DESIGN AND IN VITRO EVALUATION OF

MUCOADHESIVE MICROCAPSULES OF

ACECLOFENAC FOR ORAL CONTROLLED RELEASE

M.PHARM DISSERTATION PROTOCOL

SUBMITTED TO THE

RAJIVGANDHIUNIVERSITY OF HEALTH SCIENCES,

BANGALORE, KARNATAKA.

By

VADDE JITENDRA BABU

Under the guidance of

Dr. D. Nagendrakumar

M.Pharm., Ph.D

DEPARTMENT OF PHARMACEUTICS

S.V.E TRUST’S COLLEGE OF PHARMACY

HUMANABAD-585330

2012-2013

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA

BANGALORE

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. / Name of the candidate
(In block letters) / VADDE JITENDRA BABU
Permanent address / VADDE JITENDRA BABU,
C/O SRI SAI RAM AGRO AGENCIES,
X – ROAD, PINAPAKA MANDAL,
KHAMMAM DIST.
ANDHRA PRADESH.
2. / Name of the institution / S.V.E TRUST’S COLLEGE OF PHARMACY
HUMANABAD-585330
3. / Course of study and subjects / M.PHARM
(PHARMACEUTICS)
4. / Date of admission to the course / 16.06.2012
5. / Title of the topic / designAND IN VITROEVALUATION OF MUCOADHESIVE MICROCAPSULES OF ACECLOFENAC FOR ORAL CONTROLLED RELEASE
6 / Brief resume of the intended work
6.1 / Need for the study:
Oral ingestion is the traditionally preferred route of drug administration, providing a convenient method of effectively achieving both local and systemic effect. Oral controlled release products are formulations that release active drug compounds into the body gradually and predictably over a 12 to 24 hr period and that can be taken once or twice a day. Typically these products provide numerous benefits compared with immediate release drugs, including greater effectiveness in the treatment of chronic conditions, reduced side effects, greater convenience and higher level of patients compliance due to simplified dosing schedule. Because of the above advantages, such systems form the major segment of the drug delivery market. A number of techniques are used to achieve controlled release of drugs through g.i. tract.Microcapsules are one of the most interesting modes of drug delivery systems. Recently, dosage forms that can precisely control the release rates and target drugs to a specific body site have made an enormous impact in the formulation and development of novel drug delivery systems. Microparticles are defined as spherical polymeric particles. These microparticles constitute an important part of these drug delivery systems, by virtue of their small size and efficient carrier characteristics. However, the success of these novel microparticles is limited due to their short residence time at the site of absorption. It would, therefore, be advantageous to have means for providing an intimate contact of the drug delivery system with the absorbing membranes. Mucoadhesion is a topic of current interest in the design of drug delivery systems to prolong the residence time of the dosage form at the site of application or absorption and to facilitate intimate contact of the dosage form with the underlying absorption surface to improve and enhance the bioavailability of drugs.
NSAIDs have become widely used in the treatment of Rheumatoid Arthritis for their pain-relieving and anti-inflammatory properties. Since long-term NSAID treatment is indicated for osteoarthritis, rheumatoid arthritis the ideal agent should have good efficiency and a low propensity to cause adverse events. Aceclofenac appears to be particularly well tolerated among the NSAIDs, with a lower incidence of gastrointestinal adverse effects. Recommended dose of Aceclofenac is 100 mg twice daily, due to short biological half-life of the drug 4 h makes it suitable candidate for the modified release dosage forms. To reduce the dose frequency of administrations and to improve patient compliances, aceclofenac can be made into a sustained release dosage form.
In this study an attempt was made to prepare oral controlled release mucoadhesive microcapsules of Aceclofenac by employing Sodium alginate as coat material in combination with mucoadhesive polymers like Sodium carboxy methyl cellulose (NaCMC), Methyl cellulose (MC), Carbopol-934P, Hydroxy propyl methyl cellulose (HPMC), employing Orifice ionic gelation technique and Emulsification ionic gelation technique.
6.2 / Review of Literature
Literature review shows that very few works has been published on the mucoadhesive microcapsules of aceclofenac. Some of the published reports of similar work for various medicinal agents are:
S. K. Prajapati,Purnima Tripathi,Udhumansha Ubaidulla,andVikas Anand; Design and Development of Gliclazide Mucoadhesive Microcapsules:In VitroandIn VivoEvaluation;AAPS PharmSciTech, Vol. 9, No. 1, March 2008:224-230.
K.P.R. Chowdary and Y. Srinivasa Rao; Preparation And Evaluation Of Mucoadhesive Microcapsules Of Indomethacin; Indian J.Pharm. Sci., 2003, 65(1): 49-52.
Mradul R. Gupta, Rahul Kapoor, M. S. Sudheesh, Umesh K. patil; An Applauded Novel Drug Delivery System For Arthritis Using NSAIDS By Microencapsulation Technique -A Review;Scholars Research Library,Der Pharmacia Lettre, 2010, 2(4): 335-354.
A. Ashok Kumar, Putta Rajesh Kumar, A. Anil Kumar, K. Lokeswara Reddy, T.E.G.K. Murthy and T. Venkateswara rao, Formulation Design Of Aceclofenac Microcapsules By Ionotropic Gelation Technique, Characterization Studies And Release Kinetics, Journal Of Applied Pharmaceutical Science 01 (06); 2011: 127-132.
Santhosh kumar, Chowdary K.A and Sammaiah G; Controlled Release Formulation And Evaluation Of Aceclofenac By Microencapsulation; Pharmanest – An International Journal Of Advances In Pharmaceutical Sciences: Vol.2 (2-3) March – June ~ 2011.
Kim CK, Lee E J. The controlled release of blue dextran from alginate beads. Int J Pharm. 1992; 79: 11-19.
Hari, P C, Chandy T, Sharma C P. Chitosan/calcium alginate microcapsules for intestinal delivery of nitrofurantoin. J Microencapsul. 1996; 13: 319-329.
Dua K., Ramana M.V., Sara U.V.S., Himaja M., Garg V., Agarwal A., Dissolution enhancement of aceclofenac through solid dispersion, Indian pharmacist., 2006,48,70-72.
M. A. Altaf,*Sreedharan,and N. Charyulu; Ionic Gelation Controlled Drug Delivery Systems for Gastric-Mucoadhesive Microcapsules of Captopril : Indian J Pharm Sci.2008 Sep-Oct;70(5): 655–658.
K.P.R. Chowdary, B.L.R. Madhavi ; Preparation And Evaluation Of HPMC-Alginate Mucoadhesive Microcapsules Of Aceclofenac For Controlled Release: IJPRD,2011;Vol 3(12) : February-2012 (72-78).
6.3 / Objectives of the study
The specific objectives of the investigation are as follows.
(i)To design mucoadhesive microcapsules suitable for oral controlled release employing sodium CMC, methyl cellulose, Carbopol and HPMC alone and in combination with sodium alginate.
(ii)To develop mucoadhesive microcapsules of the selected medicament (aceclofenac) and to evaluate them for controlled release by in vitro methods.
(iii)To characterize and evaluate the mucoadhesive microcapsules for size and size distribution, drug content and microencapsulation efficiency and surface characteristics by SEM.
(iv)To evaluate the kinetics and mechanisms of drug release from the mucoadhesive microcapsules developed.
(v)To evaluate the stability characteristics of selected mucoadhesive microcapsules designed.
Extensive experimentation, in vitro has been carried to fulfill the objectives of the investigation and the results obtained are presented and discussed in the subsequent chapters.
7.0 / Materials and Methods
7.1 / Source of Data:
  • Internet
  • S.V.E.T college library
  • International Pharmaceutical Abstract

