UHPLC Analyis of Protein Standards on TSKgel® UP-SW3000: Column Performance as a Function of Flow Rate
Materials
related product
CONDITIONS
column
TSKgel UP-SW3000, 15 cm x 4.6 mm I.D., 2 μm particles (80023449)
mobile phase
0.2 M sodium phosphate, dibasic, pH 7.0 with phosphoric acid
flow rate
0.1, 0.3 and 0.5 mL/min
pressure
710, 1865, and 2980 (49, 129, and 205 bar)
column temp.
25 °C
detector
UV, 215 nm
injection
2 μL
sample
Waters BEH200 SEC protein standard
Description
Analysis Note
In considering the capabilities of modern analytical HPLC and UHPLC instrumentation and columns, column performance (as measured by peak efficiency) signifcantly declines as flow rate increases, in the case of size exclusion chromatography of proteins. Despite this universal phenomenon, due to the slow diffusion coefficients biomacromolecules, it is remarkable that resolution can remain more than adequate at the higher flow rates, and thus allow for dramatic gains in throughput, particularly with small-particle columns.
Legal Information
TSKgel is a registered trademark of Tosoh Corporation