Asian J. Res. Pharm. Sci. 2013; Vol.

3: Issue 1, Pg 19-24

[AJPSci.]

ISSN- 22315640 (Print) ISSN- 22315659 (Online)

www.asianpharmaonline.org

RESEARCH ARTICLE

Design Formulation and Evaluation of Reservoir Type Controlled Released Moxifloxacin Hydrochloride Ocular Insert
Ramesh B. Parmar1*, Dr. H. M. Tank2
1 2

S. J. Thakkar Pharmacy College, Opp NRI bungalow, Avadh Club Road, Munjka, Kalawad Road, Rajkot. Matushree V B Manvar Pharmacy College, Dumiyani, Upleta, Rajkot. *Corresponding Author E-mail: rbparmar82@yahoo.co.in

ABSTRACT
The present studies were mainly focus to developed ocular controlled release formulation of Moxifloxacin Hydrochloride. Reservoir type of ocular insert was developed by solvent casting method. Total nine formulations was developed using different ratio of Eudragit RS 100 and Eudragit RL100 in combination as a rate controlling membrane and reservoir was prepared by using sodium CMC. All the prepared formulation were subjected for evaluation of physicochemical parameter like thickness, weight variation, percentage moisture absorption, percentage moisture loss, surface pH, sterility, drug content and anti-microbial activity. Evaluated results were shown that all the prepared formulation was suitable for patient compliance. In-vitro release study was carried out by using commercial semipermeable membrane with the help of modified standard cylindrical tube method and best formulation F7 found 98.21 % at the end of 24 hrs. Formulated ocular inserts also passed the test for sterility. The above in vitro release studies revealed that the best ocular inserts formulation followed near to zero-order release kinetics. Higuchis plot and Peppas plot revealed that the mechanism of drug release involved in all the formulations was super case II transport diffusion. The antimicrobial study was shown that formulation was able to inhibit the microbial growth for extended period of time. The controlled release ocular insert was more suitable as compared to conventional dosage form.

KEYWORDS: Reservoir drug delivery system, ocular insert, Moxifloxacin Hydrochloride, Eudragit RS100,
Eudragit RL 100

INTRODUCTION:
The eye is a one of the most, after oral route, interesting organ for local drug delivery of the medicaments. The eye is generally used for local therapy against systemic therapy in order to avoid the risk of eye damage from high blood concentrations of the drug.1 The physiological constraints imposed by the protective mechanisms of the eye lead to low absorption of drugs and a short duration of the therapeutic effect on ocular drug delivery. Upon instillation of the eye drops only 110% of the drug is bioavailable while the rest is drained out of the eye through lacrimal secretions. 2 To overcome this problem various approaches have been reported, such as ointments, inserts and aqueous gels, to increase the ocular residence time of topically applied medication. Controlled drug delivery to the eye offer several advantages over conventional therapies like drug solutions or suspensions as eye drops. Ophthalmic inserts offer many advantages over conventional dosage forms, like increased ocular residence, possibility of releasing drugs at a slow and constant rate, accurate dosing, and exclusion of preservatives, increased shelf life and reduced systemic absorption. Newer ocular drug delivery systems are being explored to develop extended duration and controlled release strategy. Some of the newer, sensitive and successful ocular delivery systems like inserts, biodegradable polymeric systems, and collagen shields are being developed in order to attain better ocular bioavailability and sustained action of ocular drugs. 3,4 Moxifloxacin HCL is (4aS - cis) - 1 - Cyclopropyl - 6 fluoro - 1,4 - dihydro - 8 - methoxy - 7 - (octahydro - 6H pyrrolol [3,4 - b]pyridin - 6 - yl) - 4 - oxo - 3 quinolinecarboxylic acid monohydrochloride. Moxifloxacin hydrochloride (HCl) is a fourth-generation fluoroquinolone with a new 8-methoxy derivate of fluoroquinolones with enhanced activity in vitro against gram positive bacteria and maintenance of activity against gram negative bacteria.32 It is an anti-infective agent useful in the treatment of eye

Received on 29.12.2012 Accepted on 27.01.2013 Asian Pharma Press All Right Reserved
Asian J. Res. Pharm. Sci. 3(1): Jan.-Mar. 2013; Page 19-24

