OraChrom, Inc.

  
       

 

 

Home
Up

APPLICATION NOTE

AN071306-42


Reversed Phase Polymeric: Comparison of STYROS™  2R with Commercial Non Porous Polymeric.

 

A non altered set of proteins found in egg white, were separated on a STYROS™ 2R/XH 4.6 x 50 mm column (volume 0.83 ml) at a linear flow rate of 1,450 cm/hr (4 ml/min volumetric flow) and compared with the performance of a non porous polymeric column of 4.1 mm x 30 mm (volume 0.4 ml) run at 900 cm/hr of linear flow rate (2 ml/min volumetric flow).

The results show a clear advantage with the STYROS™ column both in performance and operating conditions.

 

Chromatogram 1

Separation of egg white proteins on STYROS™  2R/XH

 

Table 1. Operating parameters.

HPLC System.

Agilent 1100 with thermostatted column compartment.

Columns

STYROS™ 2R/XH 4.6 X 50 mm

Mobile phase.

A: 0.075 % TFA in H2O.
B: 0.075 % TFA in ACN:H2O (95:5)

Flow rate

4 ml/min (1,450 cm/hr )

Gradient

30 to 50 % B in 1 min. 50% B to 2 min.

Temperature

30°C

Detection

215 nm

Injection volume

10 ml

Sample:

Egg white in buffer A. Mixture of 1:9 .
1- Lysozyme, 2-Conalbumin, 3-Ovalbumin.

The three major proteins are separated at high flow rates in less than 2 minutes.

 

Chromatogram 2

Egg white proteins separated on a commercial non porous polymeric midia

 

Table 2. Operating parameters.

Columns

Commercial non porous polymeric 4.1 x 30 mm

Mobile phase.

A: 0.1% TFA in H2O
B: 0.1 % TFA in Acetonitrile

Flow rates

2 ml/min (900 cm/hr )

Gradient

0 to 60 % B in 20 min

Temperature

Room temperature

Detection

215 nm

Sample:

Egg white in buffer A. Mixture of 1:9

 

The two major peaks have been identified as Conalbumin and Ovalbumin.

This clearly shows the importance of the proper manufacture of porous polymeric media in order to have a narrow distribution of throughpores. The stability of the media and the absence of fines and sub-micron particles is also important as they result in clogging of throughpores that would operate as long diffusive pores.

[ Home ] [ Up ]

Copyright © 1997-2009 OraChrom, Inc.