Preparation and Evaluation of Ocular Inserts For Controlled Delivery of Ciprofloxacin Hydrochloride

Karthikeyan D, 1* Vijayakumar S, 1 Pandey V.P, 2Nandha Kumar J, 1 Sengottuvelu S,and   T. Sivakumar T. 1

¹Department of Pharmaceutics, Nandha College of Pharmacy Perundurai main road, Erode-52, Tamilnadu, India.

2Department of Pharmacy, Annamalai University, Chidambaram, Tamilnadu, India.  
*Corresponding Author E-mail:



Topical administration of drug to treat ocular diseases is accomplished primarily by means of solutions, ointments, and suspensions. These are relatively inefficient as drug delivery systems often only 1% of the available drug is absorbed by the eye. Ciprofloxacin hydrochloride is a fluroquinolone anti-bacterial drug effective in the treatment of bacterial infections.  Ciprofloxacin hydrochloride ocuserts were prepared using different polymers in various proportions. The in-vitro release of the drug from the formulations was studied using a commercial semipermeable membrane. The objective of the present work was to prepare ocular inserts with the target of increasing the contact time, reducing the frequency of administration, improving patient compliance and greater therapeutic efficacy. Ocular inserts were prepared by solvent casting technique using different polymers such as polyvinyl alcohol alone (4 %, 5 %, 6 %) and polyvinyl alcohol with polyvinyl pyrrolidone K-30, (4:0.5, 4:1, 4:1.5, 4:2, 4:2.5, 4:3) as a drug reservoir and ethyl cellulose (4 %) as a rate controlling membrane. Ocular inserts were evaluated for their physicochemical parameters like thickness, weight uniformity, drug content, percent moisture loss, and percent moisture absorption. The in-vitro drug release studies were carried out using Bi-chambered donar, receptor compartment model. Since targeted prolong release was observed in formulation    F-7. The optimized formulation F-7 was subjected to in-vivo drug release study using rabbits as an animal model, Sterility testing and accelerated stability studies to access the effectiveness of the formulation. In-vitro drug release kinetic data was treated according to Higuchi and Peppas kinetics to access the mechanism of drug release. It can be concluded that the formulation F-7 provided the desired drug release for      24 hrs and remained stable and intact at ambient conditions.   The results indicated a good correlation between in-vitro and in-vivo studies.


KEY WORDS       Ciprofloxacin hydrochloride, ocular inserts, in-vitro release studies, in-vivo studies.  




Current methods of drug delivery to the eyes in the treatment of ocular infections include application of commercially prepared antibiotic drops, ointments, fortified topical drops and subconjunctival antibiotic injections. Continuous delivery of drugs to the eye offers major advantages over conventional therapies that involve administration of drug solutions or suspensions as eye drops. Eye drug administration often results in poor bioavailability and therapeutic response due to rapid precorneal elimination of the drug and associated with patient compliance problems1,2. One of the major problems of ocular drug delivery is to provide and maintain an adequate concentration of drug in the precorneal area. In solutions many drugs display poor penetration through the corneal barrier. Rapid nasolacrimal drainage of the instilled drug from tear fluid and non-productive absorption through the conjunctiva may lead to a short duration of action. Tear turnover and


drug binding to tear fluid proteins are additional precorneal factors that contribute to the poor ocular bioavailability of many drugs. The rate of release of drug from the tear fluid to ocular tissues is initially high when instilled as solution, but rapidly declines. This may result in a transient period of overdose and the associated risk of side effects, followed by an extended period of subtherapetic levels before the next dose is administrated. This indicates the need for an ocular drug delivery system which has the convenience of sustained release effect.