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Eudragit RL 100 in different ratio as per (as per Table 1) was dissolved in 1/3rd quantity of acetone and the plasticizer (15% dry weight of polymer) dibutyl phthalate was dissolved in remaining acetone, then both the solutions were mixed together thoroughly to get the uniform In the present study, it was aimed to prepare ocular films dispersion. This solution was poured on mercury substrate containing Moxifloxacin Hydrochloride to overcome and dried at room temperature for 24 hours. After drying 8 limitations associated with convectional dosage, an attempt mm diameter were cut using stainless steel borer. has been made to formulate ocular inserts that may not only improve the efficiency of the therapy but also patient The medicated reservoir film cut with the help of a stainless steel die. These ocular inserts were placed on a ratecompliance. controlling membrane and another rate controlling membrane was kept over it. The two rate-controlling MATERIAL AND METHODS: membranes containing the reservoir film between them Material Moxifloxacin Hydrochloride was obtained from Torrent were sealed with the help of acetone. This procedure Pharm Pvt. Ltd. Ahmadabad. Eudragit RS 100 and Eudragit resulted in sealing the two rate-controlling membranes RL 100 were gifted by Evonik Degussa India Private containing the medicated reservoir film between them. The Limited, Mumbai. Sodium CMC, Dibutyl phthalate, ocular inserts were stored in an airtight container under glycerin and other reagent was commercial purchased from ambient conditions for further use. SD Fine Chem. and Merck Pvt. Ltd. Evaluation of Ocular Inserts Thickness8 Methodology 7 The thickness of the ocular insert was measured using Preparation of ocular inserts: The preparation of ocular inserts involved three steps: (i) micrometer screw gauge. The thickness was measured at preparation of the drug-containing reservoir film of Sodium five different spots of the patch and average was taken. CMC, (ii) preparation of rate controlling films of Eudragit, 8 (iii) placing rate controlling films around the drug reservoir Weight variation From every batch, three ocular inserts were taken and their and sealing them to obtain ocular inserts. individual weights were determined by using electronic For preparation of the drug containing reservoir film, balance. The mean weight of insert was noted accurately weighed quantity of sodium CMC was soaked in 9 the 1/3rd volume of the distilled water for 24 hours. Moisture absorption Weighed Calculated amount of Moxifloxacin The percentage moisture uptake test was carried out to Hydrochloride was dispersed in the polymeric solution, check physical stability or integrity of ocular inserts. after the complete dispersion of the drug; glycerin Ocular inserts were weighed and placed in a desecrator (Plasticizer: 30% dry weight of polymer) was added and containing 100ml. of saturated solution of Aluminum stirred to form a uniform dispersion. The dispersion was Chloride and 79.5% humidity was maintained. After three casted onto mercury substrate kept in the hot air oven at days the ocular insert were taken out and reweighed, the 40C for 24 hours. The patches thus formed were cut into percentage moisture uptake was calculated by using diameter of 6mm. Each ocular insert containing 2 mg of formula. Moxifloxacin Hydrochloride. The composition of the polymeric patches containing Moxifloxacin Hydrochloride is % Moisture absorption = Final weight Initial weight given in (Table 1). (1) x100 Initial weight For preparation of the rate controlling film, the Eudragit RS 100 and Eudragit RL 100 rate controlling membrane was prepared by solvent casting technique. Eudragit RS 100 and infection such as bacterial conjunctivitis, keratitis and keratoconjunctivitis. It is presently available as eye drops (0.5%). It is administered at dosing interval of 1 drop in the affected eye 3 times a day for 7 days. 5,6
Table 1: Different Formulation of Moxifloxacin Hydrochloride ocular insert Formulation Drug Reservoir Rate Controlling Membrane Film Former Plasticizer Film Former (% w/v) (% w/w) Eudragit RS 100 (%w/v) F1 2 30 1 F2 2 30 1.5 F3 2 30 2 F4 2 30 1 F5 2 30 1.5 F6 2 30 2 F7 2 30 1 F8 2 30 1.5 F9 2 30 2

