PHYSICAL PROPERTIES OF DRUG MOLECULES

M.Pharmacy 1st year

DEPARTMENT OF PHARMACEUTICS

Submitted by
K.Vasanthi.

ADVANCED PHYSICAL PHARMACEUTICS

 CONTENTS

1) PHYSICAL PROPERTIES OF DRUG MOLECULES.

2) DIFFERENTIAL THERMAL ANALYSIS.

3) DIFFERENTIAL SCANNING CALORIMETRY.

4) DIFFUSIVE REFLECTIVE SPECTROPHOTOMETRY.

5) X-RAY DIFFRACTION ANALYSIS.

ADVANCED PHYSICAL PHARMACEUTICS

 PHYSICAL PROPERTIES OF DRUG MOLECULES

The study of these properties is essential to develop a decent formulation for a novel chemical
entity, right from the beginning to the end of drug development.

The following reasons for the evaluation of the physical properties of early developmental
candidates could be furnished:

 Reducing the time and cost of introducing a molecule into the market.
 Selection of an appropriate form of the drug substance, such as salt form, prodrugs etc.
 Selection of application type (e.g.: oral, dermal, and injectable).
 Selection of the form of delivery (e.g.: quick acting or slow release).
 Increasing the ease of product development.
 Reducing undesirable findings during clinical phases.
 Release of best dug into the market.

PHYSICAL PROPERTIES:
Specific surface area, hygroscopicity, bulk density, flow properties, crystallization are the
physical properties to be investigated for new drug substances, whether flexible or stubborn.

1.Specific surface area:

Surface area properties of a drug particle affect the dissolution and chemical reactivity of
a drug substance. These properties include size, shape and surface morphology of a drug
substance. The smaller the particles, the better are the bulk flow and formulation homogeneity.
The simplest way to measure the particle size is to use a microscope. However it is tedious to
measure the average particle size with such techniques. The best way is to use photomicrographs
and hemocytometer slides. Particles with a larger specific area are good absorbents for the
absorption of gases and of solutes from solution. The other factor that is also important is the
particle shape. Generally a sphere has minimum surface area per unit volume. The more
asymmetric a particle, is the greater surface area per unit volume. Since these surface properties
affect their homogeneity, content uniformity and dissolution properties of a tablet form, which
ultimately affect the bioavailability, these properties have to be thoroughly evaluated during
toxicological stages before clinical trials are preceded so that perfect correlation is obtained
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between the bioavailability data with a formulation. When the studies are transferred from
toxicology studies to clinical studies. Accordingly, sophisticated methods are currently used.
These include adsorption methods and air permeability methods.

Quantasorb, an instrument used to obtain specific surface area measurements. A mixture

of helium and nitrogen is passed through the sample; helium is inert and is not absorbed on the
powder surface while nitrogen is absorbed on the powder. A thermal conductivity instruments
attached to the instrument measures the conductivity associated with the absorption, which in
turn indicates the size of the particles.

In air permeability technique, the resistance to the flow of a fluid, such as air through a
plug of compacted powder is used to determine the surface area of the powder. The greater the
surface area of the powder the greater is the resistance offered to the flow of the air.

2. Hygroscopicity:
The amount of water absorbed on the surface of drug particle influences the solid state stability
as well as the flow properties and compactibiliy of a drug substance.

Most drugs are partially hygroscopic. Hygroscopicity is one such character, provided the
opportunity, the first property to be determined for a new drug characterization is to measure its
hygroscopicity. Hygroscopicity depends on the synthetic techniques and the recrystallization
methods. Judicious selection of a suitable crystal form for further development is the essential
step in the development of solid dosage forms. The stability of a solid drug depends on the
hygroscopicity of a particular solid state of a drug, which in turn depends on the type of the
crystal or physical form of the drug that in turn depends on the synthetic techniques or the
recrystallization method for that particular drug the hygroscopicity of a substance is determined
by exposing the compound to different humidity conditions for a specific time intervals and then
assaying for water content using Karl fisher reagent etc. other methods that could be used to
measure the hygroscopicity is the gas chromatography.

Dynamic water sorption (DWS) that requires very little amount of compound for handling is also
used in the hygroscopicity measurements at above ± 25ºc.

Hygroscopicity most of the times affects the compatibility of new drug substances. Compatibility
as a property is affected by compressibility, adhesive/cohesive interactions and mechanical
properties of the components. Water content also influences the compactibility, suggesting that
hygroscopicity is one of the key issues in the development of tablet dosage forms. The
mechanism of water absorption in most of the cases is either hydrate formation or site specific
adsorption. The greater the compactibility, the better are the tablet properties. Many attempts
were tried to increase the compactibility of tablet substance. In this regard the reduction of
hygroscopicity of drug substance is very crucial. This can be achieved by obtaining drug crystals
by using altered synthesis or recrystallization techniques.

ADVANCED PHYSICAL PHARMACEUTICS

3.Bulk density and porosity:
Bulk density is an essential pharmaceutical property to be thoroughly investigated for a new
chemical entity .this is because of its important in capsule filling and tablet compression.
Apparatus high bulk density will not allow a capsule to be filled in the specific volume and in
addition during tablet compression, the tablets would not be compressed either because of the
rebounded effect or because of the bulk volume occupied by the tablet powder in the die. Bulk
density along with flow properties of a drug substance occupied major investigation problems,
which have to be sorted out as early as possible in new drug chemical entity investigations.

Experimentally, the true density is determined by suspending drug particles in solvents of

various densities and in which the compound is insoluble. In these measurements, wetting and
pore penetration are enhanced by the addition of a small quantity of surfactant to the solvent
mixtures. After vigorous shaking, the sample are centrifuged briefly and then lift to stand
undisturbed until flocculation or settling has reached equilibrium the sample that retains
suspended corresponds to the true density of the material. One way of avoiding this density
problem for a new chemical entity is to use wet granulation and then punch the tablet or fill the
granules in a capsule. If a drug has very high bulk density, it may not be used in a direct
compression process. The drug has to be modified so as to obtain bulk drug with good
compressibility properties. In modern solid dosage form technology, the current practice is to
prepare dosage forms with reduced excipient content. Technology that reduces the size of the
dosage form, improve the compressibility of the solid drug, its flowability and enhances the
aesthetics as desirable.

4. Crystallization:
Crystallization is a common phenomenon in pharmaceutical processing right from the
manufacturing of active pharmaceutical ingredient to the storage of final formulation approved.
Crystallization process can be termed as a Meta stable thermo dynamic state. This occurs
because any substance or events tend to stabilize to reach the lowest possible thermodynamic
state. This state of any substance is termed as a metastable state. This metastable state is either
intentionally or unintentionally created either by supersaturating, in the crystallization of desired
solid state modifications and in the control of solid phase conventions during isolation,
manufacturing, storage and dissolution. Examples of metastable state include solid solutions,
freeze concentrated solutions, solutions of weak acids/bases exposed to a PH changes, solutions
prepared by dissolving a solid state modification with a higher solubility, residual solutions
during filtration, granulation and drying. The factors that can appear in the affect crystallization
include molecular or ionic transport, viscosity, super saturation, solubility, solid liquid interfacial
tension and temperature. Nuclear kinetics is experimentally, determined from measurement of
nucleation rates, induction time and metastability zone width as a function of initial
supersaturation. Currently, molecular simulations from the data obtained from the solution and
crystal structure of drug substance is used in establishing the crystal structure of new chemical
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entity. Molecular association process in super saturated systems is obtained by laser Raman
spectroscopy and laser light scattering is used in the identification of pre-nucleation clusters and
growth units well defined experimental conditions. Raman fluorescence spectroscopic technique
used is capable of providing information about the solution structures are the species present in
the solutions.

