ORGANOGELS AS CARRIERS FOR TOPICAL DELIVERY OF MICONAZOLE NITRATE
By MD.MUQTADAR AHMED Reg. No. 04PU254
Dissertation Submitted to the Rajiv Gandhi University of Health Sciences, Karnataka, Bangalore
In partial fulfillment of the requirements for the degree of
MASTER OF PHARMACY in PHARMACEUTICS
Under the Guidance of Dr.MOHAMED HASSAN DEHGHAN M.Pharm., Ph.D.
DEPARTMENT OF PHARMACEUTICS LUQMAN COLLEGE OF PHARMACY, GULBARGA-585 102 APRIL 2006 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) ii RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA, BANGALORE
Declaration By The Candidate
I hereby declare that this dissertation/ thesis entitled STUDIES ON LECITHIN-MICROEMULSION BASED ORGANOGELS AS CARRIERS FOR TOPICAL DELIVERY OF MICONAZOLE NITRATE is a bonafide and genuine research work carried out by me under the guidance of Dr.Mohamed Hassan Dehghan.
Date: Place: GULBARGA MD.MUQTADAR AHMED
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Certificate By The Guide
This is to certify that the dissertation entitled STUDIES ON LECITHIN-MICROEMULSION BASED ORGANOGELS AS CARRIERS FOR TOPICAL DELIVERY OF MICONAZOLE NITRATE is a bonafide research work done by Mr.MD.MUQTADAR AHMED in partial fulfillment of the requirement for the degree of MASTER OF PHARMACY in PHARMACEUTICS.
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RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA, BANGALORE
ENDORSEMENT BY THE HOD, PRINCIPAL/ HEAD OF THE INSTITUTION
This is to certify that the dissertation entitled STUDIES ON LECITHIN-MICROEMULSION BASED ORGANOGELS AS CARRIERS FOR TOPICAL DELIVERY OF MICONAZOLE NITRATE is a bonafide research work done by Mr.MD.MUQTADAR AHMED under the guidance of DR.MOHAMED HASSAN DEHGHAN.
Date: Place: GULBARGA Prof.Syed Sanaullah Principal, Luqman College of Pharmacy, Gulbarga-585102
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COPYRIGHT
Declaration By The Candidate
I here by declare that the Rajiv Gandhi University of Health Sciences, Karnataka shall have the rights to preserve, use and disseminate this dissertation/ thesis in print or electronic format for academic/ research purpose.
Date: Place: GULBARGA Mr.MD.MUQTADAR AHMED
Rajiv Gandhi University of Health Sciences, Karnataka
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ACKNOWLEDGEMENT
I am especially deeply indebted to my honourable research guide Dr.Mohammed Hassan Dehghan for the constant guidance, encouragement and support, which I have received from him whole- heartedly. I substituted for the feeling of gratitude, indebtedness and appreciation for him who brought me from darkness to lightness in my life, which I would never forge Thank You Sir.
It is my privilege to express my heartfelt thanks to Prof.Syed Sanaullah, Principal, Luqman College of Pharmacy for allowing me to use all the facilities of the college and support me like a pillar for constant inspiration and guidance as environment required.
I am most thankful to Dr.Syed Rahmatullah, General Secretary, Vocational Education Society, Gulbarga for his silent encouragement.
My sincere thanks to Dr.Mujeeb and Mr.Abdul Majeed, President, Vocational Education Society, Gulbarga for providing me all the facilities.
I honestly acknowledge Prof.M.A.Saleem, Prof.S.S.Bushetti, Prof.Syeda Humera, Prof.Divakar, Mr.M.H.Hugar, Mr.Jafar, Mr.Najmuddin, Prof.Satyanandam and Mr.Omar Khan and other teaching staff of Luqman College of Pharmacy, Gulbarga for their timely encouragement during my entire P.G. course.
A very special thanks to Mr.Zahid, Lecturer, Y.B.Chauhan College of Pharmacy, Aurangabad.
My heartfelt gratitude and sincere thanks to my teachers Mr.Rajesh Patwari, Mr.Shaikh Bahadur, Dr.Mazhar Farooqui and Mr.Sadat Ali for their timely suggestions and perspective guidance in enriching my knowledge.
I place on record a respectful thanks to Dr.M.G.Purohit, Department of Pharmaceutical Chemistry, Gulbarga University, Gulbarga for his guidance in my analytical work.
