Summary
An improved method for the quantitative determination of amino acids using HPLC, after preliminary derivatisation with o-phthalaldehyde, is described. Separation was carried out on a LiChrospher RP-18 column, using gradient elution and a fluorescence detector. Hydrolysis was performed with 6M HCl or with 3M p-toluenesulphonic acid depending on the amount of carbohydrates in the sample. In addition, the time required for total hydrolysis was shortened from 24 hours to 15 minutes by the use of a microwave oven. The results with these two hydrolytic methods are compared with regard to the extent of peptide bond cleavage and the percentage of decomposition of amino acids in a test mixture.
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References
H. S. Sista, J. of Chromatogr.359, 231 (1986).
P. Lindroth, K. Mooper, Anal. Chem.51, (11) (1979).
D. Roach, C. W. Gherke, J. of Chromatogr.52, 393 (1970).
S. Hunt, G. C. Barrett (eds.), “Chemistry and Biochemistry of the Amino Acids”, Chapman and Hall, (1985) p. 376.
S. T. Chen, S. H. Chiou, K. T. Wang, Int. J. Pep. Prot. Res.30, 572 (1987).
H. M. Yu, S. T. Chen, S. H. Chiou, K. T. Wang, J. of Chromatogr.456, 357 (1988).
R. J. Simpson, M. R. Neuberger, T. Y. Liu, J. Biol. Chem.251, 1936 (1976).
B. Penke, R. Ferenzi, K. Kovacz, Anal. Biochem.246, 2842 (1974).
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Pecavar, A., Prosek, M., Fercej-Temeljotov, D. et al. Quantitative evaluation of amino acids using microwave accelerated hydrolysis. Chromatographia 30, 159–162 (1990). https://doi.org/10.1007/BF02274539
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DOI: https://doi.org/10.1007/BF02274539