SALIENT FEATURES:

INTRODUCTION
AIMS AND OBJECTIVES
LITERATURE REVIEW
PROFILE OF DRUG
LIST OF MATERIALS AND
EQUIPMENTS
EXPERIMENTAL WORK
RESULT AND DISCUSSION
SUMMARY AND CONCLUSION

11/20/15

IT IS A BCS
CLASS IV
DRUG
LOOP DIURETIC
AGENT

FUROSEMIDE HAS
LOW
PERMEABILITY
AND SOLUBILITY

FUROSEMIDE
USED ORALLY IN THE
TREATMENT OF
EDEMATOUS STATES
ASSOCIATED WITH
CARDIAC, RENAL, AND
HEPATIC FAILURE AND
THE TREATMENT OF
HYPERTENSION.

11/20/15

ATTEMPTS HAVE BEEN

MADE TO DEVELOP
SYSTEMS WHICH
INCREASE BOTH THE
SOLUBILITY AND
PERMEABILITY OF
FUROSEMIDE.

11/20/15

HIGHER
PERMEABILITY
THAN SKIN

ORAL TRANSMUCOSAL
DELIVERY IS AN
ALTERNATE TO
SYSTEMIC DELIVERY OF
DRUGS

BUCCOADHESIVE
DRUG DELIVERY
BUCCAL MUCOSA
HAS HIGHER
BLOOD SUPPLY,
AVOIDS FIRST
PASS
METABOLISM
11/20/15

BUCCAL PATCHES AND FILMS HAVE

BEEN FORMULATED FOR DRUGS
LIKE
LIDOCAINE
MICONAZOLE
ACYCLOVIR
CLOTRIMAZOLE
SUMATRIPTAN
FOR EITHER LOCAL OR SYSTEMIC
DELIVERY.
4

HYPOTHESIS
It is hypothesized that a buccoadhesive film of
furosemide will help in improving bioavailability*
* Avoidance of first pass metabolism
*Through incorporation of chemical permeation
enhancer
*Through
improving
the
dissolution
by
complexation with hydroxypropyl--cyclodextrin

11/20/15

AIMS

11/20/15

OBJECTIVES
To prepare inclusion complex of furosemide.
To develop a buccoadhesive film dosage form for
furosemide.
To evaluate the buccoadhesive film for permeation
enhancement via transbuccal route.
To study the effect of chemical permeation enhancer on
the bioavailability enhancement of furosemide via
transbuccal route.
To study the effect of inclusion complex of furosemide
on the bioavailability enhancement of furosemide via
transbuccal route.

11/20/15

LITERATURE REVIEW

18.5.11 correction\W CORR\3. Literature review.

docx
18.5.11 correction\W CORR\REFERNCES.docx

11/20/15

PROFILE OF DRUG
Generic Name and Additional Name:
Furosemide / Frusemide
Emperical formula: C 12H11ClN2O5S
Structural formula:
Molecular weight: 330.77
Solubility: practically
Insoluble in water
Sparingly soluble in alcohol, freely soluble
acetone

11/20/15

BIOPHARMACEUTICS
DATA
PHARMACOKINETIC DATA

11/20/15

HALF LIFE

2 hours (30120 min )

PROTEIN BINDING

95 % (ALBUMIN)

ABSORPTION WINDOW

UPPER GIT

METABOLISM

HIGHER FIRST PASS EFFECT

BIOAVAILABILITY

60-70% (ERRATIC)

10

LIST OF MATERIALS
Sr. No
1

Material
Furosemide (gift sample)

Suppliers / Manufacturers
Sanofi aventis, Ankleshwar

Chitosan

Himedia Laboratories Pvt. Ltd., Mumbai (India)

Ethylcellulose

Loba chemie, Mumbai, India

PVP K 30 (poly vinyl pyrollidone)

Himedia Laboratories Pvt. Ltd., Mumbai (India)

Sodium glycocholate

Himedia Laboratories Pvt. Ltd., Mumbai (India)

Acetone

Sulab Reagents, Suvidhinath Laboratories, Baroda

(India)

Acetic Acid

Merck, India

Absolute alcohol

Shree Chalthan Vibhag Khand Udyog Sahakari

Mandli Ltd., Surat (India)

Potassium dihydrogen phosphate

Loba chemie, Mumbai, India

10

Sodium hydroxide

Loba chemie, Mumbai, India

11

Sodium chloride

Loba chemie, Mumbai, India

12

Calcium chloride

Loba chemie, Mumbai, India

*All other reagents and chemicals used were of laboratory grade.