A special thanks to M/s.Adelphi A/c Sigma Labs, Goa for providing drug sample of Miconazole Nitrate.
I also thanks Mr.Sridhar, Sipra Labs, Hyderabad for IR analytis of all formulations, the Librarians of NIN, O.U., IICT, Hyderabad for their kind help during literature survey. Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) vii
I could never forget (Mrs).Ayesha, (Mrs).Florence for the inspiration, encouragement and moral support during the course of my studies.
It gives me immense pleasure to record my sincere thanks to my colleagues and friends Abdul Muqtadir, Asif, Aleem, Vinod Singh, Taher, Imran, Anant Kulkarni, Nagashesha R, Mohan VK, Masood, Rizwan, Saleem, Mir Imran, Sajid, Ilyaz, Vijay, Imtiyaz, Manish Kumar Mital, Fazil, Shad, Noor, Shahid, Sarim, Abhisek, Wali, Areefulla H. for helping me in carrying out this work.
It gives me immense pleasure to record my thanks to my seniors Faisal, Azim, Riyaz, Ismail Mauzam for their guidance during work.
I express my thnaks to non-teaching staff Asad, Pasha, Naveed, Narender, Hassan and Librarian Rubina Anjum and Pratibha for their help and cooperation.
I am thankful to Micro Computers, Gulbarga for their cooperation during the time of typing of this research work.
In closing I have reserved my special and most grateful thanks to one who has not only initiated my interest in this work but has also led me through the dark alley and abyssess to the brighten path. I am referring to none other than my seniors Mr.Abdullah Khan and Mr.Md.Jafar who worked for my success thank you.
The presence of the Almighty God was felt by me during my research work. I am thankful to the Supreme energy for manifesting Himself through the various helpful people, I came across in my life. I bow my head to Him and ask for His blessings to be with me forever.
And above all words fails to express my feelings to my Parents and my Family whose initiation, constant source of inspiration and encouragement throughout my life.
Date: Place: Gulbarga Md.Muqtadar Ahmed
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LIST OF ABBREVIATIONS USED
..................... micro %.................... Percentage BP................... British Pharmacopoeia cm................... centimeter CPs................. centipoise F 127 ................. Pluronic fig................... figure gm.................. gram hrs................... hour ICH................. International Conference for Hormonization IP.................... Indian Pharmacopoeia IPM................. Isopropyl myristate IR.................... infrared Km.................. weight ratios LOs................. Lecithin organogels MBGs............ Microemulsion based organogels mcg/g............ microgram(s) mg.................. milligram(s) min................. minute(s) ml ................... milliliter mol-wt............ Molecular weight nm.................. nanometer NTU ............... Nephaloturbidity unit PLO................ Pluronic lecithin organogels rpm................. Revolution per minute sec.................. Second(s) Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) ix Sq-cm............. Square centimeter Std.................. Standard(s) USP................ United States Pharmacopoeia UV.................. Ultraviolet w/w................. Weight per weight Wt................... Weight P..................... Penicillin chrysogenum C..................... Candida albicans A..................... Aspergillus niger T..................... AM 1 Formulation F..................... FM 1 Formulation I ...................... SR 1 Formulation P..................... MN 1 Formulation Mkd................ Marketed
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AM 1 ....................Lecithin 7.54%, tween-80 3.77%, isopropyl myristate 30.16%, water 56.56%, miconazole nitrate 2% AM 2 ....................Lecithin 20.63%, tween-80 10.31%, isopropyl myristate 30.95%, water 36.11%, miconazole nitrate 2% AM 3 ....................Lecithin 35.43%, tween-80 1.77%, isopropyl myristate 11.81%, water 21.26%, miconazole nitrate 2% AM 4 ....................Lecithin 56.02%, tween-80 28.01%, isopropyl myristate 14.00%, water 28.01%, miconazole nitrate 2% FM 1 .....................Lecithin 7.54%, isopropyl myristate 30.16%, pluronic 30% in water 60.32, miconazole nitrate 2% FM 2 .....................Lecithin 20.63%, isopropyl myristate 26.73%, pluronic 30% in water 53.47%, miconazole nitrate 2% FM 3 .....................Lecithin 35.43%, isopropyl myristate 24.50%, pluronic 30% in water 