Direct sensitive quantitative LC/MS analysis of C-peptide from human urine by two dimensional reverse phase/reverse phase high-performance liquid chromatography

Eduard Rogatsky, Vlad Tomuta, Greg Cruikshank, Louis Vele, Harsha Jayatillake, Daniel Stein

Research output: Contribution to journalArticle

19 Scopus citations


We describe for the first time the direct quantitative analysis of human C-peptide from urine by isotope dilution assay. Implementation of 2-D reverse phase-reverse phase chromatography (2-D RP-RP) with SIM detection resulted in high assay sensitivity (LOQ= 5 pg on column), accuracy, and method ruggedness. Our experiments demonstrate the strong resolving capability of our proposed 2-D RP-RP platform which significantly reduces strong matrix effects and their resulting quantitation error over a wide range of urine concentrations. In contrast, single column methods (both SIM and multiple reaction monitoring) were found acceptable only for strongly diluted urine samples.

Original languageEnglish (US)
Pages (from-to)529-537
Number of pages9
JournalJournal of Separation Science
Issue number4
Publication statusPublished - Mar 1 2006



  • 2-D chromatography
  • C-peptide
  • Column switching
  • Isotope dilution assay
  • Matrix effects

ASJC Scopus subject areas

  • Analytical Chemistry
  • Filtration and Separation

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