1. Formulation and Evaluation of Ophthalmic Ciprofloxacin Hydrochloride In-situ Gel Ngu Wah Aung, Thein May Saw, Cho Yi Myint, Maung Maung, Wai Wai Lwin, Mya Mya Aye Univeristy of Pharmacy (Mandalay)
Professor Dr. Ye Ye Tin
Professor Dr Shin Nawn Lwin

2. INTRODUCTION
Many eye diseases such as conjunctivitis, uveitis, glaucoma etc can affect the eye that may even lead to loss of vision
The microorganisms which cause eye infections are Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Klebsiella pneumonia1
1. Akhlaghi MR, Dehghani AR, Fazel F, Ghanbari H, Ilanloo MR and Azad A. Cefazolin Gentamycin Versus Vancomycin-Ceftazidime Eye Drops for Bacterial Corneal Ulcers. Ophthalmic and Vision Research Journal 2009; 4(1): 19-23

3. For the treatment of ocular infections, topical administration of antibiotic eye drops is the most common method7
Among the fluoroquinolone antibiotics, ciprofloxacin is a second generation and uses for the treatment of eye infections
7. Kristiina J, Tomi J and Urtii A. Ocular Absorption following Drug Delivery. Journal of Advance Drug Delivery Review 1995; 16(1): 3-19
Introduction
According to Nema and Nema, 1998, fluoroquinolones are the most frequently used antibiotics for eye infections

4. Indicated the topical ophthalmic administration of 0
Indicated the topical ophthalmic administration of 0.3% ciprofloxacin eye drops in case of severe infection 13
Drops or ointments (the most commonly available ophthalmic preparations) % of the eye dosage formulations in the market
13. The Indian Pharmacopoeia. The Indian Pharmacopoeia Commission: Ghaziaba, India, Ministry of Health and Public Welfare, Government of India, New Delhi, 2010; 2: 1032
Introduction

5. However, the major problems encountered with the eye drops are rapid and extensive elimination, short pre-corneal residence time and poor ocular bioavailability
Use various ophthalmic vehicles to increase ocular bioavailability and duration of drug action
Introduction
Therefore, needed frequent instillation of eye drops to achieve desired drug concentration and therapeutic effect (Sreeraj, Mitra & Hughes, 2003)
i.e., viscous solutions and polymeric inserts

6. This can be achieved by in-situ gelation system 10
An ideal ophthalmic dosage form is one which can sustain the drug release and remain pre-corneal contact for an extended period of time
This can be achieved by in-situ gelation system 10
10. Nanjawade BK, Manvi FV and Manjappa AS. In-Situ Forming Hydrogels for Sustained Ophthalmic Drug Delivery. Journal of Controlled Release 2007; 122(2):
Introduction

7. Depending upon the method, there are three types of systems 2
In this delivery system, consists of polymers & exhibit sol to gel phase transition, in physiological conditions of the eye 3
Depending upon the method, there are three types of systems 2
2. Balasubramaniam J, Kant S and Pandit JK. In-vitro and In-vivo Evaluation of the Gelrite Gellan Gum-Based Ocular Delivery System for Indomethacin. Journal of ACTA Pharm 2003; 53:
3. Bolanle L. Development of A Cysteamine In-Situ Gelling System for The Treatment of Corneal Crystals in Cystinosis. [PhD thesis]. The School of Pharmacy: University of London; 2008
Introduction
(i) pH triggered systems, (ii) temperature dependent systems and (iii) ion activated systems

8. Advantages of in-situ systems
Introduction
Advantages of in-situ systems
to provide sustained and controlled drug delivery
to increase the ocular bioavailability of drug by increasing the pre-corneal contact time and
to provide comfort and better compliance to the patient

9. To deliver the drug effectively into the eye
Introduction
To deliver the drug effectively into the eye
the formulation of ciprofloxacin hydrochloride in-situ gel using an ion-activated phase transition polymer to obtain the sustained drug release

10. Methods
Identification of Ciprofloxacin hydrochloride Compatibility Test Formulation of In-situ Gel Evaluation of Prepared In-situ Gel

11. Formulation of Ciprofloxacin Hydrochloride In-situ Gels
Method
Formulation of Ciprofloxacin Hydrochloride In-situ Gels
Table 1. Formulation of Ciprofloxacin Hydrochloride In-situ Gels
Ingredients F1 F2 F3 F4
Ciprofloxacin hydrochloride (g)
0.3
Sodium alginate (g)
0.5 1
1.5
Hydroxypropyl methylcellulose (g) -
Benzalkonium chloride (ml)
0.01
Lactic acid (ml) qs
0.1 N Hydrochloric acid (ml)
Distilled water (ml) to
100

12. Evaluation of the Prepared Ciprofloxacin Hydrochloride In-situ Gels
Method
Evaluation of the Prepared Ciprofloxacin Hydrochloride In-situ Gels
pH of Ciprofloxacin Hydrochloride In-situ Gels
Clarity Test of Formulations
Viscosity of Ciprofloxacin Hydrochloride In-situ Gels
Gelling Capacity of Ciprofloxacin Hydrochloride In-situ Gels