36.76%, miconazole nitrate 2% FM 4 .....................Lecithin 56.02%, isopropyl myristate 14.00%, pluronic 30% in water 28.01%, miconazole nitrate 2% SR 1 ......................Lecithin 8.9%, IPM 35.1%, water 53.5%, miconazole 2% SR 2 ......................Lecithin 26.22%, IPM 30.33%, water 47.5%, miconazole 2% SR 3 ......................Lecithin 43.8%, IPM 28.8%, water 25.95%, miconazole 2% SR 4 ......................Lecithin 60.33%, IPM 15.08%, water 22.63%, miconazole 2% MN 1 ....................Lecithin 13.43%, paraffin 53.78%, water 30.88%, miconazole 2% MN 2 ....................Lecithin 31.6%, paraffin 47.4%, water 18.99%, miconazole 2% MN 3 ....................Lecithin 49.0%, paraffin 32.67%, water 16.3%, miconazole 2% MN 4 ....................Lecithin 67.61%, paraffin 16.90%, water 13.52%, miconazole 2%
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) xi ABSTRACT
In the present study lecithin-microemulsion based organogels were formulated as topical carrier for miconazole nitrate, an antifungal drug practically insoluble in water. The organogels were prepared by two different methods employing lecithin, lecithin in combination with Tween-80 and lecithin with pluronic, isopropyl myristate(IPM) was used as organic solvent, whereas gelation was achieved on addition of aqeuous phase. Organogels using lecithin in liquid paraffin and water were also prepared by hot-melt method. Increase in the amount of lecithin resulted in organogels with higher viscosity and lower spreadability. MN 4 formulation showed maximum viscosity 22868 CPs, whereas AM 1 formulation containing Tween-80 (3.77%) was least viscous 15726 CPs. Organogels with higher viscosity were found to be more stable. Organogels containing lecithin and Tween-80 were the most stable. AM 4 showed highest gel life of 220 hours at 25C. 28.5 gm% of water was required to produce maximum gelation in SR 1 organogels containing lecithin and IPM. Organogels produced by hot melt method required least amount of water for maximum gelation 9.09 gm%. In vitro release of miconazole nitrate from organogels showed that organogels containing lecithin and Tween-80 in IPM (AM 1 ) gave highest release. The order of release for the best releasing formulation prepared by different methods was AM 1 >FM 1 >SR 1 >MN 1 . In vitro antifungal activity of miconazole nitrate organogels against Candida albicans, P.chrysogenum, A.niger was in the order AM 1 >FM 1 Mkd>SR 1 >MN 1 . In vitro antifungal activity significantly correlated with in vitro release of miconazole nitrate. Lecithin microemulsion based organogels have good potential as carrier for topical delivery of antifungal agent such as miconazole nitrate.
Keywords: Lecithin organogels; Miconazole nitrate; In vitro release, In vitro anti-fungal activity.
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LIST OF TABLES........................................................... xiv-xv LIST OF FIGURES.............................................................. xvi
CHAPTER-1 INTRODUCTION............................................................ 01-29 1.1 Conventional Technologies.......................................... 01 1.2 Newer Technologies.................................................... 02 1.3 Topical Dosage Forms................................................. 04 1.4 Historical Overview..................................................... 06 1.5 Skin as a Route of Topical and Transdermal Drug Delivery System........................................................... 07 1.6 Fungal Infection........................................................... 11 1.7 Rational Approach to Drug Delivery to and Via Skin............................................................................. 14 1.8 Utilization of Different Gels as Topical Vehicles......... 15 1.9 Emulsion Gels as Topical Formulations....................... 15 1.10 Organogels................................................................... 16 1.11 Lecithin Organogels An Overview............................ 19 1.12 Method of Preparation................................................. 23 1.13 Characterization of Organogels.................................... 24 1.14 In Vitro Drug Release.................................................. 25 1.15 Topical Drug Delivery Applications of Lecithin Organogel Based System........................................... 26 1.16 Safety and Skin Compatibility...................................... 28 1.17 Future Prospects........................................................... 28
