FACULTY OF PHARMACY

KARPAGAM ACADEMY OF HIGHER EDUCATION

(DeemedUniversityUnderSection3ofUGCAct1956)
Coimbatore–641021.

LABORATORY MANUAL

PHARMACEUTICS PRACTICAL-I

(22MPH105P)

I-M.PHARM I SEMESTER (2022-23)

1
 Name :

Year/Semester :

LABORATORY MANUAL FOR

PHARMACEUTICS PRACTICAL-I

(2MPH105P)

SUBJECTINCHARGE

Name :

Signature :

Exp. Date Title of The Experiment Page Marks Staff

2
No. No. Sign.

PHARMACEUTICS

Calibration curve for Active Pharmaceutical

1.
Ingredients
In-vitro Dissolution studies of Aceclofenac
2.
Sustained Release marketed formulation

3. Formulation and evaluation of Metformin Hcl

Sustained Release Matrix tablets

4. Formulation and evaluation of Paracetamol

Floating tablets
Formulation and evaluation of Aceclofenac
5.
Buccal mucoadhesive tablets
Formulation and evaluation of Diclofenac
6.
Sodium Transdermal patches

7. Preformulation studies of Granules

Effect of Compressional force on tablets

8.
disintegration time
Micromeritic properties of powder by
9.
Andreasen pipette (sedimentation technique)
Determination of Particle size distribution
10.
using sieving method
Effect of Particle size on Dissolution of
11.
tablets

12. Effect of binders on Dissolution of tablets

13. Study of Similarity factor

14. Determination of drug release kinetics plot

3
PHARMACEUTICS

4
 Table 1.1: Calibration curve data of Paracetamol

S.No. Concentration (µg/ml) Absorbance at 257 nm

1. 2 0.122

2. 4 0.251

3. 6 0.399

4. 8 0.523

5. 10 0.649

0.7

0.6 f(x) = 0.0656571428571429 x − 0.00428571428571428

R² = 0.999338417576595
Absorbance at 257nm

0.5

0.4

0.3

0.2

0.1

0
0 2 4 6 8 10 12
Concentration μg/ml

Figure 1.1 Calibration curve of Paracetamol

Experiment No.: 1 Date:

5
 CALIBRATION CURVE FOR ACTIVE PHARMACEUTICAL
INGREDIENT

AIM:
To determine the Calibration Curve for active pharmaceutical ingredients namely
Paracetamol, Aceclofenac, Diclofenac sodium and Metformin HCl.
REQUIREMENTS:
APPARATUS REQUIRED:
UV spectrophotometer, beaker, 1ml pipette, standard flasks.
MATERIALS REQUIRED:
Paracetamol, Aceclofenac, Metformin Hcl, Diclofenac sodium, Phosphate buffer pH 7.4,
Phosphate buffer pH 6.8 and Acid buffer pH 1.2.
PRINCIPLE:
In analytical chemistry, a calibration curve, also known as a standard curve. It is a
general method for determining the concentration of a substance in an unknown sample by
comparing the unknown to a set of standard samples of known concentration. The calibration
curve is a plot of how the instrumental response, the so-called analytical signal, changes with the
concentration of the analyte (the substance to be measured). These curves use data points of
known substances at varying concentrations, and researchers or developers can use these curves
to find where an unknown substance plots. A spectrophotometer assists in gathering absorbance
for varying concentrations.
PROCEDURE
PREPARATION OF PHOSPHATE BUFFER pH 7.4:
Place 50.0 ml of 0.2 M potassium dihydrogen phosphate in a 200 ml volumetric flask
add 39.1ml of 0.2 M sodium hydroxide and then add water to volume.
PREPARATION OF PHOSPHATE BUFFER pH 6.8:
Place 50.0 ml of 0.2 M potassium dihydrogen phosphate in a 200ml volumetric flask add
22.4 ml of 0.2 M sodium hydroxide and then add water to volume.

Table 1.2: Calibration curve data of Aceclofenac

6
 Absorbance at 275 nm
S.No. Concentration (µg/ml)
pH 1.2 pH 7.4

1. 2 0.189 0.197

2. 4 0.374 0.367

3. 6 0.543 0.525

4. 8 0.736 0.702

5. 10 0.886 0.872

1
0.9
Absorbance at 275nm

0.8 f(x) = 0.0891428571428572 x + 0.00895238095238088

R² = 0.999001620976188
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
0 2 4 6 8 10 12

Concentration μg/ml

Figure 1.2 Calibration curve of Aceclofenac at pH 1.2

1
Absorbance at 275nm

0.9
0.8 f(x) = 0.0861857142857143 x + 0.0129047619047619
0.7 R² = 0.999184409975824
0.6
0.5
0.4
0.3
0.2
0.1
0
0 2 4 6 8 10 12

Concentration μg/ml

Figure 13.3 Calibration curve of Aceclofenac at pH 7.4

PREPARATION OF ACID BUFFER pH 1.2:

7
 Place 50.0 ml of 0.2 M potassium chloride in a 200 ml volumetric flask add 83.4 ml of 0.2
M hydrochloric acid and then add water to volume.
STANDARD CALIBRATION CURVE PROCEDURE:
● The standard graph for Paracetamol, Aceclofenac, Metformin Hcl and Diclofenac
sodium as per the following procedure.
● The concentration and the abosrbance maximum at UV spectrophotometer were
shown in Table 13.1 to 13.4 respectively.
● The aliquots were prepared in dissolution medium namely Acid buffer pH
1.2,Phosphate buffer of pH 7.4 and pH 6.8 with similar stock solution preparation
procedure as stated below.
● Take 100 mg of drug and dissolve in 100 ml of buffer solution in a standard flask
to make a stock solution (1000 µg/ml)10 ml from the primary stock has been
withdrawn and make up to 100 ml using the respective buffer (100 µg/ml).
● From the secondary stock solution aliquots were taken and dilute to 10 ml with the
buffer solution respectively.
● The above solutions were analyzed using UV spectrophotometer at respective
wavelength.
● A calibration curve was plotted against Concentration vs Absorbance respectively.
● The calibration curve was shown in Figure 13.1 to 13.6 respectivley.

Table 13.3: Calibration curve data of Metformin Hcl

8
 Absorbance at 233 nm
S.No. Concentration (µg/ml)
pH 1.2 pH 6.8

1.
2 0.072 0.199

2.
4 0.133 0.382

3.
6 0.195 0.556

4.
8 0.257 0.752

5.
10 0.321 0.911

0.35
Absorbance at 233nm

0.3 f(x) = 0.0317428571428572 x + 0.00428571428571425

R² = 0.999413786542191
0.25

0.2

0.15

0.1

0.05

0
0 2 4 6 8 10 12

Concentration μg/ml

Figure 13.4 Calibration curve of Metformin Hcl at pH 1.2

1
0.9
Absorbance at 233nm

f(x) = 0.0912571428571429 x + 0.0103809523809523

0.8 R² = 0.999196624562942
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
0 2 4 6 8 10 12

Concentration μg/ml

Figure 13.5 Calibration curve of Metformin Hcl at pH 6.8

9
Table 13.4: Calibration curve data of Diclofenac sodium
10
 S.No. Concentration (µg/ml) Absorbance at 277 nm

1. 2 0.091

2. 4 0.154

3. 6 0.233

4. 8 0.317

5. 10 0.402

0.45

0.4
f(x) = 0.0395285714285714 x + 0.00185714285714284
0.35 R² = 0.998157077345779

0.3
Absorbance at 277nm

0.25

0.2

0.15

0.1

0.05

0
0 2 4 6 8 10 12
Concentration μg/ml

Figure 13.6 Calibration curve of Diclofenac Sodium

REPORT:
11
DISCUSSION

REFERENCES:

1. Mubeen G, Noor K. Spectrophotometric method for analysis of metformin hydrochloride.

Indian J Pharm Sci. 2009 Jan;71(1):100-2. doi: 10.4103/0250-474X.51947. PMID:
20177473; PMCID: PMC2810039.
2. Amrita Natekar & Siddhi Naroji,Simultaneous Estimation And Validation Of Paracetamol,
Chlorpheniramine Maleate, Phenylephine Hydrochloride, Caffeine And Its Stress
Degradation Studies Using Uv Visible Spectroscopy, International Research Journal Of
Pharmacy, 2016, 7 (7).
3. Bose A, Dash P, Sahoo M. Simple spectrophotometric methods for estimation of aceclofenac
from bulk and formulations. Pharm Methods. 2010 Oct;1(1):57-60. doi: 10.4103/2229-
4708.72233. PMID: 23781418; PMCID: PMC3658021.
4. RE, Ugandar. (2013). Formulation And Evaluation Of Natural Palm Oil Based Diclofenac
Sodium Suppositories,International Journal of Pharmaceutical Sciences and Research. 4.
617-621.

