EXPERIMENT NO:06

TO PREPARE AND EVALUATE MULTIPLE EMULSION .

1. AIM: To prepare and evaluate multiple emulsion.

2. REFERENCES:
1. www.Aaps.pharm sci,2003,5(1),article 7
2. Multiple emulsion :Technology & application by Abraham Aserin.
3. www.wikipedia.com
4. Remington:the science & practice of pharmacy volume-1 ;326
5. Pak.J.pharm sci,vol 21,no.4 october 2008,pp 430-437
6. Bull chem..soc. ethiop .2010,24(1)1-10

3. MATERIALS:

NAME OF THE DRUG COMPANY’S NAME

Salicylic acid Aghadi Industrial Estate

Excipients COMPANY’S NAME

Span 80
Dextrose Aghadi industrial Estate
Tween 80

4. THEORY

Introduction

Spherical crystallization is a particle design technique, by which crystallization and

agglomeration can be carried out simultaneously in one step and which has been successfully
utilized for improvement of flowability, compactability and bioavailability of crystalline
drugs. General methods of spherical crystallization are spherical agglomeration, emulsion
solvent diffusion and ammonia diffusion method. The principle steps involved in the process
of spherical crystallization are flocculation zone, zero growth zone, fast growth zone and
constant size zone. Factors controlling the process of agglomeration are solubility profile,
mode and intensity of agitation, temperature of the system and residence time. Spherical
crystallization is having wide applications in pharmaceuticals like improvement of
flowability and compressibility of poorly compressible drugs, masking bitter taste of drugs

and improving the solubility and dissolution rate of poorly soluble drug.

Parul Institute of Pharmacy 1 of 15

5. EVALUATION PARAMETER

1. Type of emulsions.
2. Microscopic Test.
3. Globule size.
4. pH determination.
5. stabilityof multiple emulsion.
6 Centrifuge tests.
7. Electrical conductivity tests.

6. DRUG PROFILE

PARAMETER VALUE
dose 600mg
category Anti inflammatory,rubefacient,keratolytic
BCS classification Class I
Poorly soluble in water 0.2 g/100 mL H2O (20 °C),
solubility
Chloroform 0.19 M, ethanol 1.84 M, methanol 2.65 M .
Ionization constant 2.97 at 25Ċ
After oral administration 80-100% will be absorbed in
bioavailability
stomach and in the small intestine
Steady state conc.
Half life 2-4.5 hrs
Protein binding 50-80%
metabolism Salicylic acid is metabolized 80% in liver
excreted mainly by the kidney as salicylic acid, salicyluric
Renal excretion acid, salicylic glucuronides and gentisic acid.

Tmax 2hr
CmaX 1.2 mcg/l

7. FORMULA:-

SR INGREDIENTS QUANTITY TAKEN

NO.
A. FOR W/O : 1ST PHASE:
1 light liquid paraffin oil 30 ml
2 purified water 15 ml
3 span -80 6 ml
Dextrose 0.02gm
B. FOR W/O/W: 2ND PHASE:
1 purified water 30 ml
2 tween -80 1.8 ml
3 w/o emulsion 60 ml
4 Blood red color q.s.

Parul Institute of Pharmacy 2 of 15

8. PROCEDURE:-

1. Preparation of W/O emulsion:

For preparation of primary emulsion , oil phase consisting of paraffin oil and Span 80 ,was
heated to 55. Aqueous phase consisting of salicylic acid and dextrose was also heated to the same
temperature. Aqueous phase was added to the oil phase drop by drop . (15 min stirr under mechanical
stirrer).

2. Preparation of W/O/W emulsion:

Agitation was continued until cooling to room temperature of 25 oC. For obtaining the multiple emulsion,
primary emulsion was added to the aqueous phase containing hydrophilic surfactant (Tween 80) while
agitating for 10 min. Emulsion was then homogenized at 800 rpm for 5 min and further at 500 rpm for 5
min more.