Ophthalmic inserts offer many advantages over conventional dosage forms like increase in ocular residence, possibility of releasing drugs at a slow and constant rate, accuracy in dose, exclusion of preservatives and increased shelf-life3-6. Moreover, the use of these devices, reduces systemic absorption, which otherwise freely occurs with eye drops. It also ensures better patient compliance due to lower frequency of administration and lower incidence of side effects7-9. Baeynens et al10 developed soluble bioadhesive ophthalmic drug inserts for the treatment of external ocular diseases such as conjunctivitis, keratoconjunctivitis and superficial corneal ulcers. Sasaki and co-researches11 prepared a unique one-side-coated insert that releases drug from uncoated side only. Ocular application of the one-side-coated insert produced constant concentration of Tilisolol in the tear fluid over 180 min. The aim of this study was to prepare ocular inserts of ciprofloxacin hydrochloride and evaluate their potential for controlled ocular delivery.



Ciprofloxacin hydrochloride was obtained as gift from Dr. Reddy’s Laboratories (India), Polyvinyl alcohol (Qualigens fine chemicals, India), Polyvinyl pyrrolidone K -30 (Loba chemie, India), Ethyl cellulose (Loba chemie, India) were used.  All other solvents were of analytical grade.


Preparation of ocular inserts     

The preparation of ocular inserts involved three steps6:
(i) Preparation of the drug reservoir film. (ii) Preparation of rate controlling films.  (iii) Placing rate controlling films around the drug reservoir and sealing them to obtain ocular inserts

For the preparation of drug containing reservoir film, polymeric solutions were prepared by dissolving PVA alone in the percentage of 4,5,6 (formulation codes F-1, F-2, F-3) and PVA with PVP K-30 in the percentage of 4:0.5, 4:1, 4:1.5, 4:2, 4:2.5, and 4:3  (formulation codes F-4, F-5, F-6, F-7, F-8, F-9) along with   98.16 mg of ciprofloxacin hydrochloride in doubly distilled water. In this study the propylene glycol 30 % (w/w) was used as a plasticizer. The solutions were poured into a glass ring of 5.6 cm diameter placed in a Teflon coated Petri dish.  The solvent was allowed to evaporate by placing it inside an oven maintained at 35 ± 2 °C, 30 ± 0.5 % RH for 24 h.       

To prepare the rate controlling films, ethyl cellulose 4 % and 30 % w/w of plasticizer, dibutylphthalate, were dissolved in ethanol: chloroform (1:1) mixture. The solutions were poured into a glass ring of 5.6 cm diameter placed in a Teflon coated Petri dish.

The two-rate controlling membrane containing the reservoir film between them, were placed over a beaker saturated with ethanol / acetone vapors (60:40) for 1-2 minute.  This procedure resulted in sealing the two rate-controlling membranes containing the medicated reservoir film between them.  The final ocular inserts consisted of three films (mass: 30.70 ± 0.76 mg (n=5); thickness: 0.297 ± 0.010 mm). Each ocular inserts contained 98.16 mg of the drug. The ocular insert were stored in an airtight container under ambient conditions.

 In-vitro drug release studies

The in-vitro drug release studies were carried out using bichambered donor-receptor compartment model. The glass tube having a diameter of 2.5 cm, covered at its lower end by a soaked cellulose membrane and acted as donor compartment containing 0.7 ml of pH 7.4 isotonic phosphate buffer12, 13.  This membrane was used to simulate ocular in-vitro conditions like corneal epithelial barrier. The surface membrane is in contact with receptor compartment, which contain 25 ml of pH 7.4 isotonic phosphate buffer and stirred continuously using a magnetic stirrer.  Samples were withdrawn from the receptor compartment at periodic intervals and replaced with an equal volume of pH 7.4 phosphate buffer. The drug content was analyzed at 278 nm (Elico SL 164 Double beam UV spectrophotometer, India). All the formulations, F-1 to F-9, were subjected to in-vitro drug release studies.


Interaction Studies             

Interaction studies were conducted with formulation F-7 by comparing it with the pure drug of ciprofloxacin hydrochloride on the basis of Fourier Transform Infrared Analyses (FTIR).  The spectra for pure ciprofloxacin hydrochloride and the F-7 formulation was taken in the range of 450-3600 cm-1 by KBR pellet method using FTIR spectrophotometer (Shimadzu, Japan).