PHYSICO – CHEMICAL PROPERTIES :

Several physic-chemical properties of new leads have to be investigated very early on
these could include Pka , solubility analysis, partition co-efficient, dissolution rate , solid state
stability , solution state stability.

1.Pka:-
Pka determination is important because this controls solubility and consequently the oral
absorption of a molecule in a given solution, formulation or body fluids. In ph. range from 1-10,
the solubility and consequently oral absorption could be altered by orders of magnitude with
changing ph. Pka is the ph. at which 50% of the substance is ionized. Buffer, temperature, ionic
strength and cosolvents effect the pka values. Incorporation of cosolvents in pka measurements
instrument methods is important because of the likely poor solubility and possible precipitation
of these compounds in aqueous media.

Potentiometric and spectrophotometric methods are the popular methods used in the
determination of pka of new chemical entity. Currently, glpka instrument is in the market for the
determination of pka of new chemical entities. The instrument measures the potentiometric pka
of a compound. The advantage offered by the current glpka instrument is that, the assays are
fully automated; temperatures’ and ionic strength are monitored during the runs and four line
cosolvents options available. The advantage is that using organic solvents help in determination
ionization constants of poorly soluble compounds.

As per indications of manufacturers, the functions of the instruments include:

 Pka’s is measured from 2 to 12.

 Log p measurements from -2 to +8.
 Overlapping and multiple pka’s routinely measured.
 Easily handles protogenic counter ions.
 Sparingly soluble compounds titrated in either possible supported cosolvents.
 Typical sample concentrations of 0.25 to 0.5 m M( 1 – 2 mg of 400 MW compounds in
10ml).
 Fast ( typical titration = 25mins).
 Accurate and precise.

ADVANCED PHYSICAL PHARMACEUTICS

 In spectrophotometric method of determination, at a given PH, if the ion concentrations
are determined using beer’s law one can calculate the approximate pka of a drug. For example, if
the drug is a free acid [HA] in equilibrium with its base[A¯], then

Pka = PH + log [HA] / [A¯]

When [HA] = [A¯], as determined by their respective absorbence in the

spectrophotometric determinations, pka = PH.

2.Solubility analysis:
Solubility analysis is essential for further processing of a compound. The factors that would
effect the solubility of a new chemical entity are PH, temperature, ionic strength and buffer
concentrations. For equilibrium solubility determinations, different methods are employed.

To determine the aqueous solubility, the drug is solubilized in which it is highly soluble and this
solution is slowly added to the distilled water and agitated. At the end of agitation, the
suspension is filtered to obtain a filtrate that is then assayed using techniques like
spectrophotometry and HPLC. Usually, the solubility of drugs is more in high temperature
conditions. The principle can be used to saturate the aqueous suspension containing a drug. The
compound that is not soluble is precipitated out. This is filtered and submitted for analysis to
determine the solubility of a drug substance. The simplest technique that is routinely used to add
excess of drug to water and this is then agitated overnight to obtain maximum solubility of the
drug in the media and then filtered and assayed to obtain the desired aqueous solubility.

To determine the solubility of a poorly soluble compound in water, generally 24hrs equilibrium
time is given. During the time the drug slowly dissolves in water. It is a similar phenomenon
with the dissolution of the drug in gastric fluid or dissolution media from a solid powder or a
capsule or from a tablet dosage form. The drug is slowly dissolved and the drug dispersed by
agitation to form a uniform solution. It is then analyzed to obtain the concentration of the drug in
the dissolution medium. Drugs with limited solubility (< 1%) in the fluids of gastro intestinal
tract often exhibit poor or erratic absorption unless dosage forms are specifically tailored for the
drug.

3.Partition coefficient:
Octanol – water partition coefficient is the ratio of concentration of a chemical in Octanol and in
water at equilibrium and at a specified temperature. Octanol is an organic solvent that is used as
a surrogate for natural organic matter. The Octanol – water partition coefficient has been
correlated to water solubility; therefore the water solubility of a substance can be used to
estimate Octanol – water partition coefficient.

ADVANCED PHYSICAL PHARMACEUTICS

The Octanol – water partition coefficient ( Kow) is defined as the ratio of chemicals
concentration in the Octanol phase to its concentration in the aqueous phase of a two phase
Octanol – water system.

K ow = Concentration in Octanol phase / Concentration in aqueous phase.(I – 1)

Values of K ow are thus unit less. The parameter is measured using low solute concentrations,
values of Kow are usually measured at room temperature ( 20 - 25°c). The effect of temperature
on K ow is not great. Usually on the order of 0.001 – 0.01 log Kow / °c and may be either + or ve.

The octanol / water partition coefficient is not the same as the ratio of the chemical’s solubility in
octanol to its solubility in water, because organic and aqueous phases of the binary octanol /
water system are not pure octanol and pure water. Kow is often found to be a function of solute
concentration. The chemical in question is added to a mixture of octanol and water whose
volume ratio is adjusted according to the expected value of Kow. Very pure octanol and water
must be used, the concentration of the solute in the system should be less than 0.01 mole / litre.
The system is shaken gently until equilibrium was achieved (15mins – 1hr). centrigugation is
generally required to separate the two phases, especially if an emulsion is formed. An
appropriate analytical technique is then used to determine the solute concentration to each phase.
A rapid laboratory estimate of Kow may be obtained by measuring the retention time in HPLC ,
the logarithm or retention time and the logarithm of Kow have been found to be linearly
correlated. Conversely chemicals with high Kow(>104) are very hydrophobic.

4.Dissolution rate :
Dissolution rate is the predictable measure of time required for a given dug or active ingredient
in an oral solid dosage form to go into solution under the specified set of conditions. Since
absorption and physiological availability of any nutritional supplement is largely dependent upon
having in a dissolved state, a suitable dissolution rate is crucial. Calculating intrinsic dissolution
rate makes comparison of the individual drug substances and the effect of different conditions on
drug dissolution. The intrinsic dissolution rate is generally defined as the dissolution rate of a
pure drug substance under the conditions of constant surface area.

Intrinsic dissolution is generally determined by measuring the dissolution of a non-disintegrating

disc made by compressing pure powder drug substance under high pressure using a specially
constructed punch and die system. The test material is compressed with a bench – top punch
tablet press for 1 minute at the minimum compression pressure necessary to form a non-
disintegrating compacted tablet. Changes in the crystal form may occur during compression,
conformation of the solid form should be verified by powder x – ray diffraction technique.
Compression plays an important role in the test, if it is too low, a non-disintegrating tablet may
not be obtained and if it’s too high it may change the crystal form. It is important to study the
effect of compression pressure on intrinsic dissolution rates as it has been observed for several
drug substances that the intrinsic dissolution rate varies with compression pressure.
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Dissolution rate determines the availability of the drug for absorption when slower than the
absorption, dissolution becomes the rate limiting step. Overall selection of an appropriate
formulation can control absorption. Dissolution rate is affected by whether the drug is in salt,
crystal or hydrated form. The sodium salt of weak acids ( ex: barbiturates, salicylates ) dissolve
faster than their corresponding free acids regardless of the PH of the medium.

