Lab Physical Pharmaceutics-II

LABORATORTY MANUAL

B. Pharmacy (Semester -4th)

Subject: Physical Pharmaceutics-II

Subject code: BP-407-P

IKG PUNJAB TECHNICAL UNIVERSITY

Kapurthala, Punjab – 144601

CHANDIGARH COLLEGE OF PHARMACY

LANDRAN (MOHALI)-140307

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Table of Contents

S.No. Name of Practical Page No.

03
1. To study bulk density of a given powder i.e Calcium Carbonate.
To determine the flow properties of given sample by measurement
2 04-05
of angle of repose.
To determine the viscosity of a given sample by using Ostwald
3 06-07
viscometer

To determine particle size distribution of a powder by sieving

4 08
method
5 To prepare and identify the type of emulsion 09

6 Determination of reaction rate constant first order 10-11

7 Determination of reaction rate constant second order 12-13

Determination of viscosity of semisolid by using Brookfield

8 14-15
viscometer
Determination of particle size, particle size distribution using
9 16-17
Microscopic method
Determination sedimentation volume with effect of different
10 18-20
suspending agent
11 Determination sedimentation volume with effect of different 21-23
concentration of single suspending agent

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EXPERIMENT 1: To study bulk density of a given powder i.e Calcium Carbonate.

REQUIREMENTS: Calcium carbonate powder, Measuring Cylinder, Sieve No. 22

FORMULA USED: Bulk Density = Mass of powder/ Bulk Powder

PROCEDURE:

1. The given sample of powder i.e calcium carbonate was weighed and passed through sieve no. 22

2. The given amount of powder was weighed again and placed in 100ml of measuring cylinder.

3. The cylinder was then tapped continuously, noting the number of tappings. The volume occupied by
powder was also measured.

4. Volume obtained was noted as bulk Volume.

5. By using volume obtained, bulk density and mean bulk density was calculated.

OBSERVATION:

Final Volume/
Weight of No. of Initial Volume Bulk density
S.No. Bulk
powder (g) Tappings (ml) (g/ml)
Volume(ml)
1
2

Mean Bulk Density = Summation of Bulk density values/ No. of values

RESULT: Mean bulk density of a given powder was ________ g/ml.

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EXPERIMENT 2: To determine the flow properties of given sample by measurement of angle of repose.

REQUIREMENTS: Glass funnel, Funnel stand, cotton plug, sand and paper.

FORMULA USED: θ = tan-1 h/r

θ = Angle of repose
h = Height of pile
r = Radius of pile

PROCEDURE:

1. A glass funnel was selected and fixed on a funnel stand

2. Weigh accurately about 50g of sample and pour this into funnel by blocking the orifice of the funnel
either by thumb or with cotton plug.

3. Remove thumb or cotton plug and allow the sample to fall down onto paper and form a pile.

4. Height and radius of the pile was determined

5. Angle of repose was calculated by using the formula

6. Value of θ was calculated and compared with standard values to determine the flow properties of a
given sample.

Angle of Repose Flow

Less than 25 Excellent

25 – 30 Good

30 - 40 Passable

More than 40 Poor

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OBSERVATION:

Trials Height (h) Radius (r) Tan θ = h/r θ = tan-1h/r

1
2

Mean θ = trial 1+ trial 2+ trial 3/ 3

Angle of repose = ________◦

RESULT: The angle of repose was found as _______ ◦ . Therefore the flow was __________.

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EXPERIMENT 3: To determine the viscosity of a given sample by using Ostwald viscometer.

REQUIREMENTS: Ostwald viscometer, Measuring cylinder, beaker, weighing balance, glycerine, stop
watch.

FORMULA USED: Ƞ1 = (δ1t1/δ2t2)* ƞ2

Ƞ1 = Viscosity of an unknown sample

Ƞ2 = Viscosity of a known sample

t1 = Time of flow of unknown sample

t2 = Time of flow of known sample

δ1 = Density of unknown sample

δ2 = Density of known sample

PROCEDURE:

1. A clean and dry Ostwald viscometer was selected and fix in vertical position.
2. A fix amount of water was transferred through a wide limb using a rubber tube and brings
to a level above mark A.
3. Then water was allowed to flow down when lower meniscus reached mark A, then stop
watch was started.
4. When meniscus reached the mark B, then stop watch was stopped.
5. The difference in time represented the time of flow of water.
6. Averages of two determinations were noted and it gives a true value of time of flow.
7. Similarly, the above procedure was repeated with liquid under test.

