Centre for Pharmaceutical Sciences

Institute Of Science and Technology

JNTUH, Kukatpally, Hyderabad.

CERTIFICATE

NAME:

ROLL NO:

This is to certify that this is the bonafide work of the student in Advanced Drug Delivery
Systems during the academic year 2022-23.

No of practical certified out of in the subject of Advanced

Drug Delivery Systems.

-------------------------------------------- --------------------------------------------

Faculty Incharge Signature Examiner’s Signature

Date-------------------- Institution Rubber Stamp

Experiment No.: Date:

DETERMINATION OF LAMBDA MAX AND CONSTRUCTION OF STANDARD

GRAPH OF DRUG KETOROLAC BY UV - VISIBLE SPECTROSCOPY
Aim: To determine the Lambda max of Ketorolac in water by UV-
Visible Spectroscopy
Apparatus:
1. Volumetric flask
2. Funnel
3. Measuring Cylinder
4. 1ml pipette
5. 0.1 ml Micropipette
6. Whatmann Filter paper
Principle: Ultraviolet and visible absorption spectrophotometry is the
measurement of the absorption of monochromatic radiation by
solutions of chemical substances, in the range of 185 nm to 380
nm, and 380 nm to 780 nm of the spectrum, respectively. The
magnitude of the absorption of a solution is expressed in tens of
the absorbance, A, defined as the logarithm to base 10 of the
reciprocal of transmittance (T) for monochromatic radiation:

Where Io is the intensity of the incident radiation, I is the intensity

of the transmitted radiation. The absorbance depends on the
concentration of the absorbing substance in the solution and the
2
 thickness of the absorbing layer taken for measurement.
For convenience of reference and for ease in calculations, the
specific absorbance of a 1 per cent w/v solution is adopted in this
Pharmacopoeia for several substances unless otherwise indicated,
and it refers to the absorbance of a 1 per cent w/v solution in a 1
cm cell and measured at a defined wavelength.
Procedure: 100mg of pure drug (API) was taken and transferred to 100 ml
volumetric flask, to this 20 ml of Water was added and sonicated for
5min. the volume is made up to 100ml with Water. This is labelled as
I 000 ppm stock solution.
From the stock solution 1ml was drawn and diluted to 10 ml with
water. It is labelled as 100 ppm. Further dilutions were made and
solutions of concentrations 5-25ppm were made with water.
All solutions were scanned for absorbance from 200nm to 700nm
and spectrum graph is obtained. A overlay of spectrums was
obtained and the Amax is ascertained.

Procedure: 100mg of pure drug (API) was taken and transferred to 100 ml
volumetric flask, to this 20 ml of Water was added and sonicated for 5min. the
volume is made up to 100ml with Water. This is labelled as I 000 ppm stock
solution.
From the stock solution 1ml was drawn and diluted to 10 ml with Water. It is
labelled as 100 ppm. Further dilutions were made and solutions of
concentrations 5ppm, 10ppm, 15ppm, 20ppm and 25 ppm were made with Water.
Absorbance of the various solutions were measured at wavelength of 332nm and a
standard graph was plotted.
Report:

3
Experiment No.: Date:

PREPARATION OF TRANSDERMAL PATCHES OF KETOROLAC

TROMETHAMINE
Introduction:
Ketorolac Tromethamine is the tromethamine salt of ketorolac, is a non-steroidal anti-
inflammatory drug (NSAID), analgesic, and antipyretic properties. Transdermal patches are
topically administered medicament with systemic effect at a predetermined and controlled rate.
The active and passive design of transdermal drug delivery device, which provides an alternative
route for administering medication. Skin barrier allows pharmaceutical medicament from these
devices. From the patch, the high dosage of the drug, which is worn on the skin for an extended
period. Through the diffusion process, the drug enters the bloodstream directly through the skin.
Since there is a high concentration on the patch and a low concentration in the blood, the drug
will keep diffusing into the blood for a long period, the blood flow was maintained by the
constant concentration of drugs in the blood flow.
Aim: To prepare transdermal patch of ketorolac tromethamine
Apparatus and Chemicals required:
Ketorolac tromethamine, HPMC E15 , Eudragit RS 100, Eudragit RL 100 ,Oleic acid,
Dichloromethane AR, Methanol AR and Propylene glycol, Petridish, beaker, spatula, glass rod,
funnel, measuring cyclinder, weighing balance,Screw gauge,Franz diffusion cell.
Principle:
A patch for the transdermal delivery of ketorolac is described. The patch is capable of delivering
therapeutically effective levels of ketorolac through a patient’s skin for a period of 12 hours or
more. The patch maybe an adhesive matrix, or monolithic matrix or a liquid reservoir type
transdermal patch.
Procedure:
Matrix type transdermal patches containing Ketorolac were prepared by solvent evaporation
technique, using different ratios of HPMC E15, ERL 100 (KT1 to KT5), and HPMC E15, ERS
100 (KT6 to KT10). The polymers were weighed in requisite ratios and allowed for swelling for
about 6 hrs. in the solvent mixture (1:1 ratio of dichloromethane, methanol). 15% v/w propylene

