BPHM 806 Lab - Pharmaceutics-IX (Dosage Form Design)

1. To determine the flow properties of given sample of powder and study effect of glidants
on flow.
2. To determine the partition coefficient of given sample of drug.
3. To carry out the solubility studies of given poorly water soluble drug.
4. To perform assay of aspirin and study the effect of adverse condition on aspirin.
5. To find out E1%, 1cm of Paracetamol.
6. To study dissolution apparatus and perform validation of dissolution apparatus.
7. To perform dissolution study of marketed Paracetamol tablets .
8. To perform comparative study of dissolution of marketed products. (i) Crocin (ii)
Welset .
9. To perform dissolution study and compare data of conventional and sustained release
diclofenac sodium tablet.
10. To study Pharmacokinetic Parameters of given IV and oral data.
11. To Formulate & Evaluate Microcapsules of Paracetamol using ethyl cellulose.
12. To formulate and evaluate transdermal patch of Salbutamol Sulphate.
13. To formulate and evaluate microcapsule of diclofenac sodium.
14. To formulate and evaluate topical gel of paracetamol.
15. To enhance the solubility of Paracetamol by solid dispersion technique.
16. To formulate and evaluate sodium alginate beads.
S.No. Aim Of Experiment Page No.
1. To determine the flow properties of given sample of powder.

2. To study the effect of Glidants on the flow properties of given

sample of powder.
3. Formulation and Evaluation of calcium alginate beads of
Paracetamol.
4. To formulate and evaluate the diclofenac sodium gel by using
natural polymer.
5. To formulate and evaluate transdermal patches of diclofenac
potassium
6. To Formulate & Evaluate Microcapsules of Paracetamol using
ethyl cellulose.
7. Formulation and comparative Evaluation of marketed
Paracetamol tablets.
8. To perform comparative study of dissolution of marketed
products.
9. To study Pharmacokinetic Parameters of given IV and oral
data.
10. To formulate and evaluate microspheres of diclofenac sodium
using Bovine serum albumin.
11. To carry out the solubility studies of given poorly water
soluble drug.
12. To determine the partition coefficient of given sample of drug.
 EXPERIMENT NO. 1

AIM: To determine the flow properties of given sample of powder.

REFERENCES
1. CVS Subrahmanyam, “Textbook of physical pharmaceutics”, Vallabh Prakashan,Delhi,
2nd edition, reprint 2008, page no. 221-224.
2. S.P Aggarwal, Rajesh Khanna, “Physical Pharmacy, CBS Publishers and Distributors,
New Delhi, 2nd edition, reprint 2006, page no. 43-45.

REQUIREMENTS
CHEMICALS: Powder Sample (Magnesium Chloride and Calcium Chloride)

GLASSWARES: Beaker, Glass rod, Measuring cylinder, Mortar, Spatula, Tripod stand, Funnel,
Sieve 20, Filter paper, Graph Paper, Ruler

EQUIPMENTS: Weighing Balance, Bulk Density Apparatus

THEORY

The knowledge of flow properties of pharmaceutical powder is of critical significance in

operation such as blending, tablet compression, capsule filling, and transportation and in scale up
operation. Flow properties are significantly affected by Changes in particle size, density, shape,
and adsorbed moisture, which may arise from processing or formulation. The powder flow
properties can be characterized by the following methods:
1. Angle of Repose
2. Bulk Density
3. Tapped Density
4. Hausner’s Ratio
5. Carr’s Compressibility Index
Angle of repose:
It is defined as the mass possible angle between the surface of pile of angle & horizontal plane.
Tan 0 = h/r
0 = tan-1 (h/r)
H – Height of the pile
R – Radius of the pile
Angle of repose Flow property

<25 Excellent

25-30 Good

35-40 Possible

>40 Very poor

Bulk Density and Tapped Density:

Bulk density: It is defined as the ratio of mass of powder to the bulk volume of powder. Bulk
density is determined from the bulk volume and the weight of a dry powder in a graduated
cylinder. The bulk density is sometimes given for both loosely packed powder and tightly packed
powder.
Bulk volume = vol. of powder itself + vol. of inter particles + volume of intra particles.
Bulk density is calculated by using the formula
Bulk density = Mass of powder/Bulk volume