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OBSERVATION:
Table 1: Observation table for time of flow
S.No. Sample t1 (sec) t2 (sec) Mean t
1 Tap water
2 5% Glycerine
3 10% Glycerine
4 15% Glycerine

Table 2: Observation table for weighing of liquid and calculation of density

S. Sample Vol. of Wt of Wt of Wt of Density
No. liquid (ml) empty filled liquid (ml) (g/ml)
beaker (g) beaker (g)
1 5% Glycerine 10
2 10 %Glycerine 10
3 15% glycerine 10

Table 3: Observation table for viscosity

S.No. Sample Viscosity
1 5% Glycerine
2 10% Glycerine
3 15% Glycerine

RESULT: Viscosity of 5% Glycerine = _______ cp

Viscosity of 10% Glycerine = _______ cp
Viscosity of 15% Glycerine = _______ cp

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EXPERIMENT 4: To determine particle size distribution of a powder by sieving method

REQUIRENTS: Mortar pestle, beaker, stirrer, measuring cylinder, hot air oven, calcium carbonate,
starch solution.
PROCEDURE:

1. Set of sieves are arranged in descending order.

2. Granules of calcium carbonate was prepared by using 5% starch solution.
3. Weighed amount of granules were placed in sieve at top of the sieve set.
4. Sieving machine was started. The length time was controlled semi-automatically.
5. Powder retained on each sieve was collected
6. Cumulative percent frequency of each size of particle was calculated.

OBSERVATION:

1. Weight of granules = ________ g

2. Time of shaking = __________ min
S.No. Sieve No. Wt. retained on % wt retained Cumulative % wt
(passed/retained) sieve (g) retained
1
2
3
4
Ʃ=

% wt retained = (given wt/total wt)* 100

= __________ %

RESULT: Cumulative % frequency was found to be ___________ %

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EXPERIMENT 5: To prepare and identify the type of emulsion.

REQUIREMENTS: Gum, oil, water, mortar pestle, measuting cylinder, beaker, dye.

FORMULA USED:

Acacia Gum = 2g
Castor oil = 8ml
Water = 30 ml

PROCEDURE:

A. PREPARATION OF EMULSION
1. Thoroughly clean and dry mortar pestle. Weighed quantity of acacia gum was weighed and then
was transferred to mortar
2. Appropriate quantity of water was measured and was triturated with gum so as to form mucilage.
To this castor oil was added in small quantity with thorough trituration.
3. Then, it was triturated until a clicking sound was produced and product becomes clearly white. At
this stage, emulsion is known as primary emulsion.
4. The remaining water was added to produce the final volume of 30ml.
5. The product was finally triturated to form uniform solution.
B. IDENTIFICATION OF EMULSION
1. Dye test: Amaranth powder is sprinkled over the emulsion. If external phase is colored
then emulsion of oil in water type, if not, then emulsion is of water in oil type.
2. Dilution test: Water is added to the emulsion. If water is miscible with emulsion
then it is of oil in water type, if not, then emulsion is of water in oil type.
RESULT: The emulsion was prepared as identified as _____________.

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EXPERIMENT 6: Determination of reaction rate constant First order

REQUIREMENTS: thermostat, pipettes, burette, volumetric flasks, titrimetric flasks, stop-

clock Chemicals: solution of ethyl acetate ( c = 0,04 mol.dm-3), solution of sodium hydroxide ( c
= 0,04 mol.dm-3), solution of hydrochloric acid ( c = 0,04 mol.dm-3), phenolphthalein

Procedure: 1. Transfer 50 ml of the solution of ethyl acetate ( c = 0,04 mol.dm-3) into the
volumetric flask (V=50 ml) and 50 ml of the solution of sodium hydroxide ( c = 0,04 mol.dm-3)
into another volumetric flask (V=200 ml). Both flasks cork down and put them in the
thermostated bath (20 °C).

