EXPERIMENT NO.

1
“Determination of Particle size, particle size distribution using Sieving Method”

Aim: To determine Particle size, particle size distribution using Sieving Method.
Theory:
Sieving process is the process of separating particles by size. The process started in
ancient times in Egypt where sieves were used to separate grain. Today there are high
tech methods such as laser diffraction and image analysis used to determine particle size
distribution. Other methods such as centrifuge techniques and sedimentation are also
used. However, sieves are the most commonly used devices for particle size analysis. The
sieving process is comparatively inexpensive, simple in concept and easy to use. Our
Selection includes frame diameters from 100milimeters to 18 inches in stainless steel and
brass frames.
Sieve mesh material options include stainless steel wire mesh in aperture sizes from 125
milimeters (5 inches) to 20 microns, electroformed openings as small as 2 microns and
perforated plate of various shaped holes.
Sieving method gives sieve diameter dsieve
Sieve diameter is defined as the diameter of a sphere that passes through the sieve
aperture as the asymmetric particle. Sieving method gives directly the weight
distribution. Standard sieves of different mesh numbers are available commercially as per
the specification of IP and USP as shown in Table 1.1. The sieves are arranged in a nest
with the coarsest at the top. A sample of 50.0g of the powder (or granules) is placed on
the top sieve. This sieve set is fixed to the mechanicals shaker apparatus and shaken for a
period of time 20minutes. The powder retained on each sieve is weighed. Then the
different size distribution curves are constructed.
Constructions of Sieves
Sieves for pharmaceutical testing are constructed from wire cloth with square meshes,
woven from wire of brass, bronze, stainless steel or other suitable materials.
Factors influencing the sieving
Factors influencing sieving are weight of sample, duration of shaking and type of motion.
The types of motion influencing sieving are vibratory motion (most efficient), side-tap
motion, bottom pat motion, rotary motion with tap and rotary motion. The type of motion
(intensity) of the shaker is fixed and standardised. Care should be taken in order to get
reproducible results.
Applications
Sieving method finds its application in dosage form development of tablets and capsules.
Normally, 15 percent of fine powder (passed through mesh 100) should be present in
granulated material to get a proper flow of material and achieve good compaction in
tabletting. Therefore, percent of coarse and fine powder can be quickly estimated.
Advantages
It is inexpensive, simple and rapid with reproducible results.
Sieving method is useful when particles are having size range between 50 and 1500um.
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 Disadvantages
Lower limit of particle size is 50um
If the powder is not dry, aperture become clogged with particles, leading to improper
sieving.
During shaking, attrition (particles colliding with each other) occurs causing size
reduction of particles. This leads to errors in estimation.
Requirements:
Apparatus: Standard Sieve sets (20,40,60,80,100 & 120 and bottom pan and lid) 1
Mechanical Shaker 1
Chemicals: Granules, powder
Procedure

1. Select standard sieve set [IP or USP]. This set consists of various size.
2. Arrange them in such a manner that the coarsest at the top and finest at the bottom of
the sieve set. Keep the pan of the sieve set at the bottom of the sieve set as shown in
figure 1.
3. Weigh approximately 50.0g of sample (granule or powder).
4. Place the sample on the coarsest sieve. Keep the lid.
5. Fix the above sieve set on a mechanical shaker and clamp it tightly.
6. Switch ON the mechanical shaker and set the timer for 10 minutes.
7. When the shaker automatically stops, collect the sample retained on each sieve into a
paper.
8. Weigh all the samples. Report them in the Table no.1
9. Report the weights retained on each sieve in the Table against the corresponding
sieve number or sieve size.
10. Analyse the data for normal, Log-normal, Cumulative frequency and probability
graphs.
Observation table

Weight distribution of Granular material (powder or granules) measured by Standard

sieves
Sieve number Apertur Arithmet Log Weigh Percen Cumulati Weig
(passed/Retaine e size ic means arithmeti t t ve percent ht size
d) um size of c mean retaine weight retained
opening size of d on a retaine
um opening sieve g d
(under
size)

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 (1) (2) (3) (4) (5) (6) (7) (3)x
(5)
D n
Dxn
Pan
120
100
85
60
44

Result
Mode of the distribution = …………um
Average diameter of the powder d = ……….um

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 EXPERIMENT NO.2
“Determination of particle size, particle size distribution using Microscopic
method”
Aim: To determine particle size, particle size distribution using Microscopic method
Theory
The particle size distributions measured by the optical microscope were compared with those by
the light scattering particle counter to validate the microscopic method for particle size
distribution quantification. While the optical microscope concentrations increased with the
particle counter concentrations, the optical microscope concentrations tended to be higher than
those by the particle counter. To explain the difference between the two methods, we estimated
their relationship based on the Mie scattering theory. The calculation roughly estimated the
particle geometric diameters in theory were 1.7-2.0 times as large as the corresponding particle
counter were converted to match with the optical microscope measurements (particle counter).
Overall, the optical microscope concentrations were lower than the particle counter
concentrations. The advantage in the optical microscope method particularly for ambient
aerosols lied in its accuracy of particle sizing although the counting efficiency might be lowered
due to its intrinsic limitations such as inability of counting particles having the similar refractive
index of the filter.
Powder is considered as a collection of particles. If a powder contains particles of one size, such
powder is termed as monosize or monodisperse. Most of the pharmaceutical powders are
polydisperse, i.e., contain particles of different sizes. In the analysis of powders
- The size of individual particles
- The number of particles of the same size (size distribution)
Are important. These can be expressed in terms of number of distribution and weight
distribution.
Particle size
In a powder, the shape of the particle is normally not spherical, but asymmetrical (uneven).
Therefore, it is difficult to express the size as a meaningful diameter. However, size is expressed
as the diameter, which is related to an equivalent spherical diameter. The important expressions
for a particle are as follows:
Surface diameter, ds
Stokes diameter dst