5. Solid state stability:

This involves stability of the drug substance as a solid and stability of a drug substance
in a solid dosage form. Drug instability in pharmaceutical formulation may be detected in some
instances by a change in the physical appearance, color, odor, taste or texture of the formulation
whereas the chemical stability of the drug substance is determined by chemical analysis. The
second study is termed reaction kinetics.

A kinetic study on a drug substance is examined by subjecting an NCE in several

physical and chemical and stressed conditions. The samples are withdrawn at periodic times and
assayed for the drug content using a HPLC or other techniques. Then the active chemicals and
degrades are mathematically dissected to obtain chemical kinetics of the drug substance. This
reaction kinetics could be zero order, first order, second order and sometimes inverse reaction
kinetics. Inverse kinetics are determined when there is a transition of one impurity to other or
one degrading to the drug, which may help in long run in the formulation movement predictions
and during storage. As a standard stability protocol, the utilization of exaggerated conditions
such and high temperature and high light intensity and high humidity are investigated for the
stability determination. Accelerated temperature studies, for example, may be conducted for 6
months at 40ºc and 75% RH. If a significant change occurs in the drug or drug product under
these conditions, lesser temperature and humidity may be used such as 30ºc and 60% RH.
Product container, closures, and other packaging conditions features are also to be considered in
stability testing during this stage.

6.Solution state stability:

Solution state stability of a drug is valid for stability testing of liquid formulations and for HPLC
method development. NCE is generally mixed in aqueous media at different PH conditions. The
samples are withdrawn at regular time intervals and are submitted for analysis. Once the data is
obtained, the active amount present is mathematically fitted to obtain the reaction kinetics in the
solution state. Different PH conditions, different humidity conditions, different temperature
conditions, different packaging conditions can be used in the solution state stability
determination. The reaction kinetics is the same and is zero order, first order, second order, multi
order and inverse kinetics. In solid state characterization apart from the stability impurity,
polymorphs, racemates etc are determined as a first step in the physical characterization of a new
chemical entity.

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7. Enantiomers and racemates:

Enantiomer is one of two stereoisomers that are mirror images of each other that are non-
superimposable (not identical), much as one's left and right hands are the same except for
opposite orientation. Organic compounds that contain an asymmetric (chiral) Carbon usually
have two non-superimposable structures. These two structures are mirror images of each other
and are, thus, commonly called enantiomorphs Hence, optical isomerism is now commonly
referred to as Enantiomerism.

Enantiomers have, when present in a symmetric environment, identical chemical and physical
properties except for their ability to rotate plane-polarized light (+/−) by equal amounts but in
opposite directions (although the polarized light can be considered an asymmetric medium). A
mixture of equal parts of an optically active isomer and its enantiomer is termed racemic and has
zero net rotation of plane-polarized light because the positive rotation of each (+) form is exactly
counteracted by the negative rotation of a (−) one.
Enantiomers of each other often show different chemical reactions with other substances
that are also enantiomers. Since many molecules in the body of living beings are enantiomers
themselves, there is often a marked difference in the effects of two enantiomers on living beings.
In drugs, for example, often only one of a drug's enantiomers is responsible for the desired
physiologic effects, while the other enantiomer is less active, inactive, or sometimes even
responsible for adverse effects (unwanted side-effects).
The following table lists pharmaceuticals that have been available in both racemic and single-
enantiomer form.

Racemic mixture Single-enantiomer

Amphetamine (Benzedrine) dextroamphetamine (Dexedrine)

Bupivacaine (Marcain) levobupivacaine (Chirocaine)

Cetirizine (Zyrtec /
levocetirizine (Xyzal)
Reactine)

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8.Impurities:
Impurities in new drug substances are addressed from two perspectives:

 Chemistry aspects include classification and identification of impurities, report

generation, listing of impurities in specifications, and a brief discussion of analytical
procedures.
 Safety aspects include specific guidance for qualifying those impurities that were not
present, or were present at substantially lower levels, in batches of a new drug substance
used in safety and clinical studies.
The studies conducted to characterize the structure of actual impurities present in a new drug
substance at a level greater than 1% the identification threshold many batch manufactured by the
proposed commercial process should be identified. In addition, any degradation product
observed in stability studies at recommended storage conditions at a level greater than 1% the
identification threshold should be identified. Identification of impurities present at apparent level
of not more than 1% the identification threshold is generally not considered necessary.

9.Polymorphs:

Polymorphism is often characterized as the ability of a drug substance to exist as two or more
crystalline phases that have different arrangements and or conformations of the molecules in the
crystal lattice. Amorphous solids consist of disordered arrangements of molecules and do not
possess a distinguishable crystal lattice. Solvates are crystalline solid adducts containing either
stoichiometric or nonstoichiometric amounts of a solvent incorporated within the crystal
structure. If the incorporated solvent is water, the solvates are also commonly known as
hydrates.polymorphism refers to the occurrence of different crystalline forms of the same drug
substance.
Polymorphs and solvates of a pharmaceutical solid can have differen chemical and physical
properties such as melting point, chemical reactivity, apparent solubility, dissolution rate, optical
and electrical properties,vapour pressure, and density. The properties can have a direct impact on
the processability of drug substances and the quality / performance of drug products, such as
stability, dissolution, and bioavailability. A metastable pharmaceutical solid form can change
crystalline structure or solvate / desolvate in response to changes in environmental conditions,
processing, or over time.

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APPLICATIONS:

 Separation of analytes by precipitation, extraction, or distillation.

 Qualitative analysis by reaction of analytes with reagents that yielded products that could be
recognized by their colors, boiling or melting points, solubilities, optical activities, or
refractive indexes.
 Quantitative analysis by gravimetric or by titrimetric techniques:
1. Gravimetric Methods – the mass of the analyte or some compound produced from the analyte
was determined.

2. Titrimetric Methods – the volume or mass of a standard reagent required to react completely
with the analyte was measured.

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 DIFFERENTIAL THERMAL ANALYSIS

DTA measures the temperature difference between the sample and a reference as a function of
Temperature or time when heating at a constant rate.

PRINCIPLE

Differential thermal analysis (or DTA) is a thermo analytic technique, similar to differential
scanning calorimeter. In DTA, the material under study and an inert reference are made to
undergo identical thermal cycles, while recording any temperature difference between sample
and reference. This differential temperature is then plotted against time, or against temperature
(DTA curve or thermogram). Changes in the sample, either exothermic or endothermic, can be
detected relative to the inert reference. Thus, a DTA curve provides data on the transformations
that have occurred, such as glass transitions, crystallization, melting and sublimation. The area
under a DTA peak is the enthalpy change and is not affected by the heat capacity of the sample.

INSTRUMENTATION
A DTA consists of a sample holder comprising thermocouples, sample containers and a
ceramic or metallic block; a furnace; a temperature programmer; and a recording system. The
key feature is the existence of two thermocouples connected to a voltmeter. One thermocouple is
placed in an inert material such as Al2O3, while the other is placed in a sample of the material
under study. As the temperature is increased, there will be a brief deflection of the voltmeter if
the sample is undergoing a phase transition. This occurs because the input of heat will raise the
temperature of the inert substance, but be incorporated as latent heat in the material changing
phase.

