Elevated temperature-extended column length conventional liquid chromatography to increase peak capacity for the analysis of tryptic digests.
High efficiency separations (200 000 plates) were obtained on conventional LC equipment by coupling 8 x 25 cm x 2.1 (or 4.6) mm id x 5 microm d(p) ODS columns (total length 2 m) and operation at 60 degrees C using a dedicated LC oven. The peak capacity in this 1-D set-up was 900 for the separation of human serum tryptic peptides analyzed after depletion of six highly abundant proteins. The chromatographic performance of an elevated temperature-extended column length conventional LC is highlighted.
Sandra
Pat
P
Research Institute for Chromatography (R.I.C.), Kortrijk, Belgium. pat.sandra@richrom.com
Vanhoenacker
Gerd
G
D001798:Blood Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D002853:Chromatography, Liquid / Q000295:instrumentation* / Q000379:methods*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D010446:Peptide Fragments / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D013696:Temperature; D014357:Trypsin / Q000378:metabolism*
D001798:Blood Proteins; D010446:Peptide Fragments; D014357:Trypsin