Adsorption

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Adsorption Absorption !

Absorption a fluid phase is transferred from one medium to another (i.e. water absorbed by a sponge)

Adsorption a component (the adsorbate) concentrates on a solid surface (the adsorbent) without chemical change. This occurs when a dissolved solute is bound to a solid adsorbent.

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Steps of Adsorption process

1. A feed solution is added to the adsorbent.
2. Adsorption process occurs may be both selective and slow. 3. The spent feed solution is withdrawn.

4. The adsorbed solute is eluted using a different solvent.

Adsorbent- a porous material with high internal surface area.

Main binding forces- Vander waals forces, polarity

Adsorption capacity- depends on adsorbent, adosrbate, physicochemical conditions, and the surface properties.

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Why is Adsorption Useful?

Used in many industrial processes:

dehumidification odour/colour/taste removal gas pollutant removal (H2S) water softening and deionisation hydrocarbon fractionation pharmaceutical purification

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Nature of Adsorbents

Porous material - Large surface area per unit mass size, shape, polarity cause certain particles to be held more strongly in these pores than others Rate of mass transfer is dependent on the void fraction within the pores Granular (50m - 12 mm diameter) Suitable for packed bed use

Examples
1. 2. Carbon- Activated carbon in WWT, carbons from vegetable sources ( Saw dust, Fruit pits), from mineral sources. Resins- Synthetic polymers containing fixed charges (-SO3-, -COO-, NR3+), made from Styrene, divinyl benzene, made from acrylic esters, hydrogels
Natural and synthetic zeolites Clays

3. 4.
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5.

Silica gel/ Alumina

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Types of Adsorption

Physical Adsorption

result of intermolecular forces causing preferential binding of certain substances to certain adsorbents reversible by addition of heat (via steam, hot inert gas, oven) Attachment to the outer layer of adsorbent material

Chemisorption

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result of chemical interaction large amount heat released irreversible mainly found in catalysis
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Liquid-Solid Adsorption Equilibrium

Isotherm Equilibrium relationship between the amount adsorbed (W, also referred to as adsorbate loading) and the concentration in the fluid phase (c) Equilibrium is temperature dependant hence isotherms Adsorption amount decreases with an increase in temperature
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Liquid-Solid Adsorption Isotherms

*The analysis of adsorption is based on equilibria and on mass balance.
*3 types of adsorption isotherms

1. Linear 2. Langmuir 3. Freundlich

Freundlich

In each isotherm,
Langmuir

q
Linear

Abscissa gives the solute concentration in solution i.e., mass of solute per volume of solution Ordinate gives solute concentration on the adsorbent surface i.e., mass of solute per mass of adsorbent.

Any isotherm,

concave towards abscissa- favorable

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concave towards ordinate- unfavorable

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1. Linear Isotherm

q Ky

q- amount of solute adsorbed per amount of adsorbent y- Solute concentration in solution

K- Equilibrium constant

2. Freundlich Isotherm- Adsorption for antibiotics, steroids, hormones

q Ky

Log q Log K

n, K determined from expts.

n>1- favorable, n<1- unfavorable

Log y

3. Langmuir Isotherm- for proteins

q0 y q Ky
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q 1/q0 y

K/q0

Unlike Freundlich isotherm, the Langmuir isotherm has a theoretical basischemical reaction between solute and vacant sites on adsorbent surface.
Solute + Vacant site Filled site If this reaction is at equilibrium, then it is described by an equilibrium constant K K= [Solute] [Vacant site]/ [Filled site]

The total no. of active sites must be fixed. [Total sites] = [vacant sites] + [Filled sites] [Filled sites] [Total sites] [Solute]/ { K + [Solute]}

Since the no. of filled sites is proportional to q, the above equation resembles Langmuir isotherm equation. Thus maximum value of q0 reflects the limited no. of active sites on the adsorbent.
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Separation mechanisms
Non-covalent interactions involved in adsorption:
van der Waals forces Electrostatic interactions Hydrophobic interactions Hydrogen bonding Partitioning

Separation mechanisms:
Ion exchange: Electrostatic Affinity: van der Waals, hydrogen, hydrophobic

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Reverse phase: Partition

Hydrophobic interactions: Hydrophobic
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Ion exchange: Mechanism

Adsorbents

+
-

+
-

+
+ +
-

+
-

Bound molecules

Cation exchange

Anion exchange

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Ion exchange adsorbents

Charged groups are bound on insoluble support material
Examples of support material:
Cellulose and its derivatives Agarose Acrylic resins Cross-linked dextrans

Intrinsic charge on cellulose

Charged groups or ligands:
Cation exchange:
Carboxylic acid (C) weak Carboxymethyl (CM) weak Sulphopropyl (S) strong Anion exchange: Diethylaminoethyl (DEAE) weak
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Quaternary aminoethyl (QAE) weak GVSRK/DSP/Unit-III Quaternary ammonium (Q) strong

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Ion exchange: Adsorption and desorption

Adsorption
pH Ionic strength Buffer type Counterions

Desorption or elution
High salt concentration
Shielding of electrostatic interactions Competitive binding

pH changes
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Change in sign of charge on bound protein

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Cation exchange: Lysozyme separation

Egg white proteins
Protein Isoelectric pH

Lysozyme
Ovalbumin

11.0
4.6

Conalbumin

6.1
Lysozyme + NaCl

Egg white proteins +

Other egg white proteins

+ NaCl

Cation exchange adsorbent

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Adsorbent + lysozyme
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Cation exchange adsorbent + NaCl