4
glycol was incorporated as a plasticizer. Then the drug solution was added to the polymeric
solution, cast onto an umbra Petri plate with a surface area of about 69.24sq.cm, allowed for air-
drying overnight followed by vacuum drying for 8-10 hrs. The entire sheet was cut into small
patches with an area of 6.9 cm2 i.e. with a diameter of 2.9 cm. About 7 patches were obtained
from each sheet. All formulations carried 15% v/w polyethylene glycol as plasticizer and 12%
oleic acid as a penetration enhancer
Evaluation of Transdermal patches:
Thickness of patch:
The thickness of the Drug loaded patch is measured in different points by using a digital
micrometer and the average Thickness and standard deviation is determined to Ensure the
thickness of the prepared patch. The Thickness of transdermal film is determined by Travelling
microscope dial gauge, screw gauge or Micrometer at different points of the film.
Content uniformity test:
10 patches are selected and content is determined for individual patches. If 9 out of 10 patches
have content between 85% to 115% of the specified value, then transdermal patches pass the test
of content uniformity. If 3 patches have content in the same range of 75% to 125%, then
additional 20 patches are tested for drug content. If these 20 patches have range from 85% to
115%, then transdermal patches pass the test.
Uniformity of weight:
Weight variation is studied by individually weighing 10 randomly selected patches and
calculating the average Weight. The individual weight should not deviate Significantly from the
average weight.
Folding endurance:
Evaluation of folding endurance involves determining the Folding capacity of the films subjected
to frequent extreme Conditions of folding. Folding endurance is determined by Repeatedly
folding the film at the same place until it breaks. The number of times the films could be folded
at the same Place without breaking is folding endurance value.
Percentage moisture content:
The prepared films are weighed individually and kept in a Desiccators containing calcium
chloride at room temperature for 24 h. The films are weighed again after a specified Interval
until they show a constant weight. The percent Moisture content is calculated using following

5
formula.

% Moisture content = (Initial weight -Final weight) *100

Final weight
Moisture uptake:
The weighed films were kept in desiccators at room Temperature for 24h containing saturated
solution of potassium Chloride in order to maintain 84% RH. After 24h, the films were
Reweighed and determined the percentage moisture uptake from the below mentioned formula.
% Moisture content= (Final weight -Initial weight) *100
Initial weight
Drug content:
An accurately weighed portion of film (about 100 mg) is Dissolved in 100 mL of suitable solvent
in which drug is Soluble and then the solution is shaken continuously for 24 h In shaker
incubator. Then the whole solution is sonicated. After sonication and subsequent filtration, drug
in solution is Estimated spectrophotometrically by appropriate dilution.
Flatness:
A transdermal patch should possess a smooth surface and should not constrict with time. This
can be demonstrated with Flatness study. For flatness determination, one strip is cut from the
centre and two from each side of patches. The Length of each strip is measured and variation in
length is Measured by determining percent constriction. Zero percent Constriction is equivalent
to 100 percent flatness.

%constriction = I1 – I2 X 100
I1
I2 = Final length of each strip
I1 = Initial length of each strip

Report:

6
Experiment No.: Date:

DIFFUSION STUDIES OF KETOROLAC PATCH

Introduction:
Transdermal drug delivery system (TDDS) is a sustained drug delivery of active pharmaceutical
ingredients (API) into systemic circulation at a predetermined rate .Ketorolac is used to relieve
moderately severe pain, usually pain that occurs after an operation or other painful procedure.
Aim:
To Perform the Diffusion Studies of Ketorolac Patch
Apparatus and chemical required:
Ketorolac Patch , Franz Diffusion Cell ,test tubes ,porous synthetic membranes (e.g., cellulose
acetate, polysulfone, Egg Membrane.)
Principle:
The diffusion is based on the principle that the net flow of molecules occurs from regions of
higher concentration to regions of low concern under the influence of concentration gradient.
Procedure:
Preparation of phosphate buffer solution, pH 7.8:
Five hundred ml of 0.2M potassium dihydrogen is taken in 2000ml volumetric flask, to which
445.0ml of 0.2 N sodium hydroxide solution is added and the volumetric flask to which 445.0 ml
of 0.2 N sodium hydroxide solution is added and the volume is made up to the mark with
distilled water.
Extraction of Egg Membrane:
The egg membrane is used in the Diffusion studies, it allows several molecules and ions to pass
through it, it is extracted by the inner membrane removal. Membrane removal is achieved by
either plasma ashing (etching), sodium hypochlorite, or acid (HCl) treatment. These dissolves the
shell of the Egg leaving the membrane and the membrane should be placed in pH 7.4 Phosphate
buffer before use.
Diffusion Method:

 The permeation studies were performed in a modified Franz’s diffusion cell having receptor
compartment capacity of 75 mL and cross-sectional area of 3.14 cm2.

