Method Migration from UPLC Technology to Preparative HPLC
UltraPerformance LC (UPLC Technology) has been widely accepted due to the improvements over HPLC in sensitivity, resolution, and speed. For applications such as impurity and metabolite profiling, methods developed with UPLC Technology must be transferred to the preparative scale in order to isolate and purify desired compounds for further characterization.
Therefore, a systematic approach for transferring UPLC separations to the preparative scale was developed. Key parameters to consider include: column chemistry, particle size and dimension, injection volume, separation temperature, mobile phase composition, flow rate, gradient slope, cycle time, detector sensitivity, injector design, and system volume (both UPLC and preparative HPLC). Step-by-step calculations are included for a better understanding of evaluating method transfer validity. In addition, optimization of the preparative HPLC separation for impurity isolation and purification is discussed.
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