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  • Title: Reversed-phase liquid chromatography of peptides for direct micro-sequencing.
    Author: Reinbolt J, Hounwanou N, Boulanger Y, Wittmann-Liebold B, Bosserhoff A.
    Journal: J Chromatogr; 1983 Mar 25; 259(1):121-30. PubMed ID: 6343406.
    Abstract:
    Tryptic and cyanogen bromide peptides derived from yeast aspartyl-tRNA synthetase and from Escherichia coli ribosomal proteins were separated by reversed-phase liquid chromatography, employing volatile buffers of low ionic strength. The conditions used allow the performance of micro-sequencing without desalting or extensive lyophilization, and can therefore be applied to peptide mixtures containing hydrophobic fragments which tend to precipitate. To prevent losses of peptides, direct ultra-violet detection of the peptides was preferred, to detection by post-column derivatization with an additional stream splitting device. Preparative separations were performed with 5-10 nmol of peptide mixture; analytical runs were made with 5-10 micrograms of protein hydrolysate.
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