7
 The Egg Membrane was extracted by treating the egg with HCL acid, the film was tied up in
the donor compartment so that polymeric side facing the receiver compartment. To place the
Transdermal film of Ketorolac above it.

 The receptor compartment was filled with phosphate buffer (pH 7.4) and the receptor media
was constantly stirred with the help of a magnetic stirrer. The temperature of the diffusion
cell was maintained at 37 ± 1°C by circulating water jacket.

 The samples (1 mL each time) were withdrawn at different time intervals and an equal
amount of receptor media was replaced each time.

 Absorbance of the samples were read spectrophotometrically at 322 nm. The amount of drug
permeated per square centimeter of Egg Membrane at different time intervals was plotted
against the time.

Report:

8
Experiment No.: Date:

PREPARATION AND EVALUATION OF KETOROLAC NIOSOMES

Aim: To prepare and evaluate ketorolac Niosomes.
Apparatus and chemical required:
Rotary evaporator, round bottomed flask, Beakers, Micro Pipette, Span 60, Cholesterol.
Principle:
Dissolution is defined as “the amount of drug substance that goes into solution per unit time
under standardized conditions of liquid/solid interface, temperature and solvent composition” .
Dissolution is conducted to know the minimum time taken by the drug to dissolve itself in the
systemic circulation and starts its action in the body. There are 2 methods of to know the
dissolution rate,
1.In-vitrostudies
2. In-vivo studies

In the pharmaceutical industry, drug dissolution testing is routinely used to provide analytical in
vitro drug release erudition for both quality control scheme, i.e., to compute batch-to-batch
consistency of solid oral dosage forms such as tablets and drug reinforcement, i.e., to anticipate
in vivo drug release profiles.

Procedure:

Preparation of Ketorolac Niosomes

The Drug (Ketorolac) & surfactant (Span 60 & Span 80) were placed in a round bottomed flask.
The solvent system is then added to the mixture and the ingredients were dissolved in the solvent
(Chloroform: methanol) by hand shaking. The flask was attached to a rotary evaporator and
immersed in water bath maintained at 60˚C, rotated with 100rpm for 45min. Formation of thin
film at the bottom was observed. The thin film is hydrated using 6.8pH buffer. The resultant
solution was sonicated in Bath sonicator for 10mins. The niosomal dispersion formulations were
shown in the Table-1 with various surfactants and their composition.

9
Evaluation of Niosomes

Particle size, poly dispersity index and zeta potential the vesicle size, of the prepared Niosomes
were determined based on laser diffraction using the Malvern Master sizer by diluting the sample
using water as dispersant.

Report:

10
Experiment No.: Date:

DETERMINATION OF LAMBDA MAX AND CONSTRUCTION OF STANDARD

GRAPH OF DRUG PARACETAMOL IN WATER BY UV - VISIBLE
SPECTROSCOPY
Aim: To determine the Lambda max of Paracetamol in water by UV-
Visible Spectroscopy
Apparatus:
1. Volumetric flask
2. Funnel
3. Measuring Cylinder
4. Whatmann Filter paper
Principle: Ultraviolet and visible absorption spectrophotometry is the
measurement of the absorption of monochromatic radiation by
solutions of chemical substances, in the range of 185 nm to 380
nm, and 380 nm to 780 nm of the spectrum, respectively. The
magnitude of the absorption of a solution is expressed in tenns of
the absorbance, A, defined as the logarithm to base 10 of the
reciprocal of transmittance (T) for monochromatic radiation:

Where Io is the intensity of the incident radiation I is the intensity

of the transmitted radiation. The absorbance depends on the
concentration of the absorbing substance in the solution and the
thickness of the absorbing layer taken for measurement.
11
 For convenience of reference and for ease in calculations, the
specific absorbance of a 1 per cent w/v solution is adopted in this
Pharmacopoeia for several substances unless otherwise indicated,
and it refers to the absorbance of a 1 per cent w/v solution in a 1
cm cell and measured at a defined wavelength.
Procedure: 100mg of pure drug (API) was taken and transferred to 100 ml
volumetric flask, to this 20 ml of Water was added and sonicated for
5min. the volume is made up to 100ml with Water. This is labelled as
I 000 ppm stock solution.
From the stock solution 1ml was drawn and diluted to 10 ml with
water. It is labelled as 100 ppm. Further dilutions were made and
solutions of concentrations 2-10ppm were made with water.
All solutions were scanned for absorbance from 200nm to 700nm
and spectrum graph is obtained. A overlay of spectrums was
obtained and the Amax is ascertained.

