B. Pharm.
8th Semester
PHARMACEUTICS X
(SUB CODE: PHA-UG-P803)
Date Experiment Name of the Experiment Page No Signature Remarks
No
1. 3
PREPARATION OF CALIBRATION CURVE
FOR PRACETAMOL IN ACIDIC AND BASIC
MEDIUM
2. PREPARATION OF CALIBRATION CURVE 6
FOR DICLOFENAC SODIUM IN BASIC
MEDIUM
3. PREPARATION OF CALIBRATION CURVE 8
FOR ACECLOFENAC SODIUM IN BASIC
MEDIUM
4. DISSOLUTION STUDIES OF SUSTAINED 10
RELEASE TABLET
5. FORMULATION OF FLOATING TABLETS 12
OF DICLOFENAC SODIUM
6. EVALUATION OF FLOATING TABLETS OF 13
DICLOFENAC SODIUM
7. PREPARATION OF TRANSDERMAL 16
FLIM OF DICLOFENAC SODIUM
8. EVALUATION OF TRANSDERMAL FLIM OF 17
DICLOFENAC SODIUM
9. PREPARATION OF MUCOADHESIVE 19
TABLETS
10. EVALUATION OF MUCOADHESIVE 20
TABLETS
11. PREPARATION OF MICROSPHERE BY 22
THERMAL CONGELING METHOD
HIMALAYAN PHARMACY INSTITUTE Page 1
12. PREPARATION OF ALBUMIN E 23
MICROSPHERES BY EMULSION
POLYMERISATION METHOD.
13. EVALUALTION OF ALBUMIN *
MICROSPHERES 25
HIMALAYAN PHARMACY INSTITUTE Page 2
EXPERIMENT NO: 01 DATE:
PREPARATION OF CALIBRATION CURVE FOR
PRACETAMOL IN ACIDIC AND BASIC MEDIUM
AIM: To prepare calibration curve for paracetamol in acidic and basic medium.
PRINCIPLE: Calibration curve of a pure drug required to estimate the drug
content and to know the drug release profile of a various formulations.
Standard curve is a quantitative research tool, method assay data that is used
to determine the concentration of a substance particularly protein and DNA. It
can be used in many biological experiments.
The assay is first performed with various known concentration of a substance
similar to that being major. The assay procedure may measure absorbance,
optical density, luminescence, fluorescence , radio activity or something that
can be measured. These data are used to make standard Curve plotting
concentration on the x-axis and assay measurement on y-axis. A some assay is
then performed with samples of unknown concentration to analysis the data
and locate the measurement on y-axis corresponding to assay measurement of
unknown sample and follows a line to intersect the standard curve. The co-
responding value of x-axis concentration of substance in the unknown sample.
In this experiment calibration curve of pure drug was constructed to determine
the slope by various Methods. For this experiment λ max was determined and
after getting the λ max value, the absorbance of different working standard
curve was determined.
REQIREMENTS:
Chemicals: Disodium hydrogen phosphate(2.28gm), potassium
Dihydrogen phosphate(0.19gm), sodium chloride(8gm), conc. HCL(8.5ml),
Paracetamol(100mg).
Glassware: Beaker, volumetric flask, pipette, funnel.
HIMALAYAN PHARMACY INSTITUTE Page 3
PROCEDURE:
Preparation of acidic buffer (pH 1.2): 8.5 ml of conc.HCL taken in 1000ml
volumetric flask and the volume was made up to 1000ml with distilled water.
pH was checked and adjusted.
Preparation of phosphate buffer (pH 7.4): Start with 800 ml of distilled water
to dissolve all the salts; Disodium hydrogen phosphate (2.28gm), potassium
Dihydrogen phosphate (0.19gm), sodium chloride(8gm). Adjust the pH to 7.4
with HCL. Add distilled water to make up the volume 1000 ml.