12
Table 2.1: Invitro Dissolution Study: USP TYPE- I (BASKET)

Brand: Hifenac SR 200 mg

Tablet Formulation: 200 mg Aceclofenac SR tablet

Aceclofenac
Analyte:

200 mg
Tablet Dose:

275 nm
Wavelength:

Calibration Curve Parameters:

0.089 (pH 1.2)

Slope:
0.093 (pH 7.4)
0.009 (pH 1.2)
Intercept:
0.083 (pH 7.4)
Assay Dilution Factor: 10 X (Dilution factor: 1:10)
Volume of the Dissolution Medium: 900 ml

13
Experiment No.: 2 Date:
IN-VITRO DISSOLUTION STUDIES OF ACECLOFENAC SUSTAINED
RELEASED MARKETED FORMULATION
AIM:
To study the In-vitro Drug release of marketed Aceclofenac SR tablets.
REQUIREMENTS:
APPARATUS REQUIRED:
USP Dissolution apparatus, UV spectrophotometer, beaker, 1ml pipette and standard
flasks.
MATERIALS REQUIRED:
Aceclofenac SR tablets, Phosphate buffer pH 7.4 and Acid buffer pH 1.2 .
PRINCIPLE:
Sustained designed to slowly release a drug in the body over an extended period of time
especially to sustain therapeutic levels. Among the various routes of drug delivery, oral route is
most preferred route. Sustained release dosage form is defined as well characterized and
reproducible dosage form, which is designed to control drug release profile at a specified rate to
achieve and maintain the desired drug concentration constantly either in blood plasma or at
target site.
In-vitro dissolution testing is widely used to predict In-vivo performance of oral drug
products in the gastrointestinal (GI) tract to ensure product quality and performance, but also
assist in development and regulatory processes of the nanoparticulate delivery system.
Dissolution testing measures the extent and rate of solution formation from a dosage form such
as tablet, oinment, capsule etc. Dissolution of a drug is important for its bioavailability and
therapeutic effectiveness.

PROCEDURE:
PREPARATION OF BUFFER:
The buffers (pH 1.2 & 7.4) were prepared suitably as per the procedure mentioned in
Experiment No. 13

14
Table 2.2: In-vitro Drug Release Data of Marketed Aceclofinac SR Tablets
Amount Cumulative
Drug % Drug
S. Absorbance of Drug % Drug
Time (h) Concentration Release
No. at 275 nm Release Release
(µg/ml)
(mg)
Acid buffer pH 1.2
1. 0.50 0.089 0.8 7.2 7.2 7.2

2. 1.00 0.144 1.2 10.8 10.8 10.8

3. 1.50 0.249 1.9 17.1 17.1 17.1

4. 2.00 0.301 2.4 21.6 21.6 21.6

Phosphate buffer pH 7.4

5. 2.50 0.403 3.2 28.8 28.8 28.8

6. 3.00 0.499 3.9 35.1 35.1 35.1

7. 3.50 0.557 4.4 39.6 39.6 39.6

8. 4.00 0.609 5.2 46.8 46.8 46.8

50

45 R² = 0.996980063434527
40

35
Cumulative % Drug release

30

25

20

15

10

0
0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00

Time (Hours)

Figure 2.1: In-vitro Drug Release of Marketed Aceclofinac SR Tablets

15
INVITRO DRUG RELEASE:
● Dissolution test for the tablet samples was performed using USP Type- I rotating basket
type.
● The parameters like temperature and speed was adjusted to 37 ± 0.5 0C and 50 RPM
respectively.
● The drug dissolution studies was carried out at Acid buffer pH 1.2 for the first two h
followed by Phosphate buffer pH 7.4 with the medium of 900 ml in the hemispherical
basket.
● Then the rotating baskets were positioned into the basket containing the dissolution
medium.
● The tablet samples were placed into individual baskets and the apparatus was operated at
the above specified condition.
● Specified volume of sample was withdrawn from the zone midway between the surface of
the dissolution medium and top of the rotating paddle at different time intervals.
● Sink condition was maintained by replacing equivalent volume of fresh medium each time
of withdrawal of the sample.
● Samples withdrawn were diluted suitably and analyzed using UV spectrophotometer at
275 nm.
● The percentage drug release was calculated from the obtained data and studied for kinetic
parameters.

16
17
REPORT:

DISCUSSION:

REFERENCE:
1) Sharma, Arpit & Kumar, Manish & Malik, Anuj & Saini, Vipin & Pandurangan, Dr.
(2018). Comparative Quality Evaluation of Aceclofenac Marketed Tablet.

18
Table 3.1: Composition of 50 mg Metformin Hcl SR tablets
Working formula
S. No. Ingredients Master Formula
(25 tablets)

1. Metformin Hcl 50 mg 1.25 g

2. HPMC 30 mg 0.75 g

3. Poly vinyl pyrolidine 30 mg 0.75 g

4. Lactose 80 mg 2.00 g

5. Magnesium Stearate 5 mg 0.12 g

6. Talc 5 mg 0.12 g

7. Isopropyl alcohol 1 ml 2.5 ml

EVALUATION PARAMETERS:
Table 3.2: Weight Variation test data for the prepared tablets

S. No. 1 2 3 4 5 6 7 8 9 10

Weight of each 0.20

0.202 0.198 0.199 0.200 0.201 0.202 0.196 0.202 0.200
tablet (g) 1

S. No. 11 12 13 14 15 16 17 18 19 20

Weight of each 0.20

0.200 0.196 0.202 0.201 0.200 0.199 0.201 0.199 0.202
tablet (g) 2

Average Weight = Total weight of Tablets

Number of Tablets

= 0.200 g

19
Experiment No.:3 Date:
FORMULATION AND EVALUATION OF METFORMIN HCL
SUSTAINED RELEASE MATRIX TABLETS
AIM:
To formulate and evaluate Metformin Hcl sustained release matrix tablets by Direct
compression method.
REQUIREMENTS:
APPARATUS REQUIRED:
Mortar and Pestle, Sieve no. 40, Beaker, Weighing balance, Rotary tablet compression
machine, Monsanto tester,UV spectrophotometer and Vernier Calliper.
MATERIALS REQUIRED:
Metformin Hcl, Hydroxy Propyl Methyl Cellulose (HPMC), Poly Vinyl Pyrolidine
(PVP), Lactose, Magnesium stearate, Talc,Isopropyl alcohol and Phosphate buffer pH 6.8 and
Acid buffer pH 1.2 .
PRINCIPLE:
Sustained release dosage form are designed to slowly release a drug in the body over an
extended period of time especially to sustain therapeutic levels. Among the various routes of
drug delivery, oral route is most preferred route. Sustained release dosage form is defined as well
characterized and reproducible dosage form, which is designed to control drug release profile at
a specified rate to achieve and maintain the desired drug concentration constantly either in blood
plasma or at target site.
PROCEDURE:
DIRECT COMPRESSION TECHNIQUE:
● A calculated amount of the drug, polymer (HPMC, PVP) was triturated thoroughly ensuring
free from any grity substances.
● The ingredients were passed though a 40-mesh sieve.
● Magnesium stearate was added as lubricant and Talc was added as glidant.
● The formed granules were punched into tablet using rotary punching machine.

20
Table 3.3: Hardness test
S.NO. HARDNESS (kg/cm2) AVERAGE HARDNESS( kg/cm2)

1. 4.5

2. 4.0

3. 4.1 4.12 ± 0.23

4. 3.8

5. 4.2

Table 3.4: Thickness of the prepared tablet

S.No. Thickness (mm) Average Thickness (mm)

1. 4.48

2. 4.54

3. 4.50 4.5 ± 0.03

4. 4.52

5. 4.46

Friability:

Initial weight = 2.00

Final weight =1.99
2.00 – 1.99
= X100
2.00

% Friability = 0.50%

21
EVALUATION PARAMETERS:
WEIGHT VARIATION TEST:
Weigh 20 tablets selected at random and determined their average weight. Not more than
2 of the individual weight deviate from average weight.
HARDNESS TEST:
The hardness of the tablet was determined by using Monsanto hardness tester.
THICKNESS TEST:
The thickness of the tablet was measured by using digital vernier calliper.
INVITRO DISSOLUTION STUDY:
● Dissolution test for the tablet samples was performed using USP - Type I rotating basket
type.
● The parameters like temperature and speed was adjusted to 37 ± 0.5 ◦C and 50 RPM
respectively.
● The drug dissolution studies was carried out at Acid buffer pH 1.2 for two h followed by
Phosphate buffer pH 7.4 with the medium of 900 ml in the hemispherical basket.
● The samples were withdrawn at specified time intervals, diluted with respective buffer
and scanned for absorbance at UV spectrophotometer.
● The tablet samples were placed into individual baskets and the apparatus was operated at
the above specified condition.
● Specified volume of sample was withdrawn from the zone midway between the surface of
the disso medium and top of the rotating paddle at different time intervals.
● Sink condition was maintained by replacing equivalent volume of fresh medium each time
of withdrawal of the sample.
● Samples withdrawn were diluted if necessary and analyzed using UV spectrophotometer
at 233 nm.
● The percentage drug release was calculated from the obtained data and and the graph was
plotted.