11/20/15

11

LIST OF EQUIPMENTS
Sr. No
1

Equipments
Magnetic stirrer

Model/ Manufacturers
MS-500 Remi instruments Ltd. (India)

Digital balance

ELB300 Shimadzu, Japan

Tray dryer

DTC 303 Riddhi Trading Company, Ahmedabad,

India

Propeller stirrer

RQ 122 Remi Instruments Ltd. (India)

UV spectrophotometer

UV-1700 Shimadzu, Japan

FTIR

(fourier

transformed

infrared) NICHOLET 6700, Thermo Scientific (Germany)

spectrometer
7

X ray diffractometer

D2 Phaser, Bruker AXS Inc. (Germany)

Diffrential scanning calorimetry

Pyris 1 DSC, PerkinElmer, USA

Texture analyser

Brookefield QTS-25, Germany

11/20/15

12

EXPERIMENTAL WORK

11/20/15

Q
UI
C
K
O
VE
RV
IE
W
13

PRELIMINARY
EXPERIMENTS

11/20/15

14

Drug
Characterization

FTIR

MAX in Water,
Ethanol
0.1 N NaOH
MELTING
POINT

219-222 0 C
11/20/15

229 nm,
278 nm,
330 nm
15

-10

-20

4000

11/20/15
3500
3000
2500
2000

1 1 4 3 .2 1

1 2 4 1 .1 7

50

1500
1000

cm-1

Functional Group
Reported Frequencies (cm-1)
Observed Frequencies (cm-1)

NH stretch of C-NH group

3350-3400
3351.46

C=O stretch of COOH group

1671
1672.78

NH stretch of NH2 group

1596
1592.88

-S=O stretch of SO2 group

1322
1323.70

Cl stretch of C-Cl group

582
581.66

16

58 1. 66

53 6. 79

49 5. 05

60

43 2. 01

1 0 7 2 .6 0 1 0 5 3 .5 0
1 0 1 5 .2 9
97 9. 33
94 4. 25 91 9. 30
84 6. 83 78 9. 77
74 3. 65 65 1. 39
70 8. 3568 6. 74
61 9. 36

-30

1 2 6 5 .5 2

1 3 2 3 .7 01 4 0 9 .7 2

1 3 5 3 .7 6

1 4 4 5 .7 5

1 4 9 1 .2 0

1 5 6 4 .4 8

2 5 5 6 .4 9

40

1 5 9 2 .8 8

3 1 1 8 .9 8

20

2 9 6 0 .5 0

10

3 3 9 8 .8 8
3 3 5 1 .4 6
3 2 8 4 .0 3

30

1 6 7 2 .7 8

%T
100
90

80

70

-40

-50

-60

-70

500

 Calibration curve
at pH 6.2 and 7.4

Simulated salivary fluid

(1.632 gm KH2PO4, 2.34
gm of NaCl and 0.1257
gm of CaCl2 dissolved in
1 L of distilled water
adjusted up to pH 6.2
with 0.2 M NaOH
solution )

pH 6.2

pH 7.4
Isotonic
phosphate buffer

Pre-formulation
study
11/20/15

Drug
excipient
compatibility
study
IR
FT

17

PREPARATION OF
INCLUSION COMPLEX

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18

QUICK
OVERVIEW

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19

PHASE SOLUBILITY STUDY

Different mM concentration of HP-CD solution 5 ml with excess of drug 50
mg

INCUBATOR SHAKER

0.22
filter

Time:24hours
Filtrate

0.03

UV
analysis
at 229
nm

0.01

0.04

0.02

Calculate KS
(stability
constant) and CE
(complexation
efficiency)

0.05

RPM = 50 TEMP: 37 2 0 C
11/20/15

20

PREPARATION OF
INCLUSION COMPLEX

1. Farcas, A.; Jarroux, N.; Farcas, A.-M.; Harabagiu., V.; Guegan, P., Synthesis and characterization of
furosemide complex in -Cyclodextrin. Digest Journal of Nanomaterials and Biostructures 2006, 1 (2),
55-60.
11/20/15

21

70 0C
Drug solution
Acetone

COOLING

CYCLODEXTRIN
SOLUTION

Precipitates

Distilled water

EVALUATION
OF INCLUSION
COMPLEX
11/20/15

22

RESULTS AND DISCUSSION

0

Cocentratio
n of
furosemide
(M)

RATIONALE FOR SELECTION OF

EXCIPIENTS
HP--CD (hydroxypropyl-beta-cyclodextrin)
Non-toxic at low to moderate oral and
intravenous doses as indicated by
Brewster et al1
More water-soluble than the natural cyclodextrin
PHASE SOLUBILITY STUDY :
KS = 130.01 M-1 and CE= 0.1574

0
0
0
0
0
0
0
0
0
0

Concentration of HP--CD (M)

