Chromatography of Dinitrophenyl Amino-acids and Peptides

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SANGER1 has described the use of 1 : 2 : 4-fluoro-dinitrobenzene to identify the free amino-groups of several proteins, employing the separation of the N-dinitrophenyl amino-acids on columns of silica gel. We have had only limited success applying this method, owing to variations in different batches of the silica gel, and the difficulty of preparing evenly packed columns, which led to diffuse bands, difficult to separate. Sanger stated that with these derivatives on paper chromatograms2 long diffuse ‘tails' formed, making the method quite inferior to the silica method. We are able to report some success with the two-dimensional method, however, without apparently using any special technique which might account for the improvement. Consden et al.3 have also recently described the identification of α-δ-di- dinitrophenyl -ornithine running alone on a paper chromatogram, and Keston et al.4 using the p-I131-benzene-sulphonyl derivatives of amino-acids have been able to estimate them after their partial separation on a one-dimensional paper strip.

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  1. 1

    Sanger, F., Biochem. J., 39, 507 (1945).

  2. 2

    Consden, R., Gordon, A. H., and Martin, A. J. P., Biochem. J., 38, 224 (1944).

  3. 3

    Consden, R., Gordon, A. H., Martin, A. J. P., and Synge, R. L. M., Biochem. J., 41, 596 (1947).

  4. 4

    Keston, A. S., Udenfriend, S., and Levy, M., J. Amer. Chem. Soc., 69, 3151 (1947).

  5. 5

    Phillips, D. M. P., Nature, 161, 53 (1948).

  6. 6

    Butler, J. A. V., Dodds, E. C., Phillips, D. M. P., and Stephen, J. M. L., Biochem. J., 42, 116 (1948).

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PHILLIPS, D., STEPHEN, J. Chromatography of Dinitrophenyl Amino-acids and Peptides. Nature 162, 152 (1948) doi:10.1038/162152a0

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