2. Evaluation Parameters:

EVALAUTION OF SPHERICAL CRYSTALS

As these spherical agglomerated crystals showing significant effect on the formulation and
manufacturing of pharmaceutical dosage forms so it is necessary to evalaute them by using
different
parametrs.
FLOW PROPERTY
Flow property of the material depends on the force developed between the particle, particle
size, particle

Parul Institute of Pharmacy 3 of 15

size distribution, particle shape, surface texture or roughness and surface area. Flowability
of the
agglomerates is much improved as the agglomerate
exhibits lower angle of repose then that of single crystals. Studies on spherically
agglomerated aspirin
crystals revealed that, the angle of repose of agglomerated crystals was 31.13 while that of
unagglomerated crystals was 47.12.This improvement in the flowability of agglomerates
could be
attributed to the significant reduction in inter-particle friction, due to their spherical shape
and a lower
static electric charge
Following are the methods used to determine of flow property
ANGLE OF REPOSE
This is the common method used for determination of flow property. The angle of repose is
the angle
between the horizontal and the slop of the heap or cone of solid dropped from some
elevation. Values for
angle of repose ≤ 30 usually indicate free flowing material and angle ≥ 40 suggested a poor
flowing
material. The angle of repose can be obtained from equation
Tan θ = h/0.5d
Where h- height of the cone and d- diameter of the cone
COMPRESSIBILITY OR CARR INDEX
A simple indication of ease with which a material can be induced to flow is given by
application of
compressibility index
I = (1-V/Vo) *100
Where v = the volume occupied by a sample of powder after being subjected to a
standardized tapping
procedure and Vo = the volume before tapping.
The value below 15% indicates good flow characteristics and value above 25% indicate
poor flowability
HAUSNER RATIO
It is calculated from bulk density and tap density.
Hausner ratio = Tapped density / Bulk density
Values less than 1.25 indicate good flow (20% Carr Index) and the value greater then 1.25
indicates poor
flow ( 33% Carr Index).
DENSITY
Density of the spherical crystals is the mass per unit volume.
Density = M/V
Publication Ref No.: IJPRD/2010/PUB/ARTI/VOV-1/ISSUE-12/FEB/005 ISSN 0974 – 9446
International Journal of Pharma Research and Development – Online
www.ijprd.com
5
POROSITY
Porosity of granules affects the compressibility. Porosities are of two types “intragranular
and
intergranular and these are measured with the help of true and granular densities.
Intragranular porosity = 1- Granular density /True density.

Parul Institute of Pharmacy 4 of 15

Intergranular porosity = 1- Bulk density / Granular density
Total Porosity = 1- Bulk density/ True density
PACKABILITY:
Improve packability has been reported for agglomerates prepared by spherical
crystallization. The angle
of friction, shear cohesive stress and shear indexes are lower then that of single crystals,
which can
improve the packability of the agglomerates.
The packability of agglomerates improved compared with those of the original crystals and
that the
agglomerated crystals are adaptable to direct tabletting. The packability assessed by
analysis of the
tapping process with the Kawakita(I) and Kuno(II) method and using the parameters a,
b,1/b, k in the
equation
N/C = 1/ (ab) +N/a.....................................................I
C = (Vo-Vn)/Vo,
a =(Vo-V∞) /Vo.
ρf- ρn= (ρf- ρo) . exp. (-kn)…………………………II
Where, N =Number of tapping
C =Difference in volume (degree of volume reduction.) and a, b are constant.
COMPRESSION BEHAVIOUR ANALYSIS
Good compactibility and compressibility are essential properties of directly compressible
crystals. The
compaction behavior of agglomerated crystals and single crystals is obtained by plotting the
relative
volume against the compression pressure. Spherical
agglomerates possess superior strength characteristics in comparison to conventional
crystals. It is
suggest that the surface are freshly prepared by fracture during compression of
agglomerates, which
enhances the plastic inter particle bonding,resulting in a lower compression force required
for
compressing the agglomerates under plastic deformation compared to that of single
crystals.
Compaction behaviour of agglomerated crystals were evaluated by using following
parameters
Heckel Analysis
The following Heckel's equation used to analyze the compression process of agglomerated
crystals and
assessed their comapctibility.
In [1/(1-D)]=KP+A
Where:
D is the relative density of the tablets under compression Pressure
K is the slope of the straight portion of the Heckel Plot
The reciprocal of K is the mean yield is the mean yield pressure (Py).
The following equation gives the intercept obtained by extrapolating the straight portion of
the plots
A=1n [1/(1-D0)]+B
Where:

