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06:44
Curtain Flow Column: Optimization of Efficiency and Sensitivity
Published on: June 12, 2016
Modifying conventional high-performance liquid chromatography systems to achieve fast separations with Fused-Core
A J Alexander1, T J Waeghe, K W Himes
1Analytical Research and Development, Bristol Myers Squibb Company, 1 Squibb Drive, New Brunswick, NJ 08903, USA. Anthony.Alexander@bms.com
Journal of Chromatography. A
|July 12, 2011
Summary
High efficiency HPLC columns often underperform due to extra-column dispersion (ECD). Reducing extra-column volume (ECV) on a Waters Alliance system improved ECD, but auto-injector design limited further gains, impacting performance with superficially porous columns.
Area of Science:
- Analytical Chemistry
- Chromatography
Background:
- High efficiency columns in High-Performance Liquid Chromatography (HPLC) often fail to achieve theoretical performance gains.
- This limitation is frequently attributed to extra-column dispersion (ECD) originating from instrument design, particularly in conventional HPLC systems.
Purpose of the Study:
- To investigate the impact of extra-column dispersion (ECD) from a Waters Alliance 2695 system on the performance of 2.7 μm HALO C18 Fused-Core columns.
- To assess the effectiveness of re-configuring the Alliance system to reduce extra-column volume (ECV) and its influence on ECD.
Main Methods:
- The Waters Alliance 2695 system was modified to decrease extra-column volume (ECV).
- Extra-column dispersion (ECD) was measured across various flow rates (up to 2 mL/min) for different ECV configurations.
- Auto-injection was compared with manual injection using a low-dispersion valve to isolate auto-injector contributions to ECD.
Main Results:
- Reducing ECV led to a progressive decrease in ECD, though less than theoretically predicted for lower ECV setups.
- The auto-injector module was identified as a significant contributor to residual ECD, limiting further improvements.
- Even with auto-injection, substantial column performance (up to 70% plate count) was retained with optimized instrument configurations for specific separations.
Conclusions:
- System re-configuration can mitigate ECD, but auto-injector design poses a fundamental limitation in conventional HPLC systems.
- Accurate measurement of true ECD in chromatographic systems presents challenges.
- Optimized instrument configurations enable faster separations (3-3.5x) with maintained data quality, even with autosampler injection.
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