0

1. Brewster, M. E.; Loftsson, T., Cyclodextrins as pharmaceutical solubilizers. Advanced

Drug Delivery Reviews 2007, 59 (7), 645-666..
11/20/15

23

DSC THERMOGRAPH OF
FUROSEMIDE

FIGURE-DSC THERMOGRAPH OF FUROSEMIDE

11/20/15

24

DSC OVERLAY THERMOGRAPH

Figure- Overlay DSC thermographs of a) Furosemide; b) HP--CD; c)

physical mixture of furosemide: HP--CD (1: 1.5) d) furosemide: HP--CD
complex (1:1.5)
11/20/15

25

FTIR OVERLAY SPECTRA

Figure: FTIR spectra for (a) furosemide; (b) HP--CD; (c) physical mixture
of furosemide: HP--CD (1:1.5); (d) inclusion complex furosemide: HP--CD
(1:1.5)
11/20/15

26

XRD DIFFRACTOMETRY

Figure: X ray diffractograms of (a) furosemide; (b) HP--CD; (c) their

physical mixture (1:1.5; drug: HP--CD) and (d) inclusion complex (1:1.5;
drug: HP--CD
Similar results were found by Spamer et al1for complexation of furosemide with HP--CD.
1. Spamer, E.; Mller, D. G.; Wessels, P. L.; Venter, J. P., Characterization of the complexes of furosemide with 2-hydroxypropyl[beta]-cyclodextrin and sulfobutyl ether-7-[beta]-cyclodextrin. European Journal Of Pharmaceutical Sciences 2002, 16 (4-5), 247-253.

11/20/15

27

IN VITRO DISSOLUTION PROFILE

MEDIUM: pH 7.4 buffer; Method: USP Apparatus II (paddle); Volume

of Dissolution : 900 mL Temperature: 37 2 0C ;RPM:50
11/20/15

28

SUMMARY

11/20/15

Hydroxypropyl--cyclodextrin has better solubilizing

property.
Phase solubility study was performed to know the
stoichiometric proportions of furosemide and HP-CD
and to know stability constant of the
complexation process.
Complexation efficiency was also evaluated.
Inclusion complex in molar ratio of 1:1.5 was
prepared using co-precipitation method.
Inclusion complex was evaluated using DSC, FTIR,
XRD and in vitro dissolution study.
29

CONCLUSION
In conclusion, the formation of inclusion complex is a
good approach for enhancement of solubility of
furosemide.
The formation of inclusion complex can be evaluated
further using SEM (scanning inclusion microscopy),
H-1 NMR (nuclear magnetic resonance) to get
complete information about the complex formed.
Also, different methods for formation of inclusion
complex can be explored.
Though freeze drying of the final product is an
expensive method it can be a good alternative to get
better inclusion complex product.

11/20/15

30

DEVELOPMENT OF
BUCCOADHESIVE FILMS
OF FUROSEMIDE

11/20/15

31

QUICK
OVERVIEW

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32

PREPARATION OF
MUCOADHESIVE
LAYER

REMI MOTORS

1
4

SGC

CHITOSAN

2
PVP K30

3
5

GLYCEROL

DRUG
SOLUTION

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33

TRAY
DRYER

MIXTURE

PETRY DISH

DRIED FILMS

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34

PREPARATION
OF BACKING
ETHYLCELLULOSE 300mg
LAYER
+
0.5 mL GLYCERINE

AIR DRYING
ACETONE

PETRI DISH

11/20/15

35

EXPERIMENTAL DESIGN
A 22 factorial design was employed to study
the effect of 2 independent variables at 2
levels. Independent variables: concentration
of permeation enhancer (SGC) and
concentration of chitosan and dependent
variable: steady state flux JSS.
Table 1 indicated the actual values and coded/
transformed values of independent variables.

11/20/15

36

Table 1: Levels of independent variables for 22 factorial design

for optimization of formulation
Independent Variables

Levels
Coded Values

Actual Values
(%)

Concentration of chitosan (%)* x1

-1

55

85

Concentration

-1

of

Sodium

glycocholate (%)** x2
Dependent Variables

JSS Flux

*The concentration of chitosan constitutes as % of total polymer weight

** The concentration of Sodium glycocholate is taken as % of total polymer weight.
Y= b0 + b1x1+ b2x2 + b3 x1x2

Here,

Y is the measured response of the dependent variable

x11, x22 are the coded levels of independent variables.
x11 x22 represent the interaction terms.
b00, b11, b22, b33 are regression coefficients of the respective variables and their interaction terms
computed from the observed experimental values of Y.
11/20/15