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Instrumentation and working

 The sample is loaded into a crucible, which is then inserted into the sample well (marked
S). A reference sample is made by placing a similar quantity of inert material (such as
Al2O3) in a second crucible.
 This crucible is inserted in the reference well, marked R. The dimensions of the two
crucibles and of the cell wells are as nearly identical as possible; furthermore, the weights
of the sample and the reference should be virtually equal.
 The sample and reference should be matched thermally and arranged symmetrically with
the furnace so that they are both heated or cooled in an identical manner.
 The metal block surrounding the wells acts as a heat sink. The temperature of the heat
sink is slowly increased using an internal heater. The sink in turn simultaneously heats
the sample and reference material.
 A pair of matched thermocouples is used. One pair is in contact with the sample or the
sample container; the other pair is in contact with the reference. The output of the
differential thermocouple, Ts - Tr or DT, is amplified and sent to the data acquisition

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 system. This allows the difference in temperature between the sample and the reference
to be recorded as a function of the sample temperature, the reference temperature or time.
 If there is no difference in temperature, no signal is generated, even though the actual
temperatures of the sample and reference are both increasing.
 Operating temperatures for DTA instruments are generally room temperature to about
1600 OC; some DTA equipment’s are capable of operating from -150 OC to 2400 OC.
To reach the very low sub-ambient temperatures, a liquid nitrogen cooling accessory is
needed. Some low temperatures (but, not -150 OC) may be reached with electrical
cooling devices or with forced air-cooling.
 When a physical change takes place in the sample, heat is absorbed or generated. For
example, when a metal carbonate decomposes, CO2 is evolved. This is an endothermic
reaction; heat is absorbed and the sample temperature decreases. The sample is now at a
lower temperature than the reference. The temperature difference between the sample and
reference generates a net signal, which is recorded.
 Modern DTA instruments have the ability to change atmospheres from inert to reactive
gases, as is done in TGA. As is the case with TGA, the appearance of the DTA thermal
curve depends on the particle size of the sample, sample packing, the heating rate, flow
characteristics inside the furnace, and other factors.
 Thermal matching between the sample and the reference is often improved by diluting
the sample with the inert reference, keeping the total masses in each crucible as close to
each other as possible.
 Sample crucibles are generally metallic (Al, Pt.) or ceramic (silica) and may or may not
have a lid. Many metal pans with lids have the lid crimped on using a special tool.
 Best results are obtained when the area of contact between the sample and the pan or
crucible is maximized.
 Samples are generally in the 1–10 mg range for analytical applications.

APPLICATIONS

DTA is widely used in the pharmaceutical and food industries.

DTA may be used in cement chemistry, mineralogical research and in environmental studies.
DTA curves may also be used to date bone remains or to study archaeological materials.
 Composition of Multicomponent Systems
 Thermal Stability of Materials
 Oxidative Stability of Materials
 Estimated Lifetime of a Product
 Decomposition Kinetics of Materials
 The Effect of Reactive or Corrosive Atmospheres on Materials
 Moisture and Volatiles Content of Materials.
 To construct phase diagrams and study phase transitions.

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 To fingerprint substances.
 To determine M.Pt. ,B.Pt., decomposition temperatures of organic compounds.
 To characterize inorganic materials.
 To quantitatively analyze polymer mixtures.
 To characterize polymers.

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 DIFFERENTIAL SCANNING CALORIMETER (DSC)

DEFINITION:
Differential Scanning Calorimetry, DSC, is a thermo analytical technique in which the difference
in the amount of heat required to increase the temperature of a sample and reference are
measured as a function of temperature. Both the sample and reference are maintained at nearly
the same temperature throughout the experiment. Generally, the temperature program for a DSC
analysis is designed such that the sample holder temperature increases linearly as a function of
time.

PRINCIPLE:
The basic principle underlying this technique is that when the sample undergoes a physical
transformation such as phase transitions, more or less heat will need to flow to it than the
reference to maintain both at the same temperature. Whether less or more heat must flow to the
sample depends on whether the process is exothermic or endothermic.
For example, as a solid sample melts to a liquid it will require more heat flowing to the sample
to increase its temperature at the same rate as the reference. This is due to the absorption of heat
by the sample as it undergoes the endothermic phase transition from solid to liquid. Likewise, as
the sample undergoes exothermic processes (such as crystallization) less heat is required to raise
the sample temperature. By observing the difference in heat flow between the sample and
reference, differential scanning calorimeters are able to measure the amount of heat absorbed or
released during such transitions. DSC may also be used to observe more subtle physical changes,
such as glass transitions.

THEORY:
 DSC is a method of thermal analysis that is widely used to study thermal transitions,
i.e. ., solid- solid transitions as well as solid-liquid and various other transitions and
reactions.
 A solid-solid phase transition would be if the material had its structure altered, but
not gain enough energy to become a liquid. Using thermal analysis, it is possible to
understand what is happening in a material, even if there is no visual evidence that a
change has occurred.
 For instance, it is easy to see when an ice cube melts into water and when water boils
into steam; these are visible changes.
 There are however several different phase changes within water in a solid state. Ice at
colder and colder temperatures can have several different crystal structures and
undergo many solid-solid phase transitions, and in each of these phases, the ice has
different properties ranging from brittleness to conductivity.
 By understanding the technique and instrumentation of DSC, it is possible to
understand what the materials go through during energy gain or loss.

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  Differential scanning calorimetry is a technique we use to study what happens to
polymers when they're heated.
 We use it to study what we call the thermal transitions of a polymer.

Heat capacity
We can learn a lot from this plot. Let's imagine we're heating a polymer. When we start heating
our two pans, the computer will plot the difference in heat output of the two heaters against
temperature. That is to say, we're plotting the heat absorbed by the polymer against temperature.
The plot will look something like this at first.

The heat flow at a given temperature can tell us something. The heat flow is going to be shown
in units of heat, q supplied per unit time, t. The heating rate is temperature increase T per unit
time, t.

Temperature will go up by a certain amount, and the amount of heat it takes to get a certain
temperature increase is called the heat capacity, or Cp. We get the heat capacity by dividing the
heat supplied by the resulting temperature increases.
The glass transition temperature

Of course, we can learn a lot more than just a polymer's heat capacity with DSC. when we heat
the polymer a little more after a certain temperature, our plot will shift upward suddenly, like this

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This means we're now getting more heat flow. This means we've also got an increase in the heat
capacity of our polymer. This happens because the polymer has just gone through the glass
transition. The operative definition of glass transition temperature is that at this temperature, or
within a few degrees, the specific heat, the coefficient of thermal expansion, the free volume, and
the dielectric constant (in the case of a polar polymer) all change rapidly. Because of this change
in heat capacity that occurs at the glass transition, we can use DSC to measure a polymer's glass
transition temperature. You may notice that the change doesn't occur suddenly, but takes place
over a temperature range.

Crystallization
Above the glass transition, the polymers have a lot of mobility. They wiggle and squirm, and
never stay in one position for very long. When they reach the right temperature, they will have
gained enough energy to move into very ordered arrangements, which we call crystals, of course.
When polymers fall into these crystalline arrangements, they give off heat. You can see this drop
in the heat flow as a big dip in the plot of heat flow versus temperature

The temperature at the lowest point of the dip is usually considered to be the polymer's
crystallization temperature, or Tc. Also, we can measure the area of the dip, and that will tell us
the latent energy of crystallization for the polymer. But most importantly, this dip tells us that the
polymer can in fact crystallize. Because the polymer gives off heat when it crystallizes, we call
crystallization an exothermic transition.

