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Weak and strong cation exchanger different behaviors

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Description of cation exchanger for liquid chromatography.
HPLC media. Polymeric Stationary Phases. Non Leaching media of LC.

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Weak and strong cation exchanger different behaviors

  1. 1. WEAK AND STRONG CATIONEXCHANGER’S DIFFERENTBEHAVIOR.COMPARISON OF STRONG CATION EXCHANGERS (SULFO PROPYLAND SULFO ETHYL) WITH EACH OTHERS AND WITH WEAK CATIONEXCHANGER (CARBOXYL) IN THE SEPARATION OF 5 PROTEINS.1
  2. 2. • The 5 proteins involved areMyoglobin Ribonuclease A α-Chymotrypsinogen ACytochrome cLysozyme2
  3. 3. 3USING• THE SAME SIZE COLUMNS,• THE SAME BUFFERS,• THE SAME pH’S,• THE SAME LINEAR FLOW RATES,• THE SAME DETECTION WAVELENGTHS AND• THE SAME PROTEINS,WE NOTICE CERTAIN REVERSAL OF THE PROTEIN ELUTION AS WELLAS CERTAIN CHANGES OF THE ELUTION PROFILES.PLEASE TAKE NOTE:
  4. 4. 4Separation of5 proteins on a4.6 x 100 mmSTYROS™ SP/XH.HPLC System. HP 1100 with thermostattedcolumn compartmentColumns STYROS™ SP/XH 4.6 x 100 mmMobile Phase A: 20 mM Phosphate, pH = 6.8B: A + 1 M NaCl, pH = 6.8Flow rate 1 ml/min (360 cm/hr)Gradient 12 to 55 % B in 12 cv,Temperature 30 CDetection 214 nmInjection volume 20 lPressure Drop 9 bar (131 psi)Samples (1:3:3:3:3 mg/ml each) Myoglobin, -Chymotrypsinogen A,Ribonuclease A, Cytochrome C fromequine, Lysozyme.HPLC System. HP 1100 with thermostattedcolumn compartmentColumns STYROS™ SE/XH 4.6 x 100 mmMobile Phase A: 20 mM Phosphate, pH = 7B: A + 1 M NaCl, pH = 7Flow rate 1 ml/min (360 cm/hr)Gradient 1 to 15 % B in 9 cv,Temperature 30 CDetection 214 nmInjection volume 20 lPressure Drop 6 bar (87 psi)Samples (2.5mg/ml each) Myoglobin, -Chymotrypsinogen A,Ribonuclease A, Cytochrome C fromequine, Lysozyme.Separation of5 proteins on a4.6 x 100 mmSTYROS™ SE/XH.
  5. 5. 5THE CHARGE ON THESE SURFACES ARE BOTH SIMILARTHAT IS A SULFONYL FUNCTION ( –SO3-) . HOWEVERTHESE CHARGES ARE THETHERED TO• A PROPYL CHAIN, -CH2-CH2-CH2-SO3- IN THE CASE OFSTYROS™ SP AND TO• AN ETHYL CHAIN, -CH2-CH2-SO3- IN THE CASE OFSTYROS™ SE.THE SURFACES ARE BOTH CONSIDERED AS STRONGCATION EXCHANGERS.
  6. 6. 6THERE IS AN ELUTION REVERSAL OF RIBONUCLEASE AAND α-CHYMOTRYPSINOGEN A AS WELL AS A CHANGE INTHE ELUTION PROFILE OF CYTOCHROME C.THIS WOULD CERTAINLY REQUIRE THE UNDERSTANDINGOF A MOLECULAR BIOLOGIST TO SHED SOME LIGHT TOTHE PHENOMENON AS WELL AS EXTEND SUCHUNDERSTANDING TO THE BEHAVIOR OF PROTEINSTOWARDS NATURAL CATIONIC ENTITIES WITHIN THELIVING SPECIES.
  7. 7. 7COMPARED TO STYROS™ SE THE CHANGE IN THIS CASE IS LIMITED TO THEELUTION PROFILE OF α-CHYMOTRYPSINOGEN A ONLY.HIGHER SALTS ARE ALSO REQUIRED FOR THE ELUTION.HPLC System. HP 1100 with thermostattedcolumn compartmentColumns STYROS™ CM/XH 4.6 x 100 mmMobile Phase A: 20 mM Phosphate, pH = 6.8B: A + 1 M NaCl, pH = 6.8Flow rate 1 ml/min (360 cm/hr)Gradient 3 to 35 % B in 9 cv,Temperature 30 CDetection 214 nmInjection volume 20 lPressure Drop 6 bar (87 psi)Samples (1;3;3;3;3 mg/ml each) Myoglobin, -Chymotrypsinogen A,Ribonuclease A, Cytochrome C fromequine, Lysozyme.Separation of5 proteins on a4.6x100mmSTYROS™ CM/XHSTYROS™ CM, HOWEVER AS A WEAK CATION EXCHANGER DOESDISPLAY DIFFERENT BEHAVIOR AS WELL:MyoglobinRibonucleaseAα-ChymotrypsinogenALysozymeCytochromecmAU050010001500200025000 2 4 6 8 10 12 14 min
  8. 8. 8ADDITIONAL INFORMATION AS WELL AS A NUMBER OFDETAILED APPLICATION NOTES CAN BE FOUND ONOraChrom’s site at https://www.orachrom.com orhttps://www.orachrom.net

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