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INTRODUCTION
Preformulation study is the foundation
of developing robust formulation.
It can be defined as a phase of research
& development process for an
investigation of physical and chemical
properties of new drug substance alone
or in combination with other excepients
in order to development of safe and
effective dosage form.
OBJECTIVE
The overall objective of preformulation
testing is to generate information useful to
the formulator:

To formulate stable and effective dosage form

To increased drug stability
To improve drug bioavailability
Reduce drug excipient incompatibility
APPLICATIONS
Preformulation studies begins or shall be
updated
immediately after the synthesis and initial

toxicity screening of a new drug.

when a newly synthesized drug shows

pharmacological evidence that requires further

evaluation in man
when formulation and dosage form changes

are required
when solid form changes of DS are required
Protocol for preformulation studies
 OUTLINE OF PRINCIPAL AREAS OF
PREFORMULATION RESEARCH

Principal
areas
Stability
Physico-
analysis
chemical
properties Bulk Solution stability
Solubility
characterisatio Solid state
Organolepti analysis
n
Crystallinity and stability
c properties Ionization constant
Particle size polimorphism Bulk
pka
PH solubility
and shape Hygroscopicit stability
profile
Common ion Compatibilty
y
Purity effect ksp
Particle size
characterization Thermal
Surface effects
Bulk density Partition co-
area efficient
solubilization
Powder flow Dissolution
properties
 PREFORMULATION PARAMETERS
PREFORMULATION PARAMETERS METHOD USED
Organoleptic Properties Colour and Odour Determination
Crystallinity & Polymorphyism X-ray Diffraction Studies (Lachman, 1991)
Fine Particle Characterization Microscopic Method (Lachman, 1991)
Solubility Profile Equilibrium Solubility Method (I.P. 2007)
Solubilization (Lachman, 1991)
Analytical Method Development UV Spectroscopic Method, HPLC Method
Ionization Constant, pKa Determination of Spectral Shifts by UV Spectroscopy
(Lachman, 1991)
Partition Coefficient Using octanol / water,(Lachman, 1991)
Bulk Density Tapping Method (Lachman, 1991)
Powder Flow Properties % Compressibility Determination, Angle of Repose
(Lachman, 1991)
Compatibility With Excipients DSC (Stulzer and Rodriques et al., 2008)
Stability Solution and Solid State Stability (PCT/US03/35012)
Stability Indicating Method Forced Degradation Studies (Rao et al., 2009)
Development
7
ORGANOLEPTIC PROPRTIES
Colour:-Stability problems, improve
appearance by including dye in body or
coating
Taste:-palatability, flavours, and
excepient may be added.
Odour:-degradation products, eg.
Aspirin stable form of drug to be used ,
flavours and excepients may be used.
SUGGESTED TERMINOLOGY TO DESCRIBE ORGANOLEPTIC
PROPERTIES OF PHARMACEUTICAL POWDERS

Colour Odour Taste

Off-white Pungent acidic

Cream yellow sulfurous bitter
Tan fruity bland
Shiny Aromatic intense
odourless sweet

Tasteless
PURITY
Purity studies are essential for further
studies to be carried out safely.
Impurities may make a compound toxic
or render it unstable.
TLC,HPLC,GC and Paper
chromatography used.
HPLC-Impurity Index(II), Homogeneity
index(HI).
DTA, gravimetric analysis and melting
point by hot stage microscopy are
 Impurity index(II):defined as the ratio of all responses
(peak areas) due to components other than the main one
to the total area response.
Homogeneity index(HI): defined as the ratio of
response(peak area) due to main component to the total
response.
Eg.: main component retention time: 4.39min
-area response: 4620
Impurities 7 minor peaks ;area response: 251
- total area response : 251+4620
Impurity index : = 251/(4620+ 251)
= .0515
Homogeneity index : = 1 - .0515
= .9485
OTHER TOOLS IN ASSESMENT OF IMPURITY

Differential thermal analysis(DTA)

Thermogravimetric analysis(TGA)
Differential scanning calorimetry(DSC)
Powder X-Ray Diffraction(PXRD)
PARTICLE SIZE AND SHAPE
Various chemical and physical properties of
drug substances are affected by their particle
size distribution and shapes.
The effect is not only on physical properties as
well as biopharmaceutical behavior.
It also influence the flow and the mixing
efficacy of powders and granules.
Fine materials are relatively more open to
attack from atmospheric oxygen, humidity,
than that of coarse material.
PARTICLE SIZE DETERMINATION
Microscopy. Eg. Light microscope,
electron microscope.
Anderson Pipette
Sieving method
Instruments based on light
blockage(HIAC) and blockage of
electrical conductivity path(coulter
counter are available).
COMMON TECHNIQUES FOR MEASURING FINE
PARTICLES OF VARIOUS SIZES

Technique Particle size (m)

Microscopic 1 - 100
Sieve > 50
Sedimentation >1
Elutriation 1 - 50
Centrifugal < 50
Permeability >1
Light scattering 0.5 - 50
SHAPE DETERMINATION..
Microscopy should be caarried out to determine the ratio of longest to
shortest dimension. It is a shape factor .