100 mg of any pure drug (paracetamol, in this case), accurately weighed
and transferred in a 100 ml of volumetric flask, diluted up to 100 ml with
acidic media and basic media, which produce concentration of 1000
μg/ml. This is called stock solution.
From the above solution 10 ml was transferred into a 100 ml volumetric
flask and volume was made up to 10 ml with acidic media and basic
media, this produce a concentration of 10 μg/ml.
From the above stock solution 1,2,3,4,5,6 ml were taken and volume
was made up to 10 ml with respective media to make different
concentration of 1μg/ml, 2μg/ml,3μg/ml & 4μg/ml 5μg/ml 6μg/ml in 10
ml volumetric flasks respectively. These are the sample solutions.
The above sample solutions were subjected to UV spectroscopy and
absorbance was checked at 257nm taking 0.1N HCL and phosphate
buffer as blanks respectively.
Table: 01
AMOUNT OF DRUG (µg/ml) ABSORBANCE
HIMALAYAN PHARMACY INSTITUTE Page 4
REFERENCE: Madan PL. Biopharmaceutics and pharmacokinetics. Jaypee
Brothers medical publishers, New Delhi, 2nd Edn, 2014.
REPORT:
HIMALAYAN PHARMACY INSTITUTE Page 5
EXPERIMENT NO: 02 DATE:
PREPARATION OF CALIBRATION CURVE FOR
DICLOFENAC SODIUM IN BASIC MEDIUM
AIM: To prepare calibration curve for diclofenac sodium in basic medium.
PRINCIPLE: Calibration curve of a pure drug required to estimate the drug
content and to know the drug release profile of a various formulations.
Standard curve is a quantitative research tool, method assay data that is used
to determine the concentration of a substance particularly protein and DNA. It
can be used in many biological experiments.
The assay is first performed with various known concentration of a substance
similar to that being major. The assay procedure may measure absorbance,
optical density, luminescence, fluorescence, radio activity or something that
can be measured. These data are used to make standard Curve plotting
concentration on the x-axis and assay measurement on y-axis. A some assay is
then performed with samples of unknown concentration to analysis the data
and locate the measurement on y-axis corresponding to assay measurement of
unknown sample and follows a line to intersect the standard curve. The co-
responding value of x-axis concentration of substance in the unknown sample.
In this experiment calibration curve of pure drug was constructed to determine
the slope by various Methods. For this experiment λ max was determined and
after getting the λ max value, the absorbance of different working standard
curve was determined.
REQIREMENTS:
Chemicals: Disodium hydrogen phosphate(2.28gm), potassium
Dihydrogen phosphate(0.19gm), sodium chloride(8gm), diclofenac sodium
(100mg).
Glassware: Beaker, volumetric flask, pipette, funnel.
PROCEDURE:
HIMALAYAN PHARMACY INSTITUTE Page 6
Preparation of phosphate buffer (pH 7.4): Start with 800 ml of distilled water
to dissolve all the salts; Disodium hydrogen phosphate (2.28gm), potassium
Dihydrogen phosphate(0.19gm), sodium chloride(8gm). Adjust the pH to 7.4
with HCL. Add distilled water to make up the volume 1000 ml.
100 mg of any pure drug (diclofenac sodium, in this case), accurately
weighed and transferred in a 100 ml of volumetric flask, diluted up to
100 ml with basic media, which produce concentration of 1000 μg/ml.
This is called stock solution.
From the above solution 10 ml was transferred into a 100 ml volumetric
flask and volume was made up to 10 ml with basic media, this produce a
concentration of 10 μg/ml.
From the above stock solution 1, 2, 3, 4, 5, 6 ml were taken and volume
was made up to 10 ml with phosphate buffer to make different
concentration of 1μg/ml, 2μg/ml, 3μg/ml & 4μg/ml 5μg/ml 6μg/ml in 10
ml volumetric flasks respectively. These are the sample solutions.
The above sample solutions were subjected to UV spectroscopy and
absorbance was checked at 276nm taking phosphate buffer as blank.