CHAPTER-2 OBJECTIVES .................................................................. 30-32 2.1 Need for the Study....................................................... 30 2.2 Objective of the Study.................................................. 31 2.3 Scheme of the Work..................................................... 31
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) xiii CHAPTER-3 REVIEW OF LITERATURE.......................................... 33-58 3.1 Review of Literature.................................................... 33 3.2 Drug Profile................................................................. 42 3.3 Polymer Profile............................................................ 50
CHAPTER-4 METHODOLOGY........................................................... 59-69 4.1 Raw Material Characterization..................................... 61 4.2 Preparation of Miconazole Nitrate Loaded Lecithin Microemulsion based Organogels................................ 61 4.3 Construction of Calibration Curve of Miconazole Nitrate.......................................................................... 64 4.4 Evaluation of Lecithin-Microemulsion based Organogels................................................................... 65
CHAPTER-5 RESULTS ....................................................................... 70-115 5.1 Standardization of Raw Materials............................ 70-79 5.2 Preparation of Lecithin based Organogels.................... 80 5.3 Construction of Curve of Miconazole Nitrate............... 80 5.4 Evaluation of Lecithin-Microemulsion based Organogels............................................................ 82-115
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) xiv LIST OF TABLES Sl. No. Table No. Title Page No. 1. 1.1 Major Lipids of the Stratum Corneum 10 2. 4.2.1 Preparation of Organogels Loaded with Miconazole Nitrate 62 3. 4.2.2 Preparation of Pluronic Lecithin Organogels 63 4. 4.2.3 Preparation of Miconazole Nitrate Loaded Organogels 63 5. 4.2.4 Preparation of Hot Melt Type Organogels 64 6. 5.1.1 Standardization of Miconazole Nitrate 70 7. 5.1.2a Standardization of Lecithin 72 8. 5.1.2b Standardization of tween 80 72 9. 5.1.2c Standardization of pluronic (poloxames) 75 10. 5.1.2d Standardization of Isopropyl myristate 75 11. 5.1.2e Standardization of Lipid Paraffin 78 12. 5.3 Calibration curve of Miconazole Nitrate 80 13. 5.4 Physical Evaluation of Formulations 82 14. 5.4.1a Gelation Kinetics AM 1 83 15. 5.4.1b Gelation Kinetics AM 2 84 16. 5.4.1c Gelation Kinetics AM 3 85 17. 5.4.1d Gelation Kinetics AM 4 86 18. 5.4.2a Gelation Kinetics FM 1 88 19. 5.4.2b Gelation Kinetics FM 2 89 20. 5.4.2c Gelation Kinetics FM 3 90 21. 5.4.2d Gelation Kinetics FM 4 91 22. 5.4.3a Gelation Kinetics SR 1 93 23. 5.4.3b Gelation Kinetics SR 2 94
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Sl. No. Table No. Title Page No. 24. 5.4.3c Gelation Kinetics SR 3 95 25. 5.4.3d Gelation Kinetics SR 4 96 26. 5.4.4a Gelation Kinetics MN 1 98 27. 5.4.4b Gelation Kinetics MN 2 98 28. 5.4.4c Gelation Kinetics MN 3 99 29. 5.4.4d Gelation Kinetics MN 4 99 30. 5.4.5 Gelatin Kinetics Data 101 31. 5.4.6 Percentage drug contents of Organogels 102 32. 5.4.7a In vitro percentage drug release of formulations AM 1 , AM 2 , AM 3 and AM 4
103 33. 5.4.7b In vitro percentage drug release of formulations FM 1 , FM 2 , FM 3
and FM 4
105 34. 5.4.7c In vitro percentage drug release of formulations SR 1 , SR 2 , SR 3
and SR 4
107 35. 5.4.7d In vitro percentage drug release of formulations MN 1 , MN 2 , MN 3 and MN 4
109 36. 5.4.8 In Vitro Antifungal Activity of Miconazole Nitrate Organogels 111
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Sl. No. Figure No. Title Page No. 1. 1.1 Structure of Skin 8 2. 5.1.1 IR Spectra of Miconazole Nitrate 71 3. 5.1.2a IR Spectra of Lecithin 73 4. 5.1.2b IR Spectra of Tween-80 74 5. 5.1.2c IR Spectra of Pluronic 76 6. 5.1.2d IR Spectra of Isopropyl Myristate 77 7. 5.1.2e IR Spectra of Liquid Paraffin 79 8. 5.3 Calibration curve of miconazole nitrate 81 9. 5.4.1 Effect of addition of water on turbidity of microemulsion based organogel of formulations AM 1 , AM 2 , AM 3 & AM 4
87 10. 5.4.2 Effect of addition of water on turbidity of microemulsion based organogel of formulations FM 1 , FM 2 , FM 3 & FM 4
92 11. 5.4.3 Effect of addition of water on turbidity of microemulsion based organogel of formulations SR 1 , SR 2 , SR 3 & SR 4