ADVANCED PHYSICAL PHARMACEUTICS

Melting

If we keep heating our polymer past its Tc, eventually we'll reach another thermal transition, one
called melting. When we reach the polymer's melting temperature, or Tm, those polymer crystals
begin to fall apart, that is they melt. The chains come out of their ordered arrangements, and
begin to move around freely. When the polymer crystals melt, they must absorb heat in order to
do so. Melting is a first order transition. This means that when the melting temperature reaches,
the polymer's temperature won't rise until all the crystals have melted. This means that the little
heater under the sample pan is going to have to put a lot of heat into the polymer in order to both
melt the crystals and keep the temperature rising at the same rate as that of the reference pan.
This extra heat flow during melting shows up as a big peak on our DSC plot, like this

So let's review now: we saw a step in the plot when the polymer was heated past its glass
transition temperature. Then we saw a big dip when the polymer reached its crystallization
temperature. Then finally we saw a big peak when the polymer reached its melting temperature.
To put them all together, a whole plot will often look something like this:
Of course, not everything you see here will be on every DSC plot. The crystallization dip and the
melting peak will only show up for polymers that can form crystals. Completely amorphous
polymers won't show any crystallization, or any melting either. But polymers with both
crystalline and amorphous domains will show all the features you see above.

Putting it all together

Then we saw a big dip when the polymer reached its crystallization temperature. Then finally we
saw a big peak when the polymer reached its melting temperature. To put them all together, a
whole plot will often look something like this then we saw a big dip when the polymer reached
its crystallization temperature. Then finally we saw a big peak when the polymer reached its
melting temperature. To put them all together, a whole plot will often look something like this

ADVANCED PHYSICAL PHARMACEUTICS

If you look at the DSC plot you can see a big difference between the glass transition and the
other two thermal transitions, crystallization and melting. For the glass transition, there is no dip,
and there's no peak, either. This is because there is no latent heat given off, or absorbed, by the
polymer during the glass transition. Both melting and crystallization involve giving off or
absorbing heat. The only thing we do see at the glass transition temperature is a change in the
heat capacity of the polymer.
Because there is a change in heat capacity, but there is no latent heat involved with the glass
transition, we call the glass transition a second order transition. Transitions like melting and
crystallization, which do have latent heats, are called first order transitions.

INSTRUMENTATION AND WORKING

The calorimeter consists of a sample holder and a reference holder. Both are constructed of
platinum to allow high temperature operation. Under each holder is a resistance heater and a
temperature sensor. Currents are applied to the two heaters to increase the temperature at the
selected rate. The difference in the power to the two holders, necessary to maintain the holders at
the same temperature, is used to calculate ΔdH/dt . A schematic diagram of a DSC is shown in
Figure 1. A flow of nitrogen gas is maintained over the samples to create a reproducible and dry
atmosphere. The nitrogen atmosphere also eliminates air oxidation of the samples at high
temperatures. The sample is sealed into a small aluminum pan. The reference is usually an empty
pan and cover. The pans hold up to about 10 mg of material.

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Figure 1.Schematic of a DSC.

The triangles are amplifiers that determine the difference in the two input signals. The sample
heater power is adjusted to keep the sample and reference at the same temperature during the
scan.

Figure 2. Typical DSC scan.

The heat capacity of the sample is calculated from the shift in the baseline at the starting
transient. Glass transitions cause a baseline shift.
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Crystallization is a typical exothermic process and melting a typical endothermic process, ΔtrH
is calculated from the area under the peaks.
During the heating of a sample, for example, from room temperature to its decomposition
Temperature, peaks with positive and negative ΔdH/dt may be recorded; each peak corresponds
To a heat effect associated with a specific process, such as crystallization or melting (Fig. 2).
A special case in which the temperature of a phase transformation is of great importance in
Polymers are the glass transition temperature, Tg.
In the DSC experiment, Tg is manifested by a drastic change in the base line, indicating a change
in the heat capacity of the polymer.
No enthalpy is associated with such transition (for which reason it is also called a second order
transition); therefore, the effect in a DSC curve is slight and is observable only if the instrument
is sensitive enough.

The heat flow may be measured as exothermic or endothermic and plotted against temperature.
The slope of the curve is the rate of change of heat capacity ΔCp/dt.

During the heating of a sample, for example, from room temperature to its decomposition
temperature, peaks with positive and negative ΔdH/dt may be recorded; each peak corresponds
to a heat effect associated with a specific process, such as crystallization or melting.

The temperature scan rate is

Scan rate = DT/dt

Using the chain rule

Where dH/dt is the shift in the baseline of the thermogram and the last derivative is just the
inverse of the scan rate. For differential measurements, we determine the difference in the heat
capacity of the sample and the reference.

The units of the heat flow are mcal sec-1 and the temperature scan rate is usually expressed as
°Cmin-1. So to be consistent with units you must multiply by 60 sec min-1

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APPLICATIONS
 Differential scanning calorimetry can be used to measure a number of characteristic
properties of a sample.
 Using this technique it is possible to observe fusion and crystallization events as well as
glass transition temperatures Tg.
 DSC can also be used to study oxidation, as well as other chemical reactions. Glass
transitions may occur as the temperature of an amorphous solid is increased. These
transitions appear as a step in the baseline of the recorded DSC signal. This is due to the
sample undergoing a change in heat capacity; no formal phase change occurs.
 The technique is widely used across a range of applications, both as a routine quality test
and as a research tool. The equipment is easy to calibrate, using low melting indium at
156.5985 °C for example, and is a rapid and reliable method of thermal analysis.
Polymers
 DSC is used widely for examining polymeric materials to determine their thermal
transitions. The observed thermal transitions can be utilized to compare materials .
 Composition of unknown materials may be completed using a technique such as IR.
 Melting points and glass transition temperatures for most polymers are available from
standard compilations, and the method can show polymer degradation by the lowering of
the expected melting point, Tm.
 The percent Crystalline content of a polymer can be estimated from the
crystallization/melting peaks of the DSC graph .
 DSC can also be used to study thermal degradation of polymers using an approach.
 Impurities in polymers can be determined by examining thermo grams for anomalous
peaks, and plasticizers can be detected at their characteristic boiling points.
Liquid crystals
 DSC is used in the study of liquid crystals.
 Using DSC, it is possible to observe the small energy changes that occur as matter
transitions from a solid to a liquid crystal and from a liquid crystal to an isotropic liquid.
Oxidative stability
 Using differential scanning calorimetry to study the stability to oxidation of samples
generally requires an airtight sample chamber.
 Such analysis can be used to determine the stability and optimum storage conditions for a
material or compound.
Safety screening
 DSC makes a reasonable initial safety screening tool.
 The presence of an exothermic event can then be used to assess the stability of a
substance to heat.
 A much more accurate data set can be obtained from an adiabatic calorimeter, but such a
test may take 2–3 days from ambient at a rate of a 3 °C increment per half hour.

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Drug analysis
 DSC is widely used in the pharmaceutical and polymer industries.
 For the polymer chemist, DSC is a handy tool for studying curing processes, which
allows the fine tuning of polymer properties.
 In the pharmaceutical industry it is necessary to have well-characterized drug compounds
in order to define processing parameters.
 If it is necessary to deliver a drug in the amorphous form, it is desirable to process the
drug at temperatures below those at which crystallization can occur.
General chemical analysis
 Freezing-point depression can be used as a purity analysis tool when analyzed by
differential scanning calorimetry.
 Consequently, less pure compounds will exhibit a broadened melting peak that begins at
lower temperature than a pure compound.