Table: 01
AMOUNT OF DRUG (µg/ml) ABSORBANCE
REFERENCE: Madan PL. Biopharmaceutics and pharmacokinetics. Jaypee
Brothers medical publishers, New Delhi, 2nd Edn, 2014.
REPORT:
EXPERIMENT NO: 03 DATE:
HIMALAYAN PHARMACY INSTITUTE Page 7
PREPARATION OF CALIBRATION CURVE FOR
ACECLOFENAC SODIUM IN BASIC MEDIUM
AIM: To prepare calibration curve for Aceclofenac sodium in basic medium.
PRINCIPLE: Calibration curve of a pure drug required to estimate the drug
content and to know the drug release profile of a various formulations.
Standard curve is a quantitative research tool, method assay data that is used
to determine the concentration of a substance particularly protein and DNA. It
can be used in many biological experiments.
The assay is first performed with various known concentration of a substance
similar to that being major. The assay procedure may measure absorbance,
optical density, luminescence, fluorescence, radio activity or something that
can be measured. These data are used to make standard Curve plotting
concentration on the x-axis and assay measurement on y-axis. A some assay is
then performed with samples of unknown concentration to analysis the data
and locate the measurement on y-axis corresponding to assay measurement of
unknown sample and follows a line to intersect the standard curve. The co-
responding value of x-axis concentration of substance in the unknown sample.
In this experiment calibration curve of pure drug was constructed to determine
the slope by various Methods. For this experiment λ max was determined and
after getting the λ max value, the absorbance of different working standard
curve was determined.
REQIREMENTS:
Chemicals: Disodium hydrogen phosphate(2.28gm), potassium
Dihydrogen phosphate(0.19gm), sodium chloride(8gm), Aceclofenac sodium
(100mg).
Glassware: Beaker, volumetric flask, pipette, funnel.
PROCEDURE: Preparation of phosphate buffer (pH 7.4): Start with 800 ml of
distilled water to dissolve all the salts; Disodium hydrogen phosphate
(2.28gm), potassium Dihydrogen phosphate (0.19gm), sodium chloride (8gm).
HIMALAYAN PHARMACY INSTITUTE Page 8
Adjust the pH to 7.4 with HCL. Add distilled water to make up the volume 1000
ml.
100 mg of any pure drug (Aceclofenac sodium, in this case), accurately
weighed and transferred in a 100 ml of volumetric flask, diluted up to
100 ml with basic media, which produce concentration of 1000 μg/ml.
This is called stock solution.
From the above solution 10 ml was transferred into a 100 ml volumetric
flask and volume was made up to 10 ml with basic media, this produce a
concentration of 10 μg/ml.
From the above stock solution 1, 2, 3, 4, 5, 6 ml were taken and volume
was made up to 10 ml with phosphate buffer to make different
concentration of 1μg/ml, 2μg/ml, 3μg/ml & 4μg/ml 5μg/ml 6μg/ml in 10
ml volumetric flasks respectively. These are the sample solutions.
The above sample solutions were subjected to UV spectroscopy and
absorbance was checked at 273nm taking phosphate buffer as blank.
Table: 01
AMOUNT OF DRUG (µg/ml) ABSORBANCE
REFERENCE: Madan PL. Biopharmaceutics and pharmacokinetics. Jaypee
Brothers medical publishers, New Delhi, 2nd Edn, 2014.
REPORT:
EXPERIMENT NO: 04 DATE:
DISSOLUTION STUDIES OF SUSTAINED RELEASE TABLET
HIMALAYAN PHARMACY INSTITUTE Page 9
AIM: To determine the dissolution characteristics of sustained release
marketed Diclofenac sodium tablets.
PRINCIPLE: Sustained delivery may provide an immediate dose required for the
normal therapeutic response followed by the gradual release of drugs in
amounts sufficient to maintain the therapeutic response for a specific
extended period of time usually 8-12 hours. Controlled release in which
delivery of drug is set a predetermined rate for a local or systemic effect for a
specific period of time.