2. Fill the burette with the solution of sodium hydroxide ( c = 0,04 mol.dm-3).

3. Pipette VHCl = 5 ml solution of hydrochloric acid ( c = 0,04 mol.dm-3) into a clean and dry
titrimetric flask.

4. After 10 minutes, take out the flasks with solutions from thermostated bath and pour the
solution of ethyl acetate to the solution of sodium hydroxide, put the mixture again into the
thermostated bath and start the stop-clock.

5. 5 minutes after mixing, pipette 10 ml of reaction mixture (leave the flask in the bath!) to the
titrimetric flask (with 5 ml of HCl). Remark: HCl stops the reaction given by the Eq. (21).

6. Titrate with the solution of sodium hydroxide adding 1 drop of phenolphthalein as indicator.
When the endpoint of titration has been reached, read the used volume of NaOH from the burette
(VNaOH). Write it down to the Tab.1.

7. Repeat the step 5 and 6 every 5 minutes six times more (in the 10th, 15th, 20th, 25th, 30th and
35th min. from the moment of mixing).

8. Write down to the Tab. 1 the temperature of the bath.

9. Repeat the same experiment at 30 °C. Because the reaction is faster, the times for titrations
will be in the 5th, 10th, 15th, 20th, and 25th min. from the mixing. Write down to the Tab.1 used
volume of NaOH for each titration.

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t = 20°C t = 30°C

t [min] VNaOH c [mol dm- 1/ c [dm3 VNaOH c [mol dm- 1/ c [dm3

[ml] 3] mol-1] [ml] 3] mol-1]

Table-1

1) Calculate the concentration c (mol dm-3)

2) Calculate the 1/c values.
3) Use MS Excell to create the dependence ( ) 1 f t c = at given temperature. Fit the
experimental points with a linear function. The slope represents the value of the rate
constant k at given temperature. If the time is in minutes, the unit of the rate constants is
dm3 mol-1 min-1 .
4) Calculate the activation energy Ea according to Eq. 7 (R = 8,314 J.K-1.mol-1).

Result: The first order constant was found to be ___________

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EXPERIMENT 7: Determination of reaction rate constant second order.

REQUIREMENTS: thermostat, pipettes, burette, volumetric flasks, titrimetric flasks, stop-

clock Chemicals: solution of ethyl acetate ( c = 0,04 mol.dm-3), solution of sodium hydroxide ( c
= 0,04 mol.dm-3), solution of hydrochloric acid ( c = 0,04 mol.dm-3), phenolphthalein

Procedure: 1. Transfer 50 ml of the solution of ethyl acetate ( c = 0,04 mol.dm-3) into the
volumetric flask (V=50 ml) and 50 ml of the solution of sodium hydroxide ( c = 0,04 mol.dm-3)
into another volumetric flask (V=200 ml). Both flasks cork down and put them in the
thermostated bath (20 °C).

2. Fill the burette with the solution of sodium hydroxide ( c = 0,04 mol.dm-3).

3. Pipette VHCl = 5 ml solution of hydrochloric acid ( c = 0,04 mol.dm-3) into a clean and dry
titrimetric flask.

4. After 10 minutes, take out the flasks with solutions from thermostated bath and pour the
solution of ethyl acetate to the solution of sodium hydroxide, put the mixture again into the
thermostated bath and start the stop-clock.

5. 5 minutes after mixing, pipette 10 ml of reaction mixture (leave the flask in the bath!) to the
titrimetric flask (with 5 ml of HCl). Remark: HCl stops the reaction given by the Eq. (21).

6. Titrate with the solution of sodium hydroxide adding 1 drop of phenolphthalein as indicator.
When the endpoint of titration has been reached, read the used volume of NaOH from the burette
(VNaOH). Write it down to the Tab.1.

7. Repeat the step 5 and 6 every 5 minutes six times more (in the 10th, 15th, 20th, 25th, 30th and
35th min. from the moment of mixing).

8. Write down to the Tab. 1 the temperature of the bath.

9. Repeat the same experiment at 30 °C. Because the reaction is faster, the times for titrations
will be in the 5th, 10th, 15th, 20th, and 25th min. from the mixing. Write down to the Tab.1 used
volume of NaOH for each titration.

