Abstract
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 language | English (US) |
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Pages (from-to) | 529-537 |
Number of pages | 9 |
Journal | Journal of Separation Science |
Volume | 29 |
Issue number | 4 |
DOIs | |
State | Published - Mar 2006 |
Keywords
- 2-D chromatography
- C-peptide
- Column switching
- Isotope dilution assay
- Matrix effects
ASJC Scopus subject areas
- Analytical Chemistry
- Filtration and Separation