Lesson_02

Preformulation I
Polymorphism, Particle size/shape
-Dr Ajay Semalty
Department of Pharmaceutical Sciences,
H.N.B Garhwal University (A Central University)
Srinagar Garhwal-246174

Learning outcomes

After learning this module you will be able to understand

 Concept of Polymorphism
 Importance of polymorphism in Preformulation
 Concept of pseudopolymorphism/hydrates/solvates
 Importance of pseudopolymorphism in Preformulation
 Effect of particle size and shape on preformulation
 Characterization of particle size and shape

Lesson Plan

 Polymorphism
 Importance of polymorphism in preformulation
 Origin of polymorphism
 Types of polymorphism
 Pseudopolymorphism
 Particle size and shape: Importance in preformulation
 Characterization of size and shape

Dear learners in continuous to study of physical properties to be studied in

preformulation, let‟s move to polymorphism.

Do you remember the definition of polymorphism, we discussed in the last

module.
 Module 2: Preformulation I (Polymorphism, Particle size/shape) 2

“The ability of a substance to exist as two or more crystalline phases that have
different arrangements and/or conformations of the molecules in a crystalline
lattice is called polymorphism.”

USFDA has the stringent guidelines regarding polymorphism. If drug shows

polymorphism, every type should be completely reported.

Importance of polymorphism

Therefore, the polymorphic forms of drugs are very vital forms for drug
developers because of their thermodynamic and physicochemical properties.
E.g. energy, melting point, density, stability, and in particular solubility. By
using different polymorphs, these properties are modulated so as to improve
the original form. By a factor of 2 the properties like solubility differ in any
two polymorph of a drug.

Origin of polymorphism

The differences in molecular packing and intermolecular interactions within

the three-dimensional framework of the crystalline state lead to variation in
crystal structure. The molecular structure governs the molecular packing and
hence the different crystal forms with different stability, physical properties
like density and the effectiveness also.

The polymorphism has been well exercised since decades. However, we

understood quite later.

Let me tell you an interesting story…. You now! Napoleon army soldiers
used to wear the uniform with buttons made of tin. When Napoleon and a part
of the Grande Army reached Moscow on 14 September 1812. At subzero
temperatures of Moscow, the shinning and highly decorated buttons of
soldiers turned into dirty grey. Soldiers believed that it is some wrath of God.
There moral went so down that with the combined effect of cold, diseases and
starvation they faced a pathetic defeat at the gate of Moscow. Actually, it was
polymorphism. The metallic white tin underwent a polymorphic transition to
the stable but nonmetallic grey tin, thus reducing the decorum of the mighty
soldiers.

-Tin or „white‟ tin, stable above 18oC, Tetragonal, I41 /amd a = b = 5.832, c
= 3.182 Å; Metallic.

SWAYAM MOOC: Industrial Pharmacy-I by Dr Ajay Semalty (Course Coordinator), HNB

Garhwal University (A Central University) Srinagar (Garhwal) Uttarakhand, India
 Module 2: Preformulation I (Polymorphism, Particle size/shape) 3

-Tin or „grey‟ tin, stable below 18o C, Cubic, Fd3m, a = b = c = 6.489Å,

Non-metallic.

Types of Polymorphism

A) On the basis of mechanisms crystal lattice formation

1. Packing polymorphism

2. Conformational polymorphism.

“Packing polymorphism is the formation of different crystal lattices of

conformationally rigid molecules that can be rearranged stably into different
3D structures through different intermolecular mechanisms.”

“When a nonconformationally rigid molecule can be folded into alternative

crystal structures, the polymorphism is categorized as conformational
polymorphism.”

These are broad classes of polymorphism.

Estrone shows three polymorphic form (packing polymorphs)

B) Thermodynamic classification

Depending upon whether or not one form can transform reversibly to another
with respect to the change of temperatures and pressures, it can be classified
into two forms

SWAYAM MOOC: Industrial Pharmacy-I by Dr Ajay Semalty (Course Coordinator), HNB

Garhwal University (A Central University) Srinagar (Garhwal) Uttarakhand, India
 Module 2: Preformulation I (Polymorphism, Particle size/shape) 4

1. Monotropes

2. Enantiotropes

The temperature at which the two polymorphs have equal stability is defined
as the transition temperature (Tt ).
If

Tt is located below the melting points of both polymorphs ---enantiotropic

Or

One polymorph can be reversibly changed into another one by varying the
temperature or pressure. -- enantiotropic
Tt is located above the melting points of both polymorphs --- monotropic,

Or

The change between the two forms is irreversible- monotropic

Fig. : Thermodynamic phase diagrams of polymorphs

Ritonivir story:

In March 1996, FDA approved Ritonivir (ABT-538). It was marketed as a

semisolid formulation. In 1998, however, batches began to fail dissolution
tests, and it was found that a more stable polymorph was precipitating from
the formulation. As a result, Abbot had to withdraw the product from the
market. (Chemburkar et al., 2000). It was a case of conformational
polymorphism in which a stable enantiotropes (form II) was precipitated by
formation of a degradation product of drug , later proved.