REQUIREMENTS:
Chemicals: Disodium hydrogen phosphate(2.28gm), potassium
Dihydrogen phosphate(0.19gm), sodium chloride(8gm), diclofenac SR tablets,
Distilled water Q.S.
Glassware: Beaker, Volumetric flask, pipette, pH meter, measuring
cylinder .
PROCEDURE:
Preparation of phosphate buffer (pH 7.4): Start with 800 ml of distilled water
to dissolve all the salts; Disodium hydrogen phosphate (2.28gm), potassium
dihydrogen phosphate(0.19gm), sodium chloride(8gm). Adjust the pH to 7.4
with HCL. Add distilled water to make up the volume 1000 ml.
Preparation of standard stock solution:
Prepare pH 7.4 PBS and transfer 900 ml PBS to dissolution apparatus,
turn on the heater and set RPM .
Then the tablet was added to it.
10ml sample was withdrawn from it every ½ an hour and 10 ml fresh
solution was added. The process was continued for 4hrs.
From each of these 10ml samples 1ml solution was withdrawn and
diluted. Absorbance was measured at 276nm using UV
spectrophotometer. % cumulative release vs time graph was plotted.
TABLE: 01
HIMALAYAN PHARMACY INSTITUTE Page 10
TIME ABSORBANCE CONCENTRATION CONCENTRATION CUM. DRUG IN CDR %CDR
(µg/ml) (gm/ml) LOSS MEDIUM
Calculations:
Formula Used:
1. Dilution Factor =
2. Amount Released:
3. Percentage Drug Release =
REPORT:
EXPERIMENT NO: 05 DATE:
HIMALAYAN PHARMACY INSTITUTE Page 11
FORMULATION OF FLOATING TABLETS OF DICLOFENAC
SODIUM
Aim: To prepare formulation of floating tablets of diclofenac sodium.
PRINCIPLE: Hydrodynamically balanced tablet is a gastro retentive drug
delivery system that prolongs gastric residence time and also provides drug
release in the upper GIT for local and systemic effects. Drug that are locally
active in the stomach have narroe therapeutic absorption window, stable in git
and exhibit low solubility at higher pH are good candidates for gastro retentive
drug delivery system.
Various approaches to active gastric retention are high density system or non
floating system, floating drug delivery system, mucoadhesive system, etc. in
effervescent system, floating is activated by generation of gas bubbles which
are entrapped within the matrix of swell able polymer thereby reducing the
density of the dosage form to less than 1gm/cm 3 and thus release the drug in a
sustained manner for prolonged period of time.
REQUIREMENTS: Diclofenac sodium(100mg), sodium carboxy methyl cellulose,
microcrystalline cellulose(12gm), Hydroxypropyl methyl cellulose E 15LV
(160gm), sodium bicarbonate(5gm), citric acid, magnesium stearate(5gm).
PROCEDRE: Preparation of floatin tablet:
Floating tablet of diclofenac was prepared using Hydroxypropyl methyl
cellulose E15LV, sodium bicarbonate and citric acid as gas generating agent and
magnesium stearate as lubricant. The drugs and the excipients were crushed in
mortar with the help of pestle to reduce their size and weighing is done the
ingredients were mixed thoroughly for 10 mins to ensure uniform mixing. The
tablets were prepared by direct compression technique using 11.2mm punch
in tablet press.
REPORT:
HIMALAYAN PHARMACY INSTITUTE Page 12
EXPERIMENT NO: 06 DATE:
EVALUATION OF FLOATING TABLETS OF DICLOFENAC
SODIUM
Aim: To evaluate the prepared floating tablets.