97 12. 5.4.4 Effect of addition of water on turbidity of microemulsion based organogel of formulations MN 1 , MN 2 , MN 3 & MN 4
100 13. 5.4.7a In vitro percentage drug release of formulations AM 1 , AM 2 , AM 3 and AM 4
104 14. 5.4.7b In vitro percentage drug release of formulations FM 1 , FM 2 , FM 3
and FM 4
106 15. 5.4.7c In vitro percentage drug release of formulations SR 1 , SR 2 , SR 3
and SR 4
108 16. 5.4.7d In vitro percentage drug release of formulations MN 1 , MN 2 , MN 3 and MN 4
110 17. 5.4.8 Comparative Antifungal Activity of Selected Miconazole Nitrate Organogel Formulations with Control 111 18. 5.4.9a IR Spectra of AM 1 Organogel 112 19. 5.4.9b IR Spectra of FM 1 Organogel 113 20. 5.4.9c IR Spectra of SR 1 Organogel 114 21. 5.4.9d IR Spectra of MN 1 Organogel 115
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Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 1 CHAPTER1 INTRODUCTION
With the advent of high throughput screening techniques the discovery of biologically active molecules is taking place at a pace never seen before. Most of the chemical entities that are being discovered are lipophilic in nature and have poor aqueous solubility, there by posing problems in their formulation into delivery systems. Because of their low aqueous solubility and high permeability, dissolution and/or release rate from the delivery system forms the rate-limiting step in their absorption and systemic availability. More than 60% of potential drug products suffer from poor water solubility. This frequently results in potentially important products not reaching the market or not achieving their full potential. Pharmaceutical industry is quick in realizing the importance of solubility and dissolution rate in bioavailability and good deal of research has been done in this area. Currently a number of technologies are available to address the poor solubility, dissolution rate and bioavailability of insoluble drugs 1 .
1.1 CONVENTIONAL TECHNOLOGIES Conventionally used techniques 2 based on Noyes-Whitney equation 3 for enhancing solubility, dissolution rate and thereby bioavailability of insoluble drugs include buffered tablets, use of salts, solvates and hydrates, polymorphic forms, complexation, prodrugs, micronisation, solid dispersions and solvent deposited systems. Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 2 1.2 NEWER TECHNOLOGIES Newer and novel drug delivery technologies developed in recent years for bioavailability enhancement of insoluble drugs are described below.
1.2.1 LIPID BASED DELIVERY SYSTEMS 1. Lipid emulsion technology 4 . 2. Self-emulsifying drug delivery system 5 . 3. Micro emulsion media as novel drug delivery system 6 .
1.2.1.1 MICRO EMULSION SYSTEM 6, 7 : Microemulsions are four component mixtures composing of an oil phase, a water phase surfactant/s and a co-surfactant. The tendency towards formation of w/o or o/w microemulsions is dependent on the properties of the oil and the surfactant, the water-to-oil-ratio and the temperature. When a mixture of surfactant and co- surfactant is added to a biphasic oil-water system, a thermodynamically stable, optically transparent or translucent, low viscosity and isotropic mixture spontaneously forms. The transparency of these systems arises from their small droplets diameter(10-100 nm). Such small droplets produce only weak scattering of visible light when compared with that from the coarse droplets (0.5-100 m) of traditional or standard macroemulsions such as emollient liquids, cream, lotions, etc., Structurally, microemulsions have normal micellar solutions, reverse micelles, cores or droplets of water or oil, and, for some systems, even bicontinuous structures could solubilize large amounts of both oil and water soluble drugs within microemulsions. Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 3 There is rather confusing situation in the medical literatures, where the term microemulsion is indifferently used to indicate systems of presumably unlike structure (true microemulsions and miniemulsions). Some studies have compared the performance of different emulsified systems (macroemulsions, microemulsion, multiple emulsion and gel-emulsions) prepared with similar oils and surfactants for applications such as controlled drug release or drug protection.
The surfactants used to stabilized such systems may be (i) Non-ionic (ii) Zwitterionic (iii) Cationic (iv) Anionic surfactants. Combinations of these, particularly ionic and non-ionic, can be very effective at increasing the extent of the microemulsion region.