13. 6. In vitro Drug Release Study
Method
Evaluation (Conts:)
5. Determination of Content of Ciprofloxacin Hydrochloride in In-situ Gels by UV-visible Spectrophotometry
6. In vitro Drug Release Study
7. Sterility Test of Ciprofloxacin Hydrochloride In-situ Gels

14. Evaluation (Conts:)
Method
8. In vivo Eye Irritation Test of Ciprofloxacin Hydrochloride In-situ Gels 9. Antimicrobial Susceptibility of Ciprofloxacin Hydrochloride In-situ Gels 10. Stability Testing

15. Evaluation (Conts:) Table 2. Coding for the Gelling Capacity 9
Method
Table 2. Coding for the Gelling Capacity 9
9. Makwan SB, Patel VA and Parmar SJ. Development and Characterization of In-Situ gel for Ophthalmic Formation Containing Ciprofloxacin Hydrochloride. Journal of Results in Pharm Science 2016; 6: 1-6
Observation
Coding
No gelation _
Gelation occurred in few minutes and remained for few hour +
Gelation immediately and remained for few hour ++
Gelation immediately and remained for extended period
+++
Very stiff Gel
++++

16. RESULTS AND DISCUSSION
1. Formulation of Ciprofloxacin Hydrochloride In-situ Gels
Before formulation, preliminary formulation is important
In identification, similar between finger print of IR spectrum for standard and sample of ciprofloxacin hydrochloride

17. Results and Discussion
1. Formulation of Ciprofloxacin Hydrochloride In-situ Gels (Conts:)
The compatibility test showed that all the excipients were compatible because there were no physical changes
Ciprofloxacin is practically insoluble in water and so used hydrochloride salt form in this formulation
Results and Discussion

18. Results and Discussion
Formulation of Ciprofloxacin Hydrochloride In-situ Gels (Conts:)
The hydrochloride salt form of ciprofloxacin is freely soluble in water at acidic pH below 5
The pH of solution of sodium alginate and hydroxylpropyl methylcellulose was 9.6 and so used lactic acid to reduce the pH of polymer solution

19. Results and Discussion
2. Evaluation of Ciprofloxacin Hydrochloride In-situ Gels
Not significantly different between the pH values of four formulations and within the acceptable pH range 4 to 8
Due to the presence of cellulose derivative, the appearance of the formulations was milky but free from particulate matter

20. Results and Discussion
2. Evaluation of Ciprofloxacin Hydrochloride In-situ Gels (Conts)
The smallest viscosity value of formulation F1 among the four formulations because of the lowest amount (0.5%) of sodium alginate
The second smallest viscosity value was formulation F2 due to the second lowest amount (1%) of sodium alginate

21. Results and Discussion
2. Evaluation of Ciprofloxacin Hydrochloride In-situ Gels (Conts)
The largest viscosity value of formulation F3 because of the highest amount (1.5%) of sodium alginate
Although F3 and F4 had the same amount (1.5%) of sodium alginate, smaller the viscosity value of F4 than F3 because F4 did not contain hydroxypropyl methylcellulose

22. Results and Discussion
2. Evaluation of Ciprofloxacin Hydrochloride In-situ Gels (Conts:)
Therefore, the viscosity was directly dependent on the concentration of polymer and the presence or absence of co-polymer
The four formulations caused gelation due to the ionic cross-linking of the alginate chain by divalent cation, Ca2+ present in the simulated lacrimal fluid

23. Results and Discussion
2. Evaluation of Ciprofloxacin Hydrochloride In-situ Gels (Conts:)
But depending on the amount of sodium alginate, different the rate and extent of gelation
The more sodium alginate present, the more gelation occurred
Therefore, the gelling capacity was excellent in formulation F3 shown in figure 1

24. Results and Discussion
Figure 1. Gelling capacity of formulation F3 after placing of the formulation into simulated lacrimal fluid (SLF)
(a) 0 minutes
Gelling capacity
(+)
(b) 30 minutes
(c) 60 minutes
(d) 120 minutes
(+ + +)

25. Results and Discussion
2. Evaluation of Ciprofloxacin Hydrochloride In-situ Gels (Conts:)
In vitro drug release studied indicated that the release pattern and duration were different in all formulations
In formulation F1, time required to reach the peak drug concentration (62.85%) and duration of drug release of formulation F1 were the same as the market sample (1 hr)

26. Results and Discussion
2. Evaluation of Ciprofloxacin Hydrochloride In-situ Gels (Conts:)
In formulation F2, time required to reach the peak drug concentration (102.79%) and duration of drug release of formulation were longer than the formulation F1 and market sample
In formulation F3 (Figure 2), time required to reach the peak drug concentration (66.57%) and duration of drug release of formulation were longest among others