22
Table 3.5: Invitro Dissolution Study: USP TYPE- I (BASKET)
Tablet Formulation: 50 mg Metformin Hcl SR tablet

Analyte: Metformin Hcl

Tablet Dose: 50 mg

Wavelength: 233 nm

Calibration Curve Parameters:

0.031 (pH 1.2)

Slope:
0.091 (pH 6.8)

0.0043 (pH 1.2)

Intercept:
0.0104 (pH 6.8)

Assay Dilution Factor: 10 X (Dilution factor: 1:10)

Volume of the 900 ml

Dissolution Medium:

23
Table 3.6 In-vitro Drug Release Data of Metformin Hcl SR Tablets
Amount Cumulative
Drug % Drug
S. Absorbance of Drug % Drug
Time (h) Concentration Release
No. at 233 nm Release Release
(µg/ml)
(mg)
Acid buffer pH 1.2
1. 0.50 0.018 0.3 2.7 5.4 5.4

2. 1.00 0.031 0.8 7.2 14.4 14.4

3. 1.50 0.048 1.1 9.9 19.8 19.8

4. 2.00 0.063 1.6 14.4 28.8 28.8

Phospahte buffer pH 7.4

5. 2.50 0.191 2 18 36 36

6. 3.00 0.231 2.4 21.6 43.2 43.2

7. 3.50 0.288 2.9 26.1 52.2 52.2

8. 4.00 0.352 3 27 54 54

60
R² = 0.993754349338901
50
Cumulative % Drug release

40

30

20

10

0
0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00

Time (Hours)

Figure 3.1: In-vitro Drug Release of Metformin Hcl SR Tablets

24
REPORT:

DISCUSSION:

REFERENCE:
1) Anil Keshav Pawar, Formulation and In-vitro Drug Release of Metformin HCL Extended
Release Matrix Tablet. 159-162.

25
 Table 4.1: Composition of 250 mg Paracetamol floating tablets

Working formula
S.No. Ingredients Master formula
(10 tablets)

1 Paracetamol 0.250 g 5.0 g

2 HPMC 0.046 g 0.092 g

3 Sodium bicarbonate 0.030 g 0.60 g

5 Polyvinyl Pyrollidine 0.027 g 0.54 g

6 Magnesium Stearate 0.002 g 0.04 g

7 Talc 0.001 g 0.02 g

EVALUATION PARAMETERS:

Table 4.2: Weight Variation test data for the prepared tablets

S. No. 1 2 3 4 5 6 7 8 9 10

Weight of each
0.398 0.400 0.401 0.399 0.402 0.400 0.404 0.398 0.396 0.399
tablet (g)

S. No. 11 12 13 14 15 16 17 18 19 20

Weight of each
0.401 0.400 0.402 0.398 0.400 0.401 0.396 0.399 0.400 0.398
tablet (g)

Average Weight = Total weight of Tablets

Number of Tablets
= 0.399g

26
 Experiment No.:4 Date:
FORMULATION AND EVALUATION OF FLOATING TABLETS OF
PARACETAMOL
AIM:
To formulate and evaluate Paracetamol floating tablets by Wet granulation method.

REQUIREMENTS:
APPARATUS REQUIRED:
Mortar and Pestle, Sieve no.16 and 20, Beaker, Weighing balance, Rotary tablet
compression machine, Monsanto hardness tester,UV spectrophotometer and Vernier Calliper.
MATERIALS:
Paracetamol, HPMC, Sodium bicarbonate, Polyvinyl pyrollidine, Magnesium stearate
and Talc.
PRINCIPLE:

The primary objective of the study is to prepare a floating paracetamol tablet as the
floating tablets prolong the gastric residence time of drugs,improve bioavailability and facilitate
local drug delivery to the stomach. The floating system (or) dynamically controlled system are
low density system that have sufficiently buoyancy to float over the gastric contents and remain
buoyant in stomach without affecting the gastric emptying rate of prolonged period of time

27
Friability:

Initial weight =3.99

Final weight =3.97

3.99 – 3.97
= X100
3.99

%Friability =0.55%

Table 4.3: Hardness test

S.NO. HARDNESS (kg/cm2) Average Hardness( kg/cm2)

1. 3.8

2. 3.5

3. 4.0 3.68±0.21

4. 3.4

5. 3.7

Table 4.4: Thickness of the prepared tablet

S.No. Thickness (mm) Average Thickness (mm)

1. 3.99

2. 4.02

3. 4.00 3.99±0.02

4. 3.98

5. 3.96

PROCEDUCE :
28
WET GRANULATION METHOD:

● The drug and the polymers were weighed individually and blended by geometric dilution.
● The coherent mass is formed by adding granulating agent and it is screened though sieve no.
22.
● The granules were dried at 60°C for 60 min in hot air oven and allowed to pass though sieve
no. 40.
● Magnesium stereate and Talc were added to the granules and the were puched into tablet.

EVALUATION PARAMETERS:

WEIGHT VARIATION:
Weigh 20 tablets selected at random and determined their average weight. Not more than
2 of the individual weight deviate from average weight.
HARDNESS TEST
The hardness of the tablet was determined by using Monsanto hardness tester..
FRIABILITY:
The friability of tablets were determined by using Roche Friabilator using 20 tablets. It is
expressed in percentage.
FLOATING INDEX:

Floating lag time (FLT) and total floating time (TFT) of floating tablets were measured
visually in dissolution apparatus type II containing 900 ml 0.1N Hcl at 37±0.5℃

SWELLING INDEX:

The extent of swelling was measured in terms of percent weight gain by the tablet. One
tablet was kept in a petri dish containing phosphate buffer. At the end of 1h the tablet was
withdrawn, blotted with tissue paper and weighed. For every 1 h weight of the tablet was noted
till 5 h.

Where, M0= Weight of the tablet at time t=0

Mt= Weight of the tablet at time‘t’

Floating time:
Floating lag time was 55 sec
Total floating time was 10 h

29
Table 4.5: Swelling Index
Time Initial Weight of Final Weight
Mt-M0/M0 x100 (%)
(h) Tablet (g) of Tablet (g)

1 0.400 0.580 45.00

2 0.402 0.612 52.23

3 0.404 0.620 53.46

4 0.396 0.616 55.50

5
0.398 0.628 57.78

Table 5.6: Invitro Dissolution Study: USP TYPE- II (Paddle)

Tablet Formulation: 250 mg Paracetamol Floating tablet

Analyte: Paracetamol

Tablet Dose: 250 mg

Wavelength: 257 nm

Calibration Curve Parameters:

Slope: 0.065 (pH 1.2)

Intercept: 0.0043 (pH 1.2)

Assay Dilution Factor: 10 X (Dilution factor: 1:10)

Volume of the 900 ml

Dissolution Medium:

INVITRO DISSOLUTION STUDY:

30
● Dissolution test for the tablet samples was performed using USP - Type II paddle type.
● The parameters like temperature and speed was adjusted to 37 ±0.5 0c and 50 RPM
respectively.
● The drug dissolution studies was carried out at followed by the acid buffer pH 1.2 with
the medium of 900 ml in the hemispherical basket.
● The samples were withdrawn at specified time intervals, diluted with respective buffer
and scanned for absorbance at UV spectrophotometer.
● The tablet samples were placed into individual baskets and the apparatus was operated
at the above specified condition.
● Specified volume of sample was withdrawn from the zone midway between the surface
of the disso medium and top of the rotating paddle at different time intervals.
● Sink condition was maintained by replacing equivalent volume of fresh medium each
time of withdrawal of the sample.
● Samples withdrawn were diluted if necessary and analyzed using UV spectrophotometer
at 257 nm.
● The percentage drug release was calculated from the obtained data and studied for
kinetic parameters.

31
Table 4.7: In-vitro Drug Release Data of Paracetamol floating tablets

Drug Amount of Cumulative

% Drug
S. Absorbance Concentrati Drug % Drug
Time (h) Release
No. at 257 nm on Release Release
(µg/ml) (mg)
Acid buffer at pH 1.2
1 0.50 0.206 2 18 7.2 7.2

2 1.00 0.253 4 36 14.4 14.4

3 1.50 0.323 5.5 49.5 19.8 19.8

4 2.00 0.414 6.5 58.5 23.4 23.4

5 2.50 0.523 8 72 28.8 28.8

6 3.00 0.591 8.5 76.5 30.6 30.6

7 3.50 0.655 9.3 83.7 33.48 33.48

8 4.00 0.711 10 90 36 36

40
R² = 0.958959084868046
35
Cumulative % Drug release

30

25

20

15

10

0
0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00

Time (Hours)

Figure 4.1: In-vitro Drug Release of Paracetamol floating tablets

REPORT:
32
DISCUSSION:

REFERENCES:
1. Rahamathulla, et.al (2021). Development and in vitro evaluation of effervescent floating
matrix tablet of neritinib: An anticancer drug. Pakistan journal of pharmaceutical
sciences, 34(4), 1297–1303.
2. Namdev, A., & Jain, D. (2019). Floating Drug Delivery Systems: An Emerging Trend for
the Treatment of Peptic Ulcer. Current drug delivery, 16(10), 874–886.
3. Heikh, F. A.,et.al. (2020). Linseed hydrogel based floating drug delivery system for
fluoroquinolone antibiotics: Design, in vitro drug release and in vivo real-time floating
detection. Saudi pharmaceutical journal : SPJ : the official publication of the Saudi
Pharmaceutical Society, 28(5), 538–549.

