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Diversity in styros™ anion exchangers

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Variations in polymeric anion exchanger from OraChrom Inc,

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Diversity in styros™ anion exchangers

  1. 1. DIVERSITY IN STYROS™ANION EXCHANGERS.Strong and weak Anion Exchangers aspolymeric Simulated-Monolith™ stationaryphases for fast and non leaching liquidchromatography.1
  2. 2. SIX STYROS™ SIMULATED-MONOLITH™ ANIONEXCHANGERS ARE CONSIDERED HERE TO HIGHLIGHTTHEIR VARIABILITY.https://www.orachrom.comhttps://www.orachrom.net 2Weak Anion ExchangerSTYROS™ PA (Polyamine)STYROS™ HPA (High capacity Polyamine)STYROS™ DEAE (Diethylaminoethyl)Strong Anion ExchangerSTYROS™ Q (Quaternary amino methyl)STYROS™ HQ (High capacity Quaternaryamino methyl)STYROS™ QAE (Quaternary amino ethyl)
  3. 3. https://www.orachrom.comhttps://www.orachrom.net 3• 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, andother proteases.The following chromatograms show that the column length remains a majorfactor in identifying the majority of the components present in the sample.
  4. 4. https://www.orachrom.comhttps://www.orachrom.net4Weak Anion ExchangerSTYROS™ PA (Polyamine)Weak Anion ExchangerSTYROS™ HPA (High capacity Polyamine)mAU05001000150020002500min0 10 20 30 40 50Pancreatin Separation on STYROS PA(4.6 x 300 mm)HPLC System. Agilent 1100 with thermostatted columncompartment and quaternary pump.Columns STYROS PA/XH 4.6 X 300 mm (4.98 ml)Mobile phase. A: 20 mM Tris, pH=8.15B: A + 1 M NaCl, pH= 8.15Flow 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 CDetection 214 nmInjectionvolume100µlPressure Drop 14 bar (203 psi)Sample: Pancreatin 10 mg/ml in buffer A.mAU020040060080010001200min0 20 40 60 80 100Pancreatin Separation on STYROS HPA(4.6 x 300 mm)HPLC System. Agilent 1100 with thermostatted columncompartment and quaternary pump.Columns STYROS HPA/XH 4.6 X 300 mm (4.98 ml)Mobile phase. A: 20 mM Tris, pH=8.15B: A + 1 M NaCl, pH= 8.15Flow 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 CDetection 214 nmInjectionvolume100µlPressure Drop 10 bar (145 psi)Sample: Pancreatin 5mg/ml in buffer A.
  5. 5. 5Weak Anion ExchangerSTYROS™ DEAE (Diethylaminoethyl)https://www.orachrom.comhttps://www.orachrom.netmin0 20 40 60 80 100025050075010001250150017502000mAUPancreatin Separation on STYROSDEAE (4.6 x 300 mm)HPLC System. Agilent 1100 with thermostatted columncompartment and quaternary pump.Columns STYROS DEAE/XH 4.6 X 300 mm (4.98 ml)Mobile phase. A: 20 mM Tris, pH=8.15B: A + 1 M NaCl, pH= 8.15Flow 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 CDetection 214 nmInjectionvolume100µlPressure Drop 10 bar (145 psi)Sample: Pancreatin 5mg/ml in buffer A.
  6. 6. 6Strong Anion ExchangerSTYROS™ Q (Quaternary amino methyl)https://www.orachrom.comhttps://www.orachrom.netStrong Anion ExchangerSTYROS™ HQ (High capacity Quaternaryamino methyl)min0 10 20 30 40 50mAU04006008001000120014002001600Pancreatin Separation on STYROS Q(4.6 x 300 mm)HPLC System. Agilent 1100 with thermostatted columncompartment and quaternary pump.Columns STYROS Q/XH 4.6 X 300 mm (4.98 ml)Mobile phase. A: 20 mM Tris, pH=8.15B: A + 1 M NaCl, pH= 8.15Flow rates 1 ml/min (360 cm/hr of linear velocity)Gradient 0 to 40 % B in 60 min.Temperature 30 CDetection 214 nmInjectionvolume100µlPressure Drop 14 bar (203 psi)Sample: Pancreatin 10 mg/ml in buffer A.mAU05001000150020002500min0 10 20 30 40 50Pancreatin Separation on STYROS HQ(4.6 x 300)HPLC System. Agilent 1100 with thermostatted columncompartment and quaternary pump.Columns STYROS HQ/XH 4.6 X 300 mm (4.98 ml)Mobile phase. A: 20 mM Tris, pH=8.15B: A + 1 M NaCl, pH= 8.15Flow rates 1 ml/min (360 cm/hr of linear velocity)Gradient 0 to 40 % B in 60 min.Temperature 30 CDetection 214 nmInjectionvolume100µlPressure Drop 14 bar (203 psi)Sample: Pancreatin 10 mg/ml in buffer A.
  7. 7. 7Strong Anion ExchangerSTYROS™ QAE (Quaternary amino ethyl)https://www.orachrom.comhttps://www.orachrom.netmAU0500100015002000min0 10 20 30 40 50Pancreatin Separation on STYROS HQ(4.6 x 300 mm)HPLC System. Agilent 1100 with thermostatted columncompartment and quaternary pump.Columns STYROS QAE/XH 4.6 X 300 mm (4.98 ml)Mobile phase. A: 20 mM Tris, pH=8.15B: A + 1 M NaCl, pH= 8.15Flow rates 1 ml/min (360 cm/hr of linear velocity)Gradient 0 to 40 % B in 60 min.Temperature 30 CDetection 214 nmInjectionvolume100µlPressure Drop 14 bar (203 psi)Sample: Pancreatin 10 mg/ml in buffer A.
  8. 8. 8https://www.orachrom.comhttps://www.orachrom.netTo NOTE:The anion exchangers featured here are all part of a series of products based on “hard gel” polystyrenedivinyl-benzene cross linked products that can withstand high back pressures and therefore run at high flowrates.As polymers they are distinct from Agarose based products that are natural “soft gel” products prone tobiodegradation 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 dropsdrastically 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 wellas higher capacity.Clean in place (CIP) should also be mentioned as they can withstand harsh acidic and basic treatmentsand therefore minimize the repetitive packing of columns.

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