TLC analysis was conducted according to BP 2003 (14). Silica gel (1 mm thick, E. Merck India Ltd., India) plate was used as the stationary phase and the mobile phase consisted of water/ammonia/ butanol /acetone in the ratio of 1:2:4:13. Solutions of pure drug, medicated formulations and placebo were compared with the drug reference standard. The depth of thin layer was 2.5 cm. For visualization, a solution of bromocresol purple in 50 % (v/v) alcohol (0.4 g L–1) was used as spray reagent and the plate was observed under UV light.

The optimized formulation was powdered in a mortar and extracted with IPB (pH 7.4). The extract was transferred to the volumetric flask of 50 ml capacity and made to the volume. It was then filtered through Whatman filter paper No. 42. The absorbance of the filtered solution was determined   spectrophotometrically. The solutions of pure drug, medicated and placebo formulations were prepared in IPB of pH 7.4 and filtered through Whatman filter paper No. 42 and scanned for UV absorption between 200 and 400 nm.

Sterility Testing

The ideal batch (F-7) of ocular film was surface sterilized by exposing to ultraviolet radiation for 10 minutes and inoculated into Alternate thioglycolate media, Fluid thioglycolate media and Soyabean casein digest media and incubated at 37°C for 1-14 days to investigate the presence of aerobic and anaerobic organisms15. And to investigate the presence of fungi, Formulation F-7 was inoculated in the Soyabean casein digest media and incubated at 25°C 14 days. Observations were taken from 1- 14 days.  


Drug Content Determination

The optimized formulation F-7 was powdered in a mortar and extracted with IPB (pH 7.4).  The extract was transferred to the volumetric flask of 50 ml capacity and made to the volume. It was then filtered through Whatman filter paper No.42. The absorbance of the filtered solution was determined spectrophotometrically6.


Table I.   Physico-chemical evaluation of ciprofloxacin hydrochloride ocuserts


Formulation code

Moisture absorption*


Moisture loss* (%)




Uniformity* (mg)

Drug content* (mg)


4.85 ±0.228

16 .23 ±0.715

0.260 ±0.01

28.46 ±0.310

1.822 ±0.003


5.15 ±0.314

14.03 ±0.530

0.282 ±0.09

29.06 ±0.120

1.863 ±0.003


5.42 ±0.282

12.45 ±0.560

0.311 ±0.02

31.86 ±0.880

1.854 ±0.001


5.14 ±0.685

12.10 ±0.688

0.286 ±0.04

28.54 ±0.230

1.870 ±0.003


5.14 ±0.154

10.09 ±0.595

0.298 ±0.01

30.64 ±0.770

1.863 ±0.007


5.97 ±0.203


0.302 ±0.06

31.18 ±0.940

1.860 ±0.008


6.12 ±0.279

8.17 ±0.932

0.306 ±0.07

31.28 ±0.740

1.867 ±0.007


6.89 ±0.144

7.93 ±0.558

0.312 ±0.01

32.40 ±0.900

1.864 ±0.003


7.02 ±0.142

6.89 ±0.184

0.323 ±0.09

32.56 ±0.520

1.862 ±0.002

* Average of 5 determinants


Stability Studies     

Stability studies for the best formulation F-7 was conducted16 for 6 weeks at different temperatures like 4°C, 37°C, and 60°C. At specific time intervals (7th, 14th, 21st, 28th, 35th, 42nd day), the samples were withdrawn and analyzed by UV spectrophotometric method.     