33
 Table 5.1: Composition of 100 mg Buccal mucoadhesive Aceclofenac tablets

S.No. Ingredients Master formula Working formula of 25

tablets (g)
(mg)

1 Aceclofenac 100 2.50

2 Carbopol 934 25 0.62

4 Aerosil 5 0.12

5 Magnesium Stearte 50 1.25

6 Talc 5 0.12

EVALUATION PARAMETERS:

Table 5.2: Weight Variation test data for the prepared tablets

S. No. 1 2 3 4 5 6 7 8 9 10

Weight of each
0.252 0.254 0.256 0.257 0.248 0.246 0.250 0.252 0.253 0.254
tablet (g)

S. No. 11 12 13 14 15 16 17 18 19 20

Weight of each
0.254 0.256 0.257 0.248 0.250 0.252 0.253 0.254 0.252 0.252
tablet (g)

Average Weight = Total weight of Tablets

Number of Tablets

= 0.252 g

34
Experiment No.:5 Date :
FORMULATION AND EVALUATION OF BUCCAL MUCOADHESIVE
TABLETS OF ACECLOFENAC
AIM:
To prepare and evaluate Bucco adhesive controlled release tablets of Aceclofenac.

REQUIREMENTS:
Aceclofenac, Carbopol 934, Micro crystalline Cellulose. Aerosil, Magnesium stearate, Talc.

PRINCIPLE
The oral route of drug administration is the most common and preferred route for drug
delivery, as it enables easy ingestion, self-medication, accurate dosage. The buccal delivery thus
implies the absorption of medication though the mucosal lining of the buccal cavity. It can
remain attached to buccal mucosa and becomes capable of bypassing hepatic first-pass
metabolism to improve absorption directly into systemic circulation

PROCEDURE:
PREPARATION OF TABLET:
Accurately weighed quantities of Drug, MCC, Aerosil and Carbopol 934 were mixed
well. Then add Magnesium stearate and Talc to the above mixture and blend them. The powder
blend was then compressed into tablets by direct compression method. A backing layer of Ethyl
cellulose was compressed over the punched tablet to confirm the unidirectional movement of
drug
EVALUATION:
HARDNESS TEST:
The hardness of the tablet was determined by using Monsanto hardness tester.

Friability:

Initial weight = 2.52

Final weight = 2.50
2.52 – 2.50
= X100
2.52
35
%Friability = 0.79%

Table 5.3: Hardness test

S.NO. HARDNESS (kg/cm2) AVERAGE HARDNESS( kg/cm2)

1. 2.6

2. 2.5

3. 3.0 3.00±0.41

4. 3.3

5. 3.6

Table 5.4: Thickness of the prepared tablet

S.No. Thickness (mm) Average Thickness (mm)

1. 3.90

2. 3.94

3. 3.92 3.91±0.02

4. 3.88

5. 3.90

WEIGHT VARIATION:

Weigh 20 tablets selected at random and determined their average weight. Not more than
2 of the individual weight deviate from average weight.
FRIABILITY:
The friability of tablets were determined by using Roche Friabilator. It is expressed in
percentage.
SWELLING INDEX:
The extent of swelling was measured in terms of percent weight gain by the tablet. One
tablet was kept in a petri dish containing phosphate buffer. At the end of 1h the tablet was

36
withdrawn, blotted with tissue paper and weighed. For every 1 h weight of the tablet was noted
till 5h.

Where, M0= Weight of the tablet at time t=0

Mt= Weight of the tablet at time‘t’
INVITRO DISSOLUTION STUDY:
● Dissolution test for the tablet samples was performed using USP - Type II paddle type.
● The parameters like temperature and speed was adjusted to 37 ±0.5 0c and 50 RPM
respectively.
● The drug dissolution studies was carried out at followed by the buffer pH 6.8 with the
medium of 900 ml in the hemispherical basket.
● The samples were withdrawn at specified time intervals, diluted with respective buffer
and scanned for absorbance at UV spectrophotometer.
● The tablet samples were placed into individual baskets and the apparatus was operated
at the above specified condition.
● Specified volume of sample was withdrawn from the zone midway between the surface
of the disso medium and top of the rotating paddle at different time intervals.
● Sink condition was maintained by replacing equivalent volume of fresh medium each
time of withdrawal of the sample.
● Samples withdrawn were diluted if necessary and analyzed using UV spectrophotometer
at 275 nm.
● The percentage drug release was calculated from the obtained data and studied for
kinetic parameters.

Table 5.5: Swelling Index

Time Initial Weight of Final Weight
Mt-M0/M0 x100 (%)
(h) Tablet (g) of Tablet (g)

1 0.250 0.440 76.00

2 0.252 0.452 79.36

3 0.248 0.458 84.67

4 0.250 0.472 88.80

37
 5 0.251 0.481 91.63

Table 5.6: Invitro Dissolution Study: USP TYPE- II (Paddle)

Tablet Formulation: 100 mg Buccal Mucodhesive Aceclofenac tablet

Analyte: Aceclofenac

Tablet Dose: 100 mg

Wavelength: 275 nm

Calibration Curve Parameters:

Slope: 0.093 (pH 6.8)

Intercept: 0.083 (pH 6.8)

Assay Dilution Factor: 10 X (Dilution factor: 1:10)

Volume of the 900 ml

Dissolution Medium:

38
Table 5.7: In-vitro Drug Release Data of Buccal mucoadhesive
Aceclofenac tablets
Drug Amount of Cumulativ
% Drug
S. Absorbance Concentrati Drug e % Drug
Time (h) Release
No. at 275 nm on Release Release
(µg/ml) (mg)
At pH Buffer 6.8

1 0.50 0.121 1 9 9 9

2 1.00 0.199 1.5 13.5 13.5 13.5

3 1.50 0.272 2 18 18 18

4 2.00 0.546 4 36 36 36

5 2.50 0.612 5 45 45 45

6 3.00 0.696 6 54 54 54

7 3.50 0.712 6.9 62.1 62.1 62.1

8 4.00 0.799 7.5 67.5 67.5 67.5

80
Cumulative % Drug release

70
R² = 0.983142999006951
60

50

40

30

20

10

0
0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00

Time (Hours)

Figure 5.1 In-vitro Drug Release of Buccal mucoadhesive Aceclofenac tablets

39
REPORT:

DISCUSSION

REFERENCE:
1. Koirala S, Nepal P, Ghimire G, Basnet R, Rawat I, Dahal A, Pandey J, Parajuli-Baral K.
Formulation and evaluation of mucoadhesive buccal tablets of aceclofenac. Heliyon.
2021 Mar 15;7(3):e06439. doi: 10.1016/j.heliyon.2021.e06439. PMID: 33786387;
PMCID: PMC7988282.

40
Table 6.1: Composition of 10 mg Diclofenac Sodium Transdermal Patches

S. No. Ingredients Working Formula Working Formula Working Formula

(1) (2) (3)

1 Diclofenac 10 mg 10 mg 10 mg
Sodium

2 Chloroform 2 ml 2 ml 2 ml

3 Ethanol 2 ml 2 ml 2 ml

4 EC 400 mg 300 mg 200 mg

5 Propylene Glycol QS QS QS

Table 6.2: Physical Evaluation of Transdermal Patches

Working Folding endurance
Average Thickness (mm) Drug Content (%)
Formula (No.)

1 0.510 100.0 82.42

2 0.514 98.0 76.84

3 0.516 90.0 70.26

41
Experiment No.:6 Date:

FORMULATION AND EVALUATION OF DICLOFENAC SODIUM

TRANSDERMAL PATCHES

AIM:
To prepare and evaluate Transdermal patches of Diclofenac Sodium.
REQUIREMENTS:
Diclofenac Sodium, Chloroform, Ethanol, Ethyl cellulose, Propylene Glycol.
PRINCIPLE:
. The transdermal patches adhere to the skin as a way to deliver drugs.They provide a
specific ,predetermined dose of medication that is absorbed through the skin and into the blood
stream.Thus they offer a non-invasive and painless method of drug delivery.The transdermal
patch acts as a carrier for a drug ,holding it until it is applied.The patch will continue to
administer its API until either they are fully absorbed (or) the patch is removed.
PROCEDURE:
PREPARATION OF CASTING SOLUTION
● The casting solution were prepared by dissolving weighed quantities of polymer in a mixture
of chloroform and methanol in 1:1 ratio.
● The drug, plasticizer and permeation enhancer were then added to the polymer solution
separately and systematically mixed to form a homogenous mixture.
● The resultant solution is sonicated for degassing.
PREPARATION OF TRANSDERMAL PATCHES
● About 10 ml of the above prepared casting solution were transfered into circular glass
moulds especially designed to hold contents.
● The glass mould containing the casting solution was allowed to dry at room temperature for
24 h and the patches are dried in oven at 40-45°C for about 30minute to remove the residual
solvents. The patches were removed and cut into the circular disc with cm.
● These patches were wrapped in aluminum foil and stored in designator for evaluation.

42
Table 6.3: Physical Evaluation of Transdermal Patches

Working
Drug Release (%)
Formula

1 32.18

2 35.00

3 40.12

Figure 6.1 Comparitive Graph of Drug release

43
EVALUATION:
Thickness Test:
Patches were taken and their thickness was measured by using screw gauge and the
average was calculated.
Folding endurance
The patch was folded several times (100 folds) at a same place until it breaks and the
endurance was noted in terms of number of folds.
Drug Content:
The formulation were cut into pieces and allowed to dissolve in the medium by stirring
with a mechanical stirrer for about 3 h. Then the solution is iltered and analyzed for the drug
content by UV Visible spectrophotometer after suitable dilution.
Franz Diffusion Cell
● The Franz diffusion cells consist of a donor chamber and a receptor chamber,
separated by a membrane.