Tapped Density: It is defined as the ratio of mass of powder to the tap volume of powder.
Tapped density provides information on how closely the particles in the powder have packed
together as a result of tapping or compression.
Tapped density is calculated by
Tapped density = mass of powder/tap volume.
Hausner’s Ratio:
This is also one of the methods which are used for the determination of flow property of the
powder. It is determined by the formula
Hausner’s ratio = Tap density/Bulk density
Hausner’s ratio Flow property
0 -1.2 Free flowing
1.2 – 1.6 Cohesive powder

Compressibility Index:
This compressibility index also denotes the flow property of powder. It is denoted by “I” and is
given by the formula
I = (I – Db/Dt) X 100
Db = Bulk density
Dt = Tapped density
A known amount of granules was taken in measuring cylinder. Measure the initial volume before
& after tapping the measuring cylinder & also measure the volume after 50 tappings. It is also
known as Carr’s consolidation index.
Carr’s index Flow property
5 – 12 Free flowing
12 – 16 Good
18 – 21 Fair
23 - 25 Poor
33 – 38 Very poor
>40 Extremely poor

PROCEDURE
Determination of Angle of repose
1. Select a glass funnel having round stem of 15-30mm in diameter with a flat edge.
2. Fix the funnel with tripod stand.
3. Place a glass plate on the table and arrange it below the glass funnel and keep one graph
paper on the glass plate.
 4. A known quantity of powder is allowed to flow through a funnel while blocking the
orifice of the funnel by thumb and adjusting the tip at 2 cm form a horizontal surface
beneath , so that the apex of the heap just touch the lower tip of the funnel.
5. Mark the base of heap.
6. Remove the thumb and allow the powder to flow down and form a cone shaped pile.
7. Measure the diameter of the formed circle (take the average of two diameters).
8. Repeat the process three times and calculate the average diameter (d), and the radius r = d
/ 2.
9. The height of the heap (the distance between the horizontal surface and the lower tip of
the funnel is called (h).
10. Tan the angle of repose (Ø) = h / r, get Ø, and tabulate your results.

Determination of Bulk Density and Tapped Density

Bulk Density: A weighed amount of powder is taken in a 100ml graduated measuring cylinder
and initial volume occupied by the powder is noted which gives the bulk volume of the powder.
Bulk volume = vol. of powder itself + vol. of inter particles + volume of intra particles.
Bulk density is calculated by using the formula
Bulk density = Mass of powder/Bulk volume
Tapped density: Weighed volume of powder is taken in a 100ml graduated measuring cylinder.
Fit a measuring cylinder in a tap density apparatus and let to 100 tappings. The volume occupied
by the powder is noted further another 50 tapping’s can be continued until concurrent value is
achieved this volume is final tapped volume.
Tapped density is evaluated by
Tapped density = Mass of powder/Tapped volume.

Determination of Hausner’s Ratio

The powder content (W) is weighed, and the bulk density is calculated as Mass of powder/Bulk
volume. Bulk density may be used as an indication of flow properties. Tapped Density is
calculated as Mass of powder/Tapped Volume. The ratio of tapped density and bulk density is
known as the Hausner’s ratio.
Determination of Compressibility of Powder
Powders used for tabulating should possess good compressibility percent less than 22 %
considered to have good flow ability.
The compressibility percent (c) was calculated according to the following equation
 EXPERIMENT NO. 2
AIM: To study the effect of Glidants on the flow properties of given sample of powder.
REFERENCE:
1. CVS Subrahmanyam, “Textbook of physical pharmaceutics”, Vallabh Prakashan,Delhi,
2nd edition, reprint 2008, page no. 221-224.
2. S.P Aggarwal, Rajesh Khanna, “Physical Pharmacy, CBS Publishers and Distributors,
New Delhi, 2nd edition, reprint 2006, page no. 43-45.