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t = 20°C t = 30°C

t [min] VNaOH c [mol dm- 1/ c [dm3 VNaOH c [mol dm- 1/ c [dm3

[ml] 3] mol-1] [ml] 3] mol-1]

Table-1

5) Calculate the concentration c (mol dm-3)

6) Calculate the 1/c values.
7) Use MS Excell to create the dependence ( ) 1 f t c = at given temperature. Fit the
experimental points with a linear function. The slope represents the value of the rate
constant k at given temperature. If the time is in minutes, the unit of the rate constants is
dm3 mol-1 min-1 .
8) Calculate the activation energy Ea according to Eq. 7 (R = 8,314 J.K-1.mol-1).

Result: The first order constant was found to be ___________

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Experiment 8: Determination of viscosity of semisolid by using Brookfield viscometer

REQUIREMENTS: carbopol 934, triethanolamine, disodium edentate, propylene glycol

Beaker, glass rod, Measuring cylinder and Weighing balance

PROCEDURE:

1. Dissolve 200 mg Carbopol 934 with stirring in 60 mL of demineralized water for 1 h

2. Then add 0.5 mg of disodium edetate and 5 ml of triethanolamine in 10 mL of
demineralized water separately and stir for 10 min
3. Then Mix 4.83 mL of propylene glycol in 12 mL of demineralized water with stirring for
10 min
4. Disodium edetate and triethanolamine solution were added to carbopol solution and the
pH was then adjusted to 7.4 by stirring the solution for 10 min
5. Then propylene glycol solution was added with stirring for 10 min until a clear consistent
gel base was obtained.

OPERATION OF BROOKFIELD VISCOMETER

1) Mount the guard leg, if used, (spindle protector) on the viscometer.

2). Attach the spindle (left-hand thread) to the viscometer lower shaft by lifting the
coupling screw slightly. Hold it firmly with one hand while screwing the spindle on with
the other (note left-hand thread).
3) Begin by immersing the spindle in a diagonal path, slowly drag the spindle across the
Gel surface, and bring the spindle to an upright position and thread onto screw.
4). Lower and center spindle in the test material (600 ml beaker) until the "meniscus" of
the fluid is at the center of the immersion groove on the spindle’s shaft.
5). To make a viscosity measurement, turn the motor switch “ON”. This energizes the
viscometer drive motor. Allow time for the indicated reading to stabilize.
6) The time required for stabilization will depend on the speed at which the viscometer is
running and the characteristics of the sample fluid. When making a viscosity

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measurement, the reading should be noted and multiplied by the factor appropriate to the
viscometer model/spindle/speed combination being used.

Formulae used
Dial reading x Factor = Viscosity in cP (mPa•s)
Example: LVT Viscometer with #1 spindle at 6 rpm Dial Reading: 75 Factor: 10 75 x 10
= 750 cP (mPa•s)
Full scale viscosity range for any speed and spindle combination is equal to the factor x
100. Factor x 100 = Full scale range Example: LVT Viscometer with #1 spindle at 6
RPM Full Scale Range: 10 x 100 = 1,000 cP

Result: The viscocity of gel was found to be _____ cp

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Experiment 9: Determination of particle size, particle size distribution using Microscopic

Method

Requirements: Microscope,Eye piece micrometer,Stage micrometer, Beaker, slide,cover slip

Castor oil,tragacanth and gum acacia
Introduction

Microscopy can be used to estimate the size of particles and their distribution. In addition, through the
microscope, one may be able to observe the shape of the particle. This however, cannot be said when
using the sieve method. However, the sieve method is by far the cheapest technique available for the
determination of particle size. When the size distribution is great and when the shape of particle is
inconsistent, the results from both methods can be expected to be vastly different. Microscopy will
give the number average particle size whilst the sieve method produces a weight average particle size.

1) Microscopic method

1. Observe the sample under an optical microscope.

Refer to the operation manual.

Record the type of microscope and the magnification used.