27. Results and Discussion
2. Evaluation of Ciprofloxacin Hydrochloride In-situ Gels (Conts:)
In formulation F4, time required to reach the peak drug concentration (69.56%) and duration of drug release of formulation were longer than the formulation F1, F2 and market sample
But it was shorter than the formulation F3 because the formulation F4 didn’t contain 0.5% of co-polymer which was present in F3
Results and Discussion

28. Results and Discussion
2. Evaluation of Ciprofloxacin Hydrochloride In-situ Gels (Conts:)
Thus in vitro drug release study indicated that F3 had the nature of prolong duration of action
Therefore, depending on the concentration of polymer and co-polymer present in the formulations, different in the viscosity, the gelling capacity, the extent and prolong of drug release
Results and Discussion

29. Results and Discussion
Figure 2. Cumulative percent release of ciprofloxacin hydrochloride in-situ gel formulation F3

30. Results and Discussion
2. Evaluation of Ciprofloxacin Hydrochloride In-situ Gels (Conts:)
At the end of sterility test, because of no appearance of turbidity, all formulations passed sterility tests
In vivo eye irritation test, there was no redness, swelling or watering of eyes after instillation of the formulations
Therefore, all formulations contained non-irritant ingredients to the eye

31. Results and Discussion
2. Evaluation of Ciprofloxacin Hydrochloride In-situ Gels (Conts:)
According to standard zone size provided by CLSI, all formulations were sensitive on the test organisms
Therefore, ciprofloxacin hydrochloride remained its antimicrobial efficacy after formulation and active against the selected strain of microorganisms
Results and Discussion

32. Results and Discussion
2. Evaluation of Ciprofloxacin Hydrochloride In-situ Gels (Conts:)
After 3 months of stability testing, slightly different in the pH values of all formulations from the initial pH value
But lied within acceptable pH range for the eyes
The preparations were also clear, no color changes and no precipitation
Results and Discussion

33. Results and Discussion
2. Evaluation of Ciprofloxacin Hydrochloride In-situ Gels (Conts:)
Decreased the viscosity of all formulations when compared with the initial value which may be because of temperature
After 3 months at 40 ˚C ± 2 ˚C, 18 %, 4 %, 12 % and 29 % decreased in the assays of ciprofloxacin hydrochloride in all formulations

34. Results and Discussion
2. Evaluation of Ciprofloxacin Hydrochloride In-situ Gels (Conts:)
After 3 months at 30 ˚C ± 2 ˚C, 3 % and 6 % decreased in the assay of ciprofloxacin hydrochloride in formulation F1 and F4 but in formulation F2 and F3,1 % decreased
After 3 months at 20 ˚C ± 2 ˚C, 1 % decreased in the assay of ciprofloxacin hydrochloride in formulation F2, F3 and F4 but in formulation F1, 2 % decreased
Results and Discussion

35. Results and Discussion
2. Evaluation of Ciprofloxacin Hydrochloride In-situ Gels (Conts:)
Therefore, should not exceed the storage condition of ciprofloxacin hydrochloride in-situ gel at 30 ˚C ± 2 ˚C

36. CONCLUSION
Well formulated ciprofloxacin hydrochloride as in-situ gel forming eye drops using sodium alginate and hydroxypropyl methylcellulose
Therefore, can be used this polymer and co-polymer mixture as an in-situ gelling vehicle to enhance prolong contact time and ocular bioavailability

37. Conclusion
The satisfactory results of pH, clarity, viscosity, gelling capacity, assay, sterility and eye irritation test indicated that all of the formulations were complied with the specifications mention in the pharmacopeia
Therefore, the developed formulations had acceptable pharmaceutical qualities

38. Conclusion
Besides, among the four formulations, formulation F3 was complied with the pharmaceutical parameters and more likely to be acceptable than the other formulations (F1, F2 and F4)
It had the best pharmaceutical qualities while using the 1.5% of sodium alginate and 0.5% of hydroxypropyl methylcellulose
because of more benefits of viscosity, gelling capacity and sustained drug release

39. Conclusion
The formulations also had efficacy because of successful in inhibiting the growth of the micro-organisms for 24 hr when compared with the ciprofloxacin standard disc

40. ACKNOWLEDGEMENT
I would like to express my sincere gratitude to Professor Dr. Thein May Saw, Rector, University of Pharmacy, Mandalay
I am extremely indebted to Dr. Cho Yi Myint, Professor and Head, Department of Pharmaceutics, University of Pharmacy, Mandalay, for her support and providing research facilities for this work

41. ACKNOWLEDGEMENT (Conts:)
I am deeply grateful to my supervisor, Dr. Wai Wai Lwin, Lecturer, Department of Pharmaceutics, University of Pharmacy, Mandalay for her invaluable suggestions and supervision throughout the period of study
I would like to thank my co-supervisor, Dr. Mya Mya Aye, Deputy Director and Head, Bacteriology Research Division, Department of Medical Research, Yangon for her permission to utilize the laboratory facilities and valuable guidance for microbiology research work

42. Thanks You