Procedure: 100mg of pure drug (API) was taken and transferred to 100 ml
volumetric flask, to this 20 ml of Water was added and sonicated for 5min. the
volume is made up to 100ml with Water. This is labelled as I 000 ppm stock
solution.
From the stock solution 1ml was drawn and diluted to 10 ml with Water. It is
labelled as 100 ppm. Further dilutions were made and solutions of
concentrations 2ppm, 4ppm, 6ppm, 8ppm and 10 ppm were made with Water.
Absorbance of the various solutions were measured at wavelength of 271nm and a
standard graph was plotted.
Report:

12
Experiment No.: Date:

DISSOLUTION STUDIES OF MARKETED PARACETAMOL TABLETS

Introduction:
Paracetamol is a non-steroidal anti-inflammatory drug. It is prominently used as antipyretic and
anodyne(analgesic), In treatment of fever, pain, headache and such other discomfort. Its
chemically familiar as acetaminophen i.e., (4- hydroxy acetanilide). It is also given in
combination with many cold medications, also in cancer pains and to reduce the pain after
surgery. It is predominantly safe in normal doses; higher doses cause hepatic disorders.
Aim: Determine Dissolution studies on marketed Paracetamol.
Apparatus and chemical required:
Marketed paracetamol containing 650mg of drug, potassium dihydrogen phosphate, sodium
hydroxide, distilled water, Volumetric flask, watt man filter paper, test tube, UV
spectrophotometer.
Principle:
Dissolution is defined as “the amount of drug substance that goes into solution per unit time
under standardized conditions of liquid/solid interface, temperature and solvent composition” .
Dissolution is conducted to know the minimum time taken by the drug to dissolve itself in the
systemic circulation and starts its action in the body. There are 2 methods of to know the
dissolution rate,
1.In-vitrostudies
2. In-vivo studies

In the pharmaceutical industry, drug dissolution testing is routinely used to provide analytical in
vitro drug release erudition for both quality control scheme, i.e., to compute batch-to-batch
consistency of solid oral dosage forms such as tablets and drug reinforcement, i.e., to anticipate
in vivo drug release profiles.

Procedure:
Preparation of buffers
Preparation of phosphate buffer solution, pH 7.8
500 ml of 0.2M potassium dihydrogen is taken in 2000ml volumetric flask, to which 445.0ml
13
of 0.2 N sodium hydroxide solution is added and the volumetric flask to which 445.0 ml of 0.2 N
sodium hydroxide solution is added and the volume is made up to the mark with distilled water.
Potassium dihydrogen phosphate (0.2 M) solution:
Potassium dihydrogen phosphate (13.609 gram) is added to 500 ml volumetric flask containing
distilled water and the volume is made up to the mark with distilled water.

Sodium hydroxide (0.2N) solution:

Four gram of sodium hydroxide is taken in 500 ml volumetric flask containing distilled water
and volume is made up to the mark with distilled water.

Dissolution Method:
The dissolution method is a kinetic method where periodical samples are withdrawn for the
determination
1) Phosphate buffer solution pH 7.8(900ml) is measured and transferred into the dissolution
flask.
2) The temperature is maintained at 37±0.50 C.
3) Various marketed paracetamol tablet containing 650 mg drug is placed at the bottom of the
jar.
4) The paddle is rotated at 50 rpm.
5) Five ml of sample is withdrawn at 0 min and transferred into the test tube appropriately
labelled. Immediately 5ml of phosphate buffer solution pH 7.8 is replaced into the dissolution
flask.
6) Similarly samples are collected at 5, 10, 15, 30, 45, 60 and 90 min intervals. A 5 ml of fresh
dissolution medium is replaced in the flask whenever the sample is withdrawn
7) All samples are filtered using watt man filter paper.
8) The absorbance at 249 nm is measured in UV spectrophotometer using the phosphate buffer
solution as a blank.
9) The absorbance are recorded in the table and further calculations are done
10) A graph is plotted by taking cumulative percent of drug dissolved on y-axis and time on x-
axis .

