Indo American Journal of Pharmaceutical Research, 2014 ISSN NO: 2231-6876

FORMULATION AND EVALUATION OF MUCOADHESIVE MICROSPHERES OF

VALSARTAN
Akram Khan*, Dr.S.K.Umadevi, Dr.B.Venkateswara Reddy, K.Navaneetha, R.Swetha Reddy, M.Ravi,
K.Revanth Chandra
Department of Pharmaceutics, St.Pauls College of Pharmacy, Turkayamjal (V), Hayathnagar (M), R.R.Dist-501510.

ARTICLE INFO ABSTRACT

Article history The objective of the current investigation is to reduce dosing frequency and improve patient
Received 04/09/2014 compliance by designing and systematically evaluating mucoadhesive microspheres of
Available online valsartan. Frequent administration and variable low bioavailability (20-25%) after oral
20/09/2014 administration are problems of conventional dosage forms of valsartan. This can be
attenuated by designing it in form of mucoadhesive microspheres which would prolong the
residence time at the absorption site to facilitate intimate contact with absorption surface and
there by improve and enhance bioavailability. Valsartan microspheres were formulated by
Keywords orifice–ionic gelation technique using combination of three polymers i.e. sodium alginate,
Valsartan, carbopol, and HPMC. Optimised formulation F9 which was formulated by taking a
Conventional Dosage, combination of 3 polymers sodium alginate, carbopol and HPMC in 1:1 ratio showed good
Mucoadhesive Microspheres, mucoadhesion property and entrapment efficiency of (87.21%).The scanning electron
Orifice–Ionic Gelation microscopy indicated that microspheres were spherical in shape and drug remained dispersed
Technique, in polymer matrix. The invitro drug release mechanism was non-fickian type controlled by
Bioavailability. swelling and relaxation of polymer. Thus it can be concluded that mucoadhesive
microspheres prepared by using sodium alginate, carbopol and HPMC in 1:1 ratio, were
found to be promising to reduce the dosing frequency with enhanced patient compliance.

Corresponding author
Akram Khan
M.Pharmacy,
Department of Pharmaceutics,
St.Pauls College of Pharmacy, Turkayamjal (V),
Hayathnagar (M), R.R.Dist-501510.
akkykhan78@gmail.com ,
8885840655

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Please cite this article in press as Akram Khan et al. Formulation and Evaluation of Mucoadhesive Microspheres of Valsartan.
Indo American Journal of Pharm Research.2014:4(09).

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Research, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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Vol 4, Issue 09, 2014. Akram Khan et al. ISSN NO: 2231-6876

INTRODUCTION
Mucoadhesive drug delivery system is a new system of drug delivery and has recently gained great concern in
pharmaceutical sciences. The concept of muco adhesives was introduced in the early 1980s. Muco adhesion is the relatively new
and emerging concept in drug delivery. Muco adhesion can be defined as the ability of synthetic or biological macromolecules to
adhere to mucosal tissues. The concept of muco adhesion is one that has the potential to improve the highly variable residence
times experienced by drugs and dosage forms at various sites in the gastrointestinal tract, and consequently to reduce variability and
improve efficacy [1]. These systems remain in close contact with the absorption tissue, the mucous membrane, releasing the drug at
the site of action leading to an increase in bioavailability. Mucoadhesive drug delivery system prolong the residence time of
the dosage form at the site of application or absorption and facilitate an intimate contact of the dosage form with the
underline absorption surface and thus contribute to improved and / or better therapeutic performance of the drug. In general, the
process involved in the muco adhesion phenomenon can be described in three steps: first of all, the wetting and swelling of the
polymer should allow an intimate contact with the tissue and secondly, interpenetration of the polymer chains and entanglement
between the polymer and the mucin chains should be attained and finally, the formation of weak chemical bonds [2].
Mucoadhesive microspheres provide a prolonged residence time at the site of application and facilitate an intimate contact
with the underlying absorption surface and improve the therapeutic performance of drug. Mucoadhesive polymer are used to
improving drug delivery by promoting the residence time and contact time of the mucosal cells may be enhanced or released at the site
for an extended period of time and enhanced bioavailability of the drug to high surface to volume ratio. In recent years such
mucoadhesive microspheres have been developed for oral, buccal, nasal, ocular, rectal, vaginal routes for either systemic or local
effects [3].
Mucoadhesive microspheres form an important part of novel drug delivery system. The short residence time of the
microspheres at the site of absorption can be overcome by coupling bioadhesion characters to the microspheres and developing
bioadhesive microspheres [4]. Mucoadhesive microspheres helps to overcome the relatively short GI residence time and to improve
localization of oral controlled or sustained release drug delivery system by intimate contact with the mucous layer and by specific
targeting of drug to the absorption site. Microspheres of biodegradable or non biodegradable polymers can be used for sustained
delivery [5-6].
The novel design of an oral controlled drug delivery system should primarily be aimed at a achieving more predictable and
increased bioavailability of drugs. But there are several difficulties, which include restraining/localizing the drug delivery system
within the regions of the gastro intestinal tract and the highly variable nature of gastro emptying process (few minutes to 12 hrs and
above) [7].
The major absorption zone (stomach or upper part of the intestine), can result in incomplete drug release from the drug
delivery system leading to diminished efficacy of the administered dose. Therefore, restraining a drug delivery system in a specific
region of the gastro intestinal tract due to its mucoadhesiveness increases the intimacy and duration of contact between a drug
containing polymer and mucus surface. Such a drug delivery system offers numerous advantages, especially for drugs exhibiting an
absorption window or for drugs with a stability problem [7]. Earlier research work was carried out to prepare mucoadhesive
microsphere to enhance the intranasal delivery of Valsartan [8].
Valsartan is a potent and specific competitive angiotensin II type1 receptor (AT1) antagonist. It is used orally for the
treatment of hypertension and has a low bioavailability of 23%, because of its poor absorption in lower gastro intestinal tract. It
undergoes little or no hepatic metabolism and its elimination half life is 6 hrs. Therefore, it is selected as a suitable drug for the design
of mucoadhesive microspheres with a view to improve its oral bioavailability and increase its drug release in a sustained manner [9].