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 Volume diameter dv
Projected diameter dp
Sieve diameter dsieve
Volume surface diameter dvs
In addition, there are a number of descriptions for the particle diameter. The type of diameter to
be estimated depends on the intended purpose for which it is applied.
Powder size
There is no universal way of defining the size of a powder. However, it is necessary to assign
powder with a characteristic value. Some of the descriptions are arithmetic mean, geometric
mean and harmonic mean.
Arithmetic mean = sum of particles size/ number of particles
Some arithmetic mean mathematical expression that are important in pharmacy are given below

These descriptions give gross picture about the powder.

Where d = mean diameter of the particle range, um

n = number of particles, i.e. frequency
Size distribution analysis
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Particle size distribution is expressed as
-number distribution
Weight distribution
Of the particles within a certain size ranges. The weight of drug particles and its distribution
should be same in each tablet to reproduce same therapeutic action. Secondly, for a given weight
of tablet or capsule, the number of particles and their distribution should also remain same. Thus,
number as well as weight distribution are important in the production of dosage forms, in order
to maintain reproducible physical, chemical and pharmacological properties.
The comprehensive view of powder is commonly presented in the form of graphs, because graph
is more concise and permits the easy visualization of the mean and skew distribution.
Principle
Microscopy is a useful method to directly visualize the particles. It offers several advantages
such as recognising the aggregation, contamination, etc. In this method, the size is expressed as
projected diameter dp. Projected diameter dp is defined as the diameter of a sphere having the
same area of the asymmetric particle, when viewed normal to its most stable plane. It is largely
related to length and breadth of the particle, but not depth. This method gives the number
distribution, which can be further converted to weight distribution. A calibrated eye-piece is used
to measure the size of particles. Applications: Microscopy method is used to determine the size
analysis of particles in suspensions, aerosols and globules in emulsions.
Advantages: (1) Microscopy allows the observer to view the particles directly.(2) Agglomeration
of particles and any contamination in powder can be detected. (3) Easy and simple.
Disadvantages: Diameter is obtained from only two directions, i.e.length and breadth. Depth of
the particles is not measurable. This method is slow and tedious, because a large number of
particles (625) must be measured to get good estimation.
Requirements:
Apparatus: microscope, eye piece, micrometer, stage micrometer, glass slide, cover slip
Chemicals: calcium carbonate
Procedure
Calibration of Eye-piece Micrometer
Standard stage micrometer is used to calibrate the eye piece micrometer. Stage micrometer is a
glass slide (7.5 cmx 2.5cm) which has the scale engraved on it. The scale is usually 1.0 milimetre
in length. One millimetre is divided into 100 divisions (0.1 and 0.01 parts). Thus, the smallest
division (least count) of the stage micrometer represents 0.01 mm (or 10 um) length.

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Eye piece micrometer is small disc of glass with a linear scale of 10 mm divided into 1.0mm and
0.1mm parts. (Micrometer discs are sometimes permanently fixed in the eyepiece).
In this experiment, the optical combination of 10X eye-piece and 45X objective is used.
(1) Place the stage micrometer on the stage of the microscope.
(2) Position the object to the centre of the objective.
(3) Initially, focus at the low power (10X objective).
(4) The scale of the stage micrometer is seen (100 divisions)
(5) Switch over to high power (45X). Care should be taken to avoid any damage to the stage
micrometer.
(6) Focus the object clearly. The scale of the stage micrometer will be seen, in which
divisions are farther from each other.
(7) Carefully replace eye-piece with the eye-piece micrometer, Now, small lines of the
numbered scale represent the ruling of eye-piece. These are also visible over the stage
micrometer divisions.
(8) If necessary, rotate the eye-piece in such a manner to bring two scales parallel to each
other.
(9) Select two points. One point, on the left-side, where divisions of both scales coincide, and
another, on the right-side, where the divisions coincide.
(10) Count the number of small divisions, x (eye-piece) and big divisions, y (stage
micrometer divisions). Record them.
(11) Calculations of calibration can be done as follows
X, number of eye-piece divisions = y, number of stage micrometer divisions
1 eye-piece divisions = number of stage micrometer divisions/ number of eye-piece
divisions x least count
(12) Remove the stage micrometer from the stage of the microscope.
Mounting of sample
Transfer a small portion of the given sample (talc or starch) on to a clean slide (use camel hair
brush for transfer of sample).
Add one or two drops of liquid paraffin.
Disperse the sample uniformly with the help of a brush. The particles should be independent and
their distribution should be uniform.
Place the cover-slip carefully. Avoid entrapment of air bubbles. Drain the excess liquid with a
blotting paper.
Place the slide on the stage of the microscope.
Measurement of particle size