Report

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Experiment No.: Date:

PREPARATION AND EVALUATION OF PARACETAMOL MICROSPHERES

Aim: To Prepare and Evaluate Paracetamol Microspheres.
Apparatus: Beakers, Petri dish, Glass Rod, Funnel, Syringe, Magnetic Stirrer.
Chemicals: Paracetamol, Sodium Alginate, Calcium Chloride, Distilled Water.
Principle: Microspheres are free flowing polymeric micro particles loaded with biologically
active drugs intended for providing constant and prolonged therapeutic effect thus reducing
the dosing frequency and thereby improving the patient compliance. These are not only used
for prolonged release but also for targeting drug to specific site for minimizing the side effects.
Paracetamol is a mild “aniline” analgesic with weak COX enzyme inhibiting activity. It is
used primarily as antipyretic and analgesic drug. It has a short biological half life (2.5h)
and about 98% of the drug gets eliminated after a single dose thus requiring a high dosing
frequency. Therefore, the present experiment aims to develop the alginate microspheres of
paracetamol prepared with crosslinking agent (CaCl2) modifying the dissolution profile and
providing a sustained release of drug leading to reduced dosing frequency.
Procedure: The microspheres of paracetamol were prepared by Ionotropic Gelation method
using sodium alginate as polymer and calcium chloride as crosslinking agent. A 2% sodium
alginate solution was prepared by dissolving 2gms of sodium alginate in 100 mL distilled
water with stirring on a magnetic stirrer. 500mg of paracetamol was dispersed in the sodium
alginate solution. Crosslinking solution of 10 %w/v is prepared by dissolving 10gms calcium
chloride in 100ml distilled water. Sodium alginate solution was filled in the syringe and
dropped into the solutions of crosslinking agent from a height of 6 inches with speed of about
50 drops per minute. Microspheres were prepared due to the crosslinking of the polymer
by the calcium ions. The prepared microspheres were collected by decantation followed by
centrifugation of the solutions, air dried overnight and then stored in vacuum desiccators.

Evaluation tests:
Drug content:
An amount of microspheres containing a quantity equivalent to 100 mg paracetamol were

15
weighed, crushed and dissolved into 100mL of distilled water using magnetic stirrer for 24h.
Sample was withdrawn after 24h,diluted appropriately and analyzed spectrophotometrically at
257nm for determination of the drug content.
Angle of repose:
It is a measure of resistance to flow and calculated by funnel method. A weighed quantity of
microspheres was passed through the funnel and the heap was formed on the paper. The area
of the heap was encircled and diameter of the circle and the height of the heap were
measured and the angle of repose was calculated as follows:
TanФ = 2H/D
Where
H = Height,
D = Diameter of heap formed,

2H/D = Surface area of the heap formed .

Bulk density:
A weighed amount of microspheres were filled into a measuring cylinder and the volume
(V0) occupied by the microspheres was noted and the bulk density was calculated as

Tapped density:
A weighed quantity of microspheres were filled in a measuring cylinder and the cylinder
was tapped against a wooden surface at regular interval for 100 times, then the volume
occupied by the microspheres was noted down and tapped density was calculated as follows

Flow properties :
Carr’s compressibility index and Hausner’s ratio were calculated for the uncoated
microspheres using the following equations:
16
In Vitro drug release studies:
In Vitro release studies was carried out in USP type II dissolution apparatus using 900mL of
acid buffer pH 1.2 (upto 3h) and phosphate buffer pH 6.8 (after 3h) as dissolution medium’s
maintained at 37±0.5℃ and 50 rpm. A quantity of microspheres equivalent to 100 mg of
paracetamol was used for the dissolution study. Samples (5mL) were withdrawn at
different intervals and an equal volume of the fresh dissolution medium was introduced into the
apparatus. Each sample was diluted suitably with dissolution medium and analyzed with UV
spectrophotometer at 257nm for determining the drug release.

Report: The Paracetamol Microspheres are prepared and are evaluated for determining
micromeritic properties, drug content and invitro drug release studies.

17
Experiment No.: Date:

SOLUBILITY OF OLMESARTAN IN VARIOUS OILS, SURFACTANTS AND

CO-SURFACTANTS

Aim: To determine the solubility of Olmesartan in various oils, surfactants and co surfactants.

Instruments: cyclo meter, isothermal mechanical shaker, whatmann fiter, uv spectrophotometry.

Chemicals:

Oils: Almond oil, Castor oil, Olive oil, Cotton seed oil, Arachis oil and Oleic acid

Surfactants: Cremophor S9, tween 20, tween 40, tween 60, tween 80, span 20, span 80,

Co surfactants: PEG 200, PEG 400, PEG 600, Propylene glycol and Glycerin.

Theory: Olmesartan (OLM) is a novel selective angiotensin II receptor blockers that are
approved for the treatment of hypertension. OLM is a poorly water-soluble drug and its aqueous
solubility is reported to be less than 1 mg/ml. It is a prodrug that is rapidly de-esterified during
absorption from the gastrointestinal tract to produce an active metabolite. The oral bioavailability
of OLM is only 26% in healthy humans due to low solubility in water and unfavorable breakage
of the ester drug to a poorly permeable parent molecule in the gastrointestinal fluids.

