Purohit et al Journal of Drug Delivery & Therapeutics.

2019; 9(1):79-84

Available online on 15.01.2019 at http://jddtonline.info

Journal of Drug Delivery and Therapeutics

Open Access to Pharmaceutical and Medical Research
© 2011-18, publisher and licensee JDDT, This is an Open Access article which permits unrestricted
non-commercial use, provided the original work is properly cited

Open Access Research Article

Formulation and evaluation of deflazacort loaded mucoadhesive
microsphere for colon drug delivery system
Akash Purohit*, Mithun Bhowmick, Jagdish Rathi
NRI Institute of Pharmaceutical Sciences, Bhopal (M.P), India

ABSTRACT
Irritable bowel disease is very common colon disease. Deflazacort is one of the best drug with clinical activity against Irritable bowel disease.
Microsphere system are effectively protect drugs against premature degradation, to localize drug molecules at the target site of action and to
control the time and rate of release. Mucoadhesive microspheres enhance the bioavailability of orally given drugs by lengthened contact time of
drug with the intestinal mucosa. The main disadvantage of these microspheres is adherence to the substrate by non-specific interaction. To
overcome this limitation, microspheres are prepared by emulsification method to treat irritable bowel disease. Chitosan microspheres were
prepared by Ionotropic Gelation method. Microspheres were coated with Eudragil S using solvent evaporation method.
Keywords: Deflazacort, Mucoadhesive, Microspheres, Irritable bowel disease

Article Info: Received 02 Sep 2018; Review Completed 29 Sep 2018; Accepted 18 Dec 2018; Available online 15 Jan 2019
Cite this article as:
Purohit A, Bhowmick M, Rathi J, Formulation and evaluation of deflazacort loaded mucoadhesive microsphere for colon
drug delivery system, Journal of Drug Delivery and Therapeutics. 2019; 9(1):79-84
DOI: http://dx.doi.org/10.22270/jddt.v9i1.2164

*Address for Correspondence:

Akash Purohit, NRI Institute of Pharmaceutical Sciences, Bhopal (M.P), India

INTRODUCTION mucoadhesive properties may prolong the residence time at

the site of drug absorption.3-4
Colon was considered as a part of body solely responsible for
absorption of water, electrolyte and temporary storage of MATERIAL AND METHODS
stools, but now it is accepted as an important site for drug
Material
delivery. Targeted delivery ensures the direct treatment at
the disease site, lower dosing, and minimizing side effects. Deflazacort was collected as a gift sample from Alcon bio
Drug targeting to colon is highly desirable for local treatment sciences PVT Ltd, Mumbai. Eudragit-S 100 obtained from
of a variety of bowel diseases. Moreover, dosage forms with Degussa India Pvt Ltd. Sodium Alginate and chitosan were
mucoadhesive properties may prolong the residence time at received from Loba Chemie Pvt. Ltd (Mumbai, India). All
the site of drug absorption. Mucoadhesive microparticulates other reagents and chemicals used were of analytical grade.
coated with a pH-dependent polymer are proposed to
initiate the release of the drug at the putative colonic pH 7– Methods
8.1-3 Deflazacort is a drug of choice in Ulcerative colitis with Preparation of mucoadhesive microsphere of
proven anti inflammatory and immunosuppressive effect. Deflazacort
Deflazacort is a corticosteroid that works by acting within
cells to prevent the release of certain chemicals that are Chitosan microspheres were prepared by ionotropic gelation
important in the immune system. These chemicals are method.
involved in producing immune and allergic responses, Preparation I: Chitosan stock solution (1% w/v) was
resulting in inflammation by decreasing the release of these prepared by dissolving chitosan in acetic acid (1% v/v) at
chemicals in a particular area, inflammation is reduced. room temperature.
Deflazacort has a shorter biological half life of 1.1-1.9 hr.
Thus by using deflazacort microspheres half life is increased Preparation II: The drug and sodium alginate was dissolved
and provide sustained release of drug for longer duration in 100 ml of water.
and thus bioavailability problems associated with oral
Preparation III: 1% Sodium tripolyphosphate solution was
administration is also improved and microspheres with
prepared.