2. Take a picture of a representative sample, consisting of at least 50 particles, together with a scale.
3. Measure the size of all particles captured in the picture to 1 mm.
4. Tabulate your results. Refer to Table 3 and Table 4.
5. Based on your data (Table 4), plot the size distribution: size against % number.
6. Plot the undersize distribution: size against % cumulative number.
7. From the plot (no. 5), estimate the following:
a. d(0.5) or mean
b. d(0.1)
c. d(0.9)
d. Calculate the polydispersity index: d(0.9) – d(0.1)/d(0.5)
Table 1 Microscopy data – Sample A

Size No. of Average size ∑xi ni / ∑ni

particles

x1 n1 Standard deviation

x2 n2 Minimum size

x3 n3 Maximum size

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x4 n4 Range, R size max – size min

x5 n5 Number of divisions √N (round up to ODD

no.)

x6 n6 Size interval, r R/√N

x7 n7 Smallest unit of measurement 0.1 m

Total no. particles ∑ni = N Start size, d1 size min – (0.1 m/2)

Table 2 Size (number) distribution using the microscopic method – Sample A

Size range (m) Mid size No. % number % cumulative number

(m) particles

d1- (d1+r)

d2 – d3 d2=d1+r

d3– d4 d3=d2+r

d4 – d5 d4+d5 /2

d5 – d6

d6 – d7

Total no. particles N

RESULT: 1) The arithmetic mean diameter was found to be _______

2) The weight moment mean diameter was found to be______
3) the surface mean diameter was found to be_______

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Experiment 10. Determination sedimentation volume with effect of different suspending agent

Requirement: Beaker, Mortar, Pestle, Measuring Cylinder, zinc oxide, glycerin, tragacanth and
methyl cellulose

Theory: Suspensions are defined as preparations containing finely divided drug particles (the suspensoid)
dispersed uniformly in a liquid medium in which the drug exhibits limited solubility. In most stable
pharmaceutical suspensions, the particle diameter is about 1 to 50 µm. There are many factors to be
considered when developing and preparing a pharmaceutically elegant suspension. In addition to the
therapeutic efficacy, chemical stability of the formulation components, and esthetic appearance of the
preparation, a properly prepared pharmaceutical suspension should settle slowly and be readily
resuspended upon gentle shaking without the formation of a solid cake on standing. Furthermore, the
particle size of the suspensoid should remain unchanged in the vehicle throughout the product shelf-life
and an accurate dose should be readily withdrawn from the container either by pouring or by means of a
syringe.

The various factors governing the rate of settling of a particle in a suspension are described by the
Stokes’ equation which is presented as:

2 r 2 ( 1   2 ) g
V
9

where V is the rate of settling, r is the particle radius, ρ1 is the density of the particle, ρ2 is the density of
the medium, g is the gravitational constant, and η is the viscosity of the medium.

PROCEDURE:

Preparation of suspensions of hydrophilic drugs and measurement of sedimentation volume

Table 1 Formulations of zinc oxide suspensions

No. 1 2 3 4

Zinc oxide (g) 0.5 0.5 0.5 0.5

50% glycerin (mL) — 6.0 — —

Methylcellulose (g) — — 0.1 —

Tragacanth (g) — — — 0.1

Distilled water to (mL) 10 10 10 10

Procedures

(a) Preparation of formulation 1 and 2

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Weigh the designated amount of zinc oxide and transfer it to a mortar. Add 0.3 mL of distilled water
or glycerin and levigate the mixture to form a paste. Add the remaining glycerin and continue mixing.
Further dilute the suspension with 80% of the remaining distilled water, transfer the suspension to a 10
mL graduate cylinder, and add distilled water to volume (10 mL).

(b) Preparation of formulation 3

Weigh 0.1 g of methylcellulose and transfer to a mortar. Add distilled water and mix until a solution is
formed. To this solution, add zinc oxide and levigate to form a paste. Add 80% of the remaining distilled
water and continue mixing until a uniform suspension is formed. Transfer the suspension to a 10 mL
graduate cylinder and add distilled water to volume (10 mL).

(c) Preparation of formulation 4

Weigh 0.1 g of tragacanth, transfer to a mortar, and then add several drops of alcohol as the wetting
agent. Add a small amount of distilled water to the mixture and levigate to form a paste. Add zinc oxide
and follow the procedures as described in (b).

(d) Measurement of sedimentation volume

Agitate the four suspensions contained in the stoppered graduate cylinders by turning it upside down
for five times and let the samples stand. Measure the height of the sediments (mL) at 1, 5, 10, 30, 60, 90
and 120 min after shaking and calculate the sedimentation volume. Record the results in Table 3 and plot
the sedimentation volume vs. different formulations.

RESULTS AND DISCUSSION

Fill in the sedimentation volume results in Table 2.