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 DIFFUSIVE REFLECTIVE SPECTROPHOTOMETRY

PRINCIPLE

A beam of light impinging on a flat polished surface of a crystal larger than the beam
Cross section is partly specularly reflected and partly refracted following the laws of geometric
optics (contained in the Fresnel equations). In absorbing materials, the radiant flux Is absorbed
according to the well-known Lambert Absorption Law.

I = I e-K xEq. [1]

Where I is the radiation flux transmitted from an initial flux I0 following passage through a
Layer of thickness x of a medium with an absorption (or extinction) coefficient KT measured
In transmission.

When the dimensions of the particle are small compared with the beam cross section but large
relative to the light wavelength, diffraction phenomena also occur because rays striking the
crystal and passing by it result in interferences among elementary waves. In powders of
randomly oriented particles of such size, part of the incident light goes back at all angles into the
hemisphere of provenance of the light. The phenomenon resulting from the reflection, refraction,
diffraction, and absorption by particles oriented in all directions is called diffuse (or volume)
reflection, in contrast with regular (or directional) reflection
from a plane phase boundary. For ideal diffuse reflection, the angular distribution of reflected
light is independent of the angle of incidence and obeys the Lambert Cosine Law.
This law states that the remitted radiation per unit surface and unit solid angle is proportional
to the cosine of the angle i of incident light and the cosine of the angle of observation, e. There is
no such thing as an ideal diffuse reflector, but near-Lambertian behavior is normally observed in
tightly pressed powder samples. If the dimensions of the particle are similar to, or smaller than,
the wavelength, then the contributions of reflection, refraction, and diffraction to the intensity
and angular distribution of the remitted radiation flux are comparable and impossible to separate.
The phenomenon is then designated as scattering.

Various theories have provided a reasonably solid basis to interpret single scattering by isolated
molecules of absorbing or non absorbing isotropic particles. However, as the distance between
particles decreases, single scattering gives way to multiple scattering, which logically
predominates in densely packed crystal powders and pigment mixtures.
There is no general quantitative solution to the problem of multiple scattering. Purely
phenomenological theories have thus been developed to describe the system properties. Several

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theories are based on two constants that characterize the absorption and scattering per unit layer
thickness of the medium. These so-called coefficients of absorption and scattering are generally
taken to be properties of the irradiated layer, assumed to be a continuum, and are experimentally
accessible.

THE KUBELKA–MUNK THEORY

The Kubelka-Munk theory predicts a linear relationship between spectral intensity and sample
concentration under conditions of constant scattering coefficient and infinite sample dilution in a
non absorbing matrix.

The Kubelka and Munk (1931) theory assumes that a plane-parallel layer of thickness X capable
of both scattering and absorbing radiation is irradiated in the −x direction with a diffuse
monochromatic radiation flux I. The layer is very extensive relative to X and can be split into
infinitesimal layers of thickness dx. The diffuse radiation flux in the negative and positive x
directions are designated I and J, respectively. If, in passing through dx, the downward flux I is
decreased by an amount KIdx by absorption, and increased by an amount SIdx by scattering, and
a similar reasoning is made for the upward flux J, then the following differential equations can
be derived

Eq. [2,3]

where K and S are the absorption and scattering coefficient of the sample, respectively.

The most general solution is

Eq. [4]
Where R is the reflectance of the layer over a background of reflectance Rg, cothbSX the
Hyperbolic cotangent of bSX, X the layer thickness, a = 1 + K/S, and b = (a2 − 1)0.5.
furtherincrease in thickness will fail to change the reflectance. Under these conditions, the
Reflectance is given by R∞ and Eq. [4] yields

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 Eq. [5]

The validity of Eq. [5] has been tested through carefully designed measurements on
Samples of colored glass for which the scattering coefficient was shown to be independent
Of the wavelength, and both K (Eq. [5]) and KT (the absorption coefficient in transmission,
Eq. [1]) were measured. K was found to be proportional to KT by a factor similar at all
wavelengths.
As noted, the ―typical‖ absorption spectrum constructed from reflectance measurements should
reflect the true absorption spectrum only if the scattering coefficient is independent of the
wavelength. This is only the case when the average grain size is large relative to the wavelength.
Otherwise, the scattering coefficient usually decreases with increasing wavelength.
The dependence of the K–M function or the apparent absorbance [log(1/R∞)] on particle size is
rather complex, particularly in heterogeneous mixtures such as ground soil materials, where both
particle size range and differences in absorption coefficient among minerals are wide. The
absorption coefficient (either K or KT) invariably decreases with increasing particle size
throughout the size range of interest, and the spectrum flattens. However, differences between
weak and strongly absorbing materials continue to exist as relates to the size-dependence of
absorbance. In strongly absorbing materials, absorbance increases with decreasing particle size
for sizes smaller than the wavelength through an increased absorption coefficient.
In summary, the K–M theory allows one to obtain the typical absorption spectrum from
absorbing mineral or mineral mixture, but one must consider those factors affecting the curve.

In practice, significant deviations from the theory occur at R∞ <0.6. For this reason, dilution of
the sample to be measured with a ―white‖ standard possessing a known scattering coefficient at
the different wavelengths has usually been recommended. K values can then be calculated from
the K–M function on the proven assumption that the scattering coefficient of the diluted sample
can be approximated by that of the diluent. Absolute values of S can be determined by measuring
the reflectance of several mounts consisting of thin layers of standard against a background of
reflectance Rg = 0. S can then be obtained from the slope of the plot against thickness, X, of the
following function:

Where a and b are defined as for Eq. [4], and R0 is the reflectance in front of the black
background.

ADVANCED PHYSICAL PHARMACEUTICS

INSTRUMENTATION
 Diffuse reflectance measurements are usually made by using a UV-visible
spectrophotometer equipped with a diffuse reflectance accessory capable of collecting the
reflected flux.
 Currently, many high -performance, research spectrophotometers can be fitted with an
integrating sphere/detector module, which usually replaces the cell holders used for
transmission measurements in the measuring compartment.
 An integrating sphere is essentially a hollow sphere internally coated with a white
material of diffuse reflectance close to 1.
 The sphere has apertures through which a beam of radiant energy can penetrate and ports
for mounting samples and standards and placing the appropriate detectors.
 Commercially available spheres range from 50 to 250 mm in diameter and are internally
coated with highly diffusing poly tetrafluoroethylene (PTFE) or barium sulfate (BaSO4).
 Reflectance measurements are performed under specific geometric conditions.
 Reflectance is measured by illuminating the sample with diffuse light the angle between
the normal to the sample surface and the axis of the viewing beam not exceeding 10°.
 Most commercial spectrophotometers measure directional–hemispherical reflectance,
whereby the sample is illuminated by a beam whose axis does not depart by more than
10° from the normal to the sample and the reflected radiation collected by the sphere goes
to the detector.
 Integrating spheres usually contain small baffles placed between the sample and the area
of the sphere that is illuminated or viewed.
 These baffles prevent directly reflected light from superimposing on the illuminated
sample or on the area of the surface that is being viewed.