37

Table 2: 22 full factorial design for preparation of multipolymeric

buccoadhesive bilayer films
Levels
Coded values

Batch code

Actual values

Chitosan

SGC

Chitosan

SGC

x1

x2

x1

x2

P1

+1

-1

85

P2

+1

+1

85

P3

-1

-1

55

P4

-1

+1

55

*Total weight of polymer =a= 600 mg

Weight of chitosan for +1 and -1 level, c1= 510 mg and c2= 330 mg
Weight of sodium glycocholate for -1 and +1 level, d 1= 5 mg and d2= 30 mg
Weight of PVP K30, b1= a- c1 and b2= a-c2
= % weight of chitosan = c1 100 / a and c2 100 / a
= % weight of sodium glycocholate = d1 100 / a and d2 100 / a
***Rest all parameters of formulation were kept constant viz. volume of 0.5 % acetic acid = 25 ml; volume of
glycerine= 0.5 ml; volume of ethanol= 15 ml; stirring speed of propeller stirrer= 750 rpm; diameter of Petri dish= 7072 mm.
11/20/15

38

 IN VITRO PERMEATION
STUDY

Done using ovine buccal mucosa (sheep, ovis aeries) 1.

A mucosa membrane that can cover an area above 2.54 cm 2 was cut
and supported on dialysis membranes, which was applied on the
receptor chamber of a Franz diffusion cell.
Buccoadhesive bilayer film was applied on the mucosa, in such a way
that the mucoadhesive layer facing the mucosal surface.
1 ml of simulated salivary fluid, pH 6.2 was filled in the donor
chamber and isotonic phosphate buffer pH 7.4 was used in the
receptor chamber.
At regular time interval of 15, 30, 45, 60, 90 and 120 minutes and then
every hour up to 8 hours 200 L sample was withdrawn from the
receptor chamber and replaced by isotonic phosphate buffer pH 7.4.
The samples were assayed by the UV spectrophotometer at 229 nm.

11/20/15

39

CALCULATION OF STEADY STATE FLUX

(JSS) AND ENHANCEMENT RATIO

11/20/15

The amount of drug present in the receptor compartment

was determined and plotted as a function of time. The
permeability coefficients (P) were calculated from the
linear part of the curves as follows (Mashru et al2):
P = (dQ / dT ) /A* Cd
Where, A= the surface area of diffusion = 1 cm2, dQ/dt
=Jss= steady state flux = amount of drug permeated
per unit time at steady state, and Cd = drug
concentration in the donor solution.
Here, as furosemide is an acidic drug, having pKa value of
3.8 it remains in ionized form in the pH 7.4.
So, transport of furosemide is assumed to be via paracellular path and so the steady state flux for paracellar
path is considered. Jss = P * Cd (Mashru et al2).
40

ENHANCEMENT RATIO

The permeability of furosemide in presence of sodium glycocholate

was evaluated by enhancement ratio. It was calculated as (Shojaei et
al 3) :
Enchancement Ratio = P sgc/ P fur
Where, PSGC = Permeability coefficient in presence of SGC and
PFUR = Permeability coefficient in absence of any permeation
enhancer .

1. Patel, V.; Prajapati, B.; Patel, M., Design and characterization of chitosan-containing
mucoadhesive buccal patches of propranolol hydrochloride. Acta Pharmaceutica 2007, 57 (1),
61-72.
2. Mashru, R. C.; Sutariya, V. B.; Sankalia, M. G.; Sankalia, J. M., Effect of pH on In Vitro
Permeation of Ondansetron Hydrochloride Across Porcine Buccal Mucosa. Pharm Dev Technol
2005, 10 (2), 241-247.
3. Shojaei, A. H.; Berner, B.; Li, X., Transbuccal Delivery of Acyclovir: I. In Vitro
Determination of Routes of Buccal Transport. Pharmaceutical research 1998, 15 (8), 1182-1188.
11/20/15

41

11/20/15

42

EVALUATION OF FILM
1. Physical characterization thickness and folding endurance
2. Swelling index and Surface pH1
3. Mechanical properties- Tensile strength and elongation at break2
4. Mucoadhesion studies- Mucoadhesive strength2 and
Mucoadhesion time3
5. In vitro drug release study 4and Kinetics of drug release5
1.
2.
3.
4.
5.

Yehia, S. A.; Gazayerly, O. N. E.; Basalious, E. B., Fluconazole Muccoadhesive Buccal Films : In vivo / In
vitro Performance. Curr Drug Deliv 2009, 6, 17- 27.
Mura, P.; Corti, G.; Cirri, M.; Maestrelli, F.; Mennini, N.; Bragagni, M., Development of Mucoadhesive
Films for Buccal Administration of Flufenamic Acid: Effect of Cyclodextrin Complexation. Journal of
pharmaceutical sciences 2010, 99 (7), 3019-3029.
Hassan, N.; Ali, M.; Ali, J., Development and Evaluation of Novel Buccoadhesive wafers of nimodipine
for treatment of hypertension Drug Deliv 2010, 17 (2), 59-67.
Adhikari, S.; Nayak, B.; Nayak, A.; Mohanty, B., Formulation and Evaluation of Buccal Patches for
Delivery of Atenolol. AAPS PharmSciTech 2010, 11 (3), 1038-1044.
Coasta, P.; Lobo, J. M. S., Modelling and comparison of dissolution profiles. European Journal Of
Pharmaceutical Sciences 2001, 13, 123-133.

