Tandem-column LC (TC-LC) can be run on a regular U/HPLC system by connecting two columns using a zero dead-volume kit. By using the hydrophobic subtraction model (HSM) and HSM column descriptors from hplccolumns.org, we can simulate separations with individual columns and tandem columns in order to find the best combination of two columns to use in a TC-LC experiment. We use the minimum resolution in each simulated chromatogram (critical resolution, RC) to compare the performance of different separations. With same total column length (10 cm) and same total plate number (N=8000), a C++ based simulation comparison on 200 test samples each containing 16 randomly generated analytes shows that, on average, the best TC-LC separation provides a 0.419 greater amount of resolution (RC) than the best conventional single column LC (SC-LC) , i.e. RC,average=1.439 (TC-LC) VS RC,average=1.020 (SC-LC). TC-LC was able to successfully separate (RC≥1.5) 89 out of 200 random 16-analyte samples while SC-LC could only successfully separate 9 out of the same 200 random samples. The results also show that TC-LC with a randomly picked pair of columns usually gives a better separation than conventional one-dimensional HPLC with a randomly picked column. In terms of simulation accuracy, predicted versus experimentally-measured retention factors for a representative subset of HSM test compounds differed by less than 2% (|kexperimental/kpredicted -1|≤2%) when a proper extra-column time correction was applied. The simulation can be used to find better column combinations or individual columns in method development.

Improved separation performance by using tandem-column LC with conventional U/HPLC instrumentation

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.