● The static cell option uses no circulation system of the receptor medium, instead the
medium is stirred using a magnetic stirrer .

● The receptor medium can be any fluid of interest in regard to the observed
substance. Phosphate buffered solution (PBS) is predominately used, since it closely
mimics body fluid in terms of pH and salt contents .

● Typically, the cell is held at a constant temperature of thirty-two degrees Celsius for
skin, or thirty-seven degrees Celsius for body conditions .

● This can be done by using a jacketed cell where water is circulated outside the
receptor chamber . Another option is to use a climate chamber, where humidity, as
well as temperature, can be closely monitored .

● The samples extracted from the donor or receptor medium can be estimated with
different analysis techniques such as UV-VIS spectrophotometry, High Performance
Liquid Chromatography (HPLC) and liquid scintillation counting (LSC) .

● For the first two methods, the concentration of the substance is derived from a
previously composed standard calibration curve.

● When using the liquid scintillation counting technique, the investigated substance
is radio labelled and the amount of substance is measured by a scintillator counter

44
45
REPORT:

DISCUSSION

REFERENCE:.
1. Dey, Sanjoy & Banerjee, Upashana ,Sen, Tamaghna , Shankar, Ved. (2011).
Formulation and in vitro evaluation of transdermal matrix patches of diclofenac
sodium. Journal of Pharmacy Research. 4,11_712.

46
 Table 7.1: Angle of Repose
S.No. Radius(cm) Height(cm) Ø=tan (h/r)

1. 3.7 2.0 28° .36’

2. 3.8 2.0 27°.74’

3. 3.6 2.0 29°.03’

Average 28°.38’

Table 7.2: Bulk density

S. No. Mass(g) Bulk volume(g/cc) Bulk density (g/cc)

1. 20.006 38 0.526

2. 20.000 35 0.571

3. 20.004 36 0.555

Average 0.551

Table 7.3: Tapped Density

S. No. Mass(g) Tapped volume(ml) Tapped density(g/cc)

1. 20.006 36 0.555

2. 20.000 34 0.588

3. 20.004 35 0.571

Average 0.571

47
Experiment No.:7 Date :
PREFORMULATION STUDIES OF GRANULES
AIM:
To carry out Preformulation studies of Granules.
PRINCIPLE
The primary objective of preformulation studies is to provide data and information with
regard to a drug substance and manufacturing technology prior to initiating plants for
formulation development activities and produt design for a drug product. Powder flow and
packing of granules in the die cavity are the key requirements for pharmaceutical manufacturing
process specifically in solid dosage form. Tablets are often manufactured on a rotary multistation
tablet press by filling the tablet die with powders or based on volume .thus flow of powder from
the hopper into the dies often examines,weight,hardness,amd content uniformity of tablets. They
are methods used to measure the powder flow. The compendial methods measurement of angle
of repose,tapped density,Carrs compressibility index, Hausner ratio by carrying out
preformulation studies the characterisation of physical, chemical, mechanical properties of new
drug molecule can be done in order to develop safe, effective and stable dosage form
PROCEDURE:
I) PRE-COMPRESSION EVALUATION PARAMETERS:
a) Angle of repose.
b) Bulk density.
c) Tapped density.
d) Hausner’s Ratio.
e) Compressibility index (%).
II) DRUG POLYMER INTERACTION STUDY: a) FTIR studies

48
Hausner’s Ratio:

= 0.555
0.526
= 1.055

Percentage Compressibility:

0.555-0.526
= X100
0.526
= 5.22%

FTIR Graph: Drug

49
A) ANGLE OF REPOSE (Θ):
The frictional forces in a loose powder or granules can be measured by the angle of
repose. This is the maximum angle possible between the surface of a pile of powder or granules
and the horizontal plane. tanθ = h/r θ = tan-1 (h/r) Where, θ is the angle of repose, h is the
height, r is the radius.
METHOD:
A funnel was filled to the brim and the test sample was allowed to flow smoothly though
the orifice under gravity. From the cone formed on a graph sheet was taken to measure the area
of pile, there by evaluating the flow ability of the granules. Height of the pile was also measured

B)BULK DENSITY

Bulk density is defined as the mass of a powder divided by the bulk volume. The bulk
density of a powder depends primarily on particle size distribution, particle shape and the
tendency of the particles to adhere to one another.

METHOD:
Both loose bulk density (LBD) and tapped bulk density (TBD) were determined. A
quantity of accurately weighed powder (bulk) from each formula, previously shaken to break
any agglomerates formed was introduced into a 25ml measuring cylinder. After the initial
volume was observed, the cylinder was allowed to fall under its own weight on to a
hard surface from the height of 2.5cm at 2 sec interval. The tapping was continued until no
further change in volume was noted.
Weight of the Powder LBD (Loose Bulk Density) = Volume of Packing
Weight of the Powder TBD (Tapped Bulk Density) = Tapped Volume of Packing

50
FTIR Graph: Excipient

C)TAPPED DENSITY:
The measuring cylinder containing a known mass of blend was tapped for a fixed time.
The minimum volume (Vt) occupied in the cylinder and the weight (M) of the blend was
measured. The tapped density (λt) was calculated using the following formula

D)HAUSNER’S RATIO:
It is an indirect index of ease of powder flow. It is calculated by the following formula Hausner’s
ratio = d When t is the tapped density and b is bulk density. Lower H(1.25) Hausner Ratio =Vt
Vb
E)PERCENTAGE COMPRESSIBILITY:
Percent compressibility of powder mix was determined by Carr’s compressibility index
calculated by following formula.

Where,
LBD = Loose Bulk Density
TBD = Tapped Bulk Density
Grading of the powders for their flow properties according to Carr’s Index

51
 Consolidation Index (Carr%) Type of Flow

5– 15 Excellent
12– 16 Good
18– 21 Fair to passable

FTIR Studies Drug Excipient Compatibility:

Infrared absorption spectrophotometry (FT-IR) was performed for the drug, excipients
and the drug excipient mixture and spectrum was compared with reference spectrum to identity
the drug excipient interaction

52
REPORT:

DISCUSSION:

REFERENCE:
1. C.V.S Subrahmanyam. Laboratory manual of physical pharmaceutics pg no.24.
2. Guru Prasad Mohanta. Textbook of physicla pharmacy pratical pg no.22.
53
Table 8.1: Composition of 250 mg Paracetamol tablets
Working Formula For 30
S.No. Ingredients Master Formula (mg)
tablets (g)

1. Paracetamol 0.250 7.5

2. Lactose 0.082 2.46

3. Starch 0.056 1.68

4. Talc 0.004 0.12

5. Magnesium Stearate 0.008 0.24

6. 5% Starch Mucilage QS QS

54
Experiment No.8 Date:

EFFECT OF COMPRESSIONAL FORCE ON TABLETS

DISINTEGRATION TIME
AIM:
To study the variation in disintegration time due to effect of compression forces on
tablets.
REQUIREMENTS:
APPARATUS REQUIRED:
Mortar and Pestle, Sieve #10 and 22, Beaker, Weighing balance, Tablet Punching machine and
Hot air
MATERIALS REQUIRED:
Paracetamol, Lactose, Starch, Magnesium stearate, Talc and distilled water.
PRINCIPLE:
Tablet production involves the application of pressure (i.e., compression pressure) to a
powder bed, compressing it into a cohesive compact though a process known as compaction.
Compaction is a simultaneous process of compression and consolidation of a particulate solid–
gas system as a result of applied force. While compression involves a reduction in bulk volume
as a result of reduced gaseous phase with an increase in pressure within a powder bed, thereby
resulting in closer packing of the powder particles, consolidation occurs simultaneously as force
is increased to a point where further rearrangement becomes difficult and particle deformation
occurs, leading to an increase in mechanical strength as a result of particle–particle interactions
which result in bond creation. Consolidation is mainly responsible for increasing the mechanical
strength of a powder bed under a rising compressive force.

55
Table 8.2: Hardness and Disintegration Time of Tablets Prepared by Various
Compressional Force
Hardness (kg/cm2) Disintegration time ( mins)
S.No.
7 RPM 10 RPM 15 RPM 7 RPM 10 RPM 15 RPM

1. 4.0 5.2 6.7 05’06’’ 06’10’’ 08’20’’

2. 4.2 5.5 6.8 05’09’’ 06’30’’ 08’10’’

3. 4.6 5.6 6.2 05’10’’ 06’45’’ 08’40’’

56
PROCEDURE:

WET GRANULATION METHOD:

● Paracetamol and Lactose were weighed and mixed well.
● Then it is granulated with 5% Starch mucilage.
● And the obtained blend is passed though sieve #10 and kept for drying at 55ᵒC for 1 h.
● Dried granules are then passed though sieve #22 and then to this weighed quantity of talc and
magnesium stearate is added and mixed.
● Obtained granules are compressed using tablet compression machine by varying the
compressional force.
The prepared tablets were evaluated for various parameters such as hardness and
disintegration test to determine the compresssional force in tableting process.