PRINCIPLE:
1. Thickness Uniformity: Tablets thickness should be controlled with 5% or
less of a standard value. Any variation in tablet thickness should not be
apparent to the unaided eye to maintain product acceptance by
consumer as well as to facilitate packaging. At constant compressive
load, tablet thickness varies with changes in die fill and tablet weight.
Whereas, with a constant die fill, thickness varies in variation in
compressive load.
Three set of factors influence tablet thickness and tablet thickness
control:
1. The physical properties of raw materials including crystal form and true
bulk density.
2. Control of upper and lower punch lengths, which should be
standardized.
3. The granulation prosperities including bulk density, tapped density,
particle size, and particle size distribution.
2. Hardness:
Hardness may be defined as the resistance of a solid to attrition or breakage. It
has been used in characterizing tablet as it provides a simple measure of the
effectiveness of the compression process. Both Monsanto, Pfizer, Strong Cobb
and the Stokes hardness testers are used in the pharmaceutical industry to
measure the force required to break the tablet. Hardness values obtained from
these two instruments for a given set of tablets are not equivalent but they can
be correlated. There appears to be a linear relationship between the tablet
hardness and the logarithm of the compression force. One would anticipate
that as a tablet becomes more dense, its hardness would increase. Other
relationships among hardness, compression force and pack density are
empirical and can be obtained experimentally.
The desirable tablet hardness depends on the formulation intended, and tablet
weight. Small ones have low hardness (~5 kP for 100 mg tab.) and large ones
HIMALAYAN PHARMACY INSTITUTE Page 13
have high hardness (15-20 kP for ~1000 mg tab). Tablet hardness should be
high enough to keep tablet integrity in the further processing, such as coating
and packaging, and in product transportation. The following table lists
approximate values for different formulations, and serves only as an
approximate guideline:
3. Friability
Tablet friability is related to hardness. A friability tester measures the ability of
the tablet to resist abrasion which is important to packaging, shipping and
handling. Friability tests the strength of tablets against wear.
For tablets weighing <= 650 mg each, the minimum number of tablets to reach
6.5 grams are used for the test. For tablets > 650 mg each, 10 tablets are used
in the test. The total initial weight of the tablets is determined (W0).
The tablets are loaded into a friability tester, where they are subjected to
controlled falls. The test involves rotating the drum exactly 100 times, over 4
minutes (25 rpm). The intact tablets are then removed and weighed again (W).
The measure of abrasion resistance or % Friability is expressed as a percentage
loss in tablet weight:
A tablet weight loss of 1% is the USP-defined upper limit for friability; however,
a target of less than 0.3% is desirable for tablet processing and resilience.
If the size or shape of the tablet causes irregular tumbling, adjust the drum
base so that it forms an angle of about 10º with the horizontal and the tablets
HIMALAYAN PHARMACY INSTITUTE Page 14
do not bind together when lying next to each other, which prevents them from
falling freely.
4. Weight Uniformity
Variation in processing and powder flow can lead to a variation in tablet
weight. Assessing weight uniformity provides a measure of the variability in
the tabletting process. The test is performed as follows:
5. In- vitro buoyancy test:
It was calculated from floating lag time. Floating lag time is the time
period between placing the tablet in the dissolution medium and tablet
floating. N order to determine the floating time, tablet was placed in 100ml
of 0.1 N HCL and time taken to float on the surface of HCL was noted.
6. In vitro drug release study:
In vitro drug release study of formulation was carried out in USP type II
paddle apparatus with RPM set at 50 and temperature 36±0.5°c. The
dissolution medium used was 0.1N HCL. The samples were withdrawn at
intervals of 15 minutes for a period of 2 hours withdrawn sample were
replaced with same volume of prepare buffer. Then the samples were
suitably diluted and analysed in UV spectrophotometer.
REQUIREMENTS: Conc.HCL (8.5ml), Monsanto Hardness tester, Roche
friabilator, USP Dissolution apparatus, analytical balance.