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 (1) Focus the slide in low magnification (10X). Observe the presence of individual particles
(If aggregation or lumps are present, the sample should be mounted again).
(2) Shift to high power (45X) and focus the slide.
(3) Measure the size of each particle in terms of eye-piece divisions. A total of 300 particles
should be considered for size distribution (Ideally 625 particles should be measured
according to BPC)
(4) Tabulate the particles in terms of divisions of eye-piece and number of particles.
(5) Multiply the number of eye-piece divisions by the calibrated value obtained above.
(6) Classify the diameters into size ranges (Table 2.1)
(7) Arrange the frequency of particles in terms of number distribution.
(8) Transform data in different ways as mentioned in Table 2.2
(9) Draw the size distribution curves such as normal, log normal, cumulative frequency and
log probability.
(10) Report the results in different types of mean diameters.
Observations
Table 2.1 Data generated for Size distribution (Number) Estimated by Microscopy
Method
(1) (2) (3) (4) (5) (6)
Size range Mean size Log d Number of Percent Cumulative
um range, um, particles in frequency percent
d each size number frequency
of number
particles distribution

Table 2.2 Conversion of Number Distribution to weight Distribution

(1) (2) (3) (4) (5) (6) (7)
Mean size Number of Nxd nxd2 nxd3
range um particles in Percent Cumulative
each size percent
d nxd3
frequency
n undersize

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 (weight)
undersize

1 2 3 4 5 6 7 8 9 10

Result
1) Length number diameter dln =
2) Surface number diameter dsn =
3) Volume number diameter dvn =
4) Surface length diameter dsl =
5) Volume surface diameter dvs =

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 Experiment No. 3
Determination of Bulk density, True Density and Percent Porosity –Powders
Aim: to determine the following properties of powders

I. True density

II. Bulk density

III. Percent porosity

Principle:

I. Determination of true density of a powder.

The true density is the density of the powder itself i.e material exclusive of voids and intraparticle pores.

True density is defined as

True density ρp = weight of the powder/true volume of the powder

Normally true volume is determined as the volume of liquid displaced by the powder.

True density ρp = weight of the powder/Volume of liquid it displaced by the powder.

Liquids such as water, ethyl alcohol cannot occupy the pores or cervices. If the powder is non porous,
these liquids can be used. Solid be insoluble in the solvent and heavier than the liquid. Normally the
values obtained are somewhat lower than the helium displacement method.

Requirements:

Apparatus: Sieve No. 20, bulk density apparatus, Specific gravity bottle,25 ml

Chemicals: Benzene, Calcium carbonate.

Procedure:

1. Take a clean and dry specific gravity bottle. And place it in the ground-in-stopper

2. Weigh the empty specific gravity bottle. The weight is W1.

3. Fill the bottle with water and keep the stopper. Place the bottle in the water bath for
equilibrium.

4. Remove the bottle from water bath wipe outer surface of the bottle.

5. Weigh the specific gravity bottle with water. The weight is W2.

6. Remove water and wash the bottle with acetone. Dry the bottle with the help of hot air dryer.

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 7. Fill the bottle with benzene and repeat the steps 3 and 4.

8. Weigh the specific gravity with benzene. Let the weight be W3.

9. Remove benzene and wash the bottle with water and acetone. Dry the bottle with the help of
hot-air-dryer.

10. Place about 5.0g of powder in the specific gravity bottle. In case of light powders 1 or 2 gms of
powder may be adequate.

11. Weigh the specific gravity bottle with powder. The weight is W4.

12. Add slowly benzene to the specific gravity bottle with gentle swirling. Care should be taken to
avoid the entrapment of air bubbles. Gently add benzene up to the brim of the bottle.

13. Place the bottle in the thermostat (25OC) for 15 minutes. Remove the bottle from the water
bath dry the sides.

14. Weigh the specific gravity bottle with solids and benzene. The weight is W5.

Observations:

Weight of empty specific gravity bottle =W1=..............g

Weight of empty specific gravity bottle+ water=W2=.........g

Volume of bottle = V1 = (W2-W1)/0.9961

Weight of specific gravity bottle + Benzene = W3=...........g

Density of benzene = weight of benzene/volume of the bottle.

= (W3-W1)/V1

Weight of empty specific gravity bottle+ powder= W4 =.............g

Weight of empty specific gravity bottle+ powder+ Benzene=W5=..............g

Volume of benzene displaced V2=

= (W3-W1)/ρ - (W5-W4)/ρ

True density of powder, ρp =(W4-W1)/V2

Report:

II. Determination of bulk density of a powder.

Principle:

Bulk density is defined as

Bulk density, ρb = Weight of the powder/ Bulk volume of the powder.

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 Since bulk volume includes the true volume, volume of interparticle spaces and intraparticle
pores, determining the volume of powder using a measuring cylinder may be appropriate. When
particles are packed it is possible that a large amount of gas may be present between the particles.
Therefore tapping of powder allows the particles to shift and remove the voids to the minimum volume.
The volume occupied by the powder in this condition represents the bulk volume.

Procedure:

1. Pass the given powder through US standard sieve No.20.

2. Weigh about 50 g of powder. In case of light powders one can take 5g or 10 g of powder. The
criteria is that the cylinder should be filled up to the 75ml mark

3. Fix the measuring cylinder on the table of bulk density apparatus

4. Adjust the knob for 125 tapping’s and switch on the apparatus

5. Note the volume of the powder at the end of the tapping’s

6. Adjust the knob for another 50 tapping’s.

7. Note the volume of powder at the end of tapping’s

8. If two subsequent values of volume are the same switch off the apparatus. Otherwise continue
steps 6 and 7

9. Note the final volume as bulk volume.

10. If the apparatus if not available the cylinder is dropped onto a hard wooden surface 3 times from a
height of one inch at 2 second intervals. Normally concurrent results can be obtained, if the cylinder
is dropped or tapped for 500 times.