REPORT:

DISCUSSION:

REFERENCES:
1. Adeleye OA. Relationship between compression pressure, mechanical strength and release
properties of tablets. Polim Med. 2019; 49(1):27–33.
2. Olutayo.A.Adeleye, Mbangn.Femi-Oyewo, Michaela Odeniyi. Effect of compression
pressure on mechanical and release properties of tramadol matrix tablets. Current issues in
Pharmacy and Medical sciences, 2015; 28(2): 120-125.

57
Table 9.1: Evaluation of powders for micormetric properties
∑ Wn= 0.49
Weight Cumulative
Weight of
U=h/t Cumulativ %frequency
dry Wn
Time Heigh e weight under size
samples (cm/
(t) t (h) ∑ Wn under size
per 20 ml sec) C X 100
(C)
suspension
∑ Wn

0 30 0.05 10.20 0 d= 2.57µm 0.05 10.20

10 29 0.07 14.28 2.9 0.12 24.48

20 28 0.09 18.36 1.4 0.21 42.85

30 27 0.18 36.73 0.89 0.39 79.59

40 26 0.10 20.40 0.63 0.49 100

Density of ZnO = 5.60g/cc

Density of water = 1 g/cc

Acceleration due to gravity (g) = 981 cm/s 2

Viscosity of medium = 1cp (or) 0.01g. cm-1. s-1

58
Experiment No.9 Date:

MICROMERITRIC PROPERTIES OF POWDER BY ANDREASEN

PIPETTE (SEDIMENTATION TECHNIQUE)
Aim:
To determine the average particle size and particle size distribution by Andreasen pipette
(Sedimentation techniques)

REQUIREMENTS
Andreasen pipette, weighing balance, Sample (zinc oxide), Liquid medium (water),
Deflocculating agent (sodium citrate)
PRINCIPLE
Particle size determination and distribution study using Andreasen pipette utilizes the
principle of sedimentation (Stoke’s law)
Stoke’s law Rate of settling = h/t

= ( )g
18η
Where
d = diameter of the particle

= denisty of solid which is dispersed in the liquid

= density of liquid in which the solid is dispersed

g = Acceleration due to gravity
η = viscosity of medium
h = height of falling
t = time of falling

Diameter can be calculated by the above equation after rearranging stoke’ law

59
60
PROCEDURE

● A suspension of 5.5g zinc oxide and 2.75g of sodium citrate in 50ml of water was prepared
● The suspension was diluted to 550ml with water to give a final concentration of 1%.
● Suspension were added to the vessel, stoppered and shaken to distribute the particles
uniformly thoughout the suspension.
● Apparatus fitted with the pipette in place is clamped securely, preferably in a constant
temperature bath.
● 20 ml samples are withdrawn and regular time period and measure the height of falling.
● Withdrawn samples are evaporated and weighed.

REPORT
The average particle size found using Andreasen Pipette is. μm

DISCUSSION

REFERENCE:

1. Guru Prasad Mohanta, Prabal Kumar Manna, Physical pharmacy practical text, B.S.
Publications. 2ND Edition. 2007.

61
Table 10.1: Evaluation of powders for micormetric properties
Sieve size Mean value Weight % wr Cumulative Cumulative
retained Over size under size

10(2mm) 1.59 0 0 0 99.98

16(1.18mm) 1.01 10.04 10.14 10.14 89.84

20(0.84) 0.71 56.22 56.78 66.92 33.06

30(0.59) 0.54 18.69 18.87 85.79 14.19

Pan 14.05 14.19 99.98 0

Total 99.05 99.98

Mean Opening Value =0.96

% WR = Weight retained X100

Total weight retained
Calculation:

Average diameter = ∑ (% WR X Mean opening)

100
= ∑ (99.98 X 0.96)
100
= 0.959µm

62
 Experiment No.10 Date:

DETERMINATION OF PARTICLE SIZE AND SIZE DISTRIBUTION

USING SIEVING METHOD
AIM:
To determine the average particle size and their distribution pattern of the given granules.

REQUIREMENTS
Sample granules, series of sieves and mechanical sieve shaker.

PRINCIPLE
Sieve is a perforated screen and the size of the opening depends on the number of meshes
and the diameter of the wise constituting the mesh .A sieve will classify the particle as less than
the dimension of mesh /under size and more than the dimension of mesh (over size).

PROCEDURE
● About 5 Sieves are arranged keeping one above the other in a series with the coarsest at the
top and finest at the bottom.
● Around 100g of pre weighed granules is placed on the top sieve and the sieves are shaken for
about 20 mins using sieve shaker.
● The quantity of sample retained on each sieve is weighed and the average particle size is
determined.

63
64
REPORT

DISCUSSION

REFERENCE
1. Guru Prasad Mohanta, Prabal Kumar Manna, Physical pharmacy practical text, B.S.
Publications. 2ND Edition. 2007.

65
 Granule Packing Tensile Particle Porosity Friability
size (µm) fraction (Pf) strength density (%) index (%)
(T)MNm-2 (g.cm-3)

212 0.853 1.48 1.253 9.57 1.77

500 0.877 1.44 1.249 12.95 1.15

710 0.893 1.42 1.160 14.77 0.98

850 0.958 1.37 1.159 16.75 0.97

1700 0.960 1.35 1.082 17.57 0.93

Experiment No.11 Date:

66
 EFFECT OF PARTICLE SIZE ON DISSOLUTION OF TABLETS

AIM:
To study the effect of particle size on the dissolution of Paracetamol tablets.

PRINCIPLE:
The particle size distribution (PSD) of the drug substance may have significant effects on
final drug product performance (e.g., dissolution, bioavailability, content uniformity, stability,
etc.). Methods for determination of particle size can be divided into direct and indirect [9].
Among the direct methods, optical and image analysis are of major interest. The indirect
methods are: sieving, sedimentation, fluid classification, and scanning. Dissolution testing is a
required test, currently used to demonstrate the performance of all solid oral dosage forms in
which absorption of the drug is necessary for the product to exert a therapeutical effect.
Dissolution is defined as the process by which a known amount of drug substance goes into
solution at a given time under standardized conditions. In pharmaceutical terms, dissolution is a
process by which the drug released from the oral form, dissolved in the gastrointestinal tract and
absorbed into the systemic circulation.

PROCEDURE:
PREPARATION OF PARACETAMOL TABLETS:
20% (w/v) starch mucilage was used to wet mass 200 g sample of paracetamol powder in
a planetary mixer to form a crumbly cohesive mass. The wet mass was passed though a
laboratory sieve of aperture size 1.7 mm and dried at 60°C for 1 h on a tray in the hot air oven.
The mass was passed though another sieve of aperture size 710 µm and finally dried at 60°C for
5 h to a moisture content of 2.9 ± 0.3%w/w. Moisture content was determined by a balance fitted
with a sample heater After drying, the granules were stored in an air tight desiccator activated
with dry silica gel before further evaluation. 60°C for 5 h to a moisture content of 2.9 ±
0.3%w/w. Moisture content was determined by a balance fitted with a sample heater. After
drying, the granules were stored in an air tight desiccator activated with dry silica gel before
further evaluation.

67
68
SIZE CLASSIFICATION OF THE GRANULES:

The granules obtained were fractionated into five different sizes by shaking for 10 min
with a nest of sieves mounted on a sieve shaker. The following sieve sizes were employed as
follows: 1700, 850, 710, 500 and 212 µm, respectively. The weight of granules retained on each
sieve was determined.

PREPARATION OF THE TABLETS:

The single punch tableting machine was used for the preparation of the tablets from the
various size fractions. A 1% dispersion of magnesium stearate in chloroform was used to
lubricate the die and punch surfaces prior to the compression of the tablets. A sample (500 mg)
of the granules was filled into the die and compressed into tablets .A sample (500 mg) of the
granules was filled into the die and compressed into tablets at an arbitrary pressure.

DETERMINATION OF TABLETS PACKING FRACTION

The packing fraction of the tablets from each size fractions was calculated from the
particle densities of the tablet compositions. The packing fraction Pf was computed using the
following equation (Itiola and Pilpel, 1986)

Pf = W/πr2tl

Where, W is mean weight of the tablets, r is the radius, t is the tablet thickness and is the
particle density using fluid displacement method.

EVALUATION OF THE TABLET TENSILE STRENGTH (T):

The load (P) required to cause diametral fracture in a tablet was determined using the
Monsanto hardness tester . Ten tablets randomly selected from a given size fractions were used
for the determination. The mean fracture load was used to calculate the tensile strength (T) using
the expression below (Fell and Newton, 1970).

T = 2P/πDt

Where, t and D represent the thickness and diameter of the tablet respectively. Triplicate
determinations were carried out for the various size fractions and mean determinations are
reported.

69
70
VALUES OF TABLET FRIABILITY:

This determination was carried out using the Erweka friabilator (GMBH Heusenstamin,
Germany). Ten tablets randomly selected each from a given size fraction were weighed and
placed in the friabilator. These were rotated for 10 min at 20 rev.min 1. The tablets were dusted of
adherent particles and then reweighed and the percentage friability which was taken as the
friability index was calculated.

REPORT:

DISCUSSION:

REFERENCE:

1. Eichie, F. E and Kudehinbu, A. O. Effect of particle size of granules on some mechanical

properties of paracetamol tablets African Journal of Biotechnology 2009. 8 (21), 5913-
5916.