MATERIALS AND METHODOLOGY:

Materials:
Valsartan was obtained as a gift sample from Bindu pharmaceuticals, Hyderabad. Sodium alginate was obtained from Bindu
pharmaceuticals, Hyderabad, Hydroxy propyl methyl cellulose and Carbopol from Loba chemicals pvt ltd, Mumbai and Calcium
chloride from Qualigens fine chemicals, Mumbai

Compatibility studies Fourier Transform Infrared Spectroscopy (FTIR):

FT- IR spectroscopy was employed to ascertain the compatibility between Losartan potassium and the selected excipients.
The pure drug, drug-excipient combinations, and formulations were subjected to FT- IR studies. In the presence of IR lamp, potassium
bromide was mixed with drug and/or excipient in 1:1 ratio. Grinding in smooth mortar can effect mixing. The mixtures were then
placed in the sample holder of the instrument and the spectra were taken. The spectra were run from 4000 cm-1 to 500 cm-1 wave
number. FT-IR spectrum of Losartan potassium was compared with FT-IR spectra of Losartan potassium with excipients. The pure
drug and drug with excipients were scanned separately. Disappearance of Losartan potassium peaks or shifting of peak in any of the
spectra was studied.
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Preparation of Mucoadhesive Microspheres by Orifice ionic gelation method:

Microspheres were prepared by using the Orifice ionic gelation technique. In this technique cross linking was done with
calcium chloride solution to release the drug in a controlled manner. In this method Sodium alginate, HPMC, carbopol in
different ratios as mentioned in the table 1 were added to 32ml of water with continuous stirring to form homogenous solution. Then
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sonicating was done for 20 minutes the drug Valsartan was then added to the above solution to form a clear solution. The drug
polymer mixture was poured in1.5% calcium chloride solution using 22# gauge needle by stirring at 50rpm. The microspheres thus

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Vol 4, Issue 09, 2014. Akram Khan et al. ISSN NO: 2231-6876

formed were allowed for 30 min then calcium chloride solution was decanted and the formed mucoadhesive microspheres are washed
with distilled water and air dried over night at room temperature.

Table.No:1 Formulations of Mucoadhesive microspheres of Valsartan microspheres.

Formulation Ratio Valsartan Sodium Alginate (mg) Carbopol Hpmc Cmc

Code (mg) (mg) (mg) (mg)
F1 1:1 40 20 20 _ _
F2 1:2 40 40 40 _ _
F3 1:3 40 60 60 _ _
F4 1:1 40 20 _ 20 _
F5 1:2 40 40 _ 40 _
F6 1:1 40 20 _ _ 20
F7 1:1 40 20 5 15 _
F8 1:1 40 20 10 10 _
F9 1:1 40 20 15 5 _

Figure.No:1 Microspheres prepared by orifice –ionic gelation method.