Observations:

Weight of the powder taken=................g

Concurrent volume of the powder (after tappings) =..............g

Bulk density =

III. Determination of Percentage porosity of a powder.

Whether the powder is porous or nonporous the total porosity expression for the calculation remains
the same. The percent porosity can be expressed as:

% Porosity є = (true density- bulk density)/ (true density) * 100 ; % Porosity є = ( ρp - ρb)/ ρp * 100

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 Experiment No. 4
Determination of Flow properties –Powders/Granules
Aim: To evaluate the following properties of powders/granules

IV. Measurement of angle of repose of the given powder/granules

V. Study of the effect of glidants on flow properties of granules.

Principle:

IV. Measurement of angle of repose of given powder or granules

Angle of repose is defined as the maximum angle possible between the surface of the pile of
powder and the horizontal plane.

The angle of repose is designated by θ and given by equation

Tan θ = h/r or θ= tan-1 h/r

Where h= height of the pile,cms

r= radius of the base of the pile,cms

The lower the angle of repose the better is the flow properties. When granules are placed in the
hopper and allowed to slide down on to the die for compression it forms a pile. The angle of repose
can be calculated by measuring the height and radius of the base with ruler. During the flow through
the hopper, granules exhibit internal flow and demixing. Flow of granules is hindered on account of
frictional forces. Therefore, the tangent of the angle of repose is expresses as coefficient of friction.
Particle size, nature, moisture content in the powder influences the flow of powder.

Requirements:

Apparatus: Glass funnel, iron stand, ring,

Chemicals: Granules, talc.

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 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 100 g of granules

5. Pour granules into 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
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 7. Adjust the funnel clamp so that the gap between the bottle of the funnel stem and peak of the
powder pile is about 3 mm

8. Repeat the steps 5 to 7 until appropriate gap is maintained.

9. Finally pour the granules back into the funnel and allow to flow.

10. Mark 4 points which are opposite to each other on the circular graph paper and tabulate the
readings. This value is diameter. Calculate the radius.

11. Measure the height of the pile using two rulers. Keep one ruler vertically and the other
horizontally to touch the peak of the pile. Then read the value on the vertical scale. Record the
readings. This value represents the height (h)

12. Substitute these values in the above equation to obtain angle of repose.

13. Repeat the procedure for two more trials and take an average.

14. Comment on the flow of the material by the following table.

Angle of repose Flow

<25 Excellent

25-30 Good

30-40 Passable

>40 Very poor

Observations:

Trials Height,h Diameter,d Radius, r h/r θ= tan-1 h/r

(cms)
(cms) (cms) (in degrees )

Trial 1

Trial 2

Trial 3

Report:

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 Experiment No. 5

EFFECT OF GLIDANTS ON ANGLE OF REPOSE

Principle:

Glidants are those substances which are intended to promote the flow of granules or powder
material by reducing the friction forces between the particles.

A few examples of effective glidants include talc, corn starch, magnesium stearate etc. These substances
get adsorbed on the surface of the granules or powders and machine parts thereby reduce the frictional
forces.

Since the 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 or magnesium stearate is increased
continuously the angle of repose decreases gradually. This trend continues until a particular
concentration and further increase in concentration increases the angle of repose. Plot this relationship.
The inflation point i.e the concentration at which the trend changes is considered as the optimum
concentration of the glidant required.

Requirements:

Apparatus: Glass funnel, iron stand, ring,

Chemicals: Granules, talc.

Procedure :

The angle of repose of the granules is calculated in the same manner without and with the addition of
granules. The glidant selected here is talc. The concentration is :

Talc (% w/w) 0 1 2 3 4 5

1. Determine the angle of repose of granules without the addition of glidants.

2. Add the glidant in low concentration to the granules and mix them thoroughly.

3. Estimate the angle of repose of this blend in two trials. Tabulate the values.

4. Calculate the average angle of repose.

5. Add the increments of glidants and calculate the angle of repose.

6. Repeat steps 3 and 4

7. Add further increments of glidants and determine the angle of repose

8. Repeat the procedure with further addition of 1.0 g of talc until optimum concentration is
obtained.

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 9. Plot a graph taking concentration of glidants on x axis and angle of repose on the y axis.

Concentration Trial 1 Trial 2

of glidant
Diameter,d Height,h Radius,r h/r Diameter,d Height,h Radius,r h/r

Influence of talc on the angle of repose

Concentration of talc Angle of repose

w/w Trial 1 Trial 2 Average
0
0.2
0.4
0.6
0.8
1

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 EXPERIMENT No. 06
“Determination of viscosity of liquid using Ostwald’s viscometer”

Aim: To determine viscosity of give unknown liquid

Theory:

 Viscosity is an important physicochemical property of liquids.

 Viscosity is an index of resistance of a liquid to flow.

 The higher the viscosity of a liquid the greater the resistance to flow. For example groundnut oil,
honey, syrup (sucrose solution), all resist the flow more, when compared to water or alcohol.

 Unit: Poise. It is also expressed in centipoises.

 Viscosity of the given unknown liquid can be determined by using capillary viscometer i.e.
Ostwald’s viscometer.

 When a liquid flows through the capillary tube the time required for the liquid to pass between
two marks (A and B ) is determined.

 The time of flow of test liquid is compared with the time of flow of the reference sample of
known viscosity.

 Viscosity of given unknown liquid (ղ1) may be determined using equation.