Eudragit RL 100 (%w/v) 1.5 1.5 1.5 2 2 2 2.5 2.5 2.5

Plasticizer (% w/w) 15 15 15 15 15 15 15 15 15

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Moisture Loss 9 The percentage moisture loss was carried out to check integrity of the film at dry condition. Ocular inserts were weighed and kept in a desecrator containing anhydrous calcium chloride. After 3 days, the ocular insert were taken out and reweighed, the percentage moisture loss was calculated using the formula. %Moisture loss =

solidification, the Petri plate was incubated in inverted position for 24 h at 370.5 C. After incubation, the length, width and area of zone of inhibition were measured around the ocular insert. Normal saline served as a negative control. In-vitro diffusion studies16,17 The in-vitro diffusion of drug from the different ophthalmic insert was studied using the classical standard cylindrical tube method according to literature. In brief, a simple modification of a glass tube of 12 mm internal diameter and 75 mm height. The diffusion cell membrane was tied to one end of open cylinder, which acted as a donor compartment. An ocular insert was placed inside donor compartment. The diffusion cell membrane (commercial semi-permeable membrane) acted as corneal epithelium. The entire surface of the membrane was in contact with the receptor compartment containing 50 ml of simulated tear fluid (STF) in 100 ml of beaker. The content of receptor compartment was stirred continuously using a magnetic stirrer and temperature was maintained at 3700.50C. At specific intervals of time, 3 ml of the sample solution was withdrawn from the receptor compartment and replaced with fresh simulated tear fluid (STF) solutions. The sample was analyzed for the drug content using UV-VIS spectrophotometer at 288 nm after appropriate dilutions against reference using simulated tear fluid (STF) as blank. Simulated tear fluid (STF: sodium chloride: 0.670 g, sodium bicarbonate: 0.200 g, calcium chloride.2H2O: 0.008 g, and Purified water q. S. 100 g)

Initial weight Final weight x100 Initial weight

(2)

Folding Endurance10,11 The flexibility of ocular insert can be measured quantitatively in terms of what is known as folding endurance. Folding endurance of the patches was determined by repeatedly folding a small strip of the patch (approximately 2x2 cm) at the same place till it broke. The number of times patch could be folded at the same place, without breaking gives the value of folding endurance. Surface pH 12 The inserts were allowed to swell in closed petridish at room temperature for 30 minutes in 0.1 ml of double distilled water. The swollen device was removed and placed under digital pH meter to determine the surface pH.

Drug Content Uniformity13 To check the drug content uniformity, three inserts were taken out from each film and drug content determined using the procedure of IP for Moxifloxacin Hydrochloride. Kinetic analysis18 Amount of Moxifloxacin Hydrochloride in one insert is To understand the release profile and release mechanism of given by: in-virto release of drug zero order kinetics equation and Korsemeyer's equation was used. When a graph of the C = As Cr Ar (3) cumulative percentage of the drug released from the matrix Where, As is the absorbance of sample solution, Cr is the against time is plotted, zero order release is linear in such a concentration of Moxifloxacin Hydrochloride in standard plot, indicating that the release rate is independent of solution, and Ar is the absorbance of standard solution of concentration. The rate of release of the drug can be Moxifloxacin Hydrochloride. The same procedure adopted described mathematically as follows: for all the batches and drug content was noted. Rate of release = (dCs/t) = k (4) Sterility testing14 Ultra-Violet radiation was used to sterilize the ocular inserts and sterility testing was carried out under aseptic conditions. It was found visually that the Alternate thioglycolate, Soyabean casein digest media; Fluid thioglygolate media containing sterilized ocular inserts were free from turbidity. This confirmed the absence of aerobic organism, anaerobic organism and fungi. Microbiological studies The optimized ocular insert was evaluated microbiologically for controlled drug release for 1 day. The test microorganisms E. coli and S. aureus were used. A layer of seeded agar (10 mL) was allowed to solidify in the Petri plate. An ocular insert was removed from the pack and carefully placed over the agar layer and a second layer of seeded agar (10 mL) was applied to cover the insert. After
15

Where Cs = concentration of the drug present in the matrix, k = rate constant and t = time. Since Cs is a constant, and x = amount of drug released described as dx/dt = k integration of the equation yields x = k t + constant (5) (6)

A plot of x versus t results in a straight line with the slope = k. The value of k indicates the amount of the drug released per unit of time and the intercept of the line at time zero is equal to the constant in the equation. The curves plotted may have different slopes, and hence it becomes difficult to exactly pinpoint which curve follows perfect zero order release kinetics. Therefore, to confirm the kinetics of drug release, in vitro data were also analyzed using Korsemeyers equation.

