Variations in polymeric anion exchanger from OraChrom Inc,

1. DIVERSITY IN STYROS™
ANION EXCHANGERS.
Strong and weak Anion Exchangers as
polymeric Simulated-Monolith™ stationary
phases for fast and non leaching liquid
chromatography.
1

2. SIX STYROS™ SIMULATED-MONOLITH™ ANION
EXCHANGERS ARE CONSIDERED HERE TO HIGHLIGHT
THEIR VARIABILITY.
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Weak Anion Exchanger
STYROS™ PA (Polyamine)
STYROS™ HPA (High capacity Polyamine)
STYROS™ DEAE (Diethylaminoethyl)
Strong Anion Exchanger
STYROS™ Q (Quaternary amino methyl)
STYROS™ HQ (High capacity Quaternary
amino methyl)
STYROS™ QAE (Quaternary amino ethyl)

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• Similar conditions were chosen to run the separations.
• Pancreatin as a complex dynamic mixture is used as sample.
It contains enzymes such as Amylase, Trypsin, Lipase, Ribonuclease, and
other proteases.
The following chromatograms show that the column length remains a major
factor in identifying the majority of the components present in the sample.

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4
Weak Anion Exchanger
STYROS™ PA (Polyamine)
Weak Anion Exchanger
STYROS™ HPA (High capacity Polyamine)
mAU
0
500
1000
1500
2000
2500
min0 10 20 30 40 50
Pancreatin Separation on STYROS PA
(4.6 x 300 mm)
HPLC System. Agilent 1100 with thermostatted column
compartment and quaternary pump.
Columns STYROS PA/XH 4.6 X 300 mm (4.98 ml)
Mobile phase. A: 20 mM Tris, pH=8.15
B: A + 1 M NaCl, pH= 8.15
Flow rates 1 ml/min (360 cm/hr of linear velocity)
Gradient 1 to 9 % B in 9 min. to 80 % B in 60 min.
Temperature 30 C
Detection 214 nm
Injection
volume
100µl
Pressure Drop 14 bar (203 psi)
Sample: Pancreatin 10 mg/ml in buffer A.
mAU
0
200
400
600
800
1000
1200
min0 20 40 60 80 100
Pancreatin Separation on STYROS HPA
(4.6 x 300 mm)
HPLC System. Agilent 1100 with thermostatted column
compartment and quaternary pump.
Columns STYROS HPA/XH 4.6 X 300 mm (4.98 ml)
Mobile phase. A: 20 mM Tris, pH=8.15
B: A + 1 M NaCl, pH= 8.15
Flow rates 0.5 ml/min (180 cm/hr of linear velocity)
Gradient 1 to 3% B in 22 min to 80% B in 120 min.
Temperature 30 C
Detection 214 nm
Injection
volume
100µl
Pressure Drop 10 bar (145 psi)
Sample: Pancreatin 5mg/ml in buffer A.

5. 5
Weak Anion Exchanger
STYROS™ DEAE (Diethylaminoethyl)
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min0 20 40 60 80 100
0
250
500
750
1000
1250
1500
1750
2000
mAU
Pancreatin Separation on STYROS
DEAE (4.6 x 300 mm)
HPLC System. Agilent 1100 with thermostatted column
compartment and quaternary pump.
Columns STYROS DEAE/XH 4.6 X 300 mm (4.98 ml)
Mobile phase. A: 20 mM Tris, pH=8.15
B: A + 1 M NaCl, pH= 8.15
Flow rates 0.5 ml/min (180 cm/hr of linear velocity)
Gradient 0 to 1% B in 18 min to 60% B in 120 min.
Temperature 30 C
Detection 214 nm
Injection
volume
100µl
Pressure Drop 10 bar (145 psi)
Sample: Pancreatin 5mg/ml in buffer A.

6. 6
Strong Anion Exchanger
STYROS™ Q (Quaternary amino methyl)
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Strong Anion Exchanger
STYROS™ HQ (High capacity Quaternary
amino methyl)
min0 10 20 30 40 50
mAU
0
400
600
800
1000
1200
1400
200
1600
Pancreatin Separation on STYROS Q
(4.6 x 300 mm)
HPLC System. Agilent 1100 with thermostatted column
compartment and quaternary pump.
Columns STYROS Q/XH 4.6 X 300 mm (4.98 ml)
Mobile phase. A: 20 mM Tris, pH=8.15
B: A + 1 M NaCl, pH= 8.15
Flow rates 1 ml/min (360 cm/hr of linear velocity)
Gradient 0 to 40 % B in 60 min.
Temperature 30 C
Detection 214 nm
Injection
volume
100µl
Pressure Drop 14 bar (203 psi)
Sample: Pancreatin 10 mg/ml in buffer A.
mAU
0
500
1000
1500
2000
2500
min0 10 20 30 40 50
Pancreatin Separation on STYROS HQ
(4.6 x 300)
HPLC System. Agilent 1100 with thermostatted column
compartment and quaternary pump.
Columns STYROS HQ/XH 4.6 X 300 mm (4.98 ml)
Mobile phase. A: 20 mM Tris, pH=8.15
B: A + 1 M NaCl, pH= 8.15
Flow rates 1 ml/min (360 cm/hr of linear velocity)
Gradient 0 to 40 % B in 60 min.
Temperature 30 C
Detection 214 nm
Injection
volume
100µl
Pressure Drop 14 bar (203 psi)
Sample: Pancreatin 10 mg/ml in buffer A.

7. 7
Strong Anion Exchanger
STYROS™ QAE (Quaternary amino ethyl)
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mAU
0
500
1000
1500
2000
min0 10 20 30 40 50
Pancreatin Separation on STYROS HQ
(4.6 x 300 mm)
HPLC System. Agilent 1100 with thermostatted column
compartment and quaternary pump.
Columns STYROS QAE/XH 4.6 X 300 mm (4.98 ml)
Mobile phase. A: 20 mM Tris, pH=8.15
B: A + 1 M NaCl, pH= 8.15
Flow rates 1 ml/min (360 cm/hr of linear velocity)
Gradient 0 to 40 % B in 60 min.
Temperature 30 C
Detection 214 nm
Injection
volume
100µl
Pressure Drop 14 bar (203 psi)
Sample: Pancreatin 10 mg/ml in buffer A.

8. 8
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To NOTE:
The anion exchangers featured here are all part of a series of products based on “hard gel” polystyrene
divinyl-benzene cross linked products that can withstand high back pressures and therefore run at high flow
rates.
As polymers they are distinct from Agarose based products that are natural “soft gel” products prone to
biodegradation which is the primary reason for their biodegradability and their leaching.
Although “soft gel” have low back pressures and can also be run at high flow rates, their capacity drops
drastically as the linear flow rate increases and residency time decreases.
The additional feature of STYROS™ media is their porosity. Such feature allows uniform flow-through as well
as higher capacity.
Clean in place (CIP) should also be mentioned as they can withstand harsh acidic and basic treatments
and therefore minimize the repetitive packing of columns.