REQUIREMENTS:
CHEMICALS: Magnesium Stearate, Talc, Powder Sample (Calcium Carbonate)

GLASSWARES: Beaker, Glass rod, Mortar, Spatula, Tripod stand, Funnel, Sieve 20, Filter
paper, Graph Paper, Ruler

EUIPMENTS: Weighing Balance

THEORY: The knowledge of flow properties of pharmaceutical powder is of critical

significance in operation such as blending, tablet compression, capsule filling, and transportation
and in scale up operation. Flow properties are significantly affected by Changes in particle size,
density, shape, and adsorbed moisture, which may arise from processing or formulation.

Glidants are the substances that are added to a powder to improve its flowability. A glidant will
only work at a certain range of concentrations. Above a certain concentration, the glidant will in
fact function to inhibit flowability. In Tablet manufacturing, glidants are usually added just prior
to compression.

Examples of Glidant include Magnesium stearate, Talc, colloidal silica and corn starch.

PROCEDURE:

1. Select a glass funnel, which has a round stem of 15-30mm diameter with a flat edge.
2. Fix the funnel with tripod stand.
3. Place a glass plate on the table and arrange it below the glass funnel and keep one graph
paper on the glass plate.
4. The glidant selected is talc or Magnesium stearate in the concentration of 0, 1, 2, 3, 4, 5
(%w/w).
 5. Take the powder sample (approximately 5gm) and add the glidant of different
concentration to it. Mix thoroughly and allowed the sample to pass through sieve no. 20.
6. A known quantity of powder is allowed to flow through a funnel while blocking the
orifice of the funnel by thumb and adjusting the tip at 2 cm form a horizontal surface
beneath , so that the apex of the heap just touch the lower tip of the funnel.
7. Mark the base of heap.
8. Remove the thumb and allow the powder to flow down and form a cone shaped pile.
9. Measure the diameter of the formed circle (take the average of two diameters).
10. Repeat the process three times and calculate the average diameter (d), and the radius r = d
/ 2.
11. The height of the heap (the distance between the horizontal surface and the lower tip of
the funnel is called (h).
12. Tan the angle of repose (Ø) = h / r, get Ø, and tabulate your results.
13. Plot the graph by taking concentration of glidant on X-axis and Angle of Repose on Y-
axis.
RESULT AND DISCUSSION:
The angle of repose for different concentration of glidants was found to be…………………….
Indicates that
 EXPERIMENT NO. 3
AIM: To formulate and evaluate the calcium alginate beads of Diclofenac sodium.
REFERENCES
1. Leon Lachmann, Liebermann.H.A.Kaning JL, Theory and practice of industrial pharmacy, 3rd
edition, 420-428
2. S.P.Vyas, R.K.Khar, “Targeted and Controlled drug delivery”, novel carrier system, CBS
publishers and distributors, New Delhi, page no: 122
MATERIALS REQUIRED
APPARATUS: beaker, magnetic stirrer, standard flask, dissolution apparatus
CHEMICALS: 6.8 pH phosphate buffer, sodium alginate, paracetamol, 2% solution of calcium
chloride, glutaraldehyde.
THEORY
The United States Pharmacopoeia (USP) defines the modified-release (MR) dosage form as “the
one for which the drug release characteristics of time course and/or location are chosen to
accomplish therapeutic or convenience objectives not offered by conventional dosage forms such
as solutions, ointments, or promptly dissolving dosage forms”. One class of MR dosage form is
an extended-release (ER) dosage form and is defined as the one that allows at least a 2-fold
reduction in dosing frequency or significant increase in patient compliance or therapeutic
performance when compared with that presented as a conventional dosage form (a solution or a
prompt drug-releasing dosage form). The terms “controlled release (CR)”, “prolonged release”,
“sustained or slow release (SR)” and “long-acting (LA)” have been used synonymously with
“extended release”. Nearly all of the currently marketed monolithic oral ER dosage forms fall
into one of the following two technologies:
1. Hydrophilic, hydrophobic or inert matrix systems: These consist of a rate controlling polymer
matrix through which the drug is dissolved or dispersed.
2. Reservoir (coated) systems where drug containing core is enclosed within a polymer coating.
Depending on the polymer used, two types of reservoir systems are considered.
(a) Simple diffusion/erosion systems where a drug-containing core is enclosed within
hydrophilic and/or water-insoluble polymer coatings. Drug release is achieved by diffusion of
the drug through the coating or after the erosion of the polymer coating.
(b) Osmotic systems where the drug core is contained within a semi-permeable polymer
membrane with a mechanical/laser drilled hole for drug delivery. Drug release is achieved by
osmotic pressure generated within the tablet core.
Small beads are prepared from different polymers and other
pharmaceutical additives like microcrystalline cellulose and they are used carriers for
development oral sustained release capsules. These beads are filled into the capsules and they are
termed as spansules. The release rate of drugs from these beads can be altered fast or slow
release by coating with various amounts of polymers. The time of release of drugs from the
beads can be modified by adjusting the thickness of polymer coat. Finally the beads coated with
different thickness and also mixed in different ratios can be put into capsules subsequently used
as modified release capsules.
PROCEDURE
Accurately weighed 500mg of sodium alginate and 25ml of distilled water was added. This
mixture was stirred by using a mechanical stirrer. To this solution 500mg of paracetamol was
added and it is mixed well. By using a syringe this solution was slowly added into a beaker
containing 2% solution of calcium chloride which is kept under continuous agitation at a low
speed using a magnetic stirrer and stirred for 15 mins. The beads are rigidised by adding 1ml of
25% solution of glutaraldehyde. Allow the stirring for another 1hr. the solution was filtered and
beads were collected.