SAMPLE PREPARATION
 Diffuse reflectance spectra are significantly sensitive to the manner in which the soil or
mineral mixture sample is prepared.
 The operator must thus carefully consider the factors potentially influencing those
Features of the spectrum from which useful information is to be derived.
 Particle size is the factor most strongly affecting reflectance, as shown by the
occasionally dramatic changes in soil color upon grinding.
 The best results are obtained with small particle sizes, so it is generally advisable to grind
the sample to a fine silt (<10 μm) size.
 The fast and effective grinding provided by ball mills is not always advisable because,
above a certain grinding energy, some minerals are transformed into others.

ADVANCED PHYSICAL PHARMACEUTICS

  Consistent results and preservation of the spectral features of interest can be achieved in
most instances simply by grinding the sample in an agate mortar, so that minerals are in
the volume-scattering region.
 Preferential orientation of particles of layer silicates and occasionally other minerals must
be avoided because it results in regular reflection, thus breaking the laws of diffuse
reflection.
 Dilution of the sample with a barium sulfate standard usually reduces preferential
orientation to an acceptable minimum.
 Moist soil samples have occasionally been used, in observing color changes upon
moistening.
 Moisture loss during measurement, which can be a major source of error, must be
evaluated by measuring the sample spectrum at variable intervals.

SAMPLE HOLDERS
 Many holders possess a cover glass to prevent loose powder from falling into, and
damaging, the integrating sphere when sample ports are vertical or horizontally
positioned on top of the sphere.
 The only choice available with vertical ports is to use self-supporting pressed powder
mounts.
 Rectangular or ovulated holes with a maximum size of 8 to 10 by 12 to 16 mm are
usually suitable.
 Portable spectrophotometers allow the rapid measurement of the reflectance of unaltered
soil surfaces or ground soil samples without the need for special preparation.
 However, they generally measure reflectance at relatively large wavelength steps. This
restricts detailed band analysis and quantitative calculations.

PROCEDURE

 Position the diffuse reflectance accessory in the sample compartment of the

spectrophotometer and plug it in. Align the optical system according to the manufacture’s
operating instructions and let the instrument warm up for a few minutes.
 Set up the measuring program: wavelength range(s) of interest, mode (reflectance or
absorbance), double beam, and scan speed.
 When the instrument is ready, place one of the reference disks over the sample port and
the other over the reference port.
 Alternatively, prepare two standards from barium sulfate powder. To do this, place the
holder over a frosted plate glass or a flat surface covered with unglazed paper. Add the
required amount of barium sulfate powder in several layers and distribute it uniformly in

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 the hole. Press it firmly with the plunger to obtain the required packing density and a
thickness >3 mm.
 Remove the plunger and check that the barium sulfate surface is perfectly flat—this is
required to minimize phase angle effects.
 Remove loose particles remaining on the surface with the aid of a gentle jet of filtered dry
air before placing the holder over the instrument port.
 Collect the baseline scan with the standard in place and replace the standard on the
sample port with the soil sample. Take a subsample of 200 to 500 mg and grind it for 8 to
10 min, or until any apparent grittiness disappears and further grinding causes no
apparent color change. Press the resulting powder into the holder hole, as for the barium
sulfate powder, to a minimum thickness of 3 mm.
 Place the holder on the sample port, and record the spectrum. Unless time for spectrum
acquisition is limited, it is recommended for most purposes to obtain reflectance
measurements in 0.5- to 1-nm steps for instruments with a wavelength bandwidth of 2
nm. This can take 10 to 20 min for the visible (380–750 nm) range, or 20 to 30 min for
the visible-near IR range, in most spectrophotometers.

APPLICATIONS

Identification of Mineral Species

Some common soil minerals can be identified by their characteristic bands in the absorbance or
K–M function spectra. By far, iron oxides have been the most intensively examined soil minerals
to date.

Elucidation of Crystal Properties

 The decisive influence of crystal parameters on the position and intensity of the
absorption bands of many minerals is well documented.
 The spectral features of the different minerals have usually been interpreted in light of the
crystal field and ligand field theories.
 Used to elucidate the structural properties of Fe oxides.
 Diffuse reflectance spectroscopy has also been used to characterize nonmetric surface
precipitates.

Quantitative Analysis
 Applications of IR spectroscopy to qualitative analysis are mainly for the identification of
unknown compounds.
 Quantitative analyses are also performed by measuring the intensity of the characteristic
bands for each mineral in the K–M spectral curve.

ADVANCED PHYSICAL PHARMACEUTICS

  Based on it, at any wavelength, the K–M function for a mixture of a small amount of a
colored mineral in a matrix of white or colorless soil minerals is proportional to, and
depends almost exclusively on, the concentration of that mineral provided S is constant.

Qualitative Analysis:
 It can rapidly estimate the concentration of an element with an accuracy of perhaps one
order of magnitude. The sensitivity of spectrographic methods depends upon the nature
and amount of sample, the type of excitation, and the instrument employed.

Other applications include determination of

 Assay of the compound

 Surfactant chain length determination
 Hydrogen analysis
 Iodine value
 Moisture analysis

ADVANCED PHYSICAL PHARMACEUTICS

 X-RAY DIFFRACTION ANALYSIS

INTRODUCTION

X-ray powder diffraction (XRD) is one of the most powerful technique for qualitative and
quantitative analysis of crystalline compounds. The technique provides information that cannot
be obtained any other way. The information obtained includes types and nature of crystalline
phases present, structural make-up of phases, degree of crystallinity, amount of amorphous
content, microstrain & size and orientation of crystallites.

Fundamental Principles

Max von Laue, in 1912, discovered that crystalline substances act as three-dimensional
diffraction gratings for X-ray wavelengths similar to the spacing of planes in a crystal lattice. X-
ray diffraction is now a common technique for the study of crystal structures and atomic spacing.
X-ray diffraction is based on constructive interference of monochromatic X-rays and a
crystalline sample. These X-rays are generated by a cathode ray tube, filtered to produce
monochromatic radiation, collimated to concentrate, and directed toward the sample. The
interaction of the incident rays with the sample produces constructive interference (and a
diffracted ray) when conditions satisfy Bragg's Law (nλ=2d sin θ). This law relates the
wavelength of electromagnetic radiation to the diffraction angle and the lattice spacing in a
crystalline sample. These diffracted X-rays are then detected, processed and counted. By
scanning the sample through a range of 2θ angles, all possible diffraction directions of the lattice
should be attained due to the random orientation of the powdered material. Conversion of the
diffraction peaks to d-spacings allows identification of the mineral because each mineral has a
set of unique d-spacings. Typically, this is achieved by comparison of d-spacings with standard
reference patterns.

All diffraction methods are based on generation of X-rays in an X-ray tube. These X-rays
are directed at the sample, and the diffracted rays are collected. A key component of all
diffraction is the angle between the incident and diffracted rays. Powder and single crystal
diffraction vary in instrumentation beyond this.

BRAGG’S LAW AND ITS EQUATION:

DIFFRACTION OF X-RAYS THROUGH CRYSTAL:

The nature of x-rays is electromagnetic i.e. they are electromagnetic waves. X-rays have
very short wavelength of the order of 10 x 10 -10 m. Therefore it is not possible to produce
interference fringes of x-rays by Young's double slit experiment or by thin film method. The

ADVANCED PHYSICAL PHARMACEUTICS

reason is that the fringe spacing is D x = lL/d and unless the slits are separated by a distance of
10 x 10 -10 m, the fringes so obtained will be closed together that they cannot be observed.
However it is possible to obtain x-rays diffraction by making use of crystals such as rock salt in
which the atoms are uniformly spaced in planes and separated by a distance of order of 2 A to
5A. Therefore, the diffraction of x-rays takes place when they incident on the surface of crystals.