11/20/15

43

1.
2.
3.

4.
5.

6.

11/20/15

Thickness : vernier calipers.

Swelling study: 1cm2 patch placed in 25 ml of SSF in a petri dish.
Mechanical properties: texture analyser- Film strips of dimension 3
cm x 1 cm were cut and held between two clamps positioned at
distance of 20 mm. The pulley was pulled by top clamp at a rate of
5 mm/min to a distance of 5 cm before returning to starting point.
The force and elongation was measured as the films broke.
Mucoadhesive time
Mucoadhesive strength: thin films were cut out in 2.25 cm2. A
piece of sheep mucosa was fixed to a support. The film was fixed to
the upper support and wetted with simulated salivary fluid (pH 6.2)
50 L. The upper support was lowered at speed of 1mm/ min to
contact with the tissue at a force of 1 N for a contact time of 30
seconds. It was then withdrawn at a rate of 1 mm/min upto a
distance of 5 mm. The force needed for detaching the film from the
tissue was used to evaluate the bioadhesive strength of the films.
In vitro drug release study: Franz diffusion cell

44

Brookefield
11/20/15

Texture Analyser for tensile strength study

45

Assembly for mucoadhesion study

11/20/15

46

RESULTS AND DISCUSSION

RATIONALE FOR SELECTION OF EXCIPIENTS

Chitosan :
biodegradable, natural polymer, non toxic and
mucoadhesive, biocompatible and cationic polymer.
excellent gel forming and film forming properties; so it is a
good candidate for mucoadhesive polymer.( Senel et al1)
It is used for the formulation of mucoadhesive tablets,
patches, films and gels for buccal use (Miller et al 2)

1. Senel, S.; Kremer, M. J.; Ka, S.; Wertz, P. W.; H1ncal, A. A.; Squier, C. A., Enhancing
effect of chitosan on peptide drug delivery across buccal mucosa. Biomaterials 2000, 21 (20),
2067
2. Salamat-Miller, N.; Chittchang, M.; Johnston, T. P., The use of mucoadhesive polymers in
buccal drug delivery. Advanced Drug Delivery Reviews 2005, 57,1666-1691.
11/20/15

47

SELECTION OF BLEND OF
POLYMERS

Over hydration of chitosan may lead to slippery mucilage.

So, to avoid mucoadhesion failure other polymers are
added to the films. 1
Various scientists have used PVP K30 as a drug release
modifying polymer for buccal films and patches. 2

1. El-Kamel, A.; Ashri, L.; Alsarra, I., Micromatricial metronidazole benzoate film as a
local mucoadhesive delivery system for treatment of periodontal diseases. AAPS
PharmSciTech 2007, 8 (3), E184-E194.
2. Patel, V.; Prajapati, B.; Patel, M., Effect of hydrophilic polymers on buccoadhesive
eudragit patches of propranolol hydrochloride using factorial design. AAPS PharmSciTech
2007, 8 (2), E119-E126.
11/20/15

48

SELECTION OF PERMEATION ENHANCER

AND BACKING LAYER POLYMER

Various classes of chemical

permeation enhancers are used
like bile salts, terpenes, chitosan,
cyclodextrins,
surfactants,
medium chain fatty acids, azone
etc.
Criteria:

least toxic to the buccal mucosa,

reversible type of effect on mucosa
GRAS status

Sodium glycocholate :
it has been explored as
permeation enhancer for
delivery
of
morphine
sulphate 1, acyclovir 2.
Ethylcellulose has been used as
backing layer extensively 3.

1. Senel, S.; Duchene, D.; Hincal, A. A.; apan, Y.; Ponchel, G., In vitro studies on enhancing effect of
sodium glycocholate on transbuccal permeation of morphine hydrochloride. Journal of Controlled
Release 1998, 51 (2-3), 107-113.
2. Shojaei, A. H.; Berner, B.; Li, X., Transbuccal Delivery of Acyclovir: I. In Vitro Determination of
Routes of Buccal Transport. Pharmaceutical research 1998, 15 (8), 1182-1188.
3. Shidhaye, S. S.; Saindane, N. S.; Sutar, S.; Kadam, V., Mucoadhesive Bilayered Patches For
Administration of Sumatriptan Succinate. AAPS PharmSciTech 2008, 9 (3), 909- 916.
11/20/15

49

STEPS FOR IDENTIFICATION OF FORMULATION

11/20/15

50

PHYSICAL CHARACTERIZATION
Thickness of
Batch code

mucoadhesive
layer (mm)

11/20/15

Thickness of

Folding

bilayer(mm)

endurance

P1

0.373 0.01

0.440 0.01

>100

P2

0.320 0.03

0.526 0.04

>100

P3

0.246 0.01

0.650 0.04

>100

P4

0.210 0.01

0.330 0.02

>100

51

FORMULATION OPTIMIZATION: 22
FULL FACTORIAL DESIGN

The optimized batch was selected (P2) on the basis of

higher JSS.

