The New Way in HPLC Method Development
By some estimates, there are close to 800 different HPLC stationary phases to choose from when beginning an HPLC method development project. Choosing a column usually comes down to an educated guess, and whether it will successfully separate your analytes is usually not obvious until many hours have been spent evaluating different mobile phase conditions. If the column fails, another is chosen and the process is repeated until the right combination of stationary phase and mobile phase is discovered.
The reason there are so many stationary phases to choose from is quite simple; selectivity.
In Phase Optimized Liquid Chromatography (POPLC®), a technique developed by Bischoff Chromatography, the stationary phase is first optimized for a given sample.
In POPLC, rather than making a rough estimate of the appropriate column and then optimizing the mobile phase,one does the reverse. A rough choice of mobile phase is made, and then the sample is run isocratically on short columns of three or more different bonded phases.(Figure 1) Retention times for all components on each stationary phase are input to the POPLC Optimizer software. Armed with these data, the software can determine the optimum column, consisting of various length segments of the different stationary phases used. Based on the requirements specified by the investigator, the method can be optimized for speed, resolution or both.
Once the optimum column has been indentified, the various segments are assembled using the POPLink coupling system.
The reason there are so many stationary phases to choose from is quite simple; selectivity.
In Phase Optimized Liquid Chromatography (POPLC®), a technique developed by Bischoff Chromatography, the stationary phase is first optimized for a given sample.
In POPLC, rather than making a rough estimate of the appropriate column and then optimizing the mobile phase,one does the reverse. A rough choice of mobile phase is made, and then the sample is run isocratically on short columns of three or more different bonded phases.(Figure 1) Retention times for all components on each stationary phase are input to the POPLC Optimizer software. Armed with these data, the software can determine the optimum column, consisting of various length segments of the different stationary phases used. Based on the requirements specified by the investigator, the method can be optimized for speed, resolution or both.
Once the optimum column has been indentified, the various segments are assembled using the POPLink coupling system.
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