Crystallization

Dr.Sangeetha Subramanian
CRYSTALLIZATION

 Formation of solid particles of defined shape and size from a homogeneous

liquid phase
 Oldest and most common purification cum polishing stage

 Inorganic salts (sodium & ammonium sulfates), Organic compounds

(sodium and glucose) & many Pharmaceuticals and organic fine
chemicals - marketed as crystalline products
Purpose of Crystallizer

 Used to recover pure solids from solution

 Highly desirable end product because of:

 Exceptional purity
 Ease of handling
 Long shelf life

 One of the final treatment steps in the

purification and concentration of insulin

 98% of the insulin must be crystallized

Laboratory crystallization

Lab crystallization  relatively simple operation  clear warm concentrated

solution of the solute is cooled slowly in a dust free environment  until small

crystals appear.
BASIC CONCEPTS
 Crystal –
1. Highly organized particle of non-living material

2. Orderly 3-dimensional array  space lattice

3. Lattice is made of particles  atoms/ ions/ molecules

4. Polyhedrons – sharp corners and flat sides or faces

5. Angles made by the sides of all crystals of the same

compound are constant
 Saturation –
1. Maximum concentration of solute which is
thermodynamically stable in solution

2. Saturation is a result of phase equilibria among

solutes and the surrounding solution “mother liquor”

3. Solute mass per solvent mass Vs. Temperature 

relation used to understand phase equilibria

4. In many cases; solution can contain more solute

than present at saturation  Supersaturated solution
 Metastable

5. Temperature-solubility -solute concentration diagram

(next slide) helps to understand supersaturated state
of a system
 Super-
Saturated

Un-
Saturated

Solubility
curve
Metastable zone: solute in excess of the equilibrium
concentration  deposits on existing crystals BUT no new
crystal nuclei formed

Intermediate zone: growth on existing crystals +

formation of new nuclei

Labile zone: Nuclei are formed spontaneously

 Purity –
1. Prime importance as:
i. Crystallization  Polishing + Purification step
ii. Gains more importance when dealing with ‘bio’-
separation

2. Assessed by separation factor β

3. β is large  separation is effective

 Nucleation –
1. Concerns with the genesis of new crystals.

2. Can result from several mechanisms:

a. Homogeneous Nucleation: formation of crystals
from liquid phase due to supersaturation only

b. Heterogeneous Nucleation: results from presence

of insoluble material (usually desired product)
which initiates crystal growth

c. Secondary Nucleation: crystal formation induced

by contact between different crystals

d. Attritive Nucleation: breakup of existing crystals

into new particles which grow as new crystals
 Homogeneous nucleation:
• Target of research
• Depends upon brownian motion to generate small
clusters of solute particles in a solution
• Exist for a few microseconds
• However at high supersaturation  small clusters 
form ‘embryos’  grow  initiate macroscopic
crystallization
• Such embryos which lead to crystallization  nuclei
• Usual at small scale; rare at large scale

 Heterogeneous nuceleation:
• Present in both small and large scale crystallization
• Solid matter (usually of the desired product) initiate
crystal growth  it can be dust/ small seed crystal
• Avoids steps of cluster and embryo formation
 Secondary Nucleation
• Paramount in industrial crystallizers
• Results from interaction between existing crystals +
crystal contact with agitator/ walls of the crystallizer
• Various mechanisms exist
Methods of Crystallization

Supersaturation: liquid (solvent) contains

more dissolved solids (solute) than can
ordinarily be accommodated at that
temperature

Can be achieved by several methods:

Cooling
Evaporation
Solvent addition
Precipitant Addition
CRYSTALLIZATION UNIT
Cooling Method

Concentrated
solution gradually
cooled below
saturation
temperature (50-
60°C) to generate a
supersaturated state
Yields well defined
micron-sized crystals
Shell and tube heat
exchanger is used to
cool solution
CRYSTALLIZATION UNIT
Solvent Method

Solvents are generally good protein

precipitants
Their low dielectric constants lower the
solvating power of their aqueous
solutions
Requires acidic solvent
For crystallization, an insulin protein falls
out of solution at isoelectric point
pH 5.4-5.7
Addition of Zinc Ions

In the presence of zinc ions, insulin proteins

orient to form hexamer structures

Zinc ions render insulin insoluble which

results in micro-crystallization and
precipitation

Human Insulin Hexamer with Zinc ion

 Seeding Techniques
 Primary nucleation is the first step in crystallization - growth of a new
crystal
 Can bypass primary nucleation (creation of new crystals) by "seeding" the
solution

 Secondary nucleation is crystal growth initiated by contact

 Accelerated by "seeding" adding existing insulin crystals to perpetuate
crystal growth
Mechanism
of
Crystallizatio
n
Mechanism of Crystallization (when sample is quickly
cooled)
Crystal Size and Growth Rate

 Crystal size distribution is important for the

production process; affects:
 downstream processing
 solids transport
 caking and storage properties of the material
 Correct crystal size vital for economic production
 Crystals produced in commercial crystallization
processes are usually small
 30 to 100 um in diameter