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Anion exchange: HSA separation

pI of HSA is 4.9

pI of HIgG is between 6.0 and 8.0

At a pH value of 5.5 the proteins are oppositely charged
HSA + NaCl

HSA and HIgG

HIgG

+ NaCl

Anion exchange adsorbent

Anion exchange adsorbent + NaCl

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Affinity adsorption
Stereo-specific recognition of molecules by ligands

Combination of:
Hydrogen bonding

Hydrophobic interaction
van der Waals forces

Specific and general affinity adsorption Binding of a dissolved molecule on the surface of solid material

Adsorption should be reversible

Uses:1. Separation of molecule from complex mixtures of molecules 2.Separation of solute from solvent Interfacial phenomenon Binding sites and ligands

Affinity separation is a highly selective form of adsorption

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Affinity adsorption has the following steps: 1. Equilibrium between support and ligand

2. Binding of Ligand to support

3. Adsorption of Substrate

4. Desorption of Bound substrates

Ligand Support Substrate Molecules

Lock and Key mechanism of Affinity adsorption

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Selectivity

Molecular weight or size of solute Shape of the molecule and the binding site/ligand Polarity of the molecules and the adsorbent Electrostatic charge on the molecule and the adsorbent

Physical properties Chemical properties

Advantages:
High selectivity of separation (e.g. affinity adsorption) Ability to handle very dilute solute concentrations

Disadvantages:
Adsorption is a surface phenomenon; therefore the interior of the adsorbent material is not involved Batch or semi-batch operations generally have to be used due to the adsorption/desorption cycle Adsorption may results in loss of biological activity Adsorbents can get fouled by biological material
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Applications

Protein fractionation

Industrial enzymes Therapeutic proteins Food proteins Plasmid purification RNA purification Bacitracin Streptomycin

Nucleic acid separation

Antibiotic purification

Biomedical analysis Pulse or elution chromatography

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Affinity adsorbents
Support material
Cross-linked dextran Cross-linked agarose Cross-linked cellulose Synthetic resins

Ligands
Dye ligands Antibodies Antigens Protein A and protein G Substrate analogues Enzyme inhibitors Complimentary base-pair sequence Receptors Amino acids Lectins Concanavalin A Immobilized metal ions

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Affinity binding and desorption

Binding:
Physiological conditions

Conditions that favour

Hydrophobic interactions

Hydrogen bonding
Van der Waals forces High anti-chaotropic salt concentrations

Desorption:
pH

Chaotropic salts
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Affinity separation: Monoclonal antibody

Impurities Monoclonal antibody

Cell culture supernatant +

+ low pH buffer Protein-A affinity adsorbent Adsorbent + monoclonal antibody Protein-A affinity adsorbent

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Reverse phase adsorption

Partition type behaviour Adsorption of non-polar molecules Partition into thin immobilized layer of hydrocarbon: C 4 to C18

Support: silica
Adsorption: Polar solvent e.g. water

Elution: Non-polar solvent e.g. acetonitrile and isopropanol

Polar media Non-polar media

Adsorbent

Immobilized hydrocarbon layer Adsorption

Desorption

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Reverse phase separation: Insulin

Filtered fermentation media + acetonitrile +

Impurities

Insulin

C-18 reverse phase adsorbent

Adsorbent + Insulin

80% acetonitrile + 20% isopropanol

C-18 reverse phase adsorbent

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Hydrophobic interaction
Interaction between the hydrophobic patches on molecules and those on the adsorbent

Mainly used for protein separation.

Hydrophobic amino acids shielded by structured layer of water molecules Hydrophobic surface shielded by structured layer of water Anti-chaotropic salts make water molecules in bulk solution structured Less shielding of hydrophobic interactions
Molecule Structured water layer

Adsorbent Structured water in bulk solution Hydrophobic layer Adsorption

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High salt concentration

Desorption

Low salt concentration

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Hydrophobic interaction: Adsorbent, adsorption and desorption

Adsorbent:
Support: Agarose most widely Groups: Butyl, octyl and phenyl

Adsorption:
High salt concentrations: 1 M or higher Salt concentration depends on the type of salt Less sodium sulphate than ammonium sulphate or sodium chloride

Desorption:
Lowering the salt concentration pH change

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Affinity separation: rEGF

Filtered fermentation media + Ammonium sulfate +

Impurities

rEGF

Hydrophobic interaction adsorbent

Ammonium sulfate free solution Adsorbent + rEGF

Hydrophobic interaction adsorbent

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Diffusional limitations in adsorption processes

Adsorption can be mass transport limited Morphology, size and geometry of adsorbent influences mass transfer Mechanisms:

External mass transport (controlled by diffusion from the bulk solution to the surface of the adsorbent) Internal mass transport (controlled by diffusion within a porous adsorbent)

With porous adsorbents commonly used in bioseparation, the second mechanism predominates.

Adsorbent

Porous adsorbent

Film
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Packed bed adsorption

Feed Wash liquid Eluent

Spent feed

Wash

Product

Adsorption

Washing

Elution

Adsorption

Washing

Elution

Concentration

Impurities

Target molecule

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Other adsorption devices

Limitations of packed beds: High pressure drop Inability to handle feed with particulate material Need to use low flow rates Radial dispersion problems

Stack of porous membranes Feed

Feed

Fluidised bed adsorber

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Membrane adsorber
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