Table II. Stability studies on the formulation, F-7



Time in Days







R.D.C (mg)





1.86 ±0.000

1.86 ±0.000

1.86 ±0.000


1.86 ±0.070

1.85 ±0.014

1.85 ±0.028


1.84 ±0.028

1.84 ±0.035

1.84 ±0.042


1.84 ±0.028

1.84 ±0.070

1.83 ±0.035


1.83 ±0.014

1.83 ±0.063

1.82 ±0.014


1.83 ±0.035

1.83 ±0.014

1.80 ±0.013


1.82 ±0.033

1.82 ±0.035

1.79 ±0.098

R. D. C. – Remaining Drug Content (average of 3 determinants) P. A. - Physical Appearance      ♦ - Good, Flexible      ◊ - Hard


In-vivo Studies

Approval for the use of animals in the study was obtained from the Institutional Animal Ethical Committee (NCP/IAEC/PG/05/2007). Male rabbits, 10 – 12 weeks old, weighing 2.8 to 3.5 kg were used to measure the in-vivo release of the drug in the eye. The rabbits were housed single in restraining boxes during the experiment and fed with standard rodent pellet diet and as much as required. Light and dark cycle was maintained throughout the study. The temperature, relative humidity conditions were 28 ± 2º and 60 ± 15º, respectively. A group containing 12 healthy rabbits was treated as control. Similarly another set containing same number of rabbits was used as study group. All of them were kept in hygienic conditions to avoid vulnerability to any disease including ophthalmic type. The sterilized formulation F-7 and drug free films were placed in the lower eyelid of rabbits of study group and control group respectively6. At specific time intervals, the films were removed carefully and analyzed for the remaining drug content.


The potential ocular irritation or damaging effects of the ocuserts under test were evaluated by observing them for any redness, inflammation or increased tear production. Formulation was tested on six rabbits by placing the inserts in the cul-de- sac of the left eye. Both eyes of the rabbits under test were examined for any signs of irritation before treatment and observed upto 24 hrs.



In the present study, efforts have been made to prepare ocular inserts of Ciprofloxacin hydrochloride using different polymers such as polyvinyl alcohol, polyvinyl pyrrolidone K-30 and ethyl cellulose in various proportions and evaluated for their physicochemical parameters like thickness, weight uniformity, drug content, percentage moisture loss, and percentage moisture absorption and in-vitro, in-vivo drug release study, sterility study, stability study etc.


Fig. I.  Cumulative percent drug released in vitro vs. time from ocular inserts.  Mean ± S.D are presented (n=3).


For the preparation of drug reservoir film, PVA and PVP K-30 were used and propylene glycol was used as plasticizer. Best films were obtained when the plasticizer concentration was 30 % w/w of the dry mass of the polymer. The content of the drug in ocular insert was estimated to be 1.863 ± 0.007 mg. The rate controlling films were prepared with ethyl cellulose. Uniform, flexible and transparent films were obtained and denoted as F -1, F-2, F-3, F-4, F-5, F-6, F-7,    F-8, and F-9 which were subjected to in-vitro drug release studies. The maximum, 99.11 %, drug release was observed with formulation F-7 for 24 hrs whereas formulation F-1 (77.26 %), F-2 (69.30 %), F-3 (60.40 %), F-4 (86.08 %),                F-5 (94.81 %), and F-6 (96.93 %) released the drug.  Formulations   F-8 and F-9 showed 93.91 % and 96.29 % of drug release within 18 and 16 hrs respectively.  Formulation F-7 was found to be the best since constant

Table III.  Drug release kinetic data.



Zero ordera

Higuchi’s modela

Peppa’s modela






































































amean ± SD, n=3


and complete release of drug was observed up to 24 hrs (Fig. I). Synthetic polymers are widely used in biomedical applications. Polymer blends have recently paved their way in this field. The incorporation of drug characterized by different physicochemical properties in a matrix shows that the   nature of therapeutic agent loaded into the polymers plays an important role in its release rate as well as in the degradation of the polymeric matrix.  The addition of PVP K30 to PVA, the films of PVA become resilient and do not break easily and it was ascertained that the drug diffusion might improve. The release rate was found to increase by increasing the concentration of PVP K-30.