BRAGG’S EQUATION:

Consider a set of parallel lattice planes having spacing 'd' between each other as shown.
Consider two rays 'a' and 'b' incident on the surface of crystal of NaCl. After reflection, these
rays reflected and are in phase. After reflection they interfere each other.

The path difference between the two reflected rays is given by:

ADVANCED PHYSICAL PHARMACEUTICS

 Now the X-rays will interfere constructively if the path difference is an integral multiple
of wavelengths.

This relation is known as Bragg's Law. The spacing of the atomic layers of crystals can
be found from the density and atomic weight. Both 'm' and 'q' can be measured and hence the
wave length of x-rays can be measured by using Bragg's equation.

QUANTITATIVE ANALYSIS
 XRD can be used not only for qualitative identification but also for quantitative
estimation of various crystalline phases.
 This is one of the important advantage of X-ray diffraction technique.
 Several methods have been proposed and successfully used to quantify crystalline phases
in mixtures. They include external standard methods, the reference-intensity-ratio (RIR)
method, chemical methods and the whole pattern fitting Rietveld method.
 Of the available methods, the Rietveld method is probably the most accurate and reliable
method.
 The Rietveld method is a whole-pattern fitting least squares refinement technique and
has been successfully used for quantification and characterization of inorganic and
organic compounds It has also been used for crystal structure refinement, to determine
size and strain of crystallites.

INSTRUMENTATION
 It consists of
 The X-ray tube.
 The flat specimen (labeled sample in the diagram)
 The microdiffractometer includes a goniometer with a triple-axis sample oscillation
mechanism (T, P, N), an X, Y, Z stage, and a goniometer head.
 The goniometer circle (labeled measuring circle in the diagram) which remains constant
through the analysis and is defined by the position of the target in the X-ray tube, the
center of the sample, and the position of the receiving slit (labeled detector diaphragm)
on the detector side.

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  The X-ray tube, specimen and receiving slit also lie on the arc of the focusing circle.
 Unlike the goniometer circle which remains fixed, the radius of the focusing circle is a
function of θ-2θ, with the radius decreasing as θ increases.
 The incident angle θ defined as the angle between the incident beam and the sample, and
2θ defined as the angle between the incident and diffracted beams. The detector is moved
at twice the angular rate of the sample to maintain the θ-2θ geometry.
 A filter (on the diffracted beam side) is to remove all but the desired Kα radiation from
the diffracted beam before it enters the detector.
 A slit on the incident beam side is used to narrow the beam so that it is confined within
the area of the specimen.
 The path AB=BC is the radius of the diffractometer circle.
 The tube position is fixed and the θ-2θ geometry is maintained by rotating the sample
holder at ½ the angular rate of the detector.
 There are Soller slits on both the tube and detector side, and two collimating and
receiving slits.
 Note the easy-to-read angular indicators and micrometer dials for visually reading θ and
2θ.
 The detector on this system also includes a graphite monochromator adjacent to the
scintillation detector eliminating the need for any filters in the system

Sample Preparation

The Ideal Specimen is a statistically infinite amount of randomly oriented powder with crystallite
size less than 10 μm, mounted in a manner in which there is no preferred crystallite orientation.

ADVANCED PHYSICAL PHARMACEUTICS

In this day of automated data collection and analysis, the preparation of your specimen is usually
the most critical factor influencing the quality of your analytical data.

Generate Analytical X-Rays

 A coherent beam of monochromatic X-rays of known wavelength is required for XRD

Analysis Striking a pure anode of a particular metal with high-energy electrons in a
sealed vacuum tube generates X-rays that may be used for X-ray diffraction.
 Copper (Cu) X-ray tubes are most commonly used for X-ray diffraction of inorganic
materials.
 The wavelength of the strongest Cu radiation (Kα) is approximately 1.54 angstroms (Å).
Other anodes commonly used in X-ray generating tubes include Cr (Kα 2.29 Å), Fe (Kα
1.94 Å), Co (Kα 1.79 Å), and Mo (Kα 0.71 Å).
 The radiation produced in the tube includes Kα1, Kα2, and Kβ as the highest energy X-
rays and a whole host of lower energy radiation.
 We generally use the Kα for our analytical work.
 The Kβ radiation is usually removed by use of a filter, a monochromator or an energy-
selective detector.
 The Kα2 radiation is removed from the X-ray data electronically during data processing.

Direct t he X-rays at a powdered specimen

 In most powder diffractometers systems a series of parallel plates arranged parallel to the
plane of the diffractometer circle and several scatter and receiving slits are used to create
an incident beam of X-rays that are parallel.
 Soller slits are commonly used on both the incident and diffracted beam, but this will
vary depending on the particular system.
 The scatter may be varied to control the width of the incident beam that impinges upon
the specimen and the receiving slits may be varied to control the width of the beam
entering the detector.
 Filters for removing Kβ may be located in the beam path on the generator or detector side
of the path; a monochromator, if present, is usually located on the detector side between
the receiving slit and the detector.

APPLICATIONS

 Metals have found applications in thin-film technology due to their luster and high
electrical conductivity.

ADVANCED PHYSICAL PHARMACEUTICS

 Decorative and anticorrosion coatings of chromium, zinc and derivative alloys for
armatures, metal work, kitchen fittings, automotive parts, etc., are mostly deposited by
electroplating.
 The technique of gilding of jewelry, porcelain, relics, etc., by gold leaf of only
micrometers in thickness has been continuously developed for more than 4000 years.
 X-ray powder diffraction is most widely used for the identification of unknown
crystalline materials (e.g. minerals, inorganic compounds).
 Determination of unknown solids is critical to studies in geology, environmental science,
material science, engineering and biology.

Other applications include:

 Characterization of crystalline materials.

 Identification of fine-grained minerals such as clays and mixed layer clays that are
difficult to determine optically.
 Determination of unit cell dimensions.
 Measurement of sample purity
 With specialized techniques, XRD can be used to:
 Determine crystal structures using Rietveld refinement.
 Determine of modal amounts of minerals (quantitative analysis)

Characterize thin films samples by:

 Determining lattice mismatch between film and substrate and to inferring stress and
strain.
 Determining dislocation density and quality of the film by rocking curve
measurements.
 Measuring super lattices in multilayered epitaxial structures.
 Determining the thickness, roughness and density of the film using glancing
incidence X-ray reflectivity measurements.
 Make textural measurements, such as the orientation of grains, in a polycrystalline
sample.

ADVANCED PHYSICAL PHARMACEUTICS

 References

 www.pharmainfo.net.

 www.wikipedia.com

 www.pharma-books.blogspot.com

 www.pharmamirror.com

 Skoog, Douglas A., F. James Holler and Timothy Nieman (1998). Principles of Instrumental

Analysis (5 ed.). New York. pp. 805–808. ISBN 0-03-002078-6.

 Methods of pharmaceutical analysis: Roger E Schirmer.

 Instrumental methods of chemical analysis H.Kaur.

 Text book of qualitative chemical analysis Vogel’s.

 Encyclopedia of pharmaceutical technology third edition edited by James Swarbrick.

ADVANCED PHYSICAL PHARMACEUTICS