Advantages: Advantages associated with microemulsions include their thermodynamic stability, optical clarity and ease of preparation.
Applications 6 : a. Oral delivery b. Parentral delivery c. Pulmonary delivery d. Ocular delivery e. Topical delivery
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 4 1.3 TOPICAL DOSAGE FORM: Topical dosage forms are those which are applied to the skin. These preparation are applied to the skin either for their physical effects, that is for their ability to act as skin protectants, lubricants, emollients, drying agents, etc. or for their specific effect of medicinal agents present. Preparations sold over the country frequently contain mixtures of medicinal substance used in the treatment of such condition as minor skin infection, itching, bruise, acne, psoriasis and eczema. Skin application, which require a prescription generally contain a single medicinal agent intended to counter a specific diagnosed condition 8 .
Topical dosage forms have been used since very ancient times. The application of medicinal substance to skin or to various body orifices is a concept as old as humanity. Various ointments, creams, gels, lotions, pastes, powders and plasters have been used for many years 9 .
The primary topical drug delivery systems (TDDS) is that they could provide controlled constant administration of a medicament by simple application to the skin surface.
1.3.1 Advantages of Topical Systems 10 : 1. They are of least therapeutic interest but of practical relevance is good patient compliance. The systems are easy to apply and remove. It avoids risks and inconveniences associated with intravenous therapy. Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 5 2. They eliminate the variables, which influences gastrointestinal absorption such as food intake, stomach emptying, intestinal motility and transit time. 3. Produces sustained and controlled level of drug in plasma thus reduces the chance of over or under-dosing. 4. Reduces frequency of drug dosing. 5. Topical systems are easily retractable thereby termination of drug input, if toxic effects are observed. 6. Offers an alternative route when oral therapy is not possible as in case of nausea and vomiting. 7. Helps in achievement of more constant blood levels with lower dosage of drug by continuous drug input and by by-passing hepatic first-pass metabolism and consequent degradation. 8. In certain circumstances, enzymatic transformation within epidermis may be used to improve permeability of certain hydrophilic drugs when applied to the skin in the form of prodrug.
1.3.2 Limitations of Topical Systems 11 : 1. Drugs with reasonable partition coefficient and possessing solubility both in oil and water are most ideal, as drug must diffuse through lipophilic stratum corneum and hydrophilic viable epidermis to reach the systemic circulation. Only drugs, which are effectively absorbed by the percutaneous routes as such or by using penetration promoters, can be considered. 2. The route is not suitable for drugs that irritate or sensitize the skin. Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 6 3. The route is restricted by the surface area of delivery system and the dose that needs to be administered in the chronic state of disease. 4. Topical drug delivery systems are relatively expensive compared to conventional dosage forms. They may contain a large amount of drug, of which only a small percentage may be used during the application period.
Apart from these limitations other problems include pharmacokinetics and pharmacodynamic restrictions. Thus clinical need has to be examined carefully before developing a TDDS.
1.4 HISTORICAL OVERVIEW: With the advent of scientific medicine in the last half of the 19 th century, this route of administration fell out of favour. In 1877, Fliescher declared that the skin was totally impermeable. This extreme view could not hold for long. By the turn of the century, Schwenkenbecker perceived that the skin would admit some substances much better than others. In 1957, Monash proved a superficially located barrier in skin as an obstacle to penetration. Later Stoughton used simple methods to explore the permeability behaviour of human skin. He brought to light many important facts by applying substances, which cause some rapid physiologic display such as blanching, reddening or sweating.
The most important efforts in establishing a theoretical foundation were from works by Ireger, Blank and Scheuplein.
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 7 These pioneering works were followed by extensive research ultimately proving that the stratum corneum was the main barrier to percutaneous absorption and that although it allows no substance to penetrate easily, it does allow all substances to enter slightly 12 .