Evaluation and Characterization of Microspheres

Appropriate assessment of a dispersed system requires characterization of both chemical and physical stabilities. Physical
properties are very important with respect to the performance of dispersed systems.

Particle Size Determination [10, 11]

Particle size distribution for the microspheres were measured by sieving method analysis, using set of standard sieves was
weighed. Particles having size range between 50 and 1500 μm are estimated by sieving method. This method directly gives weight
distribution. The sieving method is a useful application in dosage form development of tablets and spheres

Percentage Yield [12]

The total amount of microspheres obtained were weighed and evaluated for percentage yield.

actual yield
Percentage yield = ----------------- x 100
theoretical yield

Drug content estimation and Entrapment efficiency

Valsartan microspheres (100mg) from each batch were initially stirred in 3 ml sodium citrate solution (1%w/v) until
complete dissolution. A quantity of 7 ml of methanol was added to above solution to solubilise the Valsartan. The filtrate was
assayed for drug content by measuring the absorbance at 250 nm after suitable dilution by UV-Visible spectrophotometer and
Encapsulation efficiency was calculated using the formula
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Estimated % drug content in microspheres

Entrapment efficiency = ×100
Theoretical % drug content in microspheres
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Percentage moisture content

The Valsartan loaded microspheres was evaluated to determine the percentage moisture content which sharing an idea
about its hydrophilic nature. The microspheres weighed (w1) initially kept in desiccators containing Calcium chloride at 37º

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Vol 4, Issue 09, 2014. Akram Khan et al. ISSN NO: 2231-6876

C for 24 hours. The final weight (w2) was noted when no further change in weight of sample was observed.

Moisture Percentage = W1-W2/W2×100

Scanning electron microscopy (SEM) [10]
The microspheres were observed under a Scanning Electron Microscopy. They were mounted directly onto SEM sample
stub using double-sided sticking tape and coated with gold film with ion spillter with gold target with resolution 3 nm (30 KV HV
Mode),10 nm (30 KV HV Mode), 40 nm (30 LV Mode) and a vacuum system is fitted to it.

In -vitro wash off test for mucoadhesion

The mucoadhesive property o f t h e Valsartan microspheres w a s evaluated b y an in-vitro adhesion testing method
known as the wash – off method. Freshly excised pieces of intestinal mucosa (4×5 cm) from sheep were mounted onto glass
slides (3×1inch) with poly cyanoacrylate glue. Two glass slides were connected with a suitable each wet rinsed tissue
specimen, and i mmediately thereafter the support were hung onto the arm of a USP tablet disintegrating test machine.
When the disintegrating test machine was operated, the tissue specimen was given a slow, regular up and down movement in the
test fluid (400ml) at 37ºC contained in a 1000 ml vessel of the machine. At the end of 1 hr and at hourly interval up to 8 hr, the
machine was stopped and the number of microsphere still adhering to the tissue was counted. The test was performed in
stomach (pH1.2).

In- vitro drug release studies [10, 13]

In-vitro drug release study was carried out in USP dissolution test apparatus. A quantity of microspheres equivalent to
100 mg of Valsartan microspheres was kept in basket type apparatus and immersed in 900ml of phosphate buffer (pH 1.2) in
1000 ml dissolution flask and temperature was maintained at 37 ± 0.5ºC throughout the study. At predetermined time intervals 2 ml
of samples was withdrawn by means of a syringe fitted with prefilter and same was replaced into the dissolution flask containing
pH 1.2. The absorbance of sample was measured at 250 nm after required dilution with the fresh medium (pH.1.2).All the studies
were conducted in triplicate.

Kinetics of In-vitro drug release [14]

The dissolution profile of all the batches was fitted to Zero order, First order and Higuchi to ascertain the kinetic modeling of
the drug release.
The results obtained from in vitro release studies were plotted in four kinetics models of data treatment as follows:
 Cumulative percentage drug release Vs. Time (zero order rate kinetics)
 Log cumulative percentage drug retained Vs. Time (first order rate kinetics)
 Cumulative percentage drug release Vs. √T (Higuchi’s classical diffusion equation)
 Log of cumulative percentage drug release Vs. log Time (Peppa’s exponential equation)

RESULTS AND DISCUSSION

Compatibility Studies by FTIR:
The comparative FTIR studies of Drug and excipients combination had shown negligible variation in the values as compared
with that of only pure form of Drug. Therefore it implies good compatibility of drug and excipients. The FTIR spectra of pure drug
and drug with excipients are shown in the fig 2-5.

