Sensitivity enhancement and second-dimensional information from solid phase extraction-capillary electrophoresis of entire high-performance liquid chromatography fractions
✍ Scribed by Michael A. Strausbauch; Benjamin J. Madden; Dr. Peter J. Wettstein; James P. Landers
- Publisher
- John Wiley and Sons
- Year
- 1995
- Tongue
- English
- Weight
- 873 KB
- Volume
- 16
- Category
- Article
- ISSN
- 0173-0835
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✦ Synopsis
Sensitivity enhancement and second-dimensional information from solid phase extraction-capillary electrophoresis of entire high-performance liquid chromatography fractions
A novel separation technique is demonstrated for peptide analysis of entire 100 pL high-performance liquid chromatography (HPLC) fractions in a single, low pH, capillary zone electrophoresis (CZE) separation. The method employs a hybrid capillary consisting of a removable inlet section containing a reversephase packing and a standard CZE bare-fused silica separation capillary. The packed tip allows sample to be concentrated at flow rates greater than 20 pL/min and released in a controlled manner for CZE separation. Separations are comparable to standard CZE with minor modifications in selectivity. Using a mixture of model peptides, the hybrid capillary is shown to effect enhancement in sensitivity of 2 100-fold. Five fractions from the HPLC separation of peptides obtained from the digestion of bovine serum albumin (BSA) were analyzed by the solid phase extraction-capillary electrophoresis (SPE-CE) method which was shown to provide rapid second-dimensional information. Practical concentration limits of detection for peptides are calculated to be 1-10 ng/mL with this technology.