. The change in sedimentation volume as a function of time

Time Formulation No.

(min) 1 2 3 4

Hu Hu/H0 Hu Hu/H0 Hu Hu/H0 Hu Hu/H0

10

30

60

90

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120

Note：H0 is the height of suspension；Hu is the height of the sediment.

Result: Based on the data in Table 3, plot the sedimentation curve using Hu/H0 as the Y-axis and time as
the X axis and compare the suspending capacity of different suspending agents.

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Experiment 11 Determination sedimentation volume with effect of different concentration of

single suspending agent

Requirements: Beaker, Mortar, Pestle, Measuring Cylinder, zinc oxide, glycerin and tragacanth,

Theory: Suspensions are defined as preparations containing finely divided drug particles (the suspensoid)
dispersed uniformly in a liquid medium in which the drug exhibits limited solubility. In most stable
pharmaceutical suspensions, the particle diameter is about 1 to 50 µm. There are many factors to be
considered when developing and preparing a pharmaceutically elegant suspension. In addition to the
therapeutic efficacy, chemical stability of the formulation components, and esthetic appearance of the
preparation, a properly prepared pharmaceutical suspension should settle slowly and be readily
resuspended upon gentle shaking without the formation of a solid cake on standing. Furthermore, the
particle size of the suspensoid should remain unchanged in the vehicle throughout the product shelf-life
and an accurate dose should be readily withdrawn from the container either by pouring or by means of a
syringe.

The various factors governing the rate of settling of a particle in a suspension are described by the
Stokes’ equation which is presented as:

2 r 2 ( 1   2 ) g
V
9

where V is the rate of settling, r is the particle radius, ρ1 is the density of the particle, ρ2 is the density of
the medium, g is the gravitational constant, and η is the viscosity of the medium.

PROCEDURE:

Preparation of suspensions of hydrophilic drugs and measurement of sedimentation volume

Table 1 Formulations of zinc oxide suspensions

No. 1 2 3 4

Zinc oxide (g) 0.5 0.5 0.5 0.5

50% glycerin (mL) — 6.0 — —

Tragacanth (g) 0.1 0.2 0.3 0.4

Distilled water to (mL) 10 10 10 10

Procedures

(a) Preparation of formulation 1 and 2

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Weigh the designated amount of zinc oxide and transfer it to a mortar. Add 0.3 mL of distilled water
or glycerin and levigate the mixture to form a paste. Add 0.1 and 0.2 g of tragacanth and add the
remaining glycerin and continue mixing. Further dilute the suspension with 80% of the remaining
distilled water, transfer the suspension to a 10 mL graduate cylinder, and add distilled water to volume
(10 mL).

(b) Preparation of formulation 3

Weigh 0.3 g of Tragacanth and transfer to a mortar. Add distilled water and mix until a solution is
formed. To this solution, add zinc oxide and levigate to form a paste. Add 80% of the remaining distilled
water and continue mixing until a uniform suspension is formed. Transfer the suspension to a 10 mL
graduate cylinder and add distilled water to volume (10 mL).

(c) Preparation of formulation 4

Weigh 0.4 g of tragacanth, transfer to a mortar, and then add several drops of alcohol as the wetting
agent. Add a small amount of distilled water to the mixture and levigate to form a paste. Add zinc oxide
and follow the procedures as described in (b).

(d) Measurement of sedimentation volume

Agitate the four suspensions contained in the stoppered graduate cylinders by turning it upside down
for five times and let the samples stand. Measure the height of the sediments (mL) at 1, 5, 10, 30, 60, 90
and 120 min after shaking and calculate the sedimentation volume. Record the results in Table 3 and plot
the sedimentation volume vs. different formulations.

RESULTS AND DISCUSSION

Fill in the sedimentation volume results in Table 2.

. The change in sedimentation volume as a function of time

Time Formulation No.

(min) 1 2 3 4

Hu Hu/H0 Hu Hu/H0 Hu Hu/H0 Hu Hu/H0

10

30

60

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90

120

Note：H0 is the height of suspension；Hu is the height of the sediment.

Result: Based on the data in Table 3, plot the sedimentation curve using Hu/H0 as the Y-axis and time as
the X axis and compare the suspending capacity of different suspending agents.

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