin of opium samples by studying the pattern of theRf values of the alkaloids present and the effect on it produced by the presence of various adulterants.

In few conditions is the electrophorogram of serum proteins capable alone of deciphering the diagnosis. In conjunction with the assembled clinical data and relevant applied pathological tests the definition of the disease is often clarified. The patterns as observed by the applied pathologist are serially discussed and the importance of the variations in different fractions presented. In several instances, exemplified by liver disease, the assistance rendered toward prognosis has been considerable. Perhaps the most clearcut diagnosis is possible in the agammaglobulinaemia syndrome which is immediately evident by the technique of paper electrophoresis. Current advances are mainly in the direction of estimating the protein-bound carbohydrate and lipid constituents which are believed to reflect the abnormalities in several infections and metabolic diseases. Utilisation of non-ionic detergents has also allowed separation of subfractions of the globulins into numerous new components, including lipo-proteins, some of which had been previously observed during the application of improved buffer systems designed to restrict interference of heavy metals and to facilitate accentuation of fractions having intermediate mobility.

Separation of hypoglycemic agents as tolbutamide or similar compounds is carried out on paper chromatograms using different developers. Detection of the spot contents has been done by new colouring agents which proved to be good tools for resoltion of the spot. Among these reagnets, ninhydrin proved to be a useful tool for determination of hypoglycemic agents under investigation. Radioactivation of the dried developed chromatograms, using I131 has been applied as a tool for measuring the Rt-values of the same hypoglycemic agents. Invetigation on S35-labelled hypoglycemic agents prepared in our laboratories proved that paper chromatography accompanied by radioscanning or autoradiography of the dry chromatogram could be used for the separation, detection, and determination of these agents.

An indirect immunofluorescence test using small discs (0.4 cm in diameter) cut from blood smears on filter paper is described. Discs plus three drops of phosphate-buffered saline are laid on antigenic areas on slides, which are then incubated. During incubation, eluted antibodies react with antigen. The method was tested with 10 eluates from healthy persons and with 30 eluates from three groups of 10 patients with Chagas’ disease, schistosomiasis Mansoni and South American blastomycosis, respectively. Sera from the same blood samples were tested also by conventional indirect immunofluorescence tests. Both tests gave similar results. The method proposed is useful as a rapid qualitative screening test when dealing with a large number of samples.

— The use of paper electrophoresis for the routine examination of urinary proteins in the dog has been studied. In the majority of dogs with nephritis the A/G ratios of the urinary proteins were found to be greater than those of corresponding sera and the electropherograms were fairly well defined. In a few cases the ratios were much smaller, and they were usually associated with high relative percentages of the β- and γ-fractions. Post-renal protein due to cystitis alone produced poorly defined electropherograms. These also resulted when both conditions were present. Urinary proteins with high relative percentages of β- and γ-globulins were also investigated and their electropherograms compared with those of γ-globulin preparations and with seminal plasma from male dog ejaculate. Urinary proteins from dogs suffering from a combination of diseases, e.g. nephritis and liver disorders, gave electropherograms that were not characteristic, although low A/G ratios appeared to be associated with prostatitis, with a case of splenic haematoma, and with some cases showing severe liver damage. It is concluded that paper electrophoresis of urinary proteins, particularly when performed on small “grab” samples has only limited use in routine clinical diagnosis. Resume— L'tlectrophorere au papier a ete employee pour I'etude des proteines dam l'urine du chien. Chez la majorite des chiens le rapport A/G dans l'albumine de l'urine a ete trouve augmente. Dans peu de cas, le rapport etait plus petit, quand le pourcentage Beta et Gamma etait plus eleve. L'albuminaire due a des cystites ne donnait que des electrophograms mediocres. Il en etait de meme quand une nephrite venait compliquer une cystite. Les proteines urinaires ayant une forte teneur de beta et de gamma globulines furent egalement etudiees, et leur electropherogramme compares au liquide seminal de chiens males. Les proteines urinaires de chiens souffrant simultanement de plusieurs maladies telles que nephrite et desordres hepatiques, donnerent des cliches peu caracteristiques, bien qu'un rapport A/G diminue semble etre associe a une prostatite, a un hematome splenique, et dam quelques cas de graves leions hepatiques. La conclusion est que l'electrophorese au papier, des proteines de l'urine, est d'un interet clinique limite, particulierement quand la quantite d'urine fournie est petite. Zusammenfassung— Die Anwendbarkeit der Papierelektrophorese zur Untersuchung der Harn-proteine des Hundes wurde studiert. Es konnte festgestellt werden, dass in der Mehrzahl der Hunde, die an Nephritis litten, die A/G Quotienten der Harnproteine hoher waren als der entsprechenden Sera und die erhalten Elektro-pherogramme waren ziemlich scharf definiert. In einigen wenigen Fallen waren die Quotienten betrachtlich kleiner und dann fand sich gewohnlich auch ein relativ hoher zugehoriger Anteil der Beta und Gamma Fraktion. Postrenales Protein, das lediglich durch Zystitis hervorgerufen war, zeigte verwaschene Elektrophorogramme. Solche Resultate wurden auch dann gesehen, wenn beide Erkrankungen zusammen vorlagen. Harnproteine mit einem relativ grossen Anteil von Beta und Gammaglobulin wurden ebenfalls untersucht und die erhaltenen Elektrophorogramme wurden mit solchen verglichen, die man rnit Gammaglobulin oder mit der Samenflussigkeit mannlicher Hunde erhielt. Hamproteine von Hunden, die an einer Kombination von verschiedenen Erkrankungen litten, 2.B. von Nephritis und Leberkrankheit, ergaben uncharakteristische Elektrophorogramme, es erschien jedoch, dass niedrige A/G Quotienten bei Prostatitis, bei einem Fall von Milzhaematom und bei einigen Fallen rnit schwerer Leberschadigung gefunden wurden. Es kann abschliessend gesagt werden, dass die Papierelektrophorese der Hamproteine, desonders wenn sie nur an gelegentlichen kleinen Proben ausgefuhrt wird, nur einen beschrankt klinisch diagnostischen Wert besitzt.