ղ2 = viscosity of known liquid.Pa.s

t1= time of flow of unknown liquid, s

t2= time of flow of known liquid,s

ρ1= density of unknown liquid, kg/m3

ρ2= density of known liquid, kg/m3

Relative viscosity, ղrel = ղ/ղo

ղ= viscosity of liquid, Pa.s

ղo= viscosity of known liquid, Pa.s

Requirement:

Chemicals: Distilled water, Benzene, ethanol, methanol

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Apparatus: Ostwald’s viscometer, stopwatch, specific gravity bottle, Beaker, stand and clamp, Hot air
oven

Procedure:

1. Select an Ostwald’s viscometer and clean it with and acetone. Dry the viscometer using a hot air
dryer. Viscometer is fixed to a stand firmly in a vertical position.

2. Transfer a known quantity of water ( 8 to 10 ml or 15 ml ) accurately depending up on the

capacity of viscometer with the help of pipette through the wide limb.

3. Now suck the water through the other limb up to a level higher than the upper mark A and clamp
the rubber tube with the finger.

4. Release the rubber tube and allow water to flow down. Start the stop clock when the water level
just passes the upper mark. When the water level passes the lower mark B stop the clock. Note
the time required for the flow of the water between A and B.

5. Repeat the steps 3 and 4 thrice and calculate the average time of flow.

NOTE: Determine the density of given liquid as per the procedure mentioned in experiment no 3.

Observation:

Name of the liquid Time of flow

Trial 1 Trial 2 Trial 3 Average.

Water

Calculation:

REPORT: The viscosity of given unknown solution was found to be ............

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 EXPERIMENT No. 07
“Determination of sedimentation volume with effect of different concentrations of single
suspending agent ”

Aim: To determine the optimum concentration of tragacanth required for physical stability of calcium
carbonate suspension.

Theory:

Pharmaceutical suspension are defined as the coarse dispersions in which insoluble solids are suspended
in the liquid medium. The insoluble solids may have a size range of 10 to 1000 µm. Suspending agents
are defined as physiologically inert substances which increases the viscosity when added to suspensions.
These agents help in keeping the dispersed particles suspended. Thus, enhance the physical stability of
suspensions. These also facilitate the re-dispersion of the sediment on shaking.

Physical stability may be defined as the condition in which particle remain uniformly distributed
throughout the dispersion, without any signs of sedimentation. This condition is ideal but difficult to
achieve. In practice, physical stability may be defined as the condition in which particle should be easily
re-suspendable by a moderate shaking, if they settle.

In the evaluation of physical stability, two sedimentation parameters are employed. These are:

a) Sedimentation Volume, F

b) Degree of flocculation, β

SEDIMENTAION VOLUME:

Sedimentation volume is defined as,

F= Ultimate volume of the sediment/ Initial volume of the suspension

= Hu/H0 or Vu/Vo

When a suspension is taken in an measuring cylinder volume and height are proportional and
height can be conveniently measured. Through the term volume is included in the terminology
‘sedimentation volume’ F is a dimension less number.

Most pharmaceutical suspensions have a F value less than one. If F=1, the product has value less than
one. If F = 1, the product has no sediment and no clear supernatant on standing, which is ideal condition.
Normally F value is between 0 and 1. A plot drawn by taking Hu/Ho on y axis and time on x axis. The
plot begins at 1 at zero time and then it may move horizontally or gradually downward, as time elapses.

Requirement:

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Chemicals: Tragacanth, Calcium carbonate.

Apparatus: Beaker, Measuring cylinder, Mortar and pestle, Funnel.

Procedure:

A) PREPARATION OF THE VEHICLE

Normally, tragacanth 2% w/v dispersion is used in practice. In such cases, the evaluation needs 2 or 3
days to complete. So the concentration of tragacanth is reduced to complete the experiment in 3 hrs.

Prepare aqueous tragacanth dispersion of different concentrations viz..0.0,0.1,0.2,0.3,0.4,0.5 %

w/v. All concentrations of tragacanth vehicle are prepared using the method mentioned below.

1. Weigh the required quantity of tragacanth and transfer into motor

2. Add water about 75 ml slowly in small quantities while triturating the sample continuously.

3. Transfer the dispersion using a funnel into a measuring cylinder.

4. Rinse the motor and pestle with about 15ml of water and transfer the rinsing into the measuring
cylinder(100ml) Make-up the volume now.

B.PREPARATION OF SUSPENSION

All the suspensions are prepared by the following method.

1. Weigh 5.0g of calcium carbonate and place it in a motor.

2. Add the tragacanth dispersion about 80 ml in small quantities while Triturating the sample.

3. After the powder is suspended uniformly transfer the suspension into 100 ml measuring cylinder.

4. Rinse the motor and pestle with further 10 ml of tragacanth dispersion medium and transfer into
the measuring cylinder.

5. Make up the volume to 100 ml mark with water using a pipette.

C. EVALUATION OF SEDIMENTATIO VOLUME,F

1. Shake the suspension simultaneously and set aside.

2. Note the volume of sediment at time periods 0,5,10,15,20,25,30,40,50,60 minutes. Record the
volume.

3. Calculate the sedimentation volume and report in the table.

4. Draw a plot taking F value on y axis and time on x axis.

5. Identify the active sedimentation phase. The phase is identified by the rapid downward trend of
the plot.

Observation:

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 0.0% w/v tragacanth 0.1% w/v tragacanth 0.2% w/v tragacanth 0.3% w/v tragacanth 0.4% w/v tragacanth 0.5% w/v tragacanth

Time in min

Hu F=Hu/Ho Hu F=Hu/Ho Hu F=Hu/Ho Hu F=Hu/Ho Hu F=Hu/Ho Hu F=Hu/Ho

Calculation:

REPORT: The suspension that has ............% w/v of tragacanth is the most stable.