7.2 / Materials:
Drug: Aceclofenac
Polymers: Sodium carboxymethyl cellulose(Sodium CMC) , Hydroxypropyl methyl cellulose(HPMC), Sodiumalginate, , Carbopol and Methyl cellulose.
Equipments:
  • UV-visible spectrophotometer (Shimadzu 1700).
  • PH-meter (systronics335).
  • Electronic Balance.
  • Single Pan electronic balance.
  • USP XXIII tablet dissolution test apparatus-II.
Preparation Microcapsules:
Microcapsules containing Aceclofenac were prepared employing sodium alginate in combination with four mucoadhesive polymers namely sodium carboxy- methylcellulose (sodium CMC), methyl cellulose, Carbopol and hydroxypropylmethyl cellulose (HPMC) as coat materials. Two methods namely emulsification ionic gelation and orifice ionic gelation were developed for microencapsulation by these polymers based on the ionic gelation principlewhich has been extensively used to prepare large sized alginate beads, was employed to prepare the microcapsules.
The prepared microcapsules will be evaluated for various physicochemical characteristics such as:
  • Estimation of aceclofenac
  • Particle size analysis
  • Determination of flow properties
  • Drug content evaluation
  • Microencapsulation efficiency
  • Scanning electron microscopy
  • Invitro drug release studies
  • Mucoadhesion testing by in vitro wash-off test