SWAYAM MOOC: Industrial Pharmacy-I by Dr Ajay Semalty (Course Coordinator), HNB

Garhwal University (A Central University) Srinagar (Garhwal) Uttarakhand, India
 Module 2: Preformulation I (Polymorphism, Particle size/shape) 5

C) Pseudo polymorphs (on the basis of solvent or water incorporated in

crystal)

“The phenomenon whereby solvent or water is incorporated in the

crystal lattice or in interstitial voids, is termed as pseudopolymorphism.”

Solvates - (inclusion of the solvent of crystallization),

Hydrates (inclusion of water of crystallization)

[and amorphous forms (where no long-range order exists) may also exist.]

Solvates

Residual solvents have been classified the ICH into three classes:

1. Class I solvents: Solvents to be avoided.

e.g. Strongly suspected human carcinogens and environmental hazards, e.g.,

benzene,

carbon tetrachloride and 1,2 dichloroethane.

2. Class II solvents: Solvents to be limited.

e.g. These include non-genotoxic animal carcinogens or possible causative

agents (e.g., acetonitrile, cyclohexane, toluene, methanol and N,N-
dimethylacetamide) of irreversible toxicity such as neurotoxicity or
teratogenicity. Also included are solvents suspected of other significant but
reversible toxicities.

3. Class III solvents: Solvents with low toxic potential,

e.g., acetic acid, acetone, ethanol, ethyl acetate and ethyl ether. Also included
are solvents with low toxic potential to man are also included here; no health-
based exposure limit is needed. Class III solvents have permissible daily
exposures (PDEs) of 50 mg or more a day.

“Whilst the use of solvates is not a usual practice, because of toxicity, it is

interesting to note that according to Glaxo‟s British patent 1,429,184, the
crystal form of beclomethasone dipropionate used in the MDI is the
trichlorofluoromethane solvate. By using the solvate, it was found that crystal
growth due to solvation of the propellant chlorofluorocarbon (CFC) was
prevented.”

SWAYAM MOOC: Industrial Pharmacy-I by Dr Ajay Semalty (Course Coordinator), HNB

Garhwal University (A Central University) Srinagar (Garhwal) Uttarakhand, India
 Module 2: Preformulation I (Polymorphism, Particle size/shape) 6

Hydrates

When water molecule is present in crystal it is called hydrates. It is the most

common case of solvation and it is almost always involved in hydrogen
bonding.

The hydrogen bonding network contributes to the coherence of the

crystal and hence hydrates usually show slower dissolution rates compared
to the corresponding anhydrates.

Crystalline hydrates have been classified by structural aspects into three

classes:

 Isolated lattice sites (water molecules reside in the crystal as isolate

lattice),
 Lattice channels (water molecules fill the space but not in contact with
each other in crystal), and
 Metal-ion coordinated water (metal coordinated water in salts of weak
acids) e.g. nedocromil sodium trihydrate

Other examples of hydrates (with number of hydrates in brackets) as reported

in USP are aminophyine (2), ampicillin (3), caffeine (1), dextrose (1), sodium
acetate (3) etc.
Hydrates can also exist in various polymorphs, such as in the case of
amiloride hydrochloride. “Amiloride hydrochloride dihydrate is present in
two polymorphic forms. By milling or compressing both forms, it was shown
that form A was more stable than form B. Moreover, it was shown that the
anhydrate rapidly rehydrated to form A dihydrate on exposure to atmospheric
RH.” (Jozwiakowski et al., 1993).

During preformulation, polymorphs are selected on the basis of

 Physical and chemical stability
 Behavior to processing and formulation,
 Biopharmaceutical properties (predictive assessment of in vivo
performance).