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Purohit et al Journal of Drug Delivery & Therapeutics. 2019; 9(1):79-84

Preparation IV: Solution of preparation I was slowly added dropping rate and falling distance were kept constant. The
in preparation III with continuous starring on magnetic solution was magnetically stirred for half an hour followed
stirrer. by filtration and rinsing with distilled water. Gel like beads
were obtained which was air dried for twenty four hours
Preparation II was added in preparation IV through a
followed by oven drying for six hours at 40˚C. 5-6
disposable syringe needle into a gently agitating. The

Table 1: Formulations of the mucoadhasive microspheres

Chitosan Sod. Alginate
Sr. No Formulation Code Deflazacort (mg)
(mg) (mg)
1. F1 10 25 50
2. F2 10 25 75
3. F3 10 25 100
4. F4 10 50 50
5. F5 10 50 75
6. F6 10 50 100

Coating of mucoadhasive microspheres microspheres still adhering to the tissue was counted. The
test was performed at 0.1N hydrochloric acid solution.
Microspheres were coated with Eudragil S (ES) using solvent
evaporation method. mucoadhasive microspheres (50 mg) % Mucoadhesion = (Na-Nl) / Na × 100
were dispersed in 10 mL of coating solution prepared by
Where, Na = number of microspheres applied; Nl = number
dissolution of 500 mg of ES in ethanol:acetone (2:1) to give
of microspheres leached out.9-10
5:1 (coat:core ratio). This organic phase was then poured in
70 mL of light liquid paraffin containing 1% wt/vol Span 80. Measurement of mean particle size
The system was maintained under agitation speed of 1000
rpm at room temperature for 3 hours to allow for the The mean size of the microspheres was determined by Photo
evaporation of solvent. Finally, the coated microsphere were Correlation Spectroscopy (PCS) on a submicron particle size
filtered, washed with n-hexane, and dried in desiccator.7-8 analyzer at a scattering angle of 90°. A sample (0.5mg) of the
microspheres suspended in 5 ml of distilled water was used
Evaluation of microspheres for the measurement. 11
Percentage Yield Determination of zeta potential
The prepared microspheres with a size range of 200-300nm The zeta potential of the drug-loaded microspheres was
were collected and weighed from different formulations. The measured on a zeta sizer by determining the electrophoretic
measured weight was divided by the total amount of all non- mobility in a micro electrophoresis flow cell. All the samples
volatile components which were used for the preparation of were measured in water at 25°C in triplicate.9-11
the microspheres.5-6
Shape and Surface Characterization of Microspheres by
Scanning Electron Microscopy (SEM)
From the formulated batches of microspheres, formulations
Drug Entrapment (F3) which showed an appropriate balance between the
percentage releases were examined for surface morphology
The various formulations of the mucoadhasive microspheres
and shape using scanning electron microscope Jeol Japan
were subjected for drug content. 10 mg of mucoadhasive
6000. Sample was fixed on carbon tape and fine gold
microspheres from all batches were accurately weighed and
sputtering was applied in a high vacuum evaporator. The
crushed. The powder of microspheres were dissolved in 10
acceleration voltage was set at 10KV during scanning.
ml 7.4 pH Phosphate Buffer and centrifuge at 1000 rpm. This
Microphotographs were taken on different magnification and
supernatant solution is than filtered through whatmann
higher magnification (200X) was used for surface
filter paper No. 44. After filtration, from this solution 0.1 ml
morphology.10-12
was taken out and diluted up to 10 ml with 7.4 pH Phosphate
Buffer. The percentage drug entrapment was calculated In-vitro Release Studies
using calibration curve method.7-8
In vitro drug release in gastrointestinal fluids of
In-vitro wash off test different pH
The mucoadhesive property of microspheres was evaluated The prepared microsphere was evaluated for in vitro drug
by an in vitro adhesion testing method known as the wash- release. The drug release studies were carried out using USP
off test. Freshly excised pieces of intestinal mucosa from XXII paddle type Dissolution test apparatus. The dissolution
sheep were mounted onto glass slide. About 100 study was carried out in 900 ml dissolution medium which
microspheres were spread onto wet rinsed tissue specimen was stirred at 100 rpm maintained at 37±0.2C. The scheme
and immediately thereafter the slides were hung onto the of using the simulated fluids at different timing was as
arm of a tablet disintegrating machine. Then the machine follows:
was operated. The tissue specimen was given a slow, regular
up and down movement in the test fluid at about 37°C  1st hour: Simulated gastric fluid (SGF) of pH 1.2.
contained in a vessel of the machine. At the end of 1, 2, 3, 4, 5,
6, 7,8 hrs the machine was stopped and the number of  2nd and 3rd hour: Mixture of simulated gastric and
Intestinal fluid of pH 4.5.