EVALUATION
Determination of drug content:
100 mg of beads was transferred into 100ml of phosphate buffer 6.8 pH. The dispersion was
stirred using a magnetic stirrer for 2 hrs. The solution was filtered and diluted with suitable
solution. Then the absorbance was measured at 275nm. The drug content of the prepared beads
were determined.
Percentage Drug Entrapment (%DE)
Accurately weighed 100mg of microsphere were dissolved in 100ml PBS pH 6.8. The solution
was kept overnight and was filtered through whattman filter 0.45μm. The drug concentration was
determined by UV spectrometer at maximum wavelength of 276nm. The following equation was
used to calculate % drug entrapment.
 % Drug Entrapment = Practical drug content × 100
Theoretical drug content

Determination of Swelling Index

500mg of beads was taken in a China dish and then 10 ml of distilled water was added and the
mixture was shaken and allowed to stand for 1hour. After 1 hour the remaining water in China
dish was discarded and the weight increase of the natural suspending agent was rated.
Swelling Index % (SI) = (W2 – W1/W1) x 100
W1= Weight of tablet at time ‘0’
W2= Weight of tablet at time‘t’

Dissolution Test
100mg of prepared drug loaded beads was weighed and placed in dissolution apparatus. 900ml
of pH 6.8 phosphate buffer was used as dissolution medium. The dissolution study was
performed at 50 rpm for 3 hr. 5ml of samples were withdrawn at time interval of 15, 30, 45, 60,
and 1 hr and 2hr. After each withdrawal an equivalent amount of fresh dissolution medium was
replaced.
Temperature was maintained at 37ᵒC. the absorbance of withdrawn sample after filtration was
measured at 275nm using UV spectrophotometer. The results were plotted in a graph.

REPORT
The formulation and evaluation of calcium alginate beads of diclofenac sodium was performed.
The percentage drug release of diclofenac sodium beads was found to be
 EXPERIMENT NO. 4
AIM: To formulate and evaluate the Diclofenac sodium gel by using natural polymer.
REFERENCES
1. S.P.Vyas, R.K.Khar, “Targeted and Controlled drug delivery”, novel carrier system, CBS
publishers and distributors, New Delhi, page no: 122
2. N.K. Jain “Pharmaceutical product development” CBS publishers and distributors, New Delhi,
page no: 182.
3. Encyclopedia of pharmaceutical technology, 3rd edition vol-3 page no: 567