Parul Institute of Pharmacy 5 of 15

D0 is the relative density of the powder bed when P=0.
The following equation gives the relative densities corresponding to A and B.
DA=1-e-A
DB=DA-D0
Stress Relaxation Test
Publication Ref No.: IJPRD/2010/PUB/ARTI/VOV-1/ISSUE-12/FEB/005 ISSN 0974 – 9446
International Journal of Pharma Research and Development – Online
www.ijprd.com
6
In this test put specific quantity of spherical agglomerated crystals sample in a die specific
diameter the surface of which is coated with magnesium stearate in advance, then used the
universal
tensile compression tester to compress the samples at a constant speed. After the certain
limit of
pressure attained, the upper punch held in the same position for 20 min, during which
measured time for
the reduction amount of the stress applied on the upper punch. The result corrected by
subtracting from
this measurement the relaxation measured without powder in the die under the same
conditions.
The following equation finds the relationship between relaxation ratio Y(t) and time t,
calculated the
parameters As and Bs, and assessed relaxation behavior.
t/Y(t)=1/AsBs-t/As
Y(t)=(P0-Pt)/P0
Where:
P0 is the maximum compression pressure, and Pt is the pressure at time t.
MECHANICAL STRENGTH
Spherical crystals should posses’ good mechanical strength as that directly reflects the
mechanical strength of compact or tablet. It is determine by using the following two
methods,
Tensile strength:
Tensile strength of spherical crystals is measured by applying maximum load required to
crush the
spherical crystal. This method is a direct method to measure the tensile strength of
spherical crystals
Crushing Strength
It is measured by using 50ml glass hypodermic syringe. The modification includes the
removal of the tip
of the syringe barrel and the top end of the plunger. The barrel is then used as hallow
support and the
guide tube with close fitting tolerances to the Plunger. The hallow plunger with open end
served as load
cell in which mercury could be
added. A window cut into the barrel to facilitate placement of granule on the base platen.
The plunger
acted as movable plates and set directly on the granules positioned on the lower platen as
the rate of
loading may affect crushing load (gm). Mercury is introduced from reservoir into the upper
chamber at

Parul Institute of Pharmacy 6 of 15

the rate of 10 gm/sec until the single granule crushed; loading time should be <3 minutes.
The total
weight of the plunger and the mercury required to fracture a granule is the crushing load.
FRIABILITY TEST
The friability of the spherical crystals is the combination of the attrition and sieving process
in to a
single operation. Granules along with the plastic balls placed on a test screen. The sieve is
then
subjected to the usual motion of a test sieve shaker provided the necessary attrition on the
granules. The
weight of powder passing through the sieve is recorded as function of time. The friability
index is
determined from the slop of the plot of % weight of granules remaining on the sieve as a
function of time
of shaking.
Friability of agglomerates determined by using formula
Friability(X) = {1-W/Wo}/100
Where
Wo = Initial weight of the crystalline agglomerates placed in sieve
W = Weight of the material which does not passed through sieve after 5 min.
PARTICLE SIZE AND SIZE DISTRIBUTION
Size of the particle and their distributions can be determined by simply sieve analysis. Now
with the help
of Ro-Tap sieve shaker, particle size analysis can be determined. In advance technology
image-analyzer
is used to determined size and volume of the particle.
Publication Ref No.: IJPRD/2010/PUB/ARTI/VOV-1/ISSUE-12/FEB/005 ISSN 0974 – 9446
International Journal of Pharma Research and Development – Online
www.ijprd.com
7
MOISTURE UPTAKE STUDY
The study indicates the behavior of uptake of moisture by drug and the prepared spherical
crystals,
which affect the stability. The weighted quantity of drug and spherical crystals placed in
crucible at
accelerated condition of temperature and humidity,40 C ± 10C and 75% ± 3%respectively.
The gain in
weight of drug and spherical crystals is measured
PARTICLE SHAPE / SURFACE TOPOGRAPHY
Following methods are used
Optical Microscopy
The shape of the spherical crystals is studied by observing these under a optical
microscope. The
observations are made under the observation like 10X, 45X, 60X etc.
Electron Scanning Microscopy
The surface topography, type of crystals (polymorphism and crystal habit) of the spherical
crystals is
analyzed by using scanning electron microscopy.
X-ray Powder Diffraction

Parul Institute of Pharmacy 7 of 15

 This is an important technique for establishing batch-to-batch reproducibility of a crystalline
form. The
form of crystal in agglomerates determine by using technique. An amorphous form does not
produce a
pattern. The X-ray scattered in a reproducible pattern of peak intensities at distinct angle
(2θ) relative to
the incident beam. Each diffraction pattern is characteristics of a specific crystalline lattice
for a
compound.
SUMMARY AND CONCLUSION
The spherical

2.1 Organoleptic characteristics

.
Freshly prepared primary and multiple emulsions were investigated organoleptically (color, liquefaction
and phase separation). Organoleptic characteristics of both primary and multiple emulsions kept at different
storage conditions, i.e. color, liquefaction and phase separation were noted at various intervals.

