Evaluation of Reverse Phase Columns for Protein Separation Using a Single LC-MS/LC-UV Methodology

Library Number:
Part Number:
Kurt Yardley;Paul Rainville;Reb Russell II;Jeffrey R. Mazzeo;Uwe D. Neue [Waters]
FAll ACS 2002; 224th ACS National Meeting; Boston August 18-22
Content Type:
Content Subtype:
Protein; RNase A; Cytochrome C; Horse Myoglobin; Carbonic Anhydrase
Symmetry300(™) C18 300 Å 5 µm Steel 4.6 mm 50 mm Symmetry300(™) C4 300 Å 5 µm Steel 4.6 mm 50 mm Delta-Pak C18 300 Å 5 µm Steel 4.6 mm 50 mm Delta-Pak C4 300 Å 5 µm Steel 4.6 mm 50 mm
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HPLC separations on reversed phase columns are an important part of characterizing complex protein samples from basic research and pharmaceuticals. Recent advances in mass spectrometry have enabled rapid characterization of protein and peptide samples separated by HPLC, greatly facilitating the identification of target proteins and contaminants. Thus, there is a need for good separation techniques using MS compatible methodology for the separation of proteins by HPLC. We present here the evaluation of peak capacity and recovery in protein separations along with LC/MS.

Our findings demonstrate that both Delta-Pak® and Symmetry® 300 are the preferred columns for LC/MS protein separations.

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