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Figure. No:2 FTIR Graph of Valsartan.

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Figure.No:3 FTIR Spectra of Valsartan +sod.alginate.

Figure.No4: FTIR Spectra of Valsartan+ Carbopol.

Figure.No:5 FTIR Spectra of Valsartan+HPMC.

Evaluation of Microspheres
The prepared microspheres were evaluated for various parameters and the results are given in the table 2. The total amount
of microspheres obtained were weighed and evaluated for percentage yield. From this, formulation F9 showed maximum
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percentage yield among the formulations prepared. Average particle size of microspheres was determined for all the
formulations by sieving method analysis by using standard sieves. From the values, the formulation f9 had given the less average
particle size compared to all other formulations. The drug content of microspheres were calculated for all the formulations (F1 to F9)
and the formulation F9 was showed maximum drug content among the formulations the prepared. By comparing all the values of
moisture content for all formulations, formulation F9 was found to be the best one. The formulation F9 showed less moisture
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content. The order was F9<F8<F6<F7<F5<F4<F3<F2<F1.

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Table.No:2 Evaluation parameters of Mucoadhesive microspheres.

Formulation Percentage Average Particle Drug Content Entrapment Moisture

Code Yield (%) Size (µm) (%) efficiency(%) Content (%)
F1 75.19 649.38±1.14 42.32±0.62 70.69 8.723±0.144
F2 77.40 693.18±0.44 36.34±1.46 71.96 7.637±0.078
F3 73.20 720.64±2.54 38.82±1.56 80.75 5.876±0.080
F4 82.20 738.28±0.81 40.62±1.38 83.66 4.158±0.121
F5 82.00 760.58±1.36 41.89±0.49 78.32 3.529±0.050
F6 71.60 794.56±1.94 39.64±2.26 84.68 3.069±0.132
F7 65.60 660.85±2.77 34.06±0.72 70.92 2.516±0.040
F8 87.95 690.48±0.34 32.92±1.86 77.62 1.944±0.130
F9 88.65 642.38±1.14 46.96±0.29 87.21 1.116±0.138

Scanning electron microscopy (SEM)

The shape of the Valsartan microspheres was evidenced from the Scanning Electron Microscopy was found to be
spherical and uniformly distributed and is shown in the figure 6.

Figure.No.6: Scanning electron microscopy of Valsartan loaded sodium alginate, HPMC and carbopol.

In vitro Mucoadhesion Test:

Microspheres with a coat consisting of alginate and a mucoadhesive polymer exhibited good mucoadhesive properties
in the in- vitro wash off test and the results are given in the table 3. The wash off test was faster at a pH 1.2 (stomach pH). It was
observed that the pH of the medium was critical for the degree of hydration, solubility and mucoadhesion of the polymers. The
results of the wash off test indicated that the formulation F9 had very good mucoadhesive properties with more than 62.18%
retention for 8 hrs in pH 1.2.

Table. No: 3 Data of in- vitro wash off test to assess mucoadhesive properties of F1-F9 Formulations.

% Retention
Formulation code 1hr 2hr 4hr 6hr 8hr
1 hr 6 hr
F1 96.07 88.79 68.34 56.45 40.08
F2 94.74 84.21 78.92 60.96 41.16
F3 98.19 80.32 76.49 66.46 46.36
F4 98.52 86.41 72.37 58.37 44.29
F5 96.63 82.62 76.59 54.38 52.56
F6 97.42 90.35 82.72 66.74 54.84
F7 98.27 88.53 82.49 64.36 44.19
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F8 96.86 90.81 78.67 58.49 42.28

F9 99.84 97.68 92.48 84.39 62.18

In-Vitro Drug Release Studies:

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The in-vitro drug release studies of valsartan microspheres was carried out in pH 1.2 buffer as a diffusion medium for a
period of 10hrs, and the results are given in the table 4 and figure 7. The cumulative percentage drug release for F1, F2, F3, F4, F5,
F6, F7, F8, F9 were found to be 69.26%, 65.08%, 71.08%, 63.47%, 67.89%, 70.45%, 74.13%, 76.12% and 81.76% at the end of 10th

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hrs. From this data we can find that the polymer which were used in the formulation carbopol. HPMC, sodium alginate has controlled
the drug release to a great extent which would be helpful in reducing the number of administrations and thereby improving the patient
compliance.