Method:

Solubility of OLM was carried out in different oils (almond oil, Castor oil, Olive oil, Cotton seed
oil, Arachis oil and Oleic acid), surfactants (Cremophor S9, Tween 20, Tween 40, Tween 60,
Tween 80, Span 20 and Span 80) and cosurfactants (PEG 200, PEG 400, PEG 600, Propylene
glycol and Glycerin) using shake flask method. The excess amount of the drug (approximately
500 mg) was introduced into 2 ml of each vehicle in screw capped greiner tubes.The mixtures
were mixed well using a vortex mixer (Maximix I,USA) for 10 min to enhance the proper
mixing of the drug with the vehicles and thus facilitate the solubilization. The obtained

18
mixtures were then shaken for 72 h in an isothermal mechanical shaker (Clifton shaking water
bath, UK) maintained at 40 °C to attain equilibrium. After reaching equilibrium, the equilibrated
samples were centrifuged at 3000 r. p. m for 15 min to precipitate the undissolved OLM.
Aliquots from the supernatants were then withdrawn and filtered through a membrane filter (0.45
um, Whatman). Filtered solutions were suitably diluted with methanol, and drug concentrations
were determined using UV-Visible spectrophotometer at 256nm. All measurements were done in
triplicate, and the solubility was expressed as the mean value (mg/ml) +SD.

Report:

19
Experiment No.: Date:

PARTICLE SIZE DETERMINATION BY DYNAMIC LIGHT

SCATTERING (DLS)
Aim: To determine the particle size of the given sample by DLS

Apparatus:
1. 100 ml Volumetric flask

2. 10 ml Volumetric flask

3. 1ml pipette

4. 0.1 ml Micropipette

5. 0.4µ Nylon Filter

Principle: The particle size measured in a Dynamic Light Scattering (DLS) instrument is the
diameter of the sphere that diffuses at the same speed as the particle being measured. The
Zetasizer system determines the size by first measuring the Brownian motion of the particles
in a sample using DLS and then interpreting a size from this using established theories.
Brownian motion is defined as: "The random movement of particles in a liquid due to the
bombardment by the molecules that surround them”. The particles in a liquid move about
randomly and their speed of movement is used to determine the size of the particle. It is known
that small particles move or diffuse more quickly in a liquid than larger particles. This
movement is carrying on all the time. So if we take two 'pictures' of the sample separated by a
short interval of time we can see how much the particles have moved and therefore calculate
their size. If there has been a minimal particle movement between the two 'pictures' the
particles the sample can be inferred as large; similarly, if there has been a large amount of
movement between the two 'pictures', then the particles in the sample can be inferred as
small. Using this knowledge and the relationship between diffusion speed and size, the size can
be determined. A wide range of materials exist as molecules or particles that can be
characterized by dynamic light scattering. These include proteins, polymers, emulsions and
vesicles, as well as materials more traditionally thought of as particles, such as clays, silica,
pigments and inks.

20
Procedure:

0.1 ml of sample is diluted to 10 ml of water and mixed properly. The diluted sample is allowed
to stand for 30 min at room temperature. The sample is the filtered with 0.4µm nylon filter. The
sample is put in cuvette and particle size is determined.
Report: Particle size of the given sample was found to be

21
Experiment No.: Date:

DETERMINATION OF ZETA POTENTIAL BY DYNAMIC LIGHT

SCATTERING (DLS)
Aim: To determine the particle size of the given sample by DLS

Apparatus:

1. 100 ml Volumetric flask

2. 10 ml Volumetric flask

3. 1ml pipette

4. 0.1 ml Micropipette

5. 0.4µ Nylon Filter

Principle: Most liquids contain Ions; these are negatively and positively charged
atoms called Cations and Anions respectively. When a charged particle is
suspended in a liquid ion of an opposite charge will be attracted to the surface of
the suspended particle.

A negatively charged sample attracts positive ions from the liquid and conversely a
positive charged sample attracts negative ions from the liquid. Ions close to the
surface of the particle, will be strongly bound while ions that are further away will be
loosely bound forming what is called a Diffuse layer. Within the diffuse layer there
is a notional boundary and any ions within this boundary will move with the
particle when it moves in the liquid; but any ions outside the boundary will stay
where they are - this boundary is called the Slipping plane.

A potential exists between the particle surface and the dispersing liquid which varies
according to the distance from the particle surface - this potential at the slipping
plane is called the zeta potential. Zeta potential is measured using a combination of
the measurement techniques: Electrophoresis and Laser Doppler Velocimetry,
sometimes called Laser Doppler Electrophoresis. This method measures how fast a
particle moves in a liquid when an electrical field is applied - i.e. its velocity. Once
we know the velocity of the particle and the electrical field applied we can, by using
22
two other known constants of the sample - viscosity and dielectric constant - work
out the zeta potential.

Procedure:0.1 ml of sample is diluted to 10 ml of water and mixed properly. The

diluted sample is allowed to stand for 30 min at room temperature. The sample is the
filtered with 0.4µm nylon filter. The sample is put in cuvette and zeta potential is
determined.
Report: The zeta potential of the dispersion was found to be _____________.