The alkaloids selected were atropine, cocaine, codeine, morphine, dionine, dilaudid (dihydromorphinone), heroin, strychnine, ineperidine and papaverine. After the alkaloids (100 pig. in a 2.5% solution of appropriat.e solvent) had been applied at the bottom of rectangular sheets of filter paper, they were chromatographed in a container made from a conventional aquarium covered with a heavy glass plate. Since variations � � �

A rapid filter paper disk assay for cyclic AMP based on the activation of a cyclic AMP dependent protein kinase from rabbit skeletal muscle is described. This procedure permits determination of as little as 0.2 picomoles of cyclic AMP. In addition this procedure employs ATP-γ-33P instead of ATP- γ-32P.

The effect of the ratio of toluene to isobutanol in the mobile phase on the R f values of alkaloids occurring in poppy pods is investigated. The optimum conditions for the Chromatographie separation of morphine and codeine from accompanying alkaloids are selected. This makes possible the quantitative determination of these two alkaloids in solution, in extracting alkaloids from poppy pod by an adsorption method. The necessity of using the ether-water solvent system for the quantitative determination of narcotine, papaverine, and tebaine by paper chromatography is confirmed.

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

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.

A PAPER Chromatographie study of urinary aminoacids has been carried out in 69 affected and 565 unaffected relatives from 64 families with various types of muscular dystrophy. Only a small proportion of subjects had either generalized or specific hyperaminoaciduria.

Various birch-pollen and dog-epithelium allergen extracts were chromatographed using Sephadex G-75 and Sephadex G-200 columns. The chromatographic tractions were then tested by an in vitro paper disc radioallergosorbent test, using sera from allergen-sensitive patients. Calibrated columns were used to approximate molecular weights of some birch-pollen and dog-epithelium allergenic active components.

Birch pollen and dog epithelium allergen extracts were studied by ultracentrifugation in linear sucrose gradients. The various sucrose-gradient fractions were then tested for allergenic activity by the paper disc radioallergosorbent test, using sera from allergen sensitive patients.

Limits of detection of some pollen flavonoids are recorded. The flavonoids were detected by aluminum, beryllium, and zirconium ions. Paper chromatography revealed dactylin and isoquercitrin to be present in extracts of orchard grass pollen. Only dactylin was found in timothy pollen extracts.

