3 - UPLC_Work_Practical_Considerations

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Running U-HPLC: Practical Considerations Bottle Caps WAT062341 4 L bottle (large neck) WAT062479 1 L bottle (small neck) No Parafilm ® or other plastic films to cover solvent reservoirs Transferring Methods from HPLC to UPLC and from Binary to Quaternary UPLC Shula Levin, Waters Israel 8
Running U-HPLC: Practical Considerations Method Transfer Tools • New Columns Calculator – Now from HPLC to UPLC to HPLC – Localized – Ability to cut and paste conditions into Empower Methods Editor • Pre-injection volume and Gradient Smart Start – Useful for accommodating systems with different dwell volumes • New Method Transfer Chemistry Kits Transferring HPLC Methods between HPLC Systems of Multiple Vendors • Differences in system volume – keep the column/system volume ratio constant to ensure a similar ‘isocratic hold’ caused by the system at the beginning of the gradient. • An isocratic hold can be added to the gradient table if the system volume of the new HPLC is smaller • An injection hold may be added if the system volume of the new HPLC is larger (setting the pre-column volume to non-zero in the instrument method) • Differences in column heating – Column heaters have different heating efficiencies and the differences can impact absolute retention of the components and selectivity Shula Levin, Waters Israel 9
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