23
Experiment No.: Date:

FLOW PROPERTIES – POWDERS OR GRANULES

Aim:
To evaluate flow properties of granules/powders
I. Measurement of angle of repose of the given granules/powders
II. Study of the effect of magnesium stearate or talc on the flow properties of granules

(i) Measurement of angle of repose of the given granules/powders

Principle:
Angle of repose is defined as the maximum angle possible between the surface of the pile
surface of the pile of powder and the horizontal plane
The angle of repose is designated by Ө and given by equation
tan Ө = h/r or Ө = tan-1 h/r
Where,
h = height of pile, cm
r = radius of the base of the pile, cm
The lower the angle of repose, the better the flow properties. When granules are placed in the
hopper and allowed to slide down onto the die for compression, it form a pile. The angle of repose
may be calculated by measuring height in (h) of the pile and the radius of the base (r) with ruler.
During the flow through the hoppers, the granules exhibit internal flow and demixing. Flow
of granules is hindered on acount of frictional forces. Therefore, the tangent of the angle of repose is
expressed as the coefficient of friction (µ). Angle of repose values of some pharmaceutical
excipients is given.

Factors influencing flow:

Particle size (coarse or fine particles), nature of particles (shape, surface roughness, density,

24
porosity and cohesiveness) and moisture content in the powder influence the flow of powder.
Angle of repose of some pharmaceutical excipients
Substance Angle of repose, Ө (degrees)
Lactose 15
Lactose (spary dried) 20
Magnesium oxide 20
Microcrystalline cellulose 15
Sodium bicarbonate USP 20
Starch 15
Talc 25

Applications:
During tableting, improper flow of granules from the hopper leads to under-fill or over-fill in
the die cavity. As a result, tablets will have under-weight or over-weight. Weight variation further
affects the content uniformity and dose precision. It also problems of hardness and friability during
compression. Similarly, in the filling of capsules, irregular flow of granules/powders creates
problems of weight variation etc.
Apparatus and chemicals:
Glass funnel 1 granules
Iron stand and ring 1 talc
Weights box 1 magnesium stearate
Procedure:
1. Select a glass funnel, which has a round stem of 15 to 30 mm diameter with a flat edge.
2. Fix the funnel with a clamp ( on the ring support) to the iron stand.
3. Place a glass plate on the table and arrange it below the glass funnel. Keep one graph paper
on the glass plate.
4. Weigh approximately 100g of granules.
5. Pour granules onto funnel while blocking the orifice of the funnel by thumb.
6. Remove the thumb. The granules flow down onto the graph paper and form a cone shaped
pile.

25
 7. Adjust the funnel clamp so that the gap betweeen the bottom of the funnel stem and peak of
the powder pile is about 3mm.
8. Repeat the steps 5 to 7 until appropriate gap is maintained.
9. Finally pour the granules back into funnel and allow to flow.
10. Mark 4 points, which are opposite to each other on the circular base on the graph paper.
Record the readings , this value is the diameter (d). Calculate the radius (r).
11. Measure the height of the pile using two rulers. Keep one ruler vertically an another
horizontally to touch the peak of the pile. Then read the value on the vertical scale. Record
the reading. This value represents the height (h).
12. Substitue the value in the equation to obatin the angle of repose. Generally, the h/r ratio (log
scale) versus angle of repose ( abscissa) data were plotted on semi-log paper and copies of
the curve made available to the students for the purpose of calculating the angle.
13. Repeat this procedure for two more trials and take an average.
14. Comment on the flow of the material by consulting the relationship.

(II) EFFECT OF MAGNESIUM STEARATE OR TALC ON THE

FLOW PROPERTIES OF GRANULES:
THEORY
Improves flow properties:
Flow properties can be improved by the following methods
 Processing powder into granules of spherical shape, because the spherical granules just roll
on (better flow).
 Choosing optimum size of granules, may be about 400 to 800µm.
 Incorporating optium concentration of fines, about 15% w/w.
 Incorporating optium concentration of lubricants such as magnesium stearate and talc.