The steady state flux (JSS) was calculated by plotting a

graph of amount of drug (g) permeated per cm2 verses
time (hours); the straight line of the graph gives us the
value of JSS and Permeability coefficient (Mashru et al1).

1. Mashru, R. C.; Sutariya, V. B.; Sankalia, M. G.; Sankalia, J. M., Effect of pH

on In Vitro Permeation of Ondansetron Hydrochloride Across Porcine Buccal
Mucosa. Pharm Dev Technol 2005, 10 (2), 241-247.
11/20/15

52

Table : Steady state flux and permeability coefficients and enhancement ratio
of Bilayer films (n=3)
Concentration
of chitosan
Batch code

(x1)*

Concentration
of Sodium
glycocholate

JSS (steady

Permeability

(x2)**

state flux)

coefficient

(g/cm2/hr)

(cm/hr)

Enhancement
ratio***

Coded Values

P1

+1

-1

164.63 11.85

0.032 0.002

0.998

P2

+1

199.03 13.79

0.039 0.002

1.209

P3

-1

-1

138.73 10.61

0.027 0.002

0.842

P4

-1

143.13 10.01

0.029 0.001

0.885

*-1 and 1 levels of x1 indicate 55% and 85 % of total polymer weight **-1 and 1 levels
of x2 indicate 1% and 5 % of total polymer weight. *** Compared to film containing
only Base matrix and no permeation enhancer
11/20/15

53

The statistical analysis data through regression model is presented.

Response: JSS - Analysis of variance for selected factorial model

Coefficient

Numerical value

p- value

b0

161.38

0.0008

b1

20.45

0.0003

b2

9.70

0.0204

b3

7.50

0.0567

Regression

0.8625

Model generated, JSS = b0 + b1 x1 + b2 x2 + b3 x1 x2

Polynomial equation for the model,
JSS = 161.38 + 20.45 x1+ 9.70 x2+ 7.50 x1x2
Reduced model: JSS = 161.38 + 20.45 x1+ 9.70 x2
11/20/15

54

RESPONSE SURFACE PLOT

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55

RESPONSE SURFACE PLOT

Figure illustrates the surface response plot for the model depicted.
It shows that chitosan concentration and concentration of SGC has a significant
positive effect on the response JSS (p< 0.05).
This observation depicts that increase in concentration of chitosan as well as SGC
increases the JSS. This observation is consistent with the literature 1-6.

1. He, W.; Guo, X.; Zhang, M., Transdermal permeation enhancement of N-trimethyl chitosan for testosterone. International Journal
of Pharmaceutics 2008, 356 (1-2), 82.
2. Kerec, M.; Bogataj, M.; Verani, P.; Mrhar, A., Permeability of pig urinary bladder wall: the effect of chitosan and the role of
calcium. European Journal of Pharmaceutical Sciences 2005, 25 (1), 113.
3. Maestrelli, F.; Zerrouk, N.; Chemtob, C.; Mura, P., Influence of chitosan and its glutamate and hydrochloride salts on naproxen
dissolution rate and permeation across Caco-2 cells. International Journal of Pharmaceutics 2004, 271 (1-2), 257.
4. Sadeghi, A. M. M.; Dorkoosh, F. A.; Avadi, M. R.; Weinhold, M.; Bayat, A.; Delie, F.; Gurny, R.; Larijani, B.; Rafiee-Tehrani, M.;
Junginger, H. E., Permeation enhancer effect of chitosan and chitosan derivatives: Comparison of formulations as soluble polymers
and nanoparticulate systems on insulin absorption in Caco-2 cells. European Journal of Pharmaceutics and Biopharmaceutics 2008,
70 (1), 270.
5. Zambito, Y.; Uccello-Barretta, G.; Zaino, C.; Balzano, F.; Di Colo, G., Novel transmucosal absorption enhancers obtained by
aminoalkylation of chitosan. European Journal of Pharmaceutical Sciences 2006, 29 (5), 460.
6. Sohi, H.; Ahuja, A.; Ahmad, F. J.; Khar, R. K., Critical evaluation of permeation enhancers for oral mucosal drug delivery. Drug
Dev Ind Pharm 2010, 36 (3), 254-282.
7. Senel, S.; Kremer, M. J.; Ka, S.; Wertz, P. W.; H1ncal, A. A.; Squier, C. A., Enhancing effect of chitosan on peptide drug delivery
across buccal mucosa. Biomaterials 2000, 21 (20), 2067
8. Bonferoni, M. C.; Sandri, G.; Rossi, S.; Ferrari, F.; Caramella, C., Chitosan and its salts for mucosal and transmucosal delivery.
Expert opinion on drug delivery 2009, 6 (9), 923-939.

