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Tryptic digest analysis by comprehensive reversed phase×two reversed phase liquid chromatography (RP‐LC×2RP‐LC) at different pH's
Author(s) -
François Isabelle,
Cabooter Deirdre,
Sandra Koen,
Lynen Frédéric,
Desmet Gert,
Sandra Pat
Publication year - 2009
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200800578
Subject(s) - chromatography , chemistry , orthogonality , phase (matter) , reversed phase chromatography , two dimensional chromatography , analytical chemistry (journal) , dimension (graph theory) , high performance liquid chromatography , mathematics , organic chemistry , geometry , pure mathematics
As an alternative to the classical approach of combining strong cation exchange SCX‐LC and RP‐LC for the separation of complex proteomic samples, this essay describes the online comprehensive RP‐LC×RP‐LC separation of BSA and human blood serum. High orthogonality and peak capacity are achieved through the application of a significantly different pH in the two dimensions. The coupling of fused‐core columns in series ensures high efficiency in the first dimension, while a previously designed interface with parallel second dimension columns further enhances the separation capability of the comprehensive system.