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 EXPERIMENT No. 08

“Determination of sedimentation volume with effect of different suspending agents ”

Aim: To determine the sedimentation volume with effect of different suspending agents.

Theory:

Physical stability may be defined as the condition in which particle remain uniformly distributed
throughout the dispersion, without any signs of sedimentation. This condition is ideal but difficult to
achieve. In practice, physical stability may be defined as the condition in which particle should be easily
re-suspendable by a moderate shaking, if they settle.

SEDIMENTAION VOLUME:

Sedimentation volume is defined as,

F= Ultimate volume of the sediment/ Initial volume of the suspension

= Hu/H0 or Vu/Vo

When a suspension is taken in an measuring cylinder volume and height are proportional and
height can be conveniently measured. Through the term volume is included in the terminology
‘sedimentation volume’ F is a dimension less number.

Most pharmaceutical suspensions have a F value less than one. If F=1, the product has value less than
one. If F = 1, the product has no sediment and no clear supernatant on standing, which is ideal condition.
Normally F value is between 0 and 1. A plot drawn by taking Hu/Ho on y axis and time on x axis. The
plot begins at 1 at zero time and then it may move horizontally or gradually downward, as time elapses.

Suspending agents can be classified as :

 Synthetic

 Semi- synthetic

 Natural polysaccharide.

Examples of suspending agent include tragacanth, bentonite, guargum, sodium alginate, carboxy methyl
cellulose.

Requirement:

Chemicals: Tragacanth, Calcium carbonate, bentonite, guargum, sodium alginate, carboxy methyl
cellulose.

Apparatus: Beaker, Measuring cylinder, Mortar and pestle, Funnel.

Procedure:

A) PREPARATION OF SUSPENSION

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 1. Required quantity of calcium carbonate is weighed and transferred in to a motor.

2. 50ml of water is added and triturated to get a smooth paste

3. Bentonite (1%w/v) is added gradually with constant stirring to above solution. The mixture is
transferred to the 100 ml measuring cylinder

4. The motor and pestle are washed with 10 ml of water

5. Then the washings are transferred in to a measuring cylinder.

6. The volume is made upto the mark 100 ml by adding water.

7. The procedure is repeated using other suspending agents.

B. DETERMINATION OF SEDIMENTATIO VOLUME,F

1. The suspension in the cylinder is shaked thoroughly to make a uniform distribution.

2. The cylinder is kept undisturbed on a flat surface after shaking

3. The volume of sedimentation volume at different interval of every 10 minutes is measured.

4. The sedimentation volume is determined by using above mentioned formula.

Observation:

Tragacanth Bentonite Guar gum

Time in min

Hu F=Hu/Ho Hu F=Hu/Ho Hu F=Hu/Ho Hu F=Hu/Ho Hu F=Hu/Ho Hu F=Hu/Ho

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Calculation:

REPORT: The sedimentation volume with effect of different suspending agent was determined and
reported.

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 EXPERIMENT No. 09
“Determination of viscosity of semisolid using Brookfield viscometer”

Aim: To determine viscosity of semisolid using Brookfield viscometer

Theory:

 Brookfield viscosity usually refers to a viscosity measurement performed with a Brookfield,

Viscometer.

 There are several types of viscometer available from Brookfield but the majority operates on the
same manner.

 The viscometer motor rotates the spindle at a defined speed ( rpm) or shear rate and the
viscometer measures the resistance to rotation and reports a viscosity value.

 Various spindle designs can be employed depending on the nature of the sample and the
requirements.

 Brookfield viscometer is also a rotational viscometer.

 The construction of this instrument is similar to the cup and bob viscometer.

 This viscometer is used to evaluate the rheological properties of suspension with some
modification such as helipath arrangement.

 Brookfield viscometer mounted on helipath stand with T spindle may be ideal to study the
behaviour of settling.

 The T- bar rotates the changes its position continuously.

Requirement:

Chemicals: Sample (liquid, gel, glycerine based substances)

Apparatus: Brookfield viscometer, Beaker, Glass rod.

Procedure:

1. Prepare the gel using gel forming materials ( gelling agent )

2. Keep the gel at least 24 hours, for uniform dispersion and homogenization

3. After 24 hours, gel is completely formed, place sufficient quantity in the beaker or sample holder
provided with the instrument.

4. Set up the base level of the instrument using level indicator on the top of instrument and plug in
for constant electric supply.

5. Clean the spindle and all over to the instrument.

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 6. Rotate the spindle in the gel till to get a constant dial reading on the display of the viscometer.

7. Repeat the determination at least three times for reproductable result.

Note: If the temperature is to be studied than maintain the temperature at least 20 minutes and
determine the viscosity.

Observation and Calculations:

Amount of gel =.......g
Spindle number=......
Speed of the spindle in gel=...........rpm
Average reading of three determinations=..........

REPORT: The viscosity of the prepared gel was found to be ............cp, determined by Brookfield
viscometer .......(model) using spindle number .............. at ..........rpm speed and .........torque.

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EXPERIMENT No. 10
“Determination of reaction rate constant first order”

Aim: To determine reaction rate constant and half life of an ester in 0.5N hydrochloric acid at room
temperature

Theory:

First order reaction is defined as a reaction in which the rate of the reaction is directly
proportional to the concentration of on reacting substance.

The integral equation of the first order is :

This equation helps in estimating the reaction rate constant of the order. Secondly, it is used in
calculation of concentration of drug remaining unreacted after any time, t

Half-life is defined as the time required for the concentration of the reactant to reduce to half of its
initial concentration.

t1/2= 0.693/k1

Principle:

Pseudo first order reaction is defined as a reaction which is originally a second order, but made to follow
first order kinetics by maintaining the concentration of one of the reactants in large excess such that it
will not appreciably change during progress of the reaction.