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Korsemeyer et al. used a simple empirical equation to The weights of ocular inserts of Moxifloxacin describe general solute release behavior from controlled Hydrochloride were found in between 0.211 0.05 to 0.3510.01 (Table 2). The mean weight and standard release polymer matrices: deviation were calculated. The low standard deviation that n mt/m = k t (7) indicates that uniformity of the weights of the films means good distribution of the drug, polymer and plasticizer. Where mt/m = fraction of drug released, k = kinetic constant, t = release time and n = the diffusional exponent Percentage moisture absorption: for drug release. The slope of the linear curve gives the n The percentage moisture absorption were carried out all the value. Peppas stated that the above equation could formulation and it was found in the ranged of 4.0510.22 to adequately describe the release of solutes from slabs, 7.0230.23 (Table 2). The moisture absorption was spheres, cylinders and discs, regardless of the release continuously increased when concentration of Eudragite mechanism. The value of n gives an indication of the RL100 was increased. This is may be permeability effect of release mechanism. When n = 1, the release rate is Eudragite RL 100. Higher the concentration higher the independent of time (zero order) (case II transport); n = 0.5 moisture absorption. for Fickian diffusion; and when 0.5 < n < 1, diffusion and Non-Fickian transports are implicated. Lastly, when n > 1.0 Percentage moisture loss: super case II transport is apparent. n is the slope value of The percentage moisture loss were carried out all the log mt/m versus log time curve. formulation and it was found in the ranged of 4.2300.21to 7.2580.46 (Table 2). The moisture loss was continuously RESULTS AND DISCUSSION: increased when concentration of Eudragite RL100 was Preparation of ocular insert increased. This is may be the permeability affect of The reservoir type of the ocular insert consisted of three Eudragite RL 100, higher the concentration of polymer layers of films, the inner reservoir film containing the drug higher the moisture loss. and two-rate controlling films surrounding the reservoir. The ocular inserts are composed of a central reservoir of Folding endurance: drug enclosed in specially designed semi permeable or Folding endurance of the patches was determined by micro porous membranes that allow the drug to diffuse repeatedly folding a small strip of the patch at the same from the reservoir at a precisely determined rate. The place till it broke. Range of folding endurance was between reservoir membrane was prepared by 2 % sodium CMC 1981.023 to 2212.053 (Table 2). The values of folding polymer and glycerin was subjected as a plasticizer in endurance of the film were found to be optimum and concentration of 30 % w/w. To prepare rate controlling therefore the film exhibited good physical and mechanical films, combinations of Eudragit RS 100 and RL 100 were properties. assayed in different ratios and dibutyl phthalate was chosen as plasticizer. Flexible, uniform and transparent films were Surface pH: obtained containing 15% (w/w) of plasticizer per dry mass The prepared ocular insert was subjected for measurement of polymer. of pH and it was found in range of 6.65 to 7.20 (Table 2). Physical parameter Thickness: The prepared ocular inserts were evaluated for the thickness using micrometer screw gauge. The average of three readings was taken. It was found to be in the range of 0.212 0.05 mm to 0.316 0.01 mm (Table 2). This indicated that as the concentration of the polymers increased, there was increase in the thickness of the ocular inserts. Uniformity of weights:
Table 2: Physicochemical parameters of ocular insert Formulation Thickness Weight variation % Moisture code (mm) (g) Absorption F1 0.2120.05 0.2110.07 4.0510.22 F2 0.2350.01 0.2150.09 5.9980.05 F3 0.2420.03 0.2210.02 6.5820.35 F4 0.2750.01 0.2860.06 5.2560.21 F5 0.2810.03 0.2910.04 5.3210.53 F6 0.2820.02 0.2980.05 6.0360.21 F7 0.3010.06 0.3150.02 6.3680.22 F8 0.2980.04 0.3260.01 6.5560.05 F9 0.3160.01 0.3510.01 7.0230.23 % Moisture Loss 4.2300.21 5.0210.12 5.2310.09 4.2500.11 5.0620.23 6.2310.02 6.2560.25 6.2360.52 7.2580.46 Surface pH 6.98 6.65 6.89 7.01 6.85 6.95 7.20 7.09 7.13 Drug Content (mg) 1.950.01 1.960.06 2.010.03 1.990.01 1.980.03 1.990.01 2.000.01 1.960.03 1.990.02 Folding Endurance 1981.023 2062.035 2151.520 2533.021 2311.085 2110.890 2212.053 2121.053 2163.240

