Observations upon the application of partition chromatography to the determination of the monoamino-acids in proteins

Tristram, G. R.

doi:10.1042/bj0400721

Cited by 72 publications

(26 citation statements)

References 18 publications

(9 reference statements)

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“…For the purpose of consistency, however, these substances are hereinafter referred to as D amino acids. 18 We wish to thank Dr. E. A. Kabat of Columbia University for many helpful suggestions in regard to the chromatographic technique. 1~ The solvents are described with the terminology adopted by Tristram (18).…”

Section: Experimental and Resultsmentioning

confidence: 99%

Elicitation of Delayed Allergic Skin Reactions With Haptens: The Dependence of Elicitation on Hapten Combination With Protein

Eisen

Orris

Belman

1952

Journal of Experimental Medicine

176

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Allergic skin reactions of the delayed type may be conveniently considered to involve two distinct processes: first, induction of the sensitized state, and second, elicitation of the allergic reaction in the sensitized individual. With regard to the induction of sensitization, it is generally believed that low molecular weight substances (haptens) can function effectively in this respect if they combine in vivo with tissue constituents, presumably proteins. This belief, which had long been reasonably maintained on the basis of analogy with the conditions required for the production of antisera by haptens, achieved a high degree of plausibility as a result of the systematic studies of Landsteiner and Jacobs (1, 2).The question as to whether hapten combination with protein is involved in the second process, namely, elicitation of the reaction, has received little attention. In many studies of the elicitation process, haptens which are homologous in structure to the hapten used for inducing sensitization often have produced widely divergent reactions: some homologues have elicited skin lesions, while other have not (3)(4)(5). This diversity of responses has been interpreted quite consistently in terms of specificity arising from the configurational conformity of the elicifing compound to the sensitizing hapten, little attention being paid to other factors that might modify the responses. However, from a consideration of these earlier studies, it appeared possible that, in addition to configurational features, at least one other factor--namely, combination of the eliciting hapten with protein--might be involved in accounting for the diverse responses sometimes obtained in eliciting reactions with haptens which are structurally similar to the substance used for inducing sensitization. Accordingly, we have sought to determine with a group of 2,4-dinitrophenyl compounds whether combination with protein is important in determining the effectiveness of haptens in producing delayed allergic skin lesions in sensitized guinea pigs and men.

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Section: Experimental and Resultsmentioning

confidence: 99%

Elicitation of Delayed Allergic Skin Reactions With Haptens: The Dependence of Elicitation on Hapten Combination With Protein

Eisen

Orris

Belman

1952

Journal of Experimental Medicine

176

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“…After taking to dryness the peptide mixture is fractionated on a suitable column. To describe the various solvent systems used the method and codes of Tristram (1946) Acetone-cyclohexane: prepared from 1 vol. water, 3 vol.…”

Section: Fractionation Of Dnp Peptidesmentioning

confidence: 99%

The terminal peptides of insulin

Sanger

1949

Biochemical Journal

345

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While this work was in progress an outline of a fluorimetric method for the determination of Nmethyl-2-pyridone-5-carboxylamide in urine was reported by Rosen, Perlzweig & Handler (1948). The authors considered that their values for normal urines (5.0-13.2 mg./day) may have been somewhat low, but it is obvious that they are of the same order as those obtained by the present colorimetric method (see Table 1, and also Dean & Holman, 1949). SUMMARY 1. Methods are described for the determination of N-methyl-2-pyridone-5-carboxylamide and Nmethyl-2-pyridone-3-carboxylamideinhuman urine. The methods are based on the nitration of the compounds, after isolation from urine by adsorption on Lloyd's reagent followed by extraction with chloroform, and on their colorimetric determination by means of the yellow colours given by the nitro derivatives in alkaline solution.2. The accuracy and specificity of the methods is discussed.3. Results are shown which indicate that the 2:3amide is of no significance in nicotinic acid metabolism, but that the 2:5-amide is an important metabolite. The 2:5-amide was excreted by two normal adults in amounts of the order of 7 mg./24 hr. After the ingestion of 500 mg. ofnicotinamide, the urinary excretion of the 2:5-amide increased greatly, 47-57 % of the dose being excreted in 72 hr. as the latter compound.4. It was possible to account for 73-80 % of an orally administered dose of 500 mg. of nicotinamide in the urine of two adult subjects as N-methyl-2pyridone-5-carboxylamide, N-methylnicotinamide and the total acid-hydrolysable derivatives of nicotinic acid.

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“…Even in the initial publication on this technique, however, it was noted that separate lots of silica vary significaiitly with respect to their adsorptive power and their capacity for sorption of water. Several procedures have been reported for pre-paring and standardizing silica gel suitable for use as a support (6,(9)(10)(11)(12). Isherwood ( I S ) and Resnik, et al (14), described purification processes designed to decrease the adsorptive properties of the material.…”

Section: Journal Of Pharmuceutical Sciencesmentioning

confidence: 99%

Column Partition Chromatography in Pharmaceutical Analysis

Levine

1963

Journal of Pharmaceutical Sciences

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Observations upon the application of partition chromatography to the determination of the monoamino-acids in proteins

Cited by 72 publications

References 18 publications

Elicitation of Delayed Allergic Skin Reactions With Haptens: The Dependence of Elicitation on Hapten Combination With Protein

Elicitation of Delayed Allergic Skin Reactions With Haptens: The Dependence of Elicitation on Hapten Combination With Protein

The terminal peptides of insulin

Column Partition Chromatography in Pharmaceutical Analysis

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