SHAPE FACTOR
Commonly used shape factor converts volume of particle vto its
volumetric mean diameter av
V=v.av

Shape factor may be defined which converts the surface area sof
a particle to its surface mean diameter as
S=s.as
Fractal Dimensions are carried out by imaging techniques .
In(N)=-nIn(g)+q where:N=no.of squares.
g=length
of grid size.
SURFACE AREA DETERMINATION
It is determined based on Brunaver
Emitter Teller (BET)theory of
adsorption.
Most substances adsorb mono
molecular layer of gas (Nitrogen) and
temperature.
Air adsorption and permeability
methods.
CRYSTALLINITY AND POLYMORPHISM
Crystal habit and internal structure of a drug can affect bulk and
physiochemical properties
HABBIT: Outer
appearance of
crystal.

Internal structure

Crystalline
Amorphous
CHARACTERIZATION OF SOLID
FORMS

Solid forms

Crystalline Amorphous
Repeated spacing of atoms Randomly placed atoms
in three dimentional structure or molecules
Have less energy Have high energy
Need less energy to break Need less energy to
break
crystalline form
So solubility is less solubility is high
ANALYTICAL METHODS FOR CHARACTERISATION OF
SOLID FORMS

Method Material
reqired
per sample
Microscopy 1mg
Fusion methods 1mg
(hot stage microscopy)
Infrared spectroscopy 2-20mg
X-ray powder diffraction 500mg
Scanning electron microscopy 2mg
Thermogravimetric analysis 10mg
Dissolution/Solubility analysis mg to gm
 MICROSCOPY
All substances are transparent examined under
microscope are either isotropic or anisotropic
Isotropic substances do not transmit the light
appears black and have single refractive index.
E.g. Sodium Chloride
Anisotropic substances more than one refractive
index appear bright and brilliant color uniaxial
and biaxial
Color depends upon thickness of crystal and diff. in
refractive indices.
 THERMAL ANALYSIS
Differential Scanning Colorimetry (DSC) and
Differential Thermal Analysis (DTH) measures the
heat loss or heat gain - resulting from physical or
chemical changes.
Two types of processes
1) Endothermic : like fusion, boiling, sublimation,
vaporization, desolvation
Exothermic : like crystallization ,degradation
Quantitative measurement of these process have
many application in preformulation study including
Purity, Polymorphism, solvation, degradation
 X-RAY DIFFRACTION
Crystalline materials gives characteristics pattern
by peaks in certain position & varying intensities

different Polymorphs different x-ray diffraction

pattern due to crystal lattice.

Single crystal x-ray analysis provides precise

identification & description of a crystalline
substances.
 POLYMORPHISM

- substances can exist in more than one

crystalline form
Polymorphic forms diff. physical-chemical

properties (incl. melting pt. & solubility)

Enatiotropic
Polymorphs:

Monotropic
Determination method:

Thermodyanamically-van,t Hoff plot(solubility vs

temperature)
Directly by microscopic determination
HYGROSCOPICITY
Many substances,particularly water
soluble salt form have a tendency to
absorb atmospheric pressure.
Change in moisture level can influence
chemical stability , flowability, and
compactibility.
It can be monitored by Karl Fischer
titration,TGA
 POWDER FLOW

The pharmaceutical powders are classified as ---

FREE FLOWING
COHESIVE OR NON FREE FLOWING

The powder flow are affected by the changes in

Density
Particle Size
Shape Free flowing drug may become cohesive and
Electrostatic Charge necessitates an entirely new formulation strategy
Adsorbed Moisture
Several rates of flow (g/sec) determinations at each of variety of orifice
sizes (1/8 to inches) should be made.

Greater the standard deviation between multiple flow rate measurements

greater is the weight variation in products produced from the powder.

It was found that the dependence of flow rate (w) on the true particle
density(),gravity(g), orifice diameter(D) by

D= A (4w/60g)^1/n
A and n are constants dependent upon material and particle size

Other measures of free flowing powders is COMPRESSIBILITY.

