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IEX

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8 views

IEX

Uploaded by

f20221444
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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Ion Exchange

Chromatography
Introduction

• A chromatographic method used to separate ions


and polar molecules.
• Utilizes charged resin to attract and retain oppositely
charged ions.
• Common in pharmaceuticals for purification,
separation, and analysis.
Principle
• Relies on electrostatic interactions
between charged resin and target
ions.
• Ions in the sample replace
counterions in the resin, creating
selective retention.
• Separation occurs based on ionic
affinity and strength of the mobile
phase.
Types of IEX
Cation Exchange: Retains cations; resin has negative
charge.
Anion Exchange: Retains anions; resin has positive
charge.
• Each type has specific applications based on target
ion properties.
Anion Exchange
Chromatography
• A type of ion exchange chromatography where

negatively charged ions (anions) are attracted to a

positively charged stationary phase.

• Ideal for separating anions like chloride, phosphate,

and sulfate from mixtures.


Stationary Phase:

Resin Type: Positively charged (anion-exchange) resin,


often with quaternary ammonium groups.

Example Resins: DEAE (Diethylaminoethyl) cellulose, Q-


Sepharose.

Mobile Phase:

Buffered solution with varying ionic strength or pH,


often containing a salt like NaCl.
Cation Exchange
Chromatography
• A technique where positively charged ions (cations)
are attracted to a negatively charged stationary
phase.
• Used for separating cations such as sodium,
potassium, and proteins with positively charged
residues.
• Resin Type: Negatively charged (cation-exchange)
resin, typically with carboxylic or sulfonic acid
groups.
• Example Resins: CM (Carboxymethyl) cellulose, SP-
Sepharose
Properties of Ion
Exchange Resins
• Type of Ions: Specificity for cations or anions.
• Nature of Ions: Strong or weak ion affinity.
• Efficiency of Resin: Affects resolution and separation
quality.
• Particle Size: Smaller particles increase surface area
and resolution.
• Structural Type: Porous or non-porous, affecting flow
rate.
Types of Ion Exchange
Resins
Strong Acid/Strong Base Resins: For complete
ionization at a range of pH.
Weak Acid/Weak Base Resins: For specific pH range
applications.
• Structural forms: gel-type and macroporous resins.
• Strong cation exchange resin - SO₃H

• Weak cation exchange resin - COOH, OH, SH, PO₃H₂

• Strong anion exchange resin - N⁺R₃, NR₂

• Weak anion exchange resin - NHR, NH₂


Instrumentation
Elution Methods
• Process of removing bound ions from the stationary
phase by altering conditions in the mobile phase.
• Allows selective release and separation of ions or
molecules based on their binding affinities.
Common Elution Methods: pH change, ionic strength
adjustment, and use of competing ions.
pH Change Elution
Principle: Adjusting pH affects the charge of target molecules
and the ion exchange resin.
• Lowering pH can protonate acidic groups, weakening their
interaction with anion exchange resins.
• Increasing pH can deprotonate basic groups, reducing their
interaction with cation exchange resins.

Example: Eluting acidic proteins from an anion exchange column


by lowering the pH.
Ionic Strength Adjustment
Elution
Increasing ionic strength of the mobile phase competes
with bound ions for resin binding sites.
• Higher salt concentration (e.g., NaCl or KCl) weakens
electrostatic interactions between ions and resin.
• Allows selective elution based on the affinity of ions
to the resin.
• Example: Gradual increase in NaCl concentration to
separate ions based on affinity.
Elution by Competing Ion
Adding ions that compete with the target molecules for
binding sites on the resin.
• Competing ions, with stronger binding affinity,
displace the target ions from the resin.
• Effective when specific ion release is required,
particularly in selective separations.
• Example: Using sodium ions to elute other cations
from a cation exchange column.
Development of the
Chromatogram
• Chromatogram displays separation profile of ions

based on retention time.

• Key peaks represent specific ions; peak height and

area correlate with ion concentration.

• Importance in quantification and purity assessment.


Analysis of the Elute
• Techniques like UV-Vis spectroscopy, conductivity
detection.
• Identifying and quantifying ions in the collected
elute.
• Applications in pharmaceutical analysis for purity and
composition.
Regeneration of Ion
Exchange Resin
• Process to restore resin capacity after ion binding.

• Involves washing with concentrated acid/base or salt

solution.

• Critical for resin longevity and cost-efficiency.


Applications
• Purification of active pharmaceutical ingredients
• Removal of metal ions and inorganic impurities in
drug formulations
• Separation of peptides, proteins, and amino acids in
biopharmaceuticals
• Quality control and validation of pharmaceutical
products

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