QUANTITATIVE CHROMATOGRAPHIC METHODS: PART 7. ISOLATION OF AMINO ACIDS FROM SERUM AND OTHER FLUIDS

1961 ◽  
Vol 39 (2) ◽  
pp. 439-451 ◽  
Author(s):  
C. K. Harris ◽  
E. Tigane ◽  
C. S. Hanes

A method based on the use of miniature ion-exchange columns has been devised for isolating small amounts of amino acids from biological fluids and tissue extracts. The amino acids are isolated virtually free from proteins, sugars, and inorganic cations in a single treatment.Techniques are described for the preparation of solutions of the isolated amino acids for quantitative analysis by paper chromatography.

1955 ◽  
Vol 102 (4) ◽  
pp. 435-440 ◽  
Author(s):  
Leonard T. Skeggs ◽  
Walton H. Marsh ◽  
Joseph R. Kahn ◽  
Norman P. Shumway

A preparation of hypertensin I was purified by countercurrent distribution and was shown to migrate as a single component in starch blocks at pH 9.3 and 4.2. It had an isoelectric point of 7.7. Quantitative analysis by ion exchange column chromatography showed eight amino acids in approximately unimolar proportion: aspartic, proline, valine, isoleucine, leucine, tyrosine, phenylalanine, and arginine. There were in addition two moles of histidine.


1971 ◽  
Vol 54 (1) ◽  
pp. 61-65
Author(s):  
Arthur Russell Johnson ◽  
Richard L Corliss ◽  
Enrique Fernandez-Flores

Abstract Qualitative chromatographic methods for the separation of free amino acids in table sirups are presented to aid in the development of chemical indices of composition which may be useful in establishing the identity of sirups and detecting their adulteration. Free amino acids in 2 table sirups were isolated on ion exchange columns and eluted with dilute ammonia. The concentrated amino acid mixture in the eluate was spotted directly on silica gel G plates for TLC analysis, or the amino acids were converted to their N-trifluoroacetyl n-butyl esters for GLC analysis. As many as 16 amino acids were qualitatively separated and identified and a potential for quantitative analysis was demonstrated.


1969 ◽  
Vol 52 (5) ◽  
pp. 981-984 ◽  
Author(s):  
J E Knipfel ◽  
D A Christensen ◽  
B D Owen

Abstract Amino acid analyses were performed on samples of blood, liver tissue, loin muscle, and ham muscle by ion exchange chromatography after deproteination of the samples with picric acid or sulfosalicylic acid (SSA). Resolution of threonine and serine from the ion exchange column was poor when SSA was used as the deproteinating agent. Twelve of sixteen amino acids were higher (P < 0.05) in serum deproteinated with picric acid as compared to concentrations determined after SSA deproteination. Amino acid values for ham muscle tended to be higher after deproteination with picric acid; however, with liver and loin muscle samples, the values were somewhat higher after SSA deproteination. In both serum and tissue analyses, coefficients of variation were lower for niGSt amino acids when picric acid was utilized as the deproteinating agent. The latter observation, in particular, suggests that picric acid is preferable to SSA as a deproteinating agent before amino acid analyses of biological fluids. Standardization of methods of deproteination is needed to allow meaningful comparisons of data.


1969 ◽  
Vol 15 (9) ◽  
pp. 902-907 ◽  
Author(s):  
William J Culley

Abstract Simple and rapid thin-layer chromatographic methods, using heel or finger-tip blood, are described that are suitable for screening for inborn errors of amino acid metabolism. Serum is chromatographed without deproteinizing. A variation of this method utilizes blood impregnated on paper discs for chromatography. These methods are much faster, more sensitive, and provide better resolution of amino acids than paper chromatography. Because untreated serum- or blood-impregnated discs are placed directly on thin-layer chromatograms, these methods require considerably less working time than other thin-layer chromatographic methods.


1968 ◽  
Vol 22 (3) ◽  
pp. 437-450 ◽  
Author(s):  
J. W. G. Porter ◽  
D. R. Westgarth ◽  
A. P. Williams

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