MATERIALS REQUIRED
APPARATUS: Digital weighing balance, beaker, standard flasks
CHEMICALS: Diclofenac sodium, sodium alginate, EDTA, potassium hydrogen ortho
phosphate, methyl paraben, propyl paraben, PEG 400
THEORY
The invitro permeation of drugs can be studied by trans diffusion cells. The franz diffusion cells
are made of glass with a contact area of 4.7cm2 with solubilizing agent, the franz diffusion
consist of donar compartment(A) and receptor compartment(B). The membrane is mounted
between two cells compartments. The two cell compartments are held together with a clamp. The
receptor compartment has a volume of 37ml and is filled with diffusion medium; it is kept at
37ᵒC by circulating water through an external water jacket. After 30 mins of equilibrium of
membrane with the receptor solution specific quantity of drug or formulation, applied in the
donar compartment is the covered with paraffin to prevent the evaporation of solvent. The
receptor solution is continuously stirred by means of spinning bar magnet at 400rpm. 1ml
samples are withdrawn from the receptor compartment at various time intervals. The cells are
refilled with receptor solution to keep the release of receptor solution during experiment.

PROCEDURE
Preparation of Gel
Potassium dihydrogen phosphate, diclofenac sodium, EDTA, Sodium Alginate was dissolved in
water under agitation with mechanical stirrer. Methyl paraben, propyl paraben, PEG 400 were
added to it under continuous stirring, polymer was slowly springed on the surface of purified
water for uniform distribution. The gel was buffered at 7.4 ±0.2.
EVALUATION
pH: The pH is determined by 2.5g of gel was dispersed in 25ml of purified water and pH is
measured by pH meter.
Rheological study: Rheological study using Brookfield viscometer.
Diffusion study: Diffusion studies were carried out using Franz diffusion cell. Accurately
weighed 0.3g of gel was spread uniformly on a dialysis membrane which was in contact with
receptor medium. The receptor medium stirred continuously at 20 rpm to stimulate blinking
action of eyelids. Samples were withdrawn at periodic intervals. The drug content was analyzed
using UV spectrophotometer at 271nm, against reference standards using distilled water as
blank.
REPORT
The percentage amount of drug diffused of diclofenac gel was found to be………………
 EXPERIMENT NO. 5
AIM: To formulate and evaluate transdermal patches of Diclofenac potassium.
REFERENCES:
1. Indian Journal of Pharmaceutical Sciences, July-August 2009
2. S.P. Vyas, Roop K Khar, Controlled Drug Delivery-Concepts And Advances
MATERIALS REQUIRED:
APPARATUS: Beaker, standard flask, petridish, digital weighing balance
CHEMICALS: Diclofenac potassium, propylene glycol, span 80, polymer solution, ethyl
cellulose, methyl cellulose, polyvinyl pyrrolidine, chloroform
THEORY:
Transdermal drug delivery systems are designed to provide continuous delivery of drug directly
into the systemic circulation. It provides several benefits including avoidance of first pass effect
and also to get a constant drug delivery over a long period.
Transdermal drug delivery systems are topically administered medicaments in the form of
patches (or semisolids) that deliver drugs for systemic effects at a predetermined and controlled
rate. Some of the advantages of these systems over other controlled-release formulations are-
1. Avoidance of the first pass effect
2. Stable and controlled blood level
3. Comparable characteristics with intravenous infusion
4. Ease of termination of drug action, if necessary
5. Long duration of action (ranging from a few hours to one week)
6. No interference with gastric and intestinal fluids.
Drugs which degrade into gastro intestinal tract, inactivated by hepatic first
pass effect, drugs which are administered for a long period of time and causing adverse effects to
non-targeted tissues can be formulated as TDDS. Drugs like propranolol hydrochloride are ideal
candidate for TDDS. In this formulation diclofenac potassium is used as model drug.
PROCEDURE
Weigh the calculated quantities of ethyl cellulose, methyl cellulose and polyvinyl pyrrolidine.