Summary In 21 premature infants, the birth weights of whom ranged between 1500 and 2400 g and the ages between 1 and 56 days, the total serum protein values have been determined and the serum protein components have been electrophoretieally differentiated. All infants were fed on human milk but, in 10 infants additional protein was administered (Casein preparation Secoosan). Total protein values were found to be lower in premature infants than in full-term infants. During the first eight weeks of life total protein values decreased, from 5.63 g per cent to 4.03 g per cent. The decrease in total protein is caused by a decrease in the amount of albumin (from a little below 3 g per cent in the first week of life to a little above 2 g per cent in the eighth week) and of gamma-globulin (from about 1 g per cent to about 0.5 g per cent) while the alpha1-, alpha2-and beta-globulin values remain rather constant. If casein is added to the diet the total serum protein values and the serum albumin values will range at higher levels, but the decrease with increasing age will not be prevented. On the contrary, additional casein to the diet will cause a further reduction in the gamma-globulin concentrations (Fig. 1). Taux des proteines dans le serum des prematures. Des determinations du taux des proteines totales ainsi que des taux particuliers des differentes proteines ont ete effectuees par electrophorese chez 21 bebes nes avant terme. Tous ces enfants etaient nourris au lait maternel et 10 d'entre eux recurent des supplements de proteines. Il est apparu que le taux des proteines totales etait plus faible chez les prematures que chez les enfants nes a terme. Au cours des huit premieres semaines qui suivirent la naissance, le taux des proteines totales descendit de 5,63 g%a 4,03 g%. Cet abaissenient du taux des proteines totales provenait de la diminution de la concentration en alburnine (qui d'abord legerement inferieure a 3 g % au cours de la premiere semaine etait descendue a un niveau legerement superieur a 2 g % au bout de huit semaines) et en gamma-globuline (qui, etant d'environ 1 g %etait descendue a environ 0,5 g %), tandis que les taux des globulines alpha1, alpha2 et beta etaient restes relativement constants. L'addition de caseine a la nourriture fit monter les taux des proteines totales et de l'albumine a des niveaux plus eleves, mais ces concentrations diminuerent egalement avec le temps. D'autre part, l'addition de caseine a la nourriture entraina une diminution plus importante du taux des gamma-globulines. Serumproteinwerte bei fruhgeborenen Kmdern. Bei 21 fruhgeborenen Kindern wurden die gesamten Serumproteinwerte bestimmt und die Serumproteinbestandteile elektrophoretisch differenziert. Alle Kinder wurden nut Frauenmilch ernahrt, aber 10 Kinder erhielten einen Eiweisszusatz. Die gesamten Proteinwerte erwiesen sich bei den fruhgeborenen Kindern als niedriger als in vollreif geborenen. Wahrend der ersten 8 Lebenswochen fielen die totalen Proteinwerte von 5,63 % auf 4,03 % ab. Der Fall im Gesamtproteingehalt ist durch einen Abfall im Albumingehalt bedingt (von ein wenig unter 3 g % in der ersten Lebenswoche auf ein wenig uber 2 g % in der achten Woche) und Gammaglobulingehalt (von ca. 1 g % auf ca. 0,5 g%), wahrend die Alpha1-, Alpha2-und Betaglobulinwerte ziemlich konstant bleiben. Wenn Kasein zur Diat zugefugt wird, werd. en die gesamten Serumproteinund Serumalbuminwerte ein hoheres Niveau erreichen, aber der mit dem Alter einhergehende Abfall wird nicht verhindert werden. Im Gegenteil, der Kaseinzusatz zur Diat bewirkt eine weitere Herabsetzung der Gammaglobulinkonzentrationen. Valores seroproteicos en los prematuros. Han sido determinados, en 21 prematures, los valores seroproteicos totales y diferenciados electroforeticamente los componentes seroproteicos. Todos los ninos fueron alimentados con leche humana, pero a 10 de ellos se les administraron proteinas adicionales. Hallose que los valores proteicos totales eran inferiores en los prematuros a los de los ninos nacidos a termino. Durante las ocho primeras semanas de la vida, los valores proteicos totales disminuyeron. de 5.63 g % a 4.03 g %. La disminucion del total proteico esta causada por una disminueion de la cantidad de albumina (desde un poco menos de 3 g % en la primera semana de la vida hasta un poco mas de 2 g % en la octava semana) y de gammaglobulina (desde aproximadamente 1 g % a aproximadarnente 0.5 g %) mientras los valores globulinicos alfa1, alfa2 y beta, permanecen bastante constantes. Si se anade caseina a la dieta, los valores seroproteicos totales y los valores seroalbuminosos alcanzaran niveles mas elevados, pero la disminueion, al aumentar la edad, no podra evitarse. La caseina, contraria, adioionada a la dieta, causara todavia mas reduccion en las concentraciones gammaglobulinicas.

