UHPLC Analyis of Protein Standards on Sepax Unix™ SEC-300: Column Performance as a Function of Flow Rate

UHPLC Analyis of Protein Standards on Sepax Unix™ SEC-300: Column Performance as a Function of Flow Rate

Conditions

column Sepax Unix SEC-300, 15 cm x 4.6 mm I.D., 1.8 µm particles
column temp. 25 °C
mobile phase 0.2 M sodium phosphate, dibasic, pH 7.0 with phosphoric acid
flow rate 0.3 mL/min
flow rate 0.1 mL/min
flow rate 0.5 mL/min
pressure 1550 psi (107 bar)
pressure 2400 psi (165 bar)
pressure 500 psi (34 bar)
sample Waters BEH200 SEC protein standard
injection 2 µL
detector UV, 215 nm

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.
Categories Analytical Chromatography, Proteins
Featured Industry Life Science and Biopharma
Legal Information Unix is a trademark of Sepax Technologies
suitability application for UHPLC

Materials

     
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