11/20/15

56

SEM

11/20/15

57

AMOUNT OF DRUG PERMEATED VS TIME FOR

BATCHES P1 P4.
In vitro permeation study

Amo u n t o f d ru g re le ase mcg /cm2

2500

P1
P2

2000

P3
P4

1500

1000

500

0
0

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5
6
Time in hours

10

58

VALIDATION OF THE MODEL

Concentration Concentration
of chitosan
of SGC
70%

3%

Predicted
Jss

Actual
Jss

161.38

137.20

Student t- test (p value = 0.0514)

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59

EFFECT OF INCLUSION COMPLEX

ON PERMEATION ENHANCEMENT

A batch was prepared which contained the same formulation of

P2 but furosemide was incorporated in the form of inclusion
complex with HP-- CD. This batch was coded as P5.

JSS was compared with that of batch P2.

The difference in the JSS value of inclusion complex and free

drug containing film was found to be significant.
Cyclodextrins act as permeation enhancers for transbuccal
route 1.
The effect of cyclodextrins on enhancement of permeability via
buccal route for omeprazole has been studied by Hassan et al 1.

1. Hassan, N.; Ahad, A.; Ali, M.; Ali, J., Chemical permeation enhancers for transbuccal
drug delivery. Expert opinion on drug delivery 2010, 7 (1), 97-112.
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60

Response parameter for free drug and inclusion

complex containing buccoadhesive film
JSS (steady

Permeabilit

state flux)

y coefficient

(g/cm2/hr)

(cm/hr)

P2

199

0.0390

1.209

P5

206.5

0.0413

1.255

Batch code

Enhanceme
nt ratio

*P5= film containing drug in the form of inclusion complex

t- test p-value =0.0117 (p< 0.05)
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61

In vitro permeation study

Amount of drug release mcg/cm2

2500

P2
P5

2000

1500

1000

500

0
0

5
6
Time in hours

10

Comparison of amount of drug permeated versus time for films

containing furosemide (P2) and furosemide complex (P5).
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62

 SWELLING INDEX AND

SURFACE pH

It was also observed the profile for water hydration stops after certain time;
this is because of poor solubility of chitosan in water thus liquid uptake stops
after certain limit. Similar results were found by Rossi et al 30, 31.

1. Rossi, S.; Sandri, G.; Ferrari, F.; Bonferoni, M. C.; Caramella, C., Buccal Delivery of Acyclovir from Films Based on Chitosan
and Polyacrylic Acid. Pharm Dev Technol 2003, 8 (2), 199-208.
2. Hassan, N.; Ali, M.; Ali, J., Development and Evaluation of Novel Buccoadhesive wafers of nimodipine for treatment of
hypertension Drug Deliv 2010, 17 (2), 59-67.
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63

MECHANICAL PROPERTIES OF FILMS

CONTAINING DRUG AND INCLUSION
COMPLEX
Batch code

Tensile strength

Elongation at break

(g/mm2)

(%)

P2

0.19

31.2

P5

0.21

16.6

Student t-test

p= 0.03

p=0.11

Similar result was found by Boateng et al 1

2. Boateng, J. S.; Stevens, H. N. E.; Eccleston, G. M.; Auffret, A. D.; Humphrey, M. J.; Matthews,
K. H., Development and mechanical characterization of solvent-cast polymeric films as potential
drug delivery systems to mucosal surfaces. Drug Dev Ind Pharm 2009, 35 (8), 986-996.

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64

FILM STRENGTH BATCH P2

FILM STRENGTH BATCH P5

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65

MUCOADHESIVE TIME

Also mucoadhesive time for both the batches was

found to higher enough upto 8 hours and above.
This can be attributed to large volume of hydrating
fluid present to hydrate the film and also, the force
applied by thumb to stick the film to the mucosal
surface was higher.
Also hydration of film is good due to presence of PVP
K30 in the matrix.

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66

MUCOADHESIVE STRENGTH
Batch code

Force of
adhesiveness

Adhesiveness

P2
P5

-45 gm
-66gm

2.25 g/sec
8.51 g/sec

Student t-test

0.11

These results were consistent with the findings 1-3.

