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FYI NOTE
FYI020398
Effect of long
residency time and heat during reversed phase chromatography of some proteins
Long residency time and heat are often required when
working with narrow bore columns and small, non porous particles.
It is now possible to use
STYROS™ reversed
phase columns of different formats to find out the effects of the
chromatographic conditions on your proteins.
STYROS™
media is characterized by its full recovery of the injected proteins, the high
resolution at low and high flow rates, the pressure tolerance of up to 5000 psi,
the possibility to work on the full pH range without concerns of media
dissolution or ligand leakage, therefore providing high performances over a
considerably longer period of time.
The following chromatograms are intended to illustrate such
a concept :
Chromatogram 1
Short residency time, 20°C.
Chromatogram 2
Long residency time, 20°C.
Chromatogram 3
Short residency time, 40°C
Operating parameters:
|
HPLC System.
|
HP 1100, standard cell. |
|
Column |
STYROS™ 2R /NB
150 x 2.1mm |
|
Mobile Phase |
A: 0.1% TFA in H2O
B: 0.1% TFA in Acetonitrile/H2O (95/5)(v/v) |
|
Flow rate |
Chromatogram 1:
2.5ml/min(4,300cm/h)
Chromatogram 2: 0.2ml/min (350cm/hr)
Chromatogram 3: 2 ml/min (3,500cm/hr) |
|
Gradient |
15 to 80% B in 25 column
volumes |
|
Temperature |
20°C
and 40°C
as indicated. |
|
Detection |
214 nm |
|
Injection volume |
10
ml
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Sample
(1 mg/ml each) |
1: Soybean Trypsin
Inhibitor,
2: Cytochrome C, 3: Lysozyme,
4: Hemoglobin, 5:Ovalbumin. |
Polymeric Stationary
Phases, Setting new standards in High Speed, High Performance, Liquid
Chromatography.
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