%compressibility=(1-0)/1
Angle of repose is not much useful as lack of precision

Cohesive Powders are characterized by TENSILE TESTING and

evaluated in a SHEAR CELL
SOLUBILITY
Solubility > 1 % w/v
=> no dissolution-related absorption problem
Highly insoluble drug administered in small
doses may exhibit good absorption
Unstable drug in highly acidic environment of
stomach, high solubility and consequent
rapid dissolution could result in a decreased
bioavailability
The solubility of every new drug must be
determined as a function of pH over the
physiological pH range of 1 - 8
DETERMINATION OF SOLUBILITY
Semiquantitative determination:

Solvent Vigorously Examine

(fixed volume) shaking visually

Adding solute in small

incremental amounts
Undissolved
solute particles ?

No Yes
LAW OF MASS ACTION

Total amount
Estimated solubility
added up
 Accurately Quantitative determination:

Shaking at constant
Excess drug powder Ampul/vial temperature
150 mg/ml (15 %) (2-5 ml) (25 or 37 oC)
+ solvent 2 - 8 oC ?
The first few mls of the filtrates should be
discarded due to possible filter adsorption
48 hr

Determine the drug Membrane filter

concentration in the 0.45 m
filtrate
72 hr
Same Determine the drug Membrane filter
concentration ? concentration in the 0.45 m
filtrate
? hr
Solubility
Determine the drug Membrane filter
concentration in the 0.45 m
filtrate
 General Method of
Increasing the Solubility
Addition of co-solvent
pH change method
Reduction of particle size
Temperature change method
Hydotrophy
Addition of Surfactant
Dielectrical Constant
Complexation
UNIQUE PROBLEMS IN SOLUBILITY DETERMINATION
OF POORLY SOLUBLE COMPOUNDS

Solubility could be over estimated due to the

presence of soluble impurities
Saturation solubility is not reached in a reasonable
length of time unless the amount of solid used is
greatly in excess of that needed to saturation
Many compounds in solution degrade, thus making
an accurate determination of solubility difficult
Difficulty is also encountered in the determination
of solubility of metastable forms that transform to
more stable forms when exposed to solvents
pH-Solubility Profile

Stir in beaker Continuous

Excess drug
with distilled stirring of
powder
water suspension

Determine the
concentration Filter Measure Stirring
Add
of drug in pH of
acid/base
the filtrate suspension

SOLUBILITY pH
 Poorly-soluble weakly-acidic drugs:

pH= pKa + log [(St - So)/So] (2)

Poorly-soluble weakly-basic drugs:

pH= pKa + log [So/(St - So)] (3)

where
So = solubility of unionized free acid or base
St = total solubility (unionized + ionized)
 Process Of
Solubilization
The process of solubilization involves the breaking of
inter-ionic or intermolecular bonds in the solute, the
separation of the molecules of the solvent to provide
space in the solvent for the solute, interaction between
the solvent and the solute molecule or ion.

Step 1: Holes opens in the solvent

Step2: Molecules of the solid breaks away from the
bulk

Step 3: The free solid molecule is intergraded into

the hole in the solvent

37
 SOLUBILIZATION CAN BE ENHANCED BY:
Use more soluble metastable polymorph
Use of complexation (eg.Ribloflavin-
xanthinescomplex)
Use of high-energy co-precipitates that are
mixtures of solid solutions and solid
dispersions (eg. Griseofulvin in PEG 4000,
6000, and 20,000)
in PEG 4000 and 20,000 -> supersaturated
solutions
in PEG 6000 -> bioavailability in human twice
> micronized drug
Use of suitable surfactant
 Partition Coefficient
It is the ratio of unionized drug distributed
between organic and aqueous phase at equilibrium.

P o/w = ( C oil / C water )equilibrium

It ratio of unionised drug in organic &
aq.Phase
It measure lipophilicity
Major role in drug transport
Analytical separation
 IONIZATION CONSTANT
The unionized species are more lipid-soluble
and hence more readily absorbed.
The gi absorption of weakly acidic or basic drugs
is related to the fraction of unionized drug in
solution.
Factors affecting absorption:
- pH at the site of absorption
- Ionization constant
- Lipid solubility of unionized species
pH-partition theory
Henderson-Hasselbalch equation
For acids:
pH = pKa + log [ionized form]/[unionized form]
For bases:
pH = pKa + log [unionized form]/[ionized form]

Determination of Ionization Constant

1. Potentiometric pH-Titration
2. pH-Spectrophotometry Method
3. pH-Solubility Analysis
 Dissolution

kd << ka => dissolution rate-limited

kd ka ke
C, Vc
D Xg
Dissolution Absorption Xc

Absorption site Central compartment

(gi-tract) (blood circulation)