1. Mura, P.; Corti, G.; Cirri, M.; Maestrelli, F.; Mennini, N.; Bragagni, M., Development of Mucoadhesive
Films for Buccal Administration of Flufenamic Acid: Effect of Cyclodextrin Complexation. Journal of
pharmaceutical sciences 2010, 99 (7), 3019-3029.
2. Hassan, N.; Ali, M.; Ali, J., Development and Evaluation of Novel Buccoadhesive wafers of nimodipine for
treatment of hypertension Drug Deliv 2010, 17 (2), 59-67.
3. Morales, J. O.; McConville, J. T., Manufacture and characterization of mucoadhesive buccal films.
European Journal of Pharmaceutics and Biopharmaceutics 2011, 77 (2), 187-199.
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67

MUCOADHESION STRENGTH -P2

MUCOADHESION STRENGTH P5
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68

 IN VITRO DRUG RELEASE STUDY

KINETICS OF DRUG RELEASE
STUDY
In vitro drug release

% d r u g r e le a s e

100

P2
P5

80
60
40
20
0

10

Time in hours

In vitro drug release profiles for films containing furosemide and

furosemide-HP--CD complex.
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69

In vitro dissolution profile of films containing furosemide P 2 and furosemide- HP-CD complex P5.
Time in hours

% Drug release
P2

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0.25

7.04 0.18

0.50

12.37 0.36

0.75

15.82 0.22

35.69 0.22

38.93 0.38

42.27 0.16

54.75 0.16

68.08 0.61

68.94 0.86

69.19 0.37

69.79 0.80

P5
2.24 0.90
4.90 0.55
8.24 0.60
17.20 1.37
21.76 0.10
28.05 0.91
44.51 0.62
64.53 0.53
66.34 0.64
79.53 0.48
90.95 0.55
70

CORRELATION COEFFICIENTS FOR

DIFFERENT MODELS FOR BATCH P2 AND P5
R2 value (inclusion
Model

R2 value (furosemide) P2

Zero order release model

0.8777

0.9870

First order release model

0.7063

0.7536

Hixson Crowell model

0.9132

0.9605

Higuchis model

0.9450

0.9450

Korsmeyers Peppas model

0.9370

0.9604

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complex) P5

71

KINETICS OF DRUG RELEASE

11/20/15

The higuchis model represents two limit cases in the

transport and drug release phenomenon.
In the drug release phenomenon of the film, the drug
molecules have to diffuse across unstirred aqueous layer
on the membrane surface in the donor compartment
followed by diffusion across semi permeable membrane to
the receptor compartment.
The observed release kinetics indicates that the drug
across the membrane surface is a rate limiting step-in
overall drug release process .
The complex containing film follows Zero order kinetics.
Complex formation facilitated the diffusion of drug across
the membrane and hence increased the dissolution of
drug.
72

SUMMARY

The buccal films were developed using solvent casting method.

The formulation was identified for the backing layer and
mucoadhesive layer.
To know the effect of polymer and permeation enhancer on the
permeability of drug through buccal mucosa a 22 factorial design
was employed to study the effect of 2 independent variables at 2
levels. Independent variables: concentration of permeation
enhancer (SGC) and concentration of chitosan and dependent
variable: steady state flux JSS. Design expert software [trial
version 8.0.4 (www.statease.com)] was used to derive a
polynomial equation for the design.
It was found out that chitosan concentration and concentration of
SGC has a significant positive effect on the response JSS (p< 0.05).

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73

SUMMARY

This observation depicts that increase in concentration of chitosan as

well as SGC increases the JSS.
Next the optimized batch obtained from the above experimental design
was compared with the same formulation of film containing furosemide
in the form of inclusion complex with HP--CD.
Effect of complexation on the permeability of furosemide was evaluated
and it was found that the value of J SS was higher for films containing
inclusion complex.
Both the batches were further evaluated for swelling study, mechanical
properties, in vitro mucoadhesion study, in vitro drug release study and
release kinetics.
The swelling behaviour, mucoadhesion strength, mucoadhesion time did
not show significant difference for both the films. Whereas the tensile
strength was found to be different for both the films.
The drug release was found to be higher in the films containing
inclusion complex and it followed zero order release model.

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74

CONCLUSION
From the above experiments it can be concluded that the permeability of
furosemide can be successfully enhanced by using buccoadhesive films.
Thus the strategy of incorporation of inclusion complex and permeation
enhancer in buccal films can be upcoming new technology for buccal
drug delivery.
Due to the ease of access and avoidance of the hepatic metabolism, oral
transmucosal drug delivery offers a promising alternative over
conventional drug delivery.
For this approach, the properties of film may also be modified by use of
different plasticizers, polymers and polymeric blends.
The stability of the buccoadhesive film should require to be evaluated by
performing stability study.
In addition, the clinical trials of the buccoadhesive films should be done
using animal models and human volunteers for comparison of
pharmacodynamic and pharmacokinetic data to that of conventional
dosage form of furosemide.

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75