Fig. II.  FTIR spectra for ciprofloxacin hydrochloride

Fig. III. FTIR spectra for formulation F-7


Interaction studies were carried out to ascertain any kind of interaction of the drug with the excipients used in the formulation. For this purpose, the optimized formulation F-7, placebo formulation and the pure drug were subjected to the assay, UV, TLC and IR analyses. The principal spot in TLC obtained with the test solution was similar in position, colour and size to the chromatogram obtained with the reference standard of the drug. Rf value of 0.59 was obtained with the medicated formulation and drug reference standard. The UV absorption maximum for the pure drug and the medicated formulation was found to be at 278 nm. The spectra recorded were taken as qualitative in order to assess the change in peaks, pattern of curve, etc. No major differences were observed in the IR spectra of the pure drug and the medicated formulation. Fig II and III represent the IR spectra of ciprofloxacin and its complex, respectively. The spectrum of ciprofloxacin shows an absorption band for C = O at 1730 cm-1 as the fraction of the -COOH group, whereas the IR spectra of the complex does not show any absorption band in that region, i.e. 1650 cm-1 to 1750 cm-1. The IR spectrum of the ciprofloxacin shows a sharp peak at 2400 cm-1, which indicates only one -OH group.


Fig. IV.  Formulation F-7 placed in rabbit’s eye


The physicochemical evaluation (12) data of Table I indicates that the formulation F-1, F-2, and F-3 does not contain PVP K-30 and F-4 exhibited low moisture absorption whereas formulation F-7 to F-9 had shown maximum percent moisture absorption. This may be due to the hydrophilic nature of polymers. It is reveled that the total content of the hydrophilic polymers in each batch and nature of individual polymers have influenced on moisture absorption characteristics of formulations. The percentage moisture loss is more in formulation F-4. This may be due to the presence of hydrophobic ethyl cellulose membrane and comparatively low concentration of polymers. Though percentage of moisture absorption and the percentage of moisture loss are high, there is no change in the integrity at high humid and dry conditions. Thickness of the ocuserts varies between 0.260 and 0.323mm. The formulations are not very thicker and do not produce any irritation while placing and being in cul-de-sac. The minimum standard deviation values revealed the fact that process used in the study is capable of giving films of uniform magnitude. This fact on the reliability of the process is further confirmed by drug content analysis data.

Fig. V.  In vitro In vivo correlation

The formulation F-7 was sterilized by ultraviolet radiation.  The sterilized ocuserts were tested for their sterility.  It was found visually that the Alternate Thioglycolate media, Fluid Thioglycolate media and Soybean casein digest media containing sterilized ocuserts were free from turbidity.  This confirmed the absence of aerobic, anaerobic organism and fungi.    

Stability studies were carried out by exposing the best formulation F-7 to various temperatures like 4 ± 1 °C, 37 ± 2 ° C and 60°C ± 2 for 6 weeks.  The results indicate that they are stable both physically and chemically (Table II). The physical appearance of the formulation exposed to 60°C showed a marked change after 20 daysThe period of expiry of the formulation F-7 was determined using Free and Blythe theory15. It shows that 92 % potency with 70 days stored at 25ºC.

In-vivo drug release study for the formulation F-7 was observed for 24 hrs by placing the formulation in the rabbit eye (Fig. IV) and removing it every 2 hrs for the determination of drug content. The delivery system was found to release 90.35 % of loaded drug at the end of 24 hrs. The regression analysis was carried to establish correlation between in-vitro – in-vivo release data. The correlation value of 0.9908 ± 0.318 indicated correctness of the in-vitro method followed and adaptability of the delivery system to the biological system where it can release the drug in concentration independent manner (Fig. V).  Statistical significance was studied using paired   ‘t’ test. A value of p <0.001 is considered statically significant.

The rabbits subjected to ocular irritation test did not show any signs of irritation, inflammation or abnormal discharge. However, the animal behavior was slightly agitated from the normal animals but the intake of food and water was normal.