Characterization methods
As discussed in module 1, all the methods used for crystal characterization are
used for polymorph form characterization. In characterization the complete
profiling of all polymeric form are needed to be done.
Steps are as followed
1. Characterize the forms:
e.g. - X-ray Powder Diffraction, - DSC / Thermoanalysis, Microscopy,
Spectroscopy

SWAYAM MOOC: Industrial Pharmacy-I by Dr Ajay Semalty (Course Coordinator), HNB

Garhwal University (A Central University) Srinagar (Garhwal) Uttarakhand, India
 Module 2: Preformulation I (Polymorphism, Particle size/shape) 7

2. If the forms have different properties? (solubility, stability, melting point)

then move to next step other no further testing required
3. If drug product safety, performance or efficacy is affected, then move to
step 4, otherwise no further testing is required.
4. Set acceptance criterion for polymorph content in drug substance with
respect to control on the ratio of forms, dissolution and stability.

Now moving to next physical property to be studied in preformulation

Particle size and shape

Particle size is one of the very basic parameter of preformulation and dosage
form development.

 On the basis of dosage form to be developed the demand of particle

size varies. E.g. the particle size should be in the range of 2-5 microns
for inhalational therapy.
 Particle size directly affects the solubility and dissolution of the drug.
 Particle size ensure drug content uniformity and compressibility.
 Particle shape affects the flow property and binding in tablet
manufacturing.
 Particle shape is also related to contact points and hence the solubility.
Example: flour needed for besan laddu…. Coarse not fine…….

As some basic rules are..

“Smaller means nearer or quicker to connect”
“Smaller means more reactive” e.g. sugar crystal v/s sugar powder
“Smaller means higher surface area.”
Let‟s see with the example
 Take a dice (simple cube) of 1.7 cm side each
 Cut the cube 3 times (horizontal, vertical, and transversal)
 Divided similarly 24 times
 The resultant cubes will be 1 nanometer cubes and if spread in a single
layer could cover a football field.

SWAYAM MOOC: Industrial Pharmacy-I by Dr Ajay Semalty (Course Coordinator), HNB

Garhwal University (A Central University) Srinagar (Garhwal) Uttarakhand, India
 Module 2: Preformulation I (Polymorphism, Particle size/shape) 8

But sometime decreasing the size may result in reduction of solubility e.g.
aspirin. Aspirin is hydrophobic drug. And when we reduce its particle size, its
surface area increases and being hydrophobic its hydrophobic surface area
increases and hence the water solubility decreases more.

Particle size reduction

 Mortar pestle:
o for lab scale,
o when material is less in quantity ;
o more suitable or feasible in preformulation stage
 Ball milling:
o for batch process,
o in relatively large scale,
o when material is in large quantity;
o through a combined process of compact and attrition
o consist of a hollow cylinder (containing different sizes of
balls), rotated for grinding
o critical factors include speed, mill size, wet or dry milling and
amount of sample
o but continuous milling may be detrimental for stability and
crystallinity of the compound
 Micronization
o Routine method for large scale size reduction by air jet milling
o It involves feeding the sample into a confined circular chamber
and then exposed to high velocity stream of air
o Particles of less than 10 μm are obtained.
o Solubility and bioavailability improves with the micronized
drug e.g. felodipine
o Changes in crystallinity may occur e.g. salbutamol sulphate
SWAYAM MOOC: Industrial Pharmacy-I by Dr Ajay Semalty (Course Coordinator), HNB
Garhwal University (A Central University) Srinagar (Garhwal) Uttarakhand, India
 Module 2: Preformulation I (Polymorphism, Particle size/shape) 9

There are several other methods of micronization please do refer from text
given in further readings. However you can recall from your previous
semester lectures.

Now moving to particle size distribution

Particle size distribution
The uniform distribution of particles is also an important property. The
particle size analysis can be done by following methods
 Microscopy : Optical microscopy, Electron Microscopy
 Mechanical: Sieving method
 Electronic: Coulter counter
 Laser light scattering (DLS)
 Miscellaneous techniques: Centrifugation, Air suspension,
Sedimentation (Andreasen pipet, recording balance) etc.

Adapted from: Mullin JW, Anal. Proc. 1993; 30:455-456

Optical microscopy
Using light microscope, the particles (Suspended in nondissolving fluid))
are observed through an ocular micrometer which is previous calibrated
using a bench micrometer. The number of particle covered in one, two,
three or more parts of ocular micrometer are noted and then particle
distribution is plotted and the average particle size calculated.
Electron microscopy
We have discussed the technique in module 1. It is used for shape and
size measurement of particles with very high resolution.

Sieving Method

 Quantitative particle size distribution analysis.