2.2 Types of emulsions

Types of emulsions were analyzed by dilution with paraffin oil and water separately and observation
under microscope.

2.3 Globules size

In this study, globule sizes of the multiple emulsions prepared were determined using light
Microscope fitted with a digital camera for the freshly prepared emulsions and for the emulsions
kept at different conditions for 28 days .

2.4 pH determination

The pH value of the freshly prepared emulsions and the emulsions kept at different conditions
were determined by a digital pH-meter. pH measurements were repeated for multiple emulsions
after 28 days of preparation.

2.5 Microscopic tests

Multiple emulsions were analyzed under the microscope to confirm the multiple characters. A drop of
multiple emulsion was placed on the glass slide, diluted with water and covered by a glass cover. A drop of
immersion oil was placed on the cover slide and observed under the microscope.

2.6 Stability tests

Stability tests were performed at different storage conditions for both primary and multiple
emulsions. The tests were performed on samples kept at 4 ± 0.1 oC (in refrigerator), 25 ± 0.1oC.

Parul Institute of Pharmacy 8 of 15

 2.6 Dye test
Water soluble dye was dissolved in the aq.phase of an emulsion while an oil soluble dye taken up by the oil
phase .which help ful in determination of type of emulsion.

2.8 In vitro release of drug

In vitro release studies from O/W/O emulsions were conducted using Franz diffusion cell (at room
temperature) and UV spectrophotometry. The results showed that in the first four-hour period.

2.9 Assay of salicylic acid

Calibration curve:
It was prepared by taking 100 mg salicylic acid then dissolved in 100ml dis. water. Taken
2ml from above and diluted up to 20ml which produced 100µg/ml. then 1ml was taken & diluted to
10ml which produced 10 ug/ml. taken suitable aliquot like,2ml,4ml,6ml,8ml and 10ml dilute up
to 10ml which produce concentrations,2,4,6,8 & 10 µg/ml respectively and measured the
absorbance by U.V. at 203nm. Plot the graph of Abs v/s Conc and find out the calibration curve
equation.
Calibration curve of Aspirin

3 OBSERVATION & CALCULATION

Parul Institute of Pharmacy 9 of 15

 3.1 Organoleptic characteristics
Organoleptic characteristics of the primary and multiple emulsions formulated are presented in
Table

TIME COLOUR LIQUEFACTION PHASE SEPARATION

0hr milky white No change Not observed
1hr Milky white No change Not observed
24hr Milky white No change Slightly seperation
After 1month

3.2 Type of multiple emulsion

(1) Microscopic evaluation shows continuous pink color through out the water phase.
(2) We can find out no. of globules of w/o emulsion of various size moving in the continuous
water phase.
(3) This conforms formation of w/o/w emulsion.

3.3 Globules size

Sr.No Size range Avg.diameter(d) No.of gobules(n) nd

1 0-5 2.5 15 37.5
2 6-10 7.5 35 262.5
3 11-15 12.5 25 312.5
4 16-20 17.5 20 350
5 21-25 22.5 5 112.5
∑n=100 ∑nd=1075

Avg. Diameter = ∑nd/∑n

= 1075/100

= 10.75 µm

3.4 pH determination

pH of freshly prepared emulsion was 4.8 observed.

3.5 Microscopic tests

Here multiple emulsion type was o/w/o observed as follow;

Parul Institute of Pharmacy 10 of 15

3.6 Stability tests

In this work, both primary and w/o/w multiple emulsions were divided into two samples
separately and these samples were kept at different storage conditions, i.e. at 4 °C in refrigerator,
at 25 °C at room temp.. The samples kept at different storage conditions were observed for a period of
1 month. Samples were observed .
separation.