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Purohit et al Journal of Drug Delivery & Therapeutics. 2019; 9(1):79-84

 4th to 5th hour: Simulated intestinal fluid (SIF) of pH 6.8. Figure 1: Percentage Yield for different formulation
Drug Entrapment
 6th hour and onward: SIF pH 7.5
The drug entrapment of different formulations was in range
A weighed quantity of formulation (100 mg) was spread over of 78.05- 83.25% w/w. This is due to the permeation
the surface of dissolution media (900 ml) at 37±0.2C. characteristics of HPMC that could facilitate the diffusion of
Samples were withdrawn at different time interval and part of entrapped drug to surrounding medium during
compensated with same amount of fresh dissolution preparation of deflazacort microspheres.
medium. Volume of sample withdrawn was made up to 10ml
by continuous media. The samples withdrawn were assayed Table 3: Drug Entrapment for Different Formulation
spectrophotometrically at 242.0 nm for deflazacort and
Drug entrapment (% w/w)
using UV visible spectrophotometer. The release of Formulation of prepared microsphere
deflazacort was calculated with the help of Standard curve of
deflazacort.9-12 F1 78.05±0.25
F2 79.98±0.32
Drug release kinetic data analysis F3 82.56±0.14
Several kinetic models have been proposed to describe the F4 80.12±0.57
release characteristics of a drug from matrix. The following F5 81.14±0.54
three equations are commonly used, because of their F6 83.25±0.45
simplicity and applicability. Equation 1, the zero-order
model equation (Plotted as cumulative percentage of drug
released vs time); Equation 2, Higuchi’s square-root equation
(Plotted as cumulative percentage of drug released vs square
root of time); and Equation 3, the Korsemeyer-Peppas
equation (Plotted as Log cumulative percentage of drug
released vs Log time).10-12
RESULTS AND DISCUSSION
Evaluation of deflazacort microspheres
Percentage Yield
Percentage yield of different formulation was determined by
weighing the Microspheres after drying. The percentage
yield of different formulation was in range of 82.56 –
93.58%.
Table 2: Percentage Yield for Different Formulation

Formulation Percentage Yield

Figure 2: Drug Entrapment for Different Formulation
F1 82.56±0.45
F2 83.56±0.25 The maximum Percentage Yield, Drug Entrapment was found
F3 89.98±0.32 to be formulation F3. The optimized formulation of batches
F4 85.56±0.45 subjected to further studies.
F5 88.85±0.47 Results of In-vitro wash off test
F6 93.58±0.85
Table 4: Results of In-vitro wash off test
Formulatio 1hr 2hr 4hr 6hr 8hr
n code
F1 78 74 68 65 61
F2 81 75 69 65 60
F3 92 85 80 78 75
F4 85 78 70 65 60
F5 78 70 65 60 58
F6 75 68 60 55 45

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Purohit et al Journal of Drug Delivery & Therapeutics. 2019; 9(1):79-84

Figure 3: Graph of Results of In-vitro wash off test

Particle size analysis

The mean size of the microspheres was determined by photo
correlation spectroscopy (PCS) on a submicron particle size
analyzer (Horiba Instruments) at a scattering angle of 90°. A
sample (0.5mg) of the microspheres suspended in 5 ml of
distilled water was used for the measurement. The results of
measurement of mean particle size of optimized formulation
F3 microsphere was found 150.9 nm respectively.

Figure 5: Zeta potential data of mucoadhesive

microsphere
Scanning Electronic Microscopy
Shape and surface characteristic of Deflazacort microspheres
exam in by Scanning Electronic Microscopy analysis. Surface
morphology of formulation examines at two different
magnification 55X which illustrate the smooth surface of
mucoadhesive Microspheres.

Figure 4: Particle size data of mucoadhesive

microsphere
Zeta Potential
The zeta potential of the drug-loaded microspheres was
measured on a zeta sizer (Malvern Instruments) by
determining the electrophoretic mobility in a micro
electrophoresis flow cell. All the samples were measured in
water at 25°C in triplicate. Results of zeta potential of
optimized formulation F3 microsphere was found -10.7 mV
respectively.
Figure 6: Scanning Electronic Microscopy image of
optimized formulation F-3

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Purohit et al Journal of Drug Delivery & Therapeutics. 2019; 9(1):79-84

In Vitro drug release

Table 5: Cumulative % drug release of Deflazacort from plain and coated microsphere at different pH

S. No. Dissolution medium Time (hrs) % Cumulative Drug Release

Microsphere Coated microsphere
1 SGF (pH 1.2) 1 4.8 0.3
2 2 7.2 0.7
3 3 13.0 1.0
4 SGF+SIF(pH 4.5) 4 18.9 4.2
5 5 25.7 9.7
6 6 37.0 16.2
SIF (pH 6.8)
7 7 58.4 34.3
8 SIF (pH 7.5) 8 66.7 45.5
9 9 73.1 53.1
10 10 82.3 62.0
11 12 88.6 67.8