Dissolve the three polymers in the required quantity of chloroform. Add span 80 and propylene
glycol to the solution of polymer. Stirr well added drug to the solution and mixed well. This
solution was poured into a glass petridish containing mercury. Allow it to dry at room
temperature for about 48 hours. When the film was formed, removed it and checked for any
imperfections and air bubbles. Note down the total weight and the diameter of the film.
EVALUATION OF TRANSDERMAL PATCHES
Evaluation of transdermal patches includes various parameters like peel adhesion, tacking
property, shear strength, invitro drug release, invitro release pattern and cutaneous toxicological
studies. Evaluation of transdermal system includes
Determination of folding endurance
Folding endurance of the patch is determined by folding one patch repeatedly at one point or at
same place till it breaks and noting down the number of times it resist the folding.
Determination of drug content
Put the square patch in 10ml of 6.8pH phosphate buffer, added and stirred in a magnetic stirrer
for 12 hrs at constant rpm. Centrifuge the solution at 2000 rpm. Collect the supernatant and
diluted it properly and measure the absorbance at 273nm using 6.8 pH phosphate buffer as blank.
Calculate the drug loading efficiency of square shaped film.
Invitro diffusion study
Invitro release studies were carried out using Franz diffusion cell. Transdermal patch was soaked
in buffer and placed above the membrane. The receptor medium (6.8 pH phosphate buffer)
stirred continuously at 20 rpm. Samples were withdrawn at periodic intervals of 15,
30,45,60,90,120 minutes. The drug content was analyzed using UV spectrophotometer at 275nm.
REPORT
Transdermal patches of diclofenac potassium were prepared and evaluation tests were done. The
results are as follows
Average weight=
Average thickness=
Folding endurance=
Cumulative percent of drug release in 24 hrs. =
 EXPERIMENT NO. 6
AIM: To formulate and evaluate diclofenac microcapsules with ethyl cellulose by temperature
change method.
REFERENCES:
1. Leon Lachmann, Liebermann.H.A.Kaning JL, Theory and practice of industrial pharmacy, 3rd
edition, 420-428
2. S.P.Vyas, R.K.Khar, “Targeted and Controlled drug delivery”, novel carrier system, CBS
publishers and distributors, New Delhi, page no: 122
MATERIALS REQUIRED:
APPARATUS: Beaker, standard flask,
CHEMICALS: Ethyl cellulose, Cyclohexane, Diclofenac potassium
EQUIPMENTS: Digital weighing balance
THEORY:
Microencapsulation is a mean of applying relatively thin coating to small particles of solids or
droplets of liquids or droplets of dispersions.
The applications of microencapsulation include:
1. Sustained release
2. Taste masking
3. Multilayered tablets
4. For newer formulation concepts of creams aerosols etc.
In this experiment the microcapsules were prepared by coacervation phase
separation which consists of following three steps under continuous agitation.
1. Formulation of liquid manufacturing vehicle phase, core material phase, coating material
phase. In the temperature change method the polymer ethyl cellulose which is insoluble in
cyclohexane at room temperature. But it is soluble at elevated temperature. The polymer is
dispersed and the mixture is heated to boiling point. The core material (drug) is dispersed in the
polymer solution. Allow the mixture to cool with continuous stirring which will subsequently
allow the phase separation process to occur.
2. This step consists of depositing the liquid polymer coating upon the core material by control
physical mixing.
3. This step involves rigidizing the coating by thermal or crosslinking or desolvation technique.
PROCEDURE
Disperse ethyl cellulose in cyclohexane to yield polymer solution of 2% by weight (2gm in
100ml of cyclohexane). Heat the mixture to its boiling point to form homogenous polymer
solution. Disperse the drug in polymer solution by stirring at a coating to core material ratio of
(2:1) (polymer: drug). Allow the mixture to cool with continuous stirring to induce coacervation
phase separation. Collect the micro capsules after solidification.