 For size > 50 μm upward.
 Shape has strong influence on results.
SWAYAM MOOC: Industrial Pharmacy-I by Dr Ajay Semalty (Course Coordinator), HNB
Garhwal University (A Central University) Srinagar (Garhwal) Uttarakhand, India
 Module 2: Preformulation I (Polymorphism, Particle size/shape) 10

 Nest of standard size sieves (coarsest on top and finest on

bottom) is mounted on sieve shaker, certain weighed amount is
charged on the top, sieve shaker shaken for 10-15 minutes,
amount retained on each sieve is noted, weight size is
calculated for each sieve set and then average is calculated.

Coulter counter
Particles are forced to flow through an aperture or orifice situated between
two electrodes.
When the particle traverses the measurement zone, there is a change in
voltage, current, resistance, and capacitance, all of which could be measured
and used to count and size the particle.

Laser light scattering (Dynamic Light Scattering or DLS)

 The most advanced technique
 used exclusively for the sub-100 nm region with wide range of 0.02 to
2000 μm
 The use of this technique is based on light scattered through various
angles, which is directly related to the diameter of the particle. Thus,
by measuring the angles and intensity of scattered light from the
particles, a particle size distribution can be deduced.
 Rapid: single measurement in 0.001 seconds or quicker
 Compact instrument, table top
 But expensive
SWAYAM MOOC: Industrial Pharmacy-I by Dr Ajay Semalty (Course Coordinator), HNB
Garhwal University (A Central University) Srinagar (Garhwal) Uttarakhand, India
 Module 2: Preformulation I (Polymorphism, Particle size/shape) 11

Other methods
Outdated and time consuming.

Summary
• Polymorphism means one chemical can stay in two or more physical
form;
• It may be of various types: packing, conformational, enantiotropic,
monotropic, solvates and hydrates.
• Stable and effective polymorphic form must be identified.
• Particle size and shape influence solubility, dissolution, content
uniformity and flow properties.

Further Readings
 Guidance for industry, ANDAs: Pharmaceutical Solid Polymorphism,
https://www.fda.gov/downloads/drugs/guidances/ucm072866.pdf
 Censi R, Martino PD. Polymorph impact on the bioavailability and
stability of poorly soluble drugs, Molecules 2015, 20, 18759-18776;
doi:10.3390/molecules201018759
 Niazi SK, Handbook of Preformulation, Informa health care, 2007.
 Gibson M. (Ed), Pharmaceutical preformulation and formulation: a
practical guide from candidate drug selection to commercial dosage
form, II edn, Informa Healthcare, 60- 75.
 Qiu Y, Chang Y and Zhang GZ (Exe. Eds), Developing solid oral
dosage forms: Pharmaceutical theory and practice, Elsevier, 2009, pp
25-35.
 https://mortada8.wordpress.com/tag/drug-polymorphism/
 http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1984-
82502014000100002
 Ballmill 3d speed demonstration,
https://www.youtube.com/watch?v=gEWynTtguPo

References
 Purohit R, Venugopalan P, Polymorphism: An Overview, Resonance,
2009, 882-893.
 Jozwiakowski MJ, Williams SO, Hathaway RD. Relative physical
stability of the solid forms of amiloride, hydrochloride. Int J Pharm
1993; 91:195–207.
 Mullin JW, Anal. Proc. 1993; 30:455-456
 Censi R, Martino PD. Polymorph impact on the bioavailability and
stability of poorly soluble drugs, Molecules 2015, 20, 18759-18776;
doi:10.3390/molecules201018759
 Niazi SK, Handbook of Preformulation, Informa health care, 2007.

SWAYAM MOOC: Industrial Pharmacy-I by Dr Ajay Semalty (Course Coordinator), HNB

Garhwal University (A Central University) Srinagar (Garhwal) Uttarakhand, India
 Module 2: Preformulation I (Polymorphism, Particle size/shape) 12

 Gibson M. (Ed), Pharmaceutical preformulation and formulation: a

practical guide from candidate drug selection to commercial dosage
form, II Edn, Informa Healthcare, 60- 75.
 Qiu Y, Chang Y and Zhang GZ (Exe. Eds), Developing solid oral
dosage forms: Pharmaceutical theory and practice, Elsevier, 2009, pp
25-35.

SWAYAM MOOC: Industrial Pharmacy-I by Dr Ajay Semalty (Course Coordinator), HNB

Garhwal University (A Central University) Srinagar (Garhwal) Uttarakhand, India