The pH range of all the formulation was found near to tear fluid pH so patient compliance of ocular insert is good. Drug content: The drug content of the formulations was determined according to procedure described in methods. The drug content in all formulations was found to contain 1.950.01 to 2.010.03 (Table 2).

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Figure 1: In-vitro release profile of Moxifloxacin Hydrochloride ocular insert

In vitro Drug release study: The in vitro drug diffusion studies of all the formulation was carried out in ATF pH 7.4 using bio-chambered donor receptor compartment model described under methodology chapter. The in-vitro release data obtained from the Moxifloxacin HCL containing Na-CMC as a drug reservoir with Eudragit RS100 and Eudragit RL100 in different ratio as rate controlling membrane. In vitro drug dissolution profile of different formulations is shown in figure no. 1. The results showed that drug release was prolonged. Probably this may be due to increase in Eudragit S. Aureus E. Coli RL 100 content in rate controlling membrane. The drug is Figure No. 2: Invitro antimicrobial test of optimized formulation hydrophilic and Eudragit RL 100 is also more hydrophilic RF5 than Eudragit RS 100. The release profile of all nine formulation was shown in figure No. 1 Kinetic profile: The in vitro release profile was analyzed by various kinetic Sterility test: models (Table 3). The release constants were calculated From the sterility test, it confirms that sterility of ocular from the slope of the respective plots. It indicates that the inserts good therefore, the sterilized inserts were considered release of drug from the films might have followed zero suitable for use. order kinetics. On the basis of korsemeyers and peppas plot optimized formulation followed super case II transport Antimicrobial activity: The optimized ocular insert showed antimicrobial activity mechanisams. when tested microbiologically on solidified agar. The controlled release of the drug from ocular insert was observed for 1 day.
Table 3: Kinetic release profile of different Moxifloxacin Hydrochloride ocular inserts Zero order plot Higuchis plot Formulations Code Slope (K0 ) Correlation ( r2 ) Slope ( KH ) Correlation ( r2 ) F1 3.7440 0.9794 22.609 0.9073 F2 3.7401 0.9833 22.66 0.9172 F3 3.681 0.9854 22.384 0.9258 F4 3.8679 0.9958 23.809 0.9587 F5 3.7324 0.9966 22.854 0.9495 F6 3.5162 0.9939 21.555 0.9491 F7 4.1776 0.9985 25.807 0.9685 F8 4.0148 0.9930 24.869 0.9682 F9 4.1286 0.9913 25.446 0.9636 Korsemeyer's and Peppas plot Slope ( n ) Correlation (r2 ) 1.175 0.9762 1.793 0.9648 1.717 0.9695 1.335 0.9953 1.405 0.9961 1.409 0.9811 1.191 0.9934 1.170 0.9910 1.208 0.9920

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CONCLUSION:
From above results it can be concluded that Moxifloxacin HCl can be delivered in controlled manners for extended period of time in the form of ocular inserts. Release pattern of drug from these inserts can be altered by using different formulation variables. The said promising formulation (F7) would be able to offer benefits such as increase residence time, prolonged drug release, reduction in frequency of administration and thereby definitely prove to improve the patient compliance. Further work may be carried out to establish the therapeutic utility of this system by pharmacokinetic and pharmacodynamic studies in human beings.

17. Sultana Y, Mohammad A, Ali A, Ocular inserts for controlled delivery of pefloxacin mesylate:Preparation and evaluation. Acta Pharm. 55; 2005: 305314. 18. Patel U, Chaudhary KA, Chotai NP, Nagada C, Patel KN. Formulation and Evaluation of Indomethacin Ophthalmic Inserts. Indian Journal of Pharmaceutical Education and Research. 42(4);2008: 348-350.

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