71
 Table 12.1: Composition of 100mg Aceclofenac tablets by varying binders

Master Master Working Working Working

Master Formula Formula Formula
Formula Formula
S.No. Ingredients Formula 2 1 for 30 2 for 30 3 for 30
1 3
(mg) tablets tablets tablets
(mg) (mg) (g) (g) (g)

01 Acelclofenac 100 100 100 3 2.34 2.34

02 Mannitol 78 78 78 2.34 2.34 2.34

03 Starch mucilage - 10 - - 0.3 -

04 Acacia - - 10 - - 0.3

Polyvinyl
05 10 - - 0.3 - -
Pyrrolidine

06 Starch 4 4 4 0.12 0.12 0.12

Magnesium 0.12 0.12 0.12

07 4 4 4
Stearate

08 Talc 4 4 4 0.12 0.12 0.12

Total Weight 200 200 200

72
Experiment No.:12 Date:
EFFECT OF VARIOUS BINDERS ON DISOLUTION OF TABLETS

AIM
To study the effect of various binders on dissolution of tablets.
REQUIREMENTS:
APPARATUS REQUIRED:
Dissolution apparatus(Paddle type),Weighing balance,beaker,Pipette,10 ml standard
flask, sieves #10 and 22,Hot air oven and UV Spectrophotometer.

CHEMICALS REQUIRED:
Aceclofenac, Mannitol, PVP, Starch, Acacia, Magnesium Stearate and Talc.

PRINCIPLE:
Binders are employed to convey the cohesiveness to the granules and they are added to impact
plasticity and increase interparticulate bonding strength in the tablet and ensure tablet remains
intact after compression. Binders also give volume to low active dose tablets. Rate of dissolution
can be increased by increasing the surface area of the available drug by various methods like
micornization, complexation and solid dispersion. Dissolution of a drug can also be influenced
by disintegration time of tablets

PROCEDURE:
WET GRANULATION METHOD:
● Weighed quantity of the Aceclofenac and Mannitol were mixed thoroughly And to the
above mixture different binder solutions were added and the resulting cohesive mass is
passed though sieve #10.
● Obtained granules were dried at 45ºC for 1 h in hot air oven.
● Dried granules are then passed though sieve # 22 and then to this weighed quantity of
Magnesium stearate and Talc were added and mixed well.
Obtained granules are then compressed using tablet compression machine

73
Table 12.2: Invitro Dissolution Study: USP TYPE- II (Paddle)
Polymer: Polyvinyl Pyrrolidine
Tablet Formulation: 100mg Aceclofenac tablet
Analyte: Aceclofenac
Tablet Dose: 100 mg
Wavelength: 275 nm
Calibration Curve Parameters:
Slope: 0.080 (Enter the slope from the calibration curve here)
Intercept: 0.1743 (Enter the intercept from the calibration curve here)
Assay Dilution Factor: 10 X (Dilution factor: 1:10)
Volume of the 900 ml
Dissolution Medium:

Drug Amount of Cumulative

% Drug
S. Absorbance Concentrati Drug % Drug
Time (min) Release
No. at 275 nm on Release Release
(µg/ml) (mg)
At pH Buffer 7.4
1 30 0.071 0.6 5.4 10.8 10.8
2 1 0.138 1.3 11.7 23.4 23.4
3 1.3 0.231 2 18 36 36
4 2 0.307 2.6 23.4 46.8 46.8
5 2.3 0.396 3 27 54 54
6 3 0.445 4.2 37.8 75.6 75.6
7 3.3 0.503 5 45 90 90

100
Cumulative % Drug release

90
80
70
60
50
40
30
20
10
0
0 30 1 1.3 2 2.3 3 3.3
Time (Hours)

Figure 12.1 In-vitro Drug Release of Aceclofenac tablets

74
In-vitro Dissolution Study:
● Dissolution test for the tablet samples was performed using USP - Type II paddle type.
● The parameters like temperature and speed was adjusted to 37 ±0.5 0c and 50 RPM
respectively.
● The drug dissolution studies was carried out at followed by the Acid buffer pH 1.2 with
the medium of 900 ml in the hemispherical basket.
● The samples were withdrawn at specified time intervals, diluted with respective buffer
and scanned for absorbance at UV spectrophotometer.
● The tablet samples were placed into individual baskets and the apparatus was operated
at the above specified condition.
● Specified volume of sample was withdrawn from the zone midway between the surface
of the disso medium and top of the rotating paddle at different time intervals.
● Sink condition was maintained by replacing equivalent volume of fresh medium each
time of withdrawal of the sample.
● Samples withdrawn were diluted if necessary and analyzed using UV spectrophotometer
at 257 nm.
● The percentage drug release was calculated from the obtained data and studied for
kinetic parameters.

Table 12.3: Invitro Dissolution Study: USP TYPE- II (Paddle)

75
Polymer: Starch
Tablet Formulation: 100mg Aceclofenac tablet
Analyte: Aceclofenac
Tablet Dose: 100 mg
Wavelength: 275 nm
Calibration Curve Parameters:
Slope: 0.080 (Enter the slope from the calibration curve here)
Intercept: 0.1743 (Enter the intercept from the calibration curve here)
Assay Dilution Factor: 10 X (Dilution factor: 1:10)
Volume of the Dissolution 900 ml
Medium:

Drug Amount of Cumulativ

% Drug
S. Absorbance Concentrati Drug e % Drug
Time (min) Release
No. at 275 nm on Release Release
(µg/ml) (mg)
At pH Buffer 7.4
0.018 0.3 2.7 5.4 5.4
1 30
0.082 0.8 7.2 14.4 14.4
2 1
0.122 1.1 9.9 19.8 19.8
3 1.3
0.198 1.6 14.4 28.8 28.8
4 2
2.3 0.235 2 18 36 36
5
0.289 2.4 21.6 43.2 43.2
6 3
0.337 2.9 26.1 52.2 52.2
7 3.3

Figure 12.2 In-vitro Drug Release of Aceclofenac tablets

76
Table 12.4: Invitro Dissolution Study: USP TYPE- II (Paddle)

77
Polymer: Acacia

Tablet Formulation: 100mg Aceclofenac tablet

Analyte: Aceclofenac
Tablet Dose: 100 mg
Wavelength: 275 nm
Calibration Curve Parameters:
Slope: 0.080 (Enter the slope from the calibration curve here)
Intercept: 0.1743 (Enter the intercept from the calibration curve here)
Assay Dilution Factor: 10 X (Dilution factor: 1:10)
Volume of the Dissolution 900 ml
Medium:

Drug Amount of Cumulativ

% Drug
S. Absorbance Concentrati Drug e % Drug
Time (h) Release
No. at 275 nm on Release Release
(µg/ml) (mg)
At pH Buffer 7.4
1 30 0.028 0.2 1.8 3.6 3.6

2 1 0.041 0.5 4.5 9 9

3 1.3 0.092 0.8 7.2 14.4 14.4

4 2 0.147 1.1 9.9 19.8 19.8

5 2.3 0.189 1.5 13.5 27 27

6 3 0.226 1.8 16.2 32.4 32.4

7 3.3 0.251 2 18 36 36

Figure 12.3 In-vitro Drug Release of Aceclofenac tablets

REPORT:
78
DISCUSSION

REFERENCES:

1. Setty, Chitral & Radhika, Muthadi & Gupta, V & Reddy, M & Aukunuru, Jithan. Effect of
Tablet Processing and Formulation Factors on Disintegration and Dissolution of Aceclofenac
Tablets. International Journal of Pharmaceutical Sciences and Nanotechnology. 2011; 3(4):
1240-1251.
2. Laxmi, M. Vijaya and Vamshi T Krishna. Formulation and evaluation of aceclofenac matrix
tablets using ethyl cellulose and cellulose acetate phthalate. JGTPS/ 5(3)-(2014) 1804-1810.

Calculation:

79
 TIME
Rt Tt (Rt- Tt) (Rt- Tt)2
(MIN)
0 3.6 3.6 0 0
15 18.8 16.2 2.6 6.76
30 26.5 21.5 5.0 25.0
45 94.0 58.8 35.2 1239.04
60 34.1 32.6 1.5 2.25
Σ Rt =177 Σ(Rt- Tt)=44.3 Σ(Rt- Tt)2 = 1273.05

n = number of dissolution time points

Rt = dissolution value of the Reference drug products at time t
Tt = dissolution value of the Test drug products at time t

80
Experiment No.13 Date:

STUDY OF SIMILARITY FACTOR

AIM:
To study Similarity factor calculation for the given test and reference data.

PRINCIPLE:
Comparison of therapeutic performances of two medicinal products containing the same
active substance is a critical means of assessing the possibility of alternative using between the
innovator and any essentially similar medicinal product. Assuming that in the same subject an
essentially similar plasma concentration time course will result in essentially similar
concentration at the site of action and thus in an essentially similar effect, pharmacokinetic data
instead of therapeutic results may be used to establish equivalence and bioequivalence.
A bioequivalence study is basically a comparative study designed to establish equivalence
between test and reference products. In vivo bioequivalence studies are needed when there is a
risk that possible differences in bioavailability may result in therapeutic in equivalence. But it’s
also possible to request a waiver of in vivo bioavailability and/or bioequivalence studies for solid
oral dosage forms based on an approach termed.

The Bio pharmaceutics Classification System (BCS).