An example of pseudo first order reaction is acid hydrolysis of an ester. Ester decomposes slowly
in normal storage conditions.

CH3COOCH3+H2O↔CH3COOH+HOCH3

Methyl acetate+water↔Acetic acid+ methyl alcohol

In this reaction 10 ml of ester and 100ml of 0.5 N hydrochloric acid solution are taken and the reaction
follows first order reaction.

The kinetic method involves drawing of samples periodically over a period of time viz, 0,10,20,30,.......75
minutes and analysing the concentrations of the reactants. Generally an excess of acid is taken in the
reaction. Some quantity of acid reacts with the ester over a period of time. The acid remaining

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unreacted is estimated by titrating against 0.25 N sodium hydroxide solution using phenolphthalein
indicator. The data can be substituted in the below equation for each time period sample.

Vo = volume of titrant consumed at ‘0’ time, ml

Vt= volume of titrant consumed at‘t’ time, ml

V∞= Volume of titrant consumed at ‘∞’ time,ml

An average k1 value can be calculated. Alternatively constant can be determined by graphic method. A
graph may be plotted taking log(V∞-Vt) on y-axis and time on x-axis.The slope is equal to k1/2.303.

Requirement:

Chemicals: Hydrochloric acid,ethylacetate, sodium hydroxide, phenolphthalein, ice cold water

Apparatus: Water bath, pipette, test tube, conical flask, burette, test tube stand.

Procedure:

1. Measure 100 ml of 0.5N hydrochloric acid (use burette or pipette) into a stoppered conical flask
(250ml). Keep it in a water bath. (Do not heat) for the equilibrium.

2. Measure 10 ml of the given ester in to a clean test tube. Keep it in the same water bath for 10 min
(do not heat) for the equilibrium. Normally it takes 10 min to attain equilibrium.

3. Mix the acid solution and ester sample thoroughly and keep in the same water bath, mix
occasionally to ensure uniform mixing.

4. Immediately after mixing withdraw 5 ml of mixture with pipette and transfer into a conical flask
containing 10 ml of ice cold water.

5. Add a few drops of phenolphthalein indicator.

6. Titrate the reaction mixture against 0.25 N sodium hydroxide solution. This value of alkali
represents Vo.

7. Withdraw samples periodically at 10,20,30,40,50,60,and 75 minute timings. Repeat the steps

from 4 to 6. The volume of alkali consumed at each time interval represents Vt at the respective
timings.

8. Heat the reaction mixture on a water bath at 600C for 20 minutes.

9. Cool the reaction mixture to room temperature.

10. Withdraw a 5 ml mixture into a conical flask containing 10 ml of ice cold water.

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 11. Repeat the titration (steps 5 to 6).This value represents V∞.

12. Report the relevant data in table.

The reaction rate constant can be determined by substitution method. Substitue the data in equation
mentioned above to get k1 value for each time. Calculate the average k1. From k1 value half life can be
calculated from the equation.

Observation:
Time,min Volume of NaOH Volume of (V∞-Vt) log(V∞-Vt) k1
consumed NaOH
Vt,ml
Initial Final
volume,ml volume,ml

Calculation:

REPORT: The reaction rate constant (k1) for the given ester in 0.5 N hydrochloric acid by substitution
method was=..............

Half life was equal to .................

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 EXPERIMENT No. 11
“Determination of reaction rate constant second order”

Aim: To determine reaction rate constant and half life of an ethyl acetate in 0.025N NaOH solution at
room temperature.

Theory:

The alkali hydrolysis of an ester is irreversible and follows the second order kinetics.

CH3COOC2H5+NaOH→ CH3COONa+C2H5OH

Ethyl acetate+Sodium hydroxide→Sodium acetate+Ethyl alcohol

The molecularity of the reaction is two and the order is also two.

Second order reaction is defined as a reaction in which the rate of the reaction is directly proportional to
the concentration of two reactants with each term raised to the first power.

The determination of k2 may be simplified by considering the initial concentration of ethyl acetate and
sodium hydroxide as equal(equimolar concentration 0.05N). At the same time the volume of reactants is
also made equal(50ml each).The concentration of sodium hydroxide remaining unreacted can be
estimated by titrating against hydrochloric acid solution (0.02N) using phenolphthalein indicator. The
same concentration can be treated as the concentration of ethyl acetate solution.

The integral equation for the determination of reaction rate constant for second order(a=b) is

a= initial concentration of alkali.

t= time at which the concentration is measured

x= amount of reactant consumed.

Half-life is defined as the time required for the concentration of the reactant to reduce to half of its
initial concentration.

t1/2= 1/ak2

k2 can be obtained by substitution method and graphic method.

Requirement:

Chemicals: Hydrochloric acid,ethylacetate, sodium hydroxide, phenolphthalein, ice cold water

Apparatus: Water bath, pipette, test tube, conical flask, burette, test tube stand.

Procedure:

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 13. Measure 50 ml of 0.05N NaOH (use burette or pipette) into a stoppered conical flask (250ml).
Keep it in a water bath. (Do not heat) for the equilibrium.

14. Measure 50 ml of 0.05 N of the given ester in to a clean test tube. Keep it in the same water bath
for 10 min (do not heat) for the equilibrium. Normally it takes 10 min to attain equilibrium.

15. Mix the alkali solution and ester sample thoroughly and keep in the same water bath, mix
occasionally to ensure uniform mixing.

