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Two‐step ion‐exchange chromatographic purification combined with reversed‐phase chromatography to isolate C‐peptide for mass spectrometric analysis
Author(s) -
Kabytaev Kuanysh,
Durairaj Anita,
Shin Dmitriy,
Rohlfing Curt L.,
Connolly Shawn,
Little Randie R.,
Stoyanov Alexander V.
Publication year - 2016
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.201500989
Subject(s) - chromatography , chemistry , ion chromatography , mass spectrometry , peptide , reversed phase chromatography , ion exchange , two dimensional chromatography , ion suppression in liquid chromatography–mass spectrometry , high performance liquid chromatography , ion , tandem mass spectrometry , organic chemistry , biochemistry
A liquid chromatography with mass spectrometry on‐line platform that includes the orthogonal techniques of ion exchange and reversed phase chromatography is applied for C‐peptide analysis. Additional improvement is achieved by the subsequent application of cation‐ and anion‐exchange purification steps that allow for isolating components that have their isoelectric points in a narrow pH range before final reversed‐phase mass spectrometry analysis. The utility of this approach for isolating fractions in the desired “pI window” for profiling complex mixtures is discussed.