High-Resolution Reversed-Phase Liquid Chromatography of Peptide Separations

Library Number:
980587
Author(s):
Yung-Fong Cheng;Bonnie A. Alden;Edouard S.P. Bouvier;Judy Carmody;Ray Crowley;Uwe Neue;John O''Gara;Thomas H. Walter
Source:
19th Annual Spring Symposium - Minnesota Chromatography Forum - May 19-21
Content Type:
Orals/Lectures
Content Subtype:
Other Symposium
Compounds:
Peptides; Decapeptides [Ac-Arg-Gly-X-X-Gly-Gly-Leu-Gly-LysAmide]
Matrix:
Alberta Peptides mix; Tryptic digests of: bovine cytochrome c; bovine serum albumin
Column:
Symmetry300(™) C18 300 Å 5 µm Steel 3.9 mm 150 mm SymmetryShield(™) RP8 5 µm Steel 3.9 mm 150 mm Symmetry® C8 5 µm Steel 3.9 mm 150 mm Symmetry300(™) C18 300 Å 5 µm Steel 4.6 mm 150 mm Symmetry® C18 5 µm Steel 4.6 mm 150 mm Symmetry300(™) C18 300 Å 5 µm Steel 4.6 mm 50 mm Symmetry300(™) C18 300 Å 5 µm Steel 4.6 mm 250 mm
The development of a good and reproducible separation of peptides with small differences in the amino acid composition is difficult and challenging. In this paper, we present a strategy for the rapid development of rugged and validatable peptide maps using reversed-phase gradient HPLC. The impact of the reversed-phase column characteristics (pore size, pore volume, hydrophobicity, hydrolytic stability), the mobile phase modifier, the temperature, the type of acid and its concentration will be discussed. Also, both the column and batch-to-batch reproducibility of the packing material will be emphasized.

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