80 2.5

Log cumulative Percent Drug

% Cumulative Drug release

70
2
60
50 Remaining 1.5
40
COATED 1 COATED
30
20 0.5
10
0
0
0 1 2 3 4 5 6 7 8 9 10 11 12
0 1 2 3 4 5 6 7 8 9 10 11 12
Time (Hrs) TIme (Hrs)

Figure 7: Cumulative Percent Drug Released Vs Time Figure 8: Log Cumulative Percent Drug Remaining Vs
(Zero Order Plots) Time (First Order Plots)

2.5
Log Cumulative Percent Drug

2
Released

1.5

1 COATED

0.5

0
0 0.2 0.4 0.6 0.8 1 1.2
Log Time
Figure 9: Log Cumulative Percent Drug Released Vs Log Time (Peppas Plots)

Table 6 Regression Analysis Data of mucoadhasive microspheres Formulation

Formulation Zero order First order Pappas plot
F3 y = 6.540x - 10.65 y = -0.048x + 2.123 y = 2.406x - 0.587
R² = 0.944 R² = 0.932 R² = 0.975

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Stability studies REFERENCES

Stability studies were carried out with optimized 1. Van den Mooter G. Colon drug delivery. Expert Opin Drug Deliv
formulation which was stored for a period of 45 days at 2006;3:111-25
4±1°C, RT and 40±1°C. The particle size of formulation was 2. Sarasija S, Hota: A colon specific drug delivery system. Indian J
determined by optical microscopy using a calibrated ocular Pharmaceutical Sci.. 2000; 62:1-8.
3. Asane, G.S. “ Mucoadhesive gastrointestinal drug delivery
micrometer. The particle size of the microsphere was found
system.” Pharmainfo.net -2007, 5(6). 1-2.
to increase at RT, which may be attributed to the aggregation 4. Baumgart DC: Treatment of inflammatory bowel disease: A
of microsphere at higher temperature. At 452°C the review of medical therapy. World J.Gastroenterol. 2008;
microsphere aggregate i.e. these microsphere were unstable 21:354-377
at higher temperature like 452°C. Percent efficiency of 5. Patel, J.K. et al. “Formulation and evaluation of mucoadhesive
mucoadhesive Microspheres also decrease at higher Glipzide microspheres”. AAPS PharmsciTech-2005. 06 (1), E
temperature Like 452°C. 49-E50.
6. Das MK, Senapati PC. Evaluation of furosemide-loaded alginate
CONCLUSION microspheres prepared by ionotropic external gelation
technique. Acta Pol Pharm 2007;64:253-62.
From the result of present study, it can be concluded that 7. Hori M, Onishi H, Machida Y. Evaluation of Eudragit-coated
chitosan based deflazacort mucoadhesive microspheres offer chitosan microparticles as an oral immune delivery system. Int
a high degree of protection from premature drug release in J Pharm. 2005; 297:223–234
simulated upper GIT conditions and deliver most of the drug 8. Thakral NK, Ray AR, Majumdar DK. Eudragit S-100 entrapped
chitosan microspheres of valdecoxib for colon cancer. J Mater
load in the colon and allow drug release to occur at the Sci:Mater Med. 2010;21:2691–2699
desired site. Based on the results of the physicochemical 9. Paharia, A. et al “Eudragit-coated pectin microspheres of 5-
characterization and in vitro drug release studies, it Fluorouracil for colon targeting”. AAPS ParmsciTech-2007. 8
possessed all the required physicochemical characters and (1), E1-E7.
with drug releases up to 12 h where it released 67.8 % of the 10. Khan MZ, Stedul HP, Kurjaković N. A pH-dependent colon-
deflazacort. Thus, chitosan based microspheres are a targeted oral drug delivery system using methacrylic acid
potential system for colon delivery of deflazacort for copolymers. II. Manipulation of drug release using eudragit
Ulcerative colitis. L100 and eudragit S100 combinations. Drug Dev Ind Pharm
2000;26:549-54
11. Rahman, Z. et al, “Characterization of 5-FU microspheres for
colon delivery”. AAPS PharmSciTech-2006. 7 (2), 47-53.
12. Akbuğa J, Bergişadi N. Effect of formulation variables on cis-
platin loaded chitosan microsphere properties. J Microencapsul
1999; 16:697-703.

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