Microchip electrophoresis with hydrodynamic injection and waste-removing function for quantitative analysis

Chun Che Lin, Cheng Chuan Chen, Ching Erh Lin, Shu Hui Chen

Research output: Contribution to journalConference articlepeer-review

31 Citations (Scopus)

Abstract

Quantitative analysis is problematic for microchip electrophoresis for several reasons including chip-to-chip variation, discontinuous sample re-loading, channel reconditioning, and electrokinetic injection bias. In this study, the capability for quantitative analysis on a flow-through based microchip electrophoresis, which provides continuous sample re-loading, channel washing, reconditioning and hydrodynamic injection as well as waste removing is demonstrated to be more quantifiable and more reproducible compared to manual electrokinetic injection method. Using the flow-through microchip with waste-removing function, FITC-labeled estrogen or Rhodamine B could be continuously analyzed without significant changes (R.S.D. < 6.6%) in signal intensity for over 3 h, which is sufficient for a complete set of quantitative analysis. With the use of a phosphorylated kinase substrate as the model, a calibration curve for quantitative analysis of phosphopeptides were constructed and results indicate that both R 2 value of the linearity and R.S.D. values of the peak intensity were around 0.9961 and 3.16%, respectively, without the use of an internal standard. These values were slightly improved to be around 0.9986 and 2.27%, respectively, with the use of a non-phosphopeptide counterpart as the internal standard. The potential of this flow-through device for the development of a kinase phosphorylation assay based on the quantitative method was also briefly discussed.

Original languageEnglish
Pages (from-to)69-74
Number of pages6
JournalJournal of Chromatography A
Volume1051
Issue number1-2
DOIs
Publication statusPublished - 2004 Oct 8
Event17th International Symposium on Microscale Separations - Salzburg, Austria
Duration: 2004 Feb 82004 Feb 12

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

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