Comparison of Dissolution Profiles FDA Guidelines:
Drug absorption from a solid dosage form after oral administration depends on the release
of the drug substance from the drug product, the dissolution or solubilization of the drug under
physiological conditions,and the permeability across the gastrointestinal tract. Because of the
critical nature of the first two steps, in vitro dissolution may be relevant to the prediction of in
vivo performance.
The BCS can be used as a basis for setting in vitro dissolution specifications and can also
provide a basis for predicting the likelihood of achieving a successful in vivo-in vitro correlation
(IVIVC).

81
Calculation:

= 44.30 x 100
177
f1 = 25.02

= 50 log {[ 1+ 1/5 Σ 5t=1 (1273.05)2]-0.5 x 100}

=50 log {[ 1+ 254.61]-0.5 x 100}

= 50 log {[ 255.61]-0.5 x 100}

=50 log {[0.0625] x 100}

= 50 log 6.25

= 50 x 0.795

f2 = 39.75

82
Based on drug solubility and permeability, the BCS is recommended Case
1. Case 1: High Solubility – High Permeability.
2. Case 2: Low Solubility – High Permeability.
3. Case 3: High Solubility – Low Permeability.
4. Case 4: Low Solubility – Low Permeability
The BCS suggests that for high solubility, high permeability drugs and in some instances
for high solubility, low permeability drugs, 85 % dissolution in 0.1 N HCl in 15 minutes can
ensure that the bioavailability of the drug is not limited by dissolution. In the case of low
solubility / high permeability drugs, drug dissolution may be the rate limiting step for drug
absorption and an IVIVC may be expected. A dissolution profile in multiple media is
recommended for drugs products in this category. In the case of high solubility / low
permeability drugs, permeability is the rate controlling step and a limited IVIVC may be
possible. The dissolution profile comparison may be carried out using model independent or
model dependent methods: A simple model independent approach uses a difference factor (f1)
and a similarity factor (f2) to compare dissolution profiles.

There is a specific procedure to determine difference and similarity factors as follows:

1. Determine the dissolution profile of two products (12 units each) of the test and
reference products.
2. Using the mean dissolution values from both curves at each time interval, calculate the
difference factor (f1) and similarity factor (f2) using the above equations. For curves to be
considered similar f1 values should be close to 0 and f2 values should be close to 100. Generally,
f1 values up to 15 (0-15) and f2 values greater than 50 (50- 100) ensure sameness or equivalence
of the two curves.
Difference Factor f1 Similarity Factor f2 Interference

0 100 Dissolution Profile is Identical

≤15 ≥50 Similarity or equivalence of two profile

83
84
REPORT:

REFERENCES:
1. M. C. Gohel*, K. G. Sarvaiya, A. R. Shah and B. K. Brahmbhatt. Mathematical approach
for the assessment of similarity factor using a new scheme for calculating weight. Indian
J Pharm Sci. 2009; 71(2):142-144.
2. Shah, Vinod & Tsong, Yi & Sathe, Pradeep & Williams, Roger. Dissolution Profile
Comparison Using Similarity Factor, f2. Dissolution Technologies. 1999; 6. 15-15.

85
DIFFERENT TYPES OF KINETIC MODELS

86
Experiment No.14 Date:
DETERMINATION OF DRUG RELEASE KINETICS PLOT
AIM:
The release behaviour of dosage forms will be analysed based on in-vitro drug release
data by using DD solver excel add in software.
PRINCIPLE:
DRUG RELEASE:
It is a process of drug which leaves from a dosage form and subjected to have a
pharmacological action.
➢ Immediate release (IR)
➢ Sustained release (SR)
➢ Controlled release (CR)
DRUG RELEASE KINETICS:
It is a application of mathematical model to assess the drug release pattern.
Laws in the kinetics of drug release
Noyes-Whitney law: Noyes-Whitney developed the fundamental principle of evaluation of drug
release, which explains as
dC/dt=K (Cs -Ct )
Where dC/dt is dissolution rate of drug,
K is dissolution rate constant,
Cs is the concentration of drug in the stagnant layer (also called as the saturation or maximum
drug solubility),
Ct is the concentration of the drug in the bulk of the solution at time t.
Ficks law of diffusion : According to this law drug molecules diffuse from a region of higher
concentration to one of lower concentration until equilibrium is attained and that the rate of
diffusion is directly proportional to concentration gradient across the membrane. This can be
expressed mathematically as
dQ/dt=DAKm/w(CGIT- C)/h
Where dQ/dt is rate of drug diffusion, amount per time (rate of appearance of drug in blood), D
is the diffusion coefficient. Its dimension is area per unit time, so typical units for expressing it
would be m2 /s (area/time),
A is the surface area for absorbing membrane for drug diffusion (area),

87
Zero Order Kinetics:
USP Dissolution Test Apparatus II (Paddle)

Tablet Formulation:
Analyte:
Tablet Dose: mg
Wavelength: nm
Calibration Curve Parameters:
Slope: (Enter the slope from the calibration curve here)
Intercept: (Enter the intercept from the calibration curve here)
Assay Dilution Factor: X (Dilution factor: 1:1)
Volume of the Dissolution Medium: ml
Time Absorbance Drug Concentration Amount of Drug Release % Drug Release
(min) at nm ( µg/ml ) (mg) (mg)

88
Km/w Partition coefficient of drug between lipoidal membrane and aqueous GI fluid (no unit),
CGIT- C is the difference in the concentration of drug in the GI fluid and the plasma called
concentration gradient (amount per volume),
h is the thickness of the membrane (length)
The methods of approach to investigate the kinetics of drug release from controlled
release formulation can be classified into thee categories:

● Statistical methods (exploratory data analysis method, repeated measures design,

multivariate approach [MANOVA: multivariate analysis of variance].
● Model dependent methods (zero order, first order, Higuchi, Korsmeyer-Peppas model,
Hixson Crowell, etc.).
● Model independent methods [difference factor (f1), similarity factor].

MATHEMATICAL MODELLING OF DRUG RELEASE KINETICS:

To provide a particular drug release profile from a therapeutic system, it is necessary to
know the exact mass transport mechanisms involved in the drug release and to predict
quantitatively the resulting drug release kinetics. Many mathematical models have been used to
design a number of simple and complex drug delivery systems and to predict the overall release
behaviour. They allow the measurement of some important physical parameters and resorting to
model fitting on experimental release data. It is very important to know how to use these
equations to understand the different factors that affect the release velocity and how the
Dissolution behaviours can vary and influence the efficiency or the therapeutic regimen of
patients.
INTERPRETATION OF DIFFUSIONAL RELEASE MECHANISMS

Application of drug release data on mathematical models:

Several mathematical equations which generally define the dissolution profile. Once an
appropriate function has been selected, the evaluation of dissolution profile can be carried out
and hence the drug release profile can be correlated with drug release kinetic models. Various
mathematical models are employed to understand drug release kinetics which is explained
below.

89
First Order Kinetics:
USP Dissolution Test Apparatus II (Paddle)
Tablet Formulation:
Analyte:
Tablet Dose: mg
Wavelength: nm

Calibration Curve Parameters:

Slope: (Enter the slope from the calibration curve
here)
Intercept: (Enter the intercept from the calibration curve
here)
Assay Dilution Factor: X (Dilution factor: 1:1)
Volume of the Dissolution ml
Medium:

Amount of
Drug Cumulative % Log of Cumulative
Time Absorbance Drug
Concentration Release % Drug Release
(min) at nm Release
( µg/ml ) (mg) (mg)
(mg)

90
91
Higuchi ‘s:
USP Dissolution Test Apparatus II (Paddle)
Tablet Formulation:
Analyte:
Tablet Dose: mg
Wavelength: nm
Calibration Curve Parameters:
Slope: (Enter the slope from the calibration curve here)
Intercept: (Enter the intercept from the calibration curve
here)
Assay Dilution Factor: X (Dilution factor: 1:1)
Volume of the Dissolution ml
Medium:

Log of
Drug Amount of
Cumulative % Cumulative √ Time
Time Absorbance Concent Drug
Release % Drug (Sq.R.Time)
(min) at nm ration Release
(mg) Release (min)
( µg/ml ) (mg)
(mg)

92
93
Peppas:
USP Dissolution Test Apparatus II (Paddle)
Tablet Formulation:
Analyte:
Tablet Dose: mg
Wavelength: nm
Calibration Curve Parameters:
Slope: (Enter the slope from the calibration curve
here)
Intercept: (Enter the intercept from the calibration
curve here)
Assay Dilution Factor: X (Dilution factor: 1:1)
Volume of the Dissolution ml
Medium:

Log of
Drug Amount of Cumulative
Cumulative %
Time Absorbance Concent Drug %
Release Log Time
(min) at nm ration Release Drug( Releas
(mg)
( µg/ml ) (mg) e)
(mg)

94
REPORT:

REFERENCE :

1. Srinatha, A., Pandit, J. K., & Singh, S. (2008). Ionic Cross-linked Chitosan Beads for
Extended Release of Ciprofloxacin: In vitro Characterization. Indian journal of
pharmaceutical sciences, 70(1), 16–21.

95
2. Dash, S., Murthy, P. N., Nath, L., & Chowdhury, P. (2010). Kinetic modeling on drug
release from controlled drug delivery systems. Acta poloniae pharmaceutica, 67(3), 217–
223.

96