Table. No: 4 Data of in vitro drug release profile of Valsartan loaded mucoadhesive microspheres of F1 -F9
Formulations.

Time Percentage Drug Release

(hours) F1 F2 F3 F4 F5 F6 F7 F8 F9
1 13.89 10.68 17.42 11.58 15.46 12.65 18.35 16.93 19.84
2 23.36 16.98 21.54 17.29 19.75 20.67 25.90 22.76 26.72
3 29.07 25.37 27.65 21.45 23.67 26.74 30.05 28.23 31.36
4 36.64 32.46 31.56 28.87 29.56 33.45 39.45 35.98 38.43
5 42.34 45.36 39.78 35.47 34.78 39.87 44.89 41.87 46.64
6 49.07 47.48 48.97 41.68 40.56 43.75 52.98 48.24 51.28
7 53.03 50.67 51.87 48.89 46.78 49.56 59.56 52.76 57.14
8 61.96 56.89 58.16 52.98 54.81 53.23 64.87 59.56 66.49
9 66.76 61.87 64.89 59.89 63.46 65.56 69.90 67.87 72.38
10 69.26 65.08 71.08 63.47 67.89 70.45 74.13 76.12 81.76

Figure.No. 7: Comparison of in- vitro drug released for formulations F1 to F9.

Invitro drug release Kinetics of Drug release for formulations F1-F9

The kinetics of In-vitro drug release was determined by applying the drug released data to various kinetic models such as
zero order, first order, Higuchi and Korsmeyer- Peppas. From the kinetic data of formulations, the optimized formula F 9 shows drug
release by zero order kinetics (R2=0.9647) and Krosmeyer Peppas i.e., diffusion (R2=0.9907). The ‘n’ value of Krosmeyer-Peppas
model are in the range of 0.356- 0.485. It indicates that drug release follows Fickian diffusion mechanisms i.e., release is by diffusion.
The results of linear regression analysis are summarized in table 5.

Table.No:5 In vitro drug released kinetics studies of all formulations

Formulation Higuchi Krosmeyer Peppas

Zero order 2 First order 2
code R R 2 2 n Best fit model
R R
F1 0.9621 0.9501 0.9723 0.9908 0.432 Peppas
F2 0.9650 0.9705 0.9663 0.9909 0.456 Peppas
F3 0.9900 0.9471 0.9530 0.9985 0.402 Peppas
F4 0.9503 0.8439 0.9782 0.9908 0.485 Peppas
F5 0.9592 0.8811 0.9729 0.9903 0.365 Peppas
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F6 0.9622 0.9307 0.9684 0.9903 0.462 Peppas

F7 0.9540 0.8611 0.9770 0.9901 0.398 Peppas
F8 0.9597 0.9162 0.9756 0.9907 0.441 Peppas
F9 0.9647 0.9609 0.9663 0.9907 0.356 Peppas
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CONCLUSIONS
Mucoadhesive microspheres of Valsartan were prepared by Orifice- ionic gelation technique using calcium chloride as
cross-linking agent .The higher drug content and entrapment efficiency was observed as the concentration of alginate increased.
The drug loading efficiency greatly improved when alginate was blended with carbopol. Mucoadhesive property of formulation F9
consisting of sodium alginate 3.5% along with carbopol and HPMC 1% exhibited good mucoadhesive property. The microspheres
cross-linked with calcium showed delay in disintegration and consequently a slow release of drug was obtained. On comparing the
major criteria in evaluation such as particle size, percentage yield, drug content, entrapment efficiency, in vitro wash off test and in
vitro drug release characteristics the formulation F9 was selected as the best formulation, as it showed particle size of about
642.38±1.14,percentageyield of about 88.65%the drug content as 46.96% and entrapment efficiency was 87.21%, showed a good
mucoadhesive nature in the in vitro wash off test was nearly 62.18% up to 8 hrs and in vitro drug released 81.76% up to 10 hrs.
Based on all the above evaluation parameters it was concluded that the formulation F9 was found to be best
formulation among all the formulations. In future the research can be extended to invivo studies to prove the increased
bioavailability statistically.

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