Separation of o-phthalaldehyde derivatives of amino acids of the internal pool of yeast by reverse-phase liquid chromatography
Tóm tắt
Derivatization of primary amino acids with orthophthalaldehyde and β-mercaptoethanol forms derivatives that can be detected by absorbance at 340 nm. These were separated by reverse-phase high performance liquid chromatography (HPLC). A step- or more complex gradient (acetonitrile/phosphate buffer gradient) was used. The effects of several parameters (pH, ionic strength, etc) were characterized and used to design a rapid separation of the amino acids commonly found in physiological fluids. The method described is rapid, sensitive and precise as sensitivity limits are about 25 pmol and the separation time, injection to injection, is 16 min.
Tài liệu tham khảo
Benítez, T., del Castillo, L., Aguilera, A., Conde, J., and Cerdá-Olmedo, E. (1983) Appl. Environ. Microbiol. 45:1429–1436.
Bidlingmeyer, B. A., Cohen, S. A., and Tarvin, T. L. (1984). J. Chrom. 336:93–104.
Cooper, J. D. H., Ogden, G., McIntosh, J., and Turnell, D. C. (1984) Anal. Biochem. 142:98–102.
Cunico, R. L., and Schlabach, T. (1983). J. Chrom. 266:461–465.
Fleury, M. O., and Ashley, D. V. (1983). Anal. Biochem. 133:330–335.
Gillespie, D., and Spiegelman, S. (1965) J. Mol. Biol. 12:829–842.
Hill, D. W., Walters, F. H., Wilson, T. D., and Stuart, J. D. (1979). Anal. Chem. 51:1338–1342.
Hogan, D. L., Kraemer, K. L., and Isenberg, J. I. (1982). Anal. Biochem. 127:17–24.
Jarrett, H. W., Cooksy, K. D., Ellis, B., and Anderson, J. M. (1986). Anal. Biochem. 153: 189–198.
Johansson, B., and Isaksson, K. (1986). J. Chrom. 356:383–392.
Jones, E. W., and Fink, G. R. (1982). Regulation of amino acid and nucleotide biosynthesis in yeast. In: The molecular biology of the yeast Saccharomyces (metabolism and gene expression), Strathern et al., eds. pp. 181. Cold Spring Harbor Laboratory.
Lindroth, P., and Mopper, K. (1979). Anal. Chem. 51:1667–1674.
Maugh, T. H. II (1984). Science, 225: 42.
Schuster, R. (1980) Anal. Chem. 52:617–620.
Sherman, F., Fink, G. R., and Lawrence, C. W. (1979) Methods in yeast genetics laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
Simons, S. S., and Johnson, D. F. (1977). Anal. Biochem. 82:250–254.
Umagat, H., Kucera, P., and Wen, L. F. (1982). J. Chrom. 239:463–474.