The zero order plots of F-4,  F-5, F-6, F-7, F-8 and F-9 were found to be fairly linear as indicated by their high regression value. Therefore, it was ascertained that the drug release from above formulation could follow either near zero or zero order kinetics. In this study we obtained increasing rate constant values (k), which confirmed our theory. This is probably because the PVP K30 reduces the resistance offered by the PVA film alone, and by increasing pores and/or their diameter the drug diffuses with less resistance.

In controlled delivery, zero order is the most preferred kinetics of drug release.  The zero order plots for formulation F-7 in-vitro release found to be fairly linear as indicated by their regression value (0.9914). To confirm the exact mechanism of drug release from the films, the data was computed and graphed according to Highuchis and Peppas equation17-19. Regression analysis performed and the regression value ‘r’ suggested that the curves were fairly linear (Table III).  The “n” values suggest that the formulation F-7 follows a super case II transport mechanism (n >1.00).



All the formulations were found to be smooth, transparent and flexible. Thickness was fairly uniform as indicated by their low coefficient of variation. The formulation F-7 containing 4 % polyvinyl alcohol and 2 % polyvinyl pyrrolidone     K-30 as drug reservoir of ciprofloxacin hydrochloride and 4 % ethyl cellulose as a rate controlling membrane has shown best release with 99.11% at the end of 24 hours in concentration independent manner.  Excellent correlation was observed between in-vitro and in-vivo profiles not only affirmed selection of F-7 as best formulation along with adoptability to biological environment but also suitable for the method followed to study in-vitro characteristics.  The formulation F-7 was stable at 4 °C and 37 °C hence it can be easily stored at both temperatures with no apparent chemical interaction between the drug and the excipients. On the basis of in-vitro, microbiological, in-vivo drug release, interaction and stability studies, it can be concluded that this formulation F-7 provided the desired drug release in-vitro for 24 hrs and remained stable and intact at ambient conditions.


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Received on 18.03.2008          Modified on 25.03.2008

Accepted on 28.03.2008          © RJPT All right reserved

Research J. Pharm. and Tech. 1(1): Jan.-Mar. 2008; Page 40-45









Institutional News





A national seminar on "Research and Education in Pharmaceutical Sciences" was organized by GRY Institute of Pharmacy, Borawan, Khargone, Madhya Pradesh on April 13, 2008. About 250 delegates from different parts of the country including Punjab, Uttarakhand, Sikkim, Rajasthan, Gujarat, Maharashtra, West Bengal, Madhya Pradesh, Tamil Nadu, and Uttar Pradesh, were attended the seminar. The delegates were welcomed by Mr. Akhilesh Gupta, Vice-President, GRY Institute of Pharmacy Student Association.  The Special Guest of Honor was Mr. Subhash Yadav, Ex. Deputy Chief Minister, Govt. of Madhya Pradesh, in his inaugural address, stressed upon the need of research activities in academic field. He also quoted that a "Research Centre" is soon to be established at GRY Institute of Pharmacy. The Chief Guest, Prof. Piyush Trivedi, Dean, Pharmaceutical Sciences, RGPV, Bhopal, stressed upon the importance of literature review before any research work. He cited the importance of extensive literature review and patent in contemporary research. Dr. S. K. Yadav, Principal, VNS College of Pharmacy, Bhopal, deliberated that the research work in pharmacy profession has a bright future ahead and advised the present delegates to engage themselves in some practically useful research work. Mr. Arun Yadav, MP, Khargone Constituency, called upon the young professionals and researchers to dedicate themselves for the up-liftmen of pharmacy research and education, and assured that he will continue his support for the betterment of pharmacy profession in Nimar region which happens to be one of the most neglected areas of India. About 22 oral and more than 100 posters were presented by the delegates during the seminar. The scientific sessions were categorized discipline wise. The presentations were evaluated by a panel of judges. The quality of papers presented in each session was excellent and many of them generated meaningful discussions and interesting interactions. Winners of oral and poster presentations were awarded certificate of merit and rest of participants were given certificate of participation. Mr. Amit Roy, Convener, of the seminar expressed vote of thanks to all the educationists, professionals and participants for successful completion of the seminar