Evaluation of Aceclofenac Microcapsules
Estimation of Aceclofenac :Aceclofenac content in the microcapsules was estimated by an UV spectrophotometric methodbased on the measurement of absorbance at 275 nm in phosphate buffer of pH 6.8.
Particle size analysis: For size distribution analysis, different sizes in a batch were separated by sieving using a range of standard sieves. The amounts retained on different sieves were weighed
Determination of flow properties:
Bulk density and tapped density: Accurately weighed microcapsules (M) were transferred to a 100 ml graduated cylinder to measure the apparent volumes or bulk volume (Vb). The measuring cylinder was tapped for a fixed period of time and tapped volume (Vt) occupied in the cylinder was measured. Twenty tablets from each batch is selected at random and weighed individually. The individual weights are compared with average weight for determination of weight variation.
Angle of repose: A funnel was fixed in a stand in such a way that the top of the funnel was at a height of 6 cm from the surface. The microcapsules were passed from the funnel so that they formed a pile. The height and the radius of the heap were measured and the angle of repose was calculated using the equation.
Drug content evaluation: Aceclofenac content in the microcapsules was estimated by UV-spectrophotometric method at 275 nm in phosphate buffer pH 6.8 with 10% methanol. The method obeyed Beer’s law in concentration range 5-40 µg/ml. 25 mg of microcapsules were crushed and taken into 25 ml volumetric flask and extracted with 2.5 ml of methanol and 10 ml phosphate buffer pH 6.8 was added. Then it was sonicated for 30 min. The volume was made up to 25 ml with phosphate buffer pH 6.8. 1 ml of the sample solution was taken and made upto the 10 ml with phosphate buffer pH 6.8 and absorbance was measured at 275 nm. Drug content was calculated.
Microencapsulation efficiency: Microencapsulation efficiency is used to find the drug entrapment capability of various polymers with respect to the ionic gelation method used for formulation. Microencapsulation efficiency was calculated using the following formula.
Microencapsulation efficiency = (estimated percent drug content / theoretical percent drug content) × 100.
Scanning electron microscopy: The microcapsules were observed under a scanning electron microscope (SEM-LEICA, S430, UK). For SEM, the microcapsules were mounted directly onto the SEM sample stub, using double-sided sticking tape, and coated with gold film (thickness 200 nm) under reduced pressure (0.001 torr).
In vitro drug release studies:Release of Aceclofenac from the microcapsules was studied in phosphate buffer of pH 6.8 (900 ml) using an USP type II Dissolution Rate Test Apparatus (LABINDIA DS 8000) with a rotating paddle stirrer at 50 rpm and 37±0.5C. A sample of microcapsules equivalent to 100 mg of Aceclofenac was used in each test. Samples of dissolution fluid were withdrawn through a filter (0.45 μm) at different time intervals and were assayed at 275 nm for Aceclofenac content using a UV-Visible spectrophotometer. The drug release experiments were conducted in triplicate (n=3).
Mucoadhesion testing by in vitro wash-off test: The mucoadhesive property of the microcapsules was evaluated by an in vitro adhesion testing method known as wash-off method. The mucoadhesiveness of these microcapsules was compared with that of non-bioadhesive material, ethylene vinyl acetate microcapsules. Freshly excised pieces of intestinal mucosa (2×2 cm) from sheep were mounted onto glass slides (3×1 inch) with cyanoacrylate glue. Two glass slides were connected with a suitable support. About 50 microcapsules were spread onto each wet rinsed tissue specimen and immediately thereafter the support was hung onto the arm of a USP tablet disintegrating test machine. By operating the disintegrating test machine the tissue specimen was given a slow regular up and down movement in the test fluid at 37C taken in a 1 L vessel of the machine. At the end of 30 minutes, 1 h and later at hourly intervals up to 12 h, the machine was stopped and the number of microcapsules still adhering on to the tissue was counted. The test was performed at both gastric pH (0.1 N HCl, pH 1.2) and intestinal pH (Phosphate buffer of pH 6.8).
7.3 / Does the study require any investigation or intervention to be conducted on patients or other humans or animals? If so please describe briefly
Not under the plan of the work
7.4 / Has ethical clearance have been obtained from your institution in case of 8.4?
Not applicable
8.0 / List of References
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9. / Signatures of candidate / VADDE JITENDRA BABU
10. / Remarks of Guide / Mucoadhesive microcapsules of drugs,which prolong the residence time of dosage form and thus definitely advantageous in improving bioavailability characteristics, ultimately resulting in a reduced dosage and dose related side effects.
11. / Name and designation of
(in block letters)
11.1 Guide / Dr.D.Nagendrakumar
M.Pharm., Ph.D
11.2 Signature
11.3 Co-guide / ----
11.4 Signature
11.5 Head of the department / Dr.D.Nagendrakumar
M.Pharm., Ph.D
11.6 Signature
12. / 12.1 Remarks of and Principal
12.2 Signature

1