Sr.no parameter Room temperature Temp. 4 °c

1
2
3

3.7 Dye test

Microscopic examination had seen that a water soluble dye had been taken up by continous phase & the
inner particles present in globules,which indicated the w/o/w type of emulsion.

Parul Institute of Pharmacy 11 of 15

3.8 In vitro release of drug

Sr No. Time Absorbance(nm)

(hr)
1 1 0.234
2 2 0.178
3 3 0.293

3.9 Assay of salicylic acid

Calibration curve
Sr No. Cocentration Absorbance(nm)
((ug/ml)
1 2 0.159
2 4 0.288
3 6 0.359
4 8 0.470
5 10 0.585

10. RESULT AND CONCLUSION

SR.NO PARAMETER RESULT CONCLUSION

1 ORGANOLEPTIC CHARACTERS

(a)COLOUR (a)milky white

Parul Institute of Pharmacy 12 of 15

 (b)LIQUEFACTION (b)not observed
(c)PHASE SEPARATION (c)observed after week
2 TYPES OF EMULSION w/o/w
3 AVG.GLOBULE SIZE 10.75µm
4 PH DETERMINATION 4.8
5 MICROSCOPIC TEST w/o/w
6 STABILITY TEST
7 DYE TEST w/o/w

10. RESULTS AND DISCUSSION

Table-1 Blend Evaluation

Sr. No. Parameter F1 (1:1) F2 (1:0.5) F3 (1:1) F4 (1:0.5)

1 Angle of Repose 30.98 35.65 34.70° 33.95°

2 Bulk Density (g/ml) 0.400 0.450 0.559 0.568
3 Tapped Density (g/ml) 0.466 0.500 0.746 0.746
4 Carr’s Index (%) 14.16 10.00 25.06 23.86
5 Hausner’s Ratio (%) 1.16 1.11 1.33 1.31

Table-2 Tablet Evaluation

Sr. No Parameter F1 (1:1) F2 (1:0.5) F3 (1:1) F4 (1:0.5)

Weight Variation (maximum

1 3.97 4.10 2.59 4.43
% Deviation)
2 Friability (%) 0.555 0.700 0.968 0.682
3 Hardness ± S.D. 10.26 ± 0.21 10.5 ± 0.39 8.7 ± 0.29 7.8 ± 0.22
4 Thickness (mm) ± S.D. 4.0 ± 0.05 4.0 ± 0.05 5.0 ± 0.05 5.0 ± 0.05
5 Diameter (mm) ) ± S.D. 8.0 ± 0.05 8.0 ± 0.05 8.0 ± 0.05 8.0 ± 0.05
6 Drug Content (%) 95.65 % 93.47 % 97.28 % 91.30 %

Dissolution Profile:

Parul Institute of Pharmacy 13 of 15

 90.00
Aspirin SR Matrix Tab
80.00
70.00

% Drug Release
60.00
50.00
40.00
30.00
20.00
10.00
0.00
0 2 4 6 8
Time (Hr)
EC 1:1 EC 1:0.5 HPMC 1:1 HPMC 1:0.5

Parul Institute of Pharmacy 14 of 15

 Table- 3 Swelling Index:

% Swelling Index
Time (Hr)
F1 F2 F3 F4
0.5 26.67 23.33 30.00 25.81
1.0 40.00 43.33 46.67 41.94
2.0 46.67 50.00 53.33 48.39
3.0 60.00 63.33 56.67 51.61
4.0 63.33 66.67 63.33 58.06
5.0 63.33 66.67 63.33 58.06
6.0 26.67 23.33 30.00 25.81

% Swelling Index
80
70
% Swelling Index

60
50
40
30
20
10
0
0 1 2 3 4 5 6
Time (hr)
F1 F2 F3 F4

11. CONCLUSION

Aspirin sustained release tablets are prepared using two different rate controlling polymers, Ethyl cellulose and
HPMC with the drug: polymer ratio (1: 1and 1: 0.5) the evaluation is done both in the blend and the prepared
tablets. All the evaluation tests are found to be fall with in the range of standards specified in the pharmacopoeia.
The swelling index study indicates that as the concentration of HPMC increases the swelling of the system is
increased. And both the systems observed a sudden decrease in the weight after 5th hour indicates the
disintegrations or erosion of the matrix. The dissolution study conducted for all the formulations show the
sufficient slow down of the release of the drug achieving the desired target.

Parul Institute of Pharmacy 15 of 15