Protein Mixture

• Short Analysis Time
• Simple, Easy, Fast
• Cholesterol Bonded Capillaries

Method:
A solution of Proteins (Cytochrome-C, Lysozyme (chicken) and Ribonuclease A) was prepared and injected onto a CelerityCE™ Cholesteryl High Surface Area Column using the Beckman P/ACE HPCE system. A voltage of 25kV was applied across the column resulting in 29uA. The mobile phase (60mM Citric Acid, 50mM ß-Alanine) was prepared using Good Laboratory Practice. The Capillary was conditioned following the procedure suggested by the column manufacturer. Injection was Hydrodynamic for 5 seconds under 5” of Hg.

Discussion and Rationale:
The protein mixture was chosen to show the increased resolution of proteins by the Cholesterol Bonded Capillary. The experimental conditions for the capillary were identical yet the Cholesterol column resolved peaks 1 and 2. Minor components can be seen on the Cholesterol Bonded Capillary that cannot be seen on the Bare Fused Silica Column.

Basic compounds that are as hydrophobic as these compounds, usually require a more hydrophilic phase to be separated without tailing effects. Notice that in the separation of this mixture, not only were high efficiencies achieved but also tailing is minimized.

UV detection of 211nm was selected and sharp, well resolved peaks result. Each of these proteins are basic yet they are very well resolved with the Cholesterol Bonded Capillary.