REPORT:
HIMALAYAN PHARMACY INSTITUTE Page 15
EXPERIMENT NO: 07 DATE:
PREPARATION OF TRANSDERMAL FLIM OF DICLOFENAC
SODIUM
Aim: To prepare transdermal film of diclofenac sodium
PRINCIPLE: Transdermal drug delivery systems are self contained discrete
dosage form, which when applied to the skin at a controlled rate to the
systemic circulation. In comparison of conventional dosage form, TDDS offers
many advantages such as elimination of first pass metabolism, sustained drug
delivery reduce frequency of administration, better patient compliance etc.
REQUIREMENTS:
Chemicals: Ethyl cellulose (10% w/v)- 1gm, Diclofenac sodium- 500mg,
glycerol-1-1.5 ml, PVA, paraffin, water.
Glassware: Beaker, volumetric flask, pipette, Petri plates.
PROCEDURE:
Transdermal films are prepared by solvent casting method. Ethyl
cellulose solution was prepared in two beakers by dissolving 1gm of ethyl
cellulose in 10ml toluene solution and was mixed thoroughly. Then the drug
was added to the beaker. 1ml of glycerol was added to the beaker. Then the
solution was poured in a Petri plate, previously rinsed with glycerol and
covered with inverted funnel for uniform evaporation of solvent. Then it was
allowed to dry overnight.
REPORT:
EXPERIMENT NO: 08 DATE:
HIMALAYAN PHARMACY INSTITUTE Page 16
EVALUATION OF TRANSDERMAL FLIMS
Aim: To evaluate the transdermal film of diclofenac sodium.
PRINCIPLE: Transdermal drug delivery systems are self contained discrete
dosage form, which when applied to the skin at a controlled rate to the
systemic circulation. In comparison of conventional dosage form, TDDS offers
many advantages such as elimination of first pass metabolism, sustained drug
delivery reduce frequency of administration, better patient compliance etc.
REQUIREMENTS:
Chemicals: Disodium hydrogen phosphate(2.28gm), potassium dihydrogen
phosphate(0.19gm), sodium chloride(8gm), HCL q.s, water q.s.
Glassware: Franz diffusion cell, semi permeable membrane, rubber tube,
adequate, pipette, volumetric flask (1/10 ml), beaker.
PROCEDURE:
In vitro permeation study: In vitro drug release of two different formulation of
diclofenac sod. Transdermal films were studied using Franz diffusion cell using
a cellulose membrane. The cell consists of 2 chambers i.e. the donor and
receptor compartment was surrounded by a water jacket for maintaining
temp. at 37±2°c and it was provided with a sampling part. Phosphate buffer
was stirred using magnetic stirrer. The drug containing film was cut into sq.
shape having surface area of 1cm2; compartment by cellulose membrane
should dip on the dissolution medium throughout the operation. Te diffusion
medium was stirred with magnetic stirrer. At each sampling time, the solution I
the diffusion medium was completely withdrawn and replaced with fresh
phosphate buffer solution to maintain the sink condition. Samples are
withdrawn at an interval of 30 mins for 1 and 1/2 hours. The conc. of the drug
in the test samples are calculated using regression eq. obtained from
calibration curve of dicofenac sodium in phosphate buffer solution.
Time Abs Conc (std.) Amt Loss cdr %cdr
HIMALAYAN PHARMACY INSTITUTE Page 17
released
Conc/10mg
REPORT:
HIMALAYAN PHARMACY INSTITUTE Page 18
EXPERIMENT NO: 09 DATE:
PREPARATION OF MUCOADHESIVE TABLETS
AIM: To prepare Buccal mucoadhesive tablet of Paracetamol.
PRINCIPLE: Buccal delivery of drug provides an attractive alternative to the oral route of
drug delivery system particularly in overcoming problems relating to the later mode of
administration such as first pass metabolism drug degradation in the git, etc. thus, the Buccal
mucoadhesive tablet serve as a promising approach for localising the dosage form in a
specific organ and as well as controlled release of drug.