16. Immediately after mixing withdraw a 10 ml of mixture with pipette and transfer into a conical
flask containing 10 ml of ice cold water.

17. Add a few drops of phenolphthalein indicator.

18. Titrate the reaction mixture against 0.02 N hydrochloric acid solution. This titer value at time t=0
corresponds to the original concentration a. Report the results in the table.

19. Withdraw samples periodically at 5,10,15,20,25 and 30 minute timings. Repeat the steps from 4
to 6.

20. These titer values determines the amount of sodium hydroxide or ethyl acetate remaining
unreacted,i.e at time t. Record the results in table.

21. Substitute the values in integral equation and calculate the reaction rate constant(k2).These values
will be more or less constant.

22. Calculate the average of reaction rate constant(k2)

Observation:

Time,min Volume of HCl Volume of Conc in x=a-(a-x) x/a(a-x) k2

consumed HCl,ml mol/lit a
or (a-x)
Initial Final
volume,ml volume,ml

10

15

20

25

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30

Calculation:

REPORT: The reaction rate constant (k2) for the given ester in 0.025 N sodium hydroxide at room
temperature by substitution method was=..............

Half life was equal to ..................

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School Of Pharmacy,Parul University Page 33
 EXPERIMENT No. 12
“ACCELERATED STABILITY STUDIES”

Aim: To determine the shelf life of aspirin solution (in 0.1N HCl solution ) using accelerated stability
studies.

Theory:

Accelerated stability studies is an experimental design to evaluate the stability of a product by

accelerating the rate of a reaction.

It involves the evaluation of

1. physical changes

2. Chemical changes.

Influence of temperature on degradation :

In general the rate of a reaction increases with rise in temperature. Arrhenius established a more
quantitative relationship between temperature and rate of reaction. The exponential equation is

k= Ae –Ea/RT

where,

k=specific rate constant

A= frequency factor.

Ea= energy of activation.

R= ideal gas constant

T= absolute temperature.

Arrhenius factor or frequency factor is a measure of the frequency of collision that can be expected
between the reactant molecules in a reaction. Energy of activation is defined as the minimum energy
required by the molecules so that molecular collisions give useful products.

Taking logarithms on both sides of equation gives:

log k= log A- Ea/2.303 RT

according to equation a plot can be drawn by taking log k on y axis and reciprocal temperature(1/T) on x
axis. The slope gives the energy of activation

Ea=slope*2.3030*R

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Where R= 8.314 J/mol.K

Aspirin Degradation :

During storage conditions, aspirin undergoes hydrolysis to salicylic acid and acetic acid

Requirement:

Chemicals: salicylic acid, ferric nitrate solution, HCl solution, Aspirin,

Apparatus: Volumetric flask ,conical flask, pipette, test tubes, water bath, Colorimeter.

Procedure:

Standard plot for salicylic acid

1. Prepare the following dilutions from the stock solution as mentioned below. In 20ml test tubes

2. Add 5 ml of ferric nitrate solution to the above test tubes solutions turns to purple colour.

3. Measure the absorbance of the above solutions at 547 nm using a colorimeter. Report the
observations.

4. Plot a graph by taking concentration on x axis and absorbance on y axis. This plot is a straight
line that passes through origin.

Serial number Stock solution of Distilled Concentration of Absorbance

salicylic acid,ml water,ml salicylic acid
mg/ml

1 0.0 10 0.0

2 0.2 9.8 0.02

3 0.4 9.6 0.04

4 0.6 9.4 0.06

5 0.8 9.2 0.08

6 1.0 9 0.10

Aspirin decomposition at temperature 500C

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  Switch on the water bath and adjust temperature at 500C

 Transfer 100 ml of 0.1 N HCl into a conical flask and keep it in the water bath.

 Weigh accurately 100mg of aspirin

 Transfer the aspirin into a 250ml conical flask

 Add 1 to 2 ml of alcohol to dissolve the aspirin.

 Add slowly 90ml of 0.1N HCl and shake thoroughly .Dissolved aspirin should not get
precipitated.

 Finally add 10ml of 0.1N HCl.

 Cork the conical flask and keep in the same water bath

 Immediately after placing in the water bath withdraw a 10 ml of sample mixture with pipette and
transfer into a 20 ml test tube. This represents the zero time sample.

 Withdraw samples after 10,20,30,40,50,60 minutes periods.

 Add 5 ml of ferric nitrate solution to each test tube. The solution turn purple colour.

 Measure the absorbance of the samples at 547nm using suitable colorimeter. Report the
observation in the table.

 Process the data as per the columns in the table.

 Plot a graph taking time on x-axis and log percent aspirin undecomposed at y-axis.

Aspirin decomposition at temperature 600C

The method is same as mentioned above ( steps) except that solution should be maintained at a
temp of 600C.Periodically 10ml samples are drawn ferric nitrate solution is added to each sample and
colour is developed.

Aspirin decomposition at temperature 700C

The method is same as mentioned above ( steps) except that solution should be maintained at a
temp of 700C.Periodically 10ml samples are drawn ferric nitrate solution is added to each sample and
colour is developed.

Observation:
Aspirin decomposition stored at temp 500C /600C /700C

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Time in Measured Salicylic Aspirin Aspirin Percent aspirin Log percent
minutes absorbance acid conc decomposed undecomposed remain aspirin remain
undecomposed undecomposed

A B C D E F

10

20

30

40

50

60

Calculation:

REPORT: Shelf life of aspirin in 0.1 N HCl solution=..............

Therefore the accelerated studies are Valid/Not Valid

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