1.5 SKIN AS ROUTE OF TOPICAL AND TRANSDERMAL DRUG DELIVERY: The transdermal permeation of a chemical involves partitioning into and transport through the cutaneous layers, namely the stratum corneum, the viable epidermis (stratum basale) and the upper dermis 10 . A topical product is designed to deliver the drug into the skin to treat dermal disorders and therefore skin is the target organ. Non steady state transport generally characterizes a topical product. The skin is a barrier to topically administered drugs 13,14 . Topical formulations usually contain several excipients, which also partition into the skin according to their physicochemical properties. Certain excipients change the integrity of stratum corneum. Stratum corneum can exhibit swelling by water. Thus, the permeability of drugs depends on the degree of hydration. Cosolvents may later the barrier properties of the skin 3,15 . Some substances having considerable polarities also enhance the permeability of the horney layer. It is known that use of oleaginous vehicles enhances the skin permeation 10 .
Topical preparation applied to the skin may be designed for surface, local or systemic offers. In order to understand these effects, a brief review of the skin structure is provided 16,17 . Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 8
Figure-1.1: Structure of Skin
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 9 1.5.1 Relevant Anatomy and Microstructure: The skin is one of the most extensive and readily accessible organs of the human body. The skin of average adult covers over 20,000 cm of surface area and receives overall of all blood circulation through the body. It is a multilayered organ 18 . The skin is composed of three layers: the outer most is the epidermis, next is the dermis and the innermost layer is the subcutaneous. The epidermis itself is composed of the stratum corneum, horny layer (about 10 m thick), which is a layer of compressed, overlapping keratinized cells that form a flexible, tough, coherent membrane. This layer contain dead cells with keratin filaments in a matrix of proteins with lipids and water-soluble substances. The epidermis is more resistant to the diffusion of chemicals than other layers of the skin, infact, it forms the protective barrier for the layer.
Below the epidermis is the dermis, a matrix of connective tissue, approximately 4 mm thick, woven from fibrous proteins, which are embedded in an amorphous ground substance of mucopolysaccharide. Nerves, blood vessel and lymphatics are contained in the dermis. The innermost layer of skin is the subcutaneous tissue, which contains adipose cells and collagen fibers. The eccrine sweat glands produce sweat, empty on the skin surface and function to control heat exchange 16 .
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 10 1.5.2 Biochemical Makeup of the Stratum Corneum: Along with cellular maturation of the keratonocytes, there is also a remarkable shaft of the lipid composition of the epidermal layers. In humans, the extracellular motor consists of a structural complex containing several groups of lipids.
1.5.3 Immunological Functions of Cells found in the Skin: As the skin is increasingly understood as an immunological organ, the immunologic functions of the skin has recently become an issue of considerable concern. Termination of drug through the skin may be associated with immunologic side effect 19 .
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 11 The immune response could have serious implications. In addition to contact dermatitis, the immune response could also neutralize the drug activity.
1.6 FUNGI: Although fungi causes varity of diseases to the skin few common diseases along with their causitive organism are listed below.
Fungi and yeast constitute the eumycetes. They are eukaryotes with a differentiated nucleous and rigid chitinous cell wall were formerly regarded as plant without chlorophyll or differentiation of root stem and leaves. They are now regarded as neither plant nor animals and are grouped with protozoa, slime moulds and most algae on higher protista. They may be unicellular and multicellular. The cell show various degree of specialization 21 .
Classification of Fungi 22 Fungi can be classified in two ways. Depending upon the cell morphology, fungi can be divided into 4 classes viz., yeast, yeast like fungi< moulds and dimorphic fungi.
Yeast: Yeast are unicellular fungi, which occur as spherical or ellipsoidal cells are reproduce by simple budding. On culture they form smooth, creamy, colonies. The only pathogenic yeast is cryptococcus neoformus.
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 12 Yeast like Fungi: They grow partly as yeast and partly as elongated cells resembling hyphae. The latter form as pseudomycelium, candida albicans in a pathogenic yeast like fungus. On solid media most creamy coloured colonies are produced 23 .
Moulds (Filamentous mycelial fungi): They grow as long as filamentus or hyphae, which branch and interlace to form a meshwork or mycelium, the vegetative mycelium grows on and penetrate into substrate absorbing nutrient for growth. This may become powdery on its surface due to the abundant formation of spores e.g. ring worm fungi.
The Dimorphic Fungi: They either as filamentus or as yeast, according to the culture condition. Growth usually take place in the mycelial form on culture media at 22C and in the soil but in the yeast form on media at 37C and in the animal body, Histoplasma capsulatum is the most important of them.
Fungal Infection 23 : Fungal infections are termed mycoses and is general can be divided into superficial infections (affecting skin, nails, scalp or mucous membranes) and systemic infections (affecting deeper tissue and organ).