Diagram showing dissolution and absorption of

solid dosage form into blood circulation
 BASKET
PADDLE
TYPE
TYPE

2 types of systems to maintain uniform

hydrodynamic conditions

1.STATIC DISC DISSOLUTION APPARATUS

2.ROTATING DISC APPARATUS

INTRINSIC DISSOLUTION
FILM THEORY
The dissolution of a solid in its own solution is
adequately described by Noyes-Nernsts Film Theory
-dW = DAK (Cs - C) (9)
dt h
where
dW/dt = dissolution rate
A = surface area of the dissolving solid
D = diffusion coefficient
K = partition coefficient
h = aqueous diffusion layer
Cs = solubility of solute
C = solute concentration in the bulk medium
 Intrinsic dissolution rate (mg/cm2/min) is characteristics
of each solid compound in a given solvent under fixed
hydrodynamic conditions
Intrinsic dissolution rate helps in predicting if absorption
would be dissolution rate-limited
> 1 mg/cm2/min --> not likely to present dissolution rate-
limited absorption problems
< 0.1 mg/cm2/min --> usually exhibit dissolution rate-
limited absorption
0.1 - 1.0 mg/cm2/min --> more information is needed
before making any prediction
 Effect of particle size of phenacetin on dissolution
rate of the drug from granules
0.11 - 0.15 mm
Amount Dissolved (mg in 500 ml)

0.15 - 0.21 mm

0.21 - 0.30 mm

0.50 - 0.71 mm
SOLID STATE STABILITY
for identification of stable storage condition
also for identification of compatibale excipient for a

formulation
extent a product retains within specified limits and through its

period of storage and use

Stability studies conducted in the preformulation phase:

Solid-state of the drug alone

Solution phase

with the expected excipients

PHOTOLYTIC STABILITY

Many drugs fade or dropped on exposure

light.
Exposure of drug 400 and 900 foot-candles of
illumination for 4 and 2 week periods
respectively is adequate to provide some idea
of photosensitivity.
Resulting data may be useful in determining if
an amber colored container is required for
formulation .
STABILITY TO OXIDATION

Drugs sensitivity to oxidation can be examined by

exposing it to atmosphere of high oxygen tension. Usually
a 40% oxygen atmosphere allows for rapid evaluation
Samples are kept in desiccators equipped with three-way
stop cocks, which are alternatively evacuated and flooded
with desired atmosphere.
The process is repeated 3 or 4 times to ensure 100%
desired atmosphere. Results may be useful in predicting if
an antioxidant is required in the formulation or if the final
product should be packaged under inert atmospheric
conditions.
SOLUTION PHASE STABILITY

As compared with the dry form, the

degradation is much rapid in solution form. It is
important ascertain that the drug doesnt
degrade when exposed to GI fluid.
The pH based stability study, using different
stimulator GI condition can be designed.
A poor solution stability of drug may urge the
formulator to choose a less soluble salt form,
provided the bioavailability is not compromised
COMPATIBILITY STUDIES

The knowledge of drug excipients interaction is useful for the

formulation to select appropriate excipients.
The described preformulation screening of drug excipients interaction
requires only 5mg of drug in a 50% mixture with the excipients to
maximize the likelihood of obscuring an interaction .
Mixtures should be examined under nitrogen to ultimate oxidation and
paralytic effect at a standard heating rate on DSC, over a temperature
range, which will encompass any thermal changes due to both the
drug and appearance or disappearance one or more peaks in
themogrames of drug excipient mixtures are considered of indication
of interaction.
Flow diagram to identify
excepient compatibility
with drug No
interactio
Drug n Excepient
recomme
50%Mixture DSC
nded
of Drug &
excepient Interactio
n
Excepient

HPLC
Or TLC
Alternative
excepient Yes Drug
No
suggested breakdow
n
FORMULATION RECOMMENDATION
Upon completion of preformulation evaluation of
a new drug candidate, it recommended that a
comprehensive report be prepared highlighting
problems associated with this molecule
These Reports re extremely important in
preparing regulatory documents
CONCLUSION
Preformulation studies have a significant part to
play in anticipating formulation problems and
identifying logical path in both liquid and solid
dosage form technology.
By comparing the physicochemical properties of
each drug candidate with in a therapeutic group, the
preformulation scientist can assist:
the synthetic chemist to identify the optimum
molecule,
provide the biologist with suitable vehicles to elicit
pharmacological response and
advise the bulk chemist about the selection and
production of the best salt with appropriate particle