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Two-step ion-exchange chromatographic purification combined with reversed-phase chromatography to isolate C-peptide
Kuanysh Kabytaev1, Anita Durairaj1, Dmitriy Shin1
1University of Missouri, School of Medicine, One Hospital Drive, Columbia, MO, USA.
Abstract:
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.
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