A combination of the above measures has a synergistic effect and enhances the flow
characteristic of granules or powder.
Principle:
Glidants are those substances, which are intended to promote the flow of the granules or
powder material by reducing the friction between the particles.
A few examples of effective glidants are talc, corn starch, magnesium stearate etc. The
26
substances get adsorbed on the surface of the granules or powder and machine parts, thereby
reduce the frictional forces.
Since flow characteristics are evaluated by angle of repose, it can be used to estimate the
optimum concentration of glidant for better flow. As the concentration of talc and magnesium
stearate is increased continuously, the angle of repose decreases gradually. This trend continues
up to a particular concentration (optimum) and further increase in concentration increases the
angle repose. The inflection point i.e the concentration at which the trend changes is considered
as optimum concentration of glidant required.
Apparatus and chemicals:
Glass funnel 1 granules
Iron stand and ring 1 talc
Weights box 1 magnesium stearate
Procedure:
The angle of repose of the granules is calculated in the same manner without and with the
addition of glidants.
The glidant selected here is talc or magnesium stearate. The concentrations are:
Talc (% w/w) 0 1.0 2.0 3.0 4.0 5.0
Magnesium stearate (% w/w) 0 0.2 0.4 0.6 0.8 1.0
1) Determine the angle of repose of the granules without the addition of glidants.
2) Add the glidants in low concentration (1.0g of talc or 0.2g of magnesium stearate as the
case may be ) to the granules and mix them thoroughly.
3) Estimate the angle of repose of this blend in two trials. Report it.
4) Calculate the average angle of repose.
5) Add the increments of glidant (say 1.0g of talc or 0.2g of magnesium stearate as the case
may be ) and mix them thoroughly.
6) Repeat steps 3 and 4.
7) Add further increments of glidant and determine the angle of repose.
8) Repeat steps 3 and 4 with further additions of 1.0g of talc until optimum concentration is
obtained.
9) Plot a graph taking the concentration of glidant on x-axis and angle repose on y-axis.
Observe the optimum concentration of glidant required.
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Report:

28
Experiment No.: Date:

DRUG EXCIPIENT COMPATIBILITY STUDIES BY SOLID SAMPLING

TECHNIQUES USING IR-SPECTROSCOPY
Aim: To Perform the Drug - Excipient Compatibility studies by solid sampling using IR
spectroscopy

Apparatus Required:

1. 50 ml Beaker

2. IR Mortar and Pestle

3. Tissue paper

Chemicals Required:
1. Potassium Bromide (KBr) IR Grade

2. Acetone

3. API

4. Excipients

Theory:

IR Spectroscopy

When a broad-band source of IR energy irradiates a sample, the absorption of

IR energy by the sample results from transitions between molecular
vibrational and rotational energy levels. A vibrational transition may be
approximated by treating two atoms bonded together within a molecule as a
harmonic oscillator. Based on Hooke's law, the vibrational frequency between
these two atoms may be approximated as:

1 k
v=-
21r\ µ

Whereµ is the reduced mass of the two atoms, µ = m1m2 , and k

is the force
m1 +m2

constant of the bond (dynes/cm).

29
Transmission mode

Transmittance (1) is a measure of the decrease in radiation intensity at given

wavelengths when radiation is passed through the sample. The sample is
placed in the optical beam between the source and detector. The arrangement is
analogous to that in many conventional spectrophotometers and the result can be
presented directly in terms of transmittance (T) or/and absorbance (A).
Drug - excipient Compatibility

Unless otherwise directed in the individual monograph, prepare the substance under
examination and the Reference Substance in the form of discs dispersed in potassium
bromide IR or potassium chloride IR and record the spectra between 4000 cm-1 and
625 cm-1 (2.5 µm to 16 µm) under the same operational conditions. The absorption
maxima in the spectrum obtained with the substance under examination correspond
in position and relative intensity to those in the spectrum obtained with the Reference
Substance.
Procedure: Solid Sampling

Triturate about 1 mg of the substance with approximately 300 mg of dry, finely

powdered potassium bromide IR or potassium chloride IR, as directed. These
quantities are usually suitable for a disc 13 mm in diameter. Grind the mixture
thoroughly, spread it uniformly in a suitable die and compress under vacuum at a
pressure of about 800 MPa. Commercial dies are available and the manufacturer's
instructions should be strictly followed. Mount the resultant disc in a suitable
holder in the spectrophotometer. Several factors, such as inadequate or excessive
grinding, moisture or other impurities in the halide carrier, may give rise to
unsatisfactory discs. A disc should be rejected, if visual inspection shows lack of
uniformity or if the transmittance at about 2000 cm-1 (5 µm) in the absence of a
specific absorption band is less than 75 per cent without compensation. If the
other ingredients of tablets, injections, or other dosage forms are not completely
removed from the substance being examined, they may contribute to the spectrum.
Transmission mode. The measurement of transmittance (T) is dependent on a
background transmittance spectrum for its calculation. A background reference
can be air, an empty cell, and a solvent blank or in special cases a reference
sample. The method generally applies to liquids, diluted or undiluted,
dispersions, solutions and solids. For transmittance measurements of solids, a
suitable sample accessory is to be used. The samples are examined in a cell of
30
suitable path length (generally 0.5-4 mm), transparent to NIR radiation, or by
immersion of a fibre optic probe of a suitable configuration, which yields a
spectrum situated in a zone of transmission compatible with the specifications of
the apparatus and appropriate for the intended purpose.

Report:

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