REQUIREMENTS: PCM, Carbopol 934, MCC, Magnesium stearate.
PROCEDURE:
Preparation of mucoadhesive tablet:
All the ingredients except Magnesium stearate were mixed in mortar and pestle for 10
minutes. Finally, lubricant was mixed after few minutes. Then the mixture was compressed in
11.2mm biconcave punch in 10 station compression machine. Two formulations were
prepared one with Carbopol in the ratio of 1:1 and other with 1:2.
REPORT:
EXPERIMENT NO: 10 DATE:
HIMALAYAN PHARMACY INSTITUTE Page 19
EVALUATION OF BUCCAL MUCOADHESIVE TABLET
AIM: To evaluate Buccal mucoadhesive tablet.
PRINCIPLE: Buccal delivery of drug provides an attractive alternative to the
oral route of drug delivery system particularly in overcoming problems relating
to the later mode of administration such as first pass metabolism drug
degradation in the git, etc. thus, the Buccal mucoadhesive tablet serve as a
promising approach for localising the dosage form in a specific organ and as
well as controlled release of drug.
REQUIREMENTS: Disodium hydrogen phosphate (2.28gm), potassium
Dihydrogen phosphate (0.19gm), sodium chloride (8gm), conc. HCL (8.5ml),
water q.s.
PROCEDURE:
1. Hardness:
Hardness may be defined as the resistance of a solid to attrition or breakage. It
has been used in characterizing tablet as it provides a simple measure of the
effectiveness of the compression process. Monsanto, Pfizer, Strong Cobb and
the Stokes hardness testers are used in the pharmaceutical industry to
measure the force required to break the tablet.
2. Friability
Tablet friability is related to hardness. A friability tester measures the ability of
the tablet to resist abrasion which is important to packaging, shipping and
handling. Friability tests the strength of tablets against wear.
3. Weight variation:
20 tablets were weighed individually and then together. The average weight
was calculated by:
4. Mucoadhesive property:
HIMALAYAN PHARMACY INSTITUTE Page 20
Tablets were adhered to stomach mucosa and obseserved for mucoadhesive
property by shaking the mucosa for certain period of time.
REPORT:
HIMALAYAN PHARMACY INSTITUTE Page 21
EXPERIMENT NO: 11 DATE:
PREPARATION OF MICROSPHERE BY THERMAL
CONGELING METHOD
AIM: To prepare albumin microsphere by thermal congealing method.
PRINCIPLE: Microspheres are colloidal drug delivery system intended for
parenteral administration to achieve target specificity. Microspheres can be
prepared from a variety of carrier materials (For ex. Albumin)
Albumin microspheres are chemically and physically stable, non-immunogenic
and biodegradable, and are available to be prepared in large batches and
capable of accommodating a wide variety of drug molecules in relatively non-
specific fashion. They are rapidly removed from the vascular system by
phagocytosis, potentially localised in the reticular endothelial system and
found within cytoplasm of tumour cells.
Preparation of albumin microspheres involves either thermal denaturation at
elevated temperatures (110-165°c) or chemical cross linking in vegetable oil or
Iso-octane emulsion.
REQUIREMENTS:
Chemicals: Ethyl cellulose, Bovine serum albumin, olive oil, castor oil,
anhydrous ether, cyclohexane.
PROCEDURE: An aqueous internal phase (30ml) of an emulsion (w/o type)
containing 00 mg bovine serum albumin, ethyl cellulose, 100mg paracetamol
was emulsified with 250ml of pure olive oil under constant stirring at 1200
RPM while the temperature was maintained at 4°c. This emulsion was added
gradually (10-15 drops/min) to preheat (120°c) pure olive oil under constant
stirring at 750 rpm for 15 mins. And mixture was allowed to cool down to
room temperature.
The heat stabilised albumin microspheres were separated by centrifugation at
3000 RPM for 5 min. The separated microspheres were washed with 60ml of
anhydrous ether.