Superficial fungal infections can be classified into the dermatomycoses and candidiasis. Dermatomycoses are infection of the skin, hair and nails caused by dermatophytes. The commonest are due to the tinea organism, which cause various Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 13 types of ring worm, tenia capitis affect the scalp. Tinea pedis causes atheletes foot. In superficial candiasis, yeast like organism infect the mucous membrane of the mouth, vagina or skin.
Some of the fungal infections are as follows:
Pityriasis versicolor: Pityriasis versicolor (Tenia versicolor) is a chronic usually asymptomatic involvement of stratum corneum characterized by multiple scalum in children discrete or confluent macular areas of discoloration or depigmentation of the skin. The areas involved are mainly the chest, abdomen, upper limb and back. Facial involvement is common. The causative agent is a lipophilic yeast like fungus pityrosporum orbiculare (Malassezia furfur).
Tinea Nigra: Tinea nigra is localized infection of stratum corneum, particularly of palm, producing black or brown macular lesion. It is found mainly in the tropics and is caused by cladosporium wernickii (now designated as exophiala wernickii).
Piedra: Piedra is a fungus infection of the hair, characterized by the appearance of firm, irregular nodules along the hair shaft. Nodules are composed of fungus filaments, cemented together on the hair. Two varieties of piedra are recognized black piedra caused by piedra kortai and white piedra caused by trichosporon beigelli. Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 14
Candidiasis: Candidiasis is an infection of the skin, mucosa and rarely of the internal organ caused by a yeast like fungus candida albicans normally present in the mouth, intestine and vagina 21 .
Vulvovaginal candidiasis is the infection with Candida albicans. Approximately 75% of women have a vaginal infection with Candida strains during their life and about 40-50% of them suffer a second one, and a small percentage shows chronic cause 24 .
Sporotrichosis: Sporotrichosis is caused by fungus sporothrix schenckii and is characterized by the development of the skin in subcutaneous tissue and lymph node, of nodules which soften and breakdown to form indolent ulcers.
Rhinosporidiosis: Rhinosporidiosis is a chronic granulomatus disease characterized by the development of friable polyps usually confined to nose, mouth and eye but rarely seen in the genitalia or other mucous membrane. The causative fungus is Rhinosporidium seebri.
1.7 RATIONAL APPROACH TO DRUG DELIVERY TO & VIA SKIN 20 :
There are three main ways to solve the problem of formulating a successful topical dosage formulation: Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 15 1. We can manipulate the barrier function of the skin e.g., topical antibiotics and antibacterials help a damaged barrier to ward-off infection, sunscreen agents and the horny layer protect the viable tissue from ultraviolet radiation and emollient preparations restore palatability to a desiccated horny layer. 2. We can direct drug to the viable skin tissue without using oral, systemic or other route of therapy. 3. The third approach uses skin delivery for systemic treatment. For example, topical drug delivery systems provide systemic therapy for conditions such as motion sickness, angina and pain.
Dermatologists aim at five main target regionskin surface, horny layers, viable epidermis and upper dermis, skin glands and systemic circulation.
1.8 UTILISATION OF DIFFERENT GELS AT TOPICAL VEHICLES 17 : Gels have a variety of applications in the administration of medications orally, topically, intranasally, vaginally and rectally.
1.9 EMULSION-GELS AS TOPICAL FORMULATIONS 17 : Transdermal and topical formulations are becoming increasingly important and their use in therapy is becoming more widespread. But the skin acts as a barrier to topically administered drugs. Attempts have been made to circumvent the skin barrier by several means, emulsion-gels being one such promising technique.
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 16 1.10 ORGANOGELS: The topical administration of drugs in order to achieve optimal cutaneous and percutaneous drug delivery has recently gained an importance because of various advantages such as ease of administration, non-invasive, better tolerated and compliance, local enhanced transdermal delivery, avoidance of local gastrointestinal toxicity, avoidance of first pass metabolism.
In search of a vehicle to deliver the medicament into the skin layer (cutaneous delivery) or through the skin and into systemic circulation (percutaneous absorption) and to target the skin, varied kind of formulation systems and strategies have been evolved.
Amongst the many, the lipid-based formulations have been in use since decades. Pharmaceutically, lipid emulsions may allow the sustained release of drugs by sink mechanism 7 .
The importance of lipids has