A quantitative paper chromatographic method has been developed which is routinely used as a stability assay for estrogenic hormones in castor oil and tablet formulations. The method involves: (a) A separation of the estrogenic hormone from interfering degradation products. (b) The location of the estrogenic hormone on the chromatogram by the guide strip technique employing a chromogenic agent. (c) The elution of the estrogenic hormone from the chromatogram. (d) A quantitative spectrophotofluorometric analysis of the eluate. Recoveries in excess of 95% are obtained by simultaneously chromatographing replicates of standard and sample solutions on the same chromatogram. A detailed account of the procedure is presented using as examples the assay of estradiol valerate in castor oil formulations and ethinyl estradiol in tablets.

The author describes a method, for the determination of bacterial L-histidine, L-tyrosine, L-tryptophan, L-arginine, L-lysine, L-ornithine, L-serine, L-phenyl-alanine, L-glutamic acid and L-aspartic acid decarboxylase by means of high voltage paper electrophoresis, using electrolytes of low ionic strength. For the group of test microorganisms (Enterobacteriaceae, Achromobacteriaceae andPseudomonas), L-histidine, L-arginine, L-lysine, L-ornithine and L-glutamic acid decarboxylases were found to be of the greatest practical significance. The optimal conditions for maximum formation of these decarboxylases by some of the test strains ofEscherichia coli, Hafnia andProteus are discussed.

Conclusions1. Phagocytosis of formalin-killed E. coli was promoted by previous contact with filter paper or with para-hydroxybenzoic acid. The latter, an active principle in filter paper, was effective in concentrations of 1 to 0.001 mg per ml. Fixed tissue inhibited phagocytosis. 2. The activity of the several surfaces reported by Wood et al. to promote phagocytosis may be ascribed to the wet filter paper in their experiments. These observations open to question the phenomenon of “surface” phagocytosis.

Frozen sections are cut from the specimen until the level of interest is reached. A strip of paper (bond or similar writing paper) 5 cm long and about 1 cm wider than the specimen is moistened with water, closely applied to the surface of the specimen and frozen onto it. As the section is cut, the end of the paper strip above the knife is grasped and turned backward toward the other end of the strip. The section is then applied to an albumenized glass slide, firmed and thawed by finger pressure, and the paper removed. After thorough drying, the preparation is ready for further processing. When properly performed, mounted sections whose details coincide to those of the uncut block can be obtained. If thawing on the knife is prevented by cooling the knife, the technic can be performed without a cryostat, but it is also feasible to use a cryostat if a favorable temperature is maintained. The authors obtained 30 μ serial sections, suitable for stereotaxic mapping, from rabbit brain by this method.

SummaryA procedure has been developed whereby paper chromatograms of antiviral agents can be developed by applying them to virus infected, agar overlay, chick embryo cell tissue cultures. Plaque-free zones are produced, thus indicating the area on paper chromatogram where the active material can be located. Using antiviral substances antibiotic W-112 and isatin-monothiosemicarbazone, W. Nile and vaccinia viruses, and 4 chromato-graphic solvent systems, it was possible to localize the biological activity of these antiviral agents. As little as 1 μg of antibiotic W-112 produced large plaque-free zones when paper chromatogram was applied to the overlay culture 1 hr after virus infection. When 100 μg of same material was used, large plaque-free areas resulted even when the paper was applied 46 hr after virus infection. Isatin-monothiosemicarbazone at 50 μg level not only produced a plaque-free zone but also produced a yellow spot on paper chromatogram that coincided exactly with the zone of antiviral activity.

(1973). A COMMENT ON THE WHITE PAPER. The Slow Learning Child: Vol. 20, No. 3, pp. 170-172.