Microsequence analysis of peptides and proteins: II. Separation of amino acid phenylthiohydantoin derivatives by high-performance liquid chromatography on octadecylsilane supports

✍ Scribed by David Hawke; Pau-Miau Yuan; John E. Shively

• Publisher: Elsevier Science
• Year: 1982
• Tongue: English
• Weight: 716 KB
• Volume: 120
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(82)90351-7

✦ Synopsis

A comparison of the separation of the common phenylthiohydantoin derivatives of amino acids on DuPont octadecylsilane with that obtained with Ultrasphere octadecylsilane supports is given together with the effect of acetate, phosphate, and trifluoroacetate buffers in the elution solvents. An important change in performance for two different batches of DuPont Zorbax octadecylsilane was noted. The use of combined trifluoroacetate/acetate buffer with Ultrasphere octadecylsilane gives optimal separations and peak sharpness. Practical examples of the performance of this system in low-nanomole NH*-terminal sequence analysis are discussed with emphasis on identification of unusual amino acid derivatives and interfering background peaks.