REPORT:
HIMALAYAN PHARMACY INSTITUTE Page 22
EXPERIMENT NO: 12 DATE:
PREPARATION OF ALBUMIN MICROSPHERES BY
EMULSION POLYMERISATION METHOD.
AIM: To prepare albumin microspheres by emulsion polymerisation method.
PRINCIPLE: Microspheres are colloidal drug delivery system intended for
parenteral administration to achieve target specificity. Microspheres can be
prepared from a variety of carrier materials (For ex. Albumin)
Albumin microspheres are chemically and physically stable, non-immunogenic
and biodegradable, and are available to be prepared in large batches and
capable of accommodating a wide variety of drug molecules in relatively non-
specific fashion. They are rapidly removed from the vascular system by
phagocytosis, potentially localised in the reticular endothelial system and
found within cytoplasm of tumour cells.
REQUIREMENTS:
Chemicals: Ethyl cellulose, Bovine serum albumin, castor oil,
Glutaraldehyde, toluene, acetone, PCM, dichloromethane.
Glassware: Beaker, pipette, glass rod, mechanical stirrer.
PROCEDURE: Albumin microspheres were prepared by emulsion
polymerisation method. 3ml of 1%W/V albumin was added to a mixture of 15
ml castor oil and 10 ml toluene and 100mg PCM held in a beaker at
appropriate stirring speed in a mechanical stirrer (1200RPM).
Glutaraldehyde saturated toluene solution was prepared by mixing equal vol.
of glutaraldehyde and toluene in a test tube and shaking for 10 minutes. The
mixture was allowed to separate. The upper layer (toluene) was pipette and
added drop wise to the albumin emulsion. In the present study glutaraldehyde
concentration of approx. 0.7% v/v was employed.
The dispersion was then mixed at appropriate speed until cross linking was
completed (2.5hrs). At the end, the suspension of microspheres were washed
free of oil with 4ml of toluene for 4 times at 15000 rpm for 2 mins at high
speed centrifuge. Then the microspheres were suspended in 10ml of acetone
and transferred into a paper dish for air drying at room temperature. Upon
HIMALAYAN PHARMACY INSTITUTE Page 23
drying yellow to yellowish orange coloured free flowing fine powered was
obtained.
REPORT:
HIMALAYAN PHARMACY INSTITUTE Page 24
EXPERIMENT NO: 13 DATE:
EVALUALTION OF ALBUMIN MICROSPHERES
AIM: To Evaluate Of Albumin Microspheres.
PRINCIPLE: Microspheres are colloidal drug delivery system intended for
parenteral administration to achieve target specificity. Microspheres can be
prepared from a variety of carrier materials (For ex. Albumin)
Albumin microspheres are chemically and physically stable, non-immunogenic
and biodegradable, and are available to be prepared in large batches and
capable of accommodating a wide variety of drug molecules in relatively non-
specific fashion. They are rapidly removed from the vascular system by
phagocytosis, potentially localised in the reticular endothelial system and
found within cytoplasm of tumour cells.
REQUIREMENTS: Disodium hydrogen phosphate(2.28gm), potassium
dihydrogen phosphate(0.19gm), sodium chloride(8gm), conc. HCL(8.5ml),
water q.s.
Glassware: beaker, volumetric flask, pipette, pH meter.
PROCEDURE:
Morphological studies: The prepared microspheres were visually
inspected.
Particle size determination: The size of dried microspheres
were measured using an optical microscope (Olympus
Model HB, India). A standard stage micrometer was used to
calibrate the eyepiece micrometer. Dried microspheres were
placed in a glass slide and the number of divisions of the
calibrated eyepiece was counted. Microspheres were
randomly selected from each formulation and the individual
particle diameter was calculated based on this formula
HIMALAYAN PHARMACY INSTITUTE Page 25
REPORT:
HIMALAYAN PHARMACY INSTITUTE Page 26
