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The non-protein nitrogen composition of grass silages: I. The estimation of the basic amino acids and non-volatile amines by chromatography on a weak cation exchange resin

Published online by Cambridge University Press:  27 March 2009

A. D. Hughes
Affiliation:
The Rowett Research Institute, Bucksburn, Aberdeen, Scotland

Summary

Investigations into the non-protein nitrogen composition of grass silages using the 50 cm strong cation-exchange column of Spackman, Stein & Moore (1958) to determine the basic amino acids led to difficulties in the determination of ethanolamine in the presence of high concentrations of ammonia, and of histidine in the presence of δ amino-n-valeric acid. An alternative technique for the ion exchange chromatography and estimation of histidine, lysine, ornithine, ethanolamine, arginine and ammonia on a weak cation-exchange resin has been developed. This method enables small amounts of ethanolamine to be determined in the presence of large amounts of ammonia and values for the ethanolamine content of a number of silage samples are presented. When used in conjunction with the technique of Spackman et al. (1958) the δ-amino-n-valeric acid content of grass silages could also be determined in the presence of histidine.

The estimation of amines produced by the microbial decomposition of herbage proteins during ensiling has previously involved their initial separation from the amino acids followed by quantitative partition chromatography. An alternative method for the estimation of these amines by ion-exchange chromatography on a weak cation-exchange resin is described. This method permits the colorimetric determination of β-phenylethylamine, tyramine, tryptamine, 5-hydroxytryptamine, putrescine, cadaverine and histamine without interference from the amino acids. The efficiency of this technique has been investigated using standard solutions of the naturally occurring amines and samples of good quality and of high pH spoilt silages.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1969

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References

REFERENCES

Dustin, J. P., Czajkowska, C., Moore, S. & Blgwood, E. J. (1953). A study of the chromatographic determination of amino acids in the presence of large amounts of carbohydrates. Analytica chim. Acta 9, 256.Google Scholar
Fowler, H. D. (1962). Ethanolamine in silage. Ir. J. agric. Res. 1, 212.Google Scholar
Garratini, S. & Valzelli, L. (1965). Serotonin. London and New York: Elsevier Publishing Company, Amsterdam.Google Scholar
Harpenist, E. J. (1953). The amino acid composition of insulins isolated from beef, pork and sheep glands. J. Am. chem. Soc. 75, 5528.CrossRefGoogle Scholar
Hirs, C. W. H., Moore, S. & Stein, W. H. (1953). A chromatographic investigation of pancreatic ribonuclease. J. biol. Chem. 200, 493.CrossRefGoogle ScholarPubMed
Jacobs, S. (1960). The determination of nitrogen in organic compounds by the indanetrione hydrate method. Analyst, Lond. 85, 257.Google Scholar
Kirchmeir, O. & Kiermeler, F. (1962). Protein breakdown in silages. Z. Tierphysiol. Tierernähr. Futtermittelk. 17, 264.Google Scholar
Kirchmeir, O. & Kiermeier, F. (1964). δ-Amino-n-valeric acid a specific component of spoilt silage. Naturwissenschaften 51, 13.Google Scholar
Kunin, K. & Myers, R. J. (1950). Ion Exchange Resins, New York: Wiley; London: Chapman and Hall Ltd.Google Scholar
Macpherson, H. T. (1962). Histamine, tryptamine and tyramine in grass silage. J. Sci. Fd Agric. 13, 29.CrossRefGoogle Scholar
Macpherson, H. T. & Violante, P. (1966) Ornithine, putrescine and cadaverine in farm silages J. Sci. Fd Agric. 17, 124.CrossRefGoogle Scholar
McIlvaine, T. C. (1923). A buffer solution for colorimetric comparison. J. biol. Chem. 49, 183.CrossRefGoogle Scholar
Moore, S. & Stein, W. H. (1954 a). Procedures for the chromatographic determination of amino acids on four percent cross linked sulphonated polystyrene resins. J. biol. Chem. 211, 893.CrossRefGoogle Scholar
Moore, S. & Stein, W. H. (1954 b). A modified ninhydrin reagent for the photometric determination of amino acids and related compounds. J. biol. Chem. 211, 907.CrossRefGoogle ScholarPubMed
Neumark, H. (1961). Amines in silage. Nature, Lond. 190, 839.CrossRefGoogle Scholar
Spackman, D. H., Stein, W. H. & Moore, S. (1958). Automatic recording apparatus for use in the chromatography of amino acids. Analyt. Chem. 30, no. 7, p. 1190.CrossRefGoogle Scholar
Tristram, G. R. & Smith, R. H. (1963). The amino acid composition of some purified proteins. Adv. Protein Chem. 18, 227.CrossRefGoogle ScholarPubMed
Wall, R. A. (1968). Accelerated analysis of some amines and amino acids. J. Chromatog. 37, 549.CrossRefGoogle Scholar