EVALUATION
Invitro dissolution study
100mg of prepared microcapsules was weighed and placed in dissolution apparatus. 900ml of pH
6.8 phosphate buffer was used as dissolution medium. The dissolution study was performed at 50
rpm for 3 hr. 5ml of samples were withdrawn at time interval of 15, 30,45,60 minutes, 1, 2, 3, 4,
5, 6 hours. After each withdrawal an equivalent amount of fresh dissolution medium was
replaced. Temperature was maintained at 37ᵒC. the absorbance of withdrawn sample after
filtration was measured at 275nm using UV spectrophotometer. The results were plotted in a
graph.

REPORT
Microcapsule of diclofenac sodium was formulated from ethyl cellulose by temperature change
method and the invitro dissolution testing was done. The results are as follows.
Cumulative percent drug release was found to be
 EXPERIMENT NO. 9
AIM: To study pharmacokinetic parameters of given I.V. or oral data.
REFERENCE: Laboratory manual of “Biopharmaceutics and Pharmacokinetics” by Dr. B.S.
Bias and Dr. R.J. Bias, Irinity Publishing house;
THEORY:
For Oral Data
(1) Determine Elimination rate constant and Elimination half life: Rate constant (KE) is
represents the fraction of volume of distribution (Vd) which will be cleared of drug for unit of
times.
𝒄𝒍
KE = 𝒗𝒅

Elimination Half Life is defined as the time taken for the amount of drug in body as well as
plasma concentration to declined by one half or 50% its initial value.
𝟎.𝟔𝟗𝟑
t1/2 = 𝑲𝐄

(2) Determine total Area Under Curve (AUC): Area Under Curve represents the total
integrated area under plasma curve time profile and exposes total amount of drug that comes into
systemic circulation after its administration.
𝐶1 + 𝐶0
AUC = (t1-t0)
2

(3) Determine cmax and tmax: Peak plasma concentration (cmax) point of maximum

concentration of drug in plasma. Time of peak concentration (tmax) is time for drug to reach peak

concentration in plasma is called as tmax.

For I.V. data
(1) Determine elimination rate constant and elimination half life:
Elimination rate constant = 𝑐𝑙⁄𝑣𝑑
Elimination half life (t1/2) = 0.693/t
(2) Determine volume of distribution: It is the ratio of amount of drug in body to the plasma
drug concentration.
𝑨𝒎𝒐𝒖𝒏𝒕 𝒐𝒇 𝒅𝒓𝒖𝒈 𝒊𝒏 𝒕𝒉𝒆 𝒃𝒐𝒅𝒚
Vd = 𝑷𝒍𝒂𝒔𝒎𝒂 𝒅𝒓𝒖𝒈 𝒄𝒐𝒏𝒄𝒆𝒏𝒕𝒓𝒂𝒕𝒊𝒐𝒏 = X/C

(3) Determine clearance: Clearance is the most important in clinical drug application and is
useful in evaluating mechanism by which a drug elimination according whole particular organ.
 Cl = dx /dt /c,
𝑬𝒍𝒊𝒎𝒊𝒏𝒂𝒕𝒊𝒐𝒏 𝒓𝒂𝒕𝒆
Clearance = 𝑷𝒍𝒂𝒔𝒎𝒂 𝒅𝒓𝒖𝒈 𝒄𝒐𝒏𝒄𝒆𝒏𝒕𝒓𝒂𝒕𝒊𝒐𝒏

Procedure:
(A) For Oral data:
Time (hrs.) 0.25 0.5 1 1.5 2 3 4 6 8 12 24
Conc. (ug/ml) 51.33 74.05 85.91 85.11 75.51 62.98 52.32 36.08 24.88 11.83 1.27

(1) Determine Elimination rate constant:

𝒍𝒐𝒈 𝑪𝟐−𝒍𝒐𝒈 𝑪𝟏
Slope= 𝒕𝟐−𝒕𝟏

K= slope × 2.303
(2) Elimination Half Life:
𝟎.𝟔𝟗𝟑
t1/2 = 𝑲

(3) Determine Area Under Curve:

𝑪𝟏+𝑪𝟎
(t1 –t0)
𝟐

(B) For I.V. data:

Time (hrs.) 0 2 4 6 7.5 9 12 15
Conc.(ug/ml) 0 3.4 5.4 6.5 7. 5 4.6 2 0.9

(1) Determine the elimination rate constant:

log 𝐶2−𝐶1
Slope = K = slope × 2.303
𝑡2−𝑡1

(2) Determine the elimination half life:

0.693
t1/2 = 𝐾

(3) Determine the volume of distribution:

𝐷0
Vd = 𝐾𝐶𝑠𝑠 [D0 = Dose given: 50mg]

[Css = Steady state conc: 7 mg/L]

(4) Determine the Clearance:
Cl = KVd
 EXPERIMENT NO. 10
AIM: To formulate and evaluate microspheres of diclofenac sodium using bovine serum
albumin.
REFERENCES:
1. Leon Lachmann, Liebermann.H.A.Kaning JL, Theory and practice of Industrial pharmacy, 3rd
edition, 420-428
2. S.P.Vyas, R.K.Khar, “Targeted and Controlled drug delivery”, novel carrier system, CBS
publishers and distributors, New Delhi, page no: 122
MATERIALS REQUIRED:
APPARATUS: beaker, magnetic stirrer, standard flask, dissolution apparatus
CHEMICALS: 6.8 pH phosphate buffer, bovine serum albumin, diclofenac sodium,
glutaraldehyde, liquid paraffin
THEORY
Diclofenac sodium is one of the drugs of choice to treat arthritis because of its potential anti-
inflammatory and analgesic activity and this is the only approved NSAID available for parenteral
delivery. Because of shorter biological half-life, diclofenac sodium should be given frequently to
maintain its therapeutic activity. It also has high percentage of protein binding and it undergoes
presystemic metabolism. To overcome these problems, many authors developed sustained
release formulations with an intention to maintain effective diclofenac concentration for
prolonged period. Microspheres are sometimes referred to as microparticles. Microspheres can
be manufactured from various natural and synthetic materials. Polymer used in this experiment is
albumin. Albumin is a natural polymers obtained from sources like proteins.
Methods
1. Single emulsion technique
2. Double emulsion technique
3. Polymerization
4. Phase separation/ Coacervation
5. Spray drying
6. Solvent extraction
7. Emulsion solvent evaporation
8. Wet emulsion technique
9. Hot melt microencapsulation
These are the methods used for the preparation for microspheres.
PROCEDURE
A solution of albumin (1g in 25 ml) was prepared and drug (1g) was added to the albumin
solution. The contents were slowly added to a beaker containing 100ml of preheated (60ᵒC)
liquid paraffin containing tween 80 as emulsifying agent and stirred for 1 hr. The temperature
was reduced to 40ᵒC for hardening process and was maintained for 25min. The resulting
microspheres were stabilized using glutaraldehyde solution (25%w/v) for a period of 15min. The
microspheres were collected by decantation and washed with n-hexane and dried at room
temperature.
EVALUATION
Particle Size Analysis: Determined the average size of the particles using an optical
microscopic method.
Dissolution Study:
The study was carried out using Type-2 Paddle type USP apparatus. The set condition was
900ml of 6.8pH phosphate buffer, at 50 rpm, 37ᵒC for 45 minutes, 5ml of samples were
withdrawn at time intervals of 5, 15, 30, 45 minutes, which was replaced by fresh equal volume
of dissolution medium, the sample was diluted suitably, assayed at 275nm by using UV-Visible
spectroscopy. The calibration curve was used to determine the drug concentration per ml. The
amount of drug release was calculated using calibration curve method.
Amount of drug release = 𝑐𝑜𝑛𝑐. ×𝑣𝑜𝑙.𝑜𝑓.𝑑𝑖𝑠𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 𝑚𝑒𝑑𝑖𝑢𝑚×𝑑𝑖𝑙 𝑓𝑎𝑐𝑡𝑜𝑟 1000
Percentage drug release = 𝑎𝑚𝑜𝑢𝑛𝑡 𝑜𝑓 𝑑𝑟𝑢𝑔 𝑟𝑒𝑙𝑒𝑎𝑠𝑒 𝑠𝑡𝑟𝑒𝑛𝑔𝑡ℎ x 100
Determination of Drug Content:
100mg of drug loaded microspheres were added to 100ml of 6.8pH phosphate buffer. Stir the
dispersion using magnetic stirrer for 3hrs.Filterthe dispersion using whatmann filter paper and
dilute it according to need (take 1ml dilute to 100ml) and measure the absorbance at 276nm.
Percentage Drug Entrapment (%DE)
Accurately weighed 100mg of microsphere were dissolved in 100ml PBS pH 6.8. The solution
was kept overnight and was filtered through whatman filter 0.45μm. The drug concentration was
determined by UV spectrometer at maximum wavelength of 276nm. The following equation was
used to calculate % drug entrapment:
 % Drug Entrapment = Practical drug content × 100
Theoretical drug content

REPORT
The formulation and evaluation of bovine serum albumin microspheres of diclofenac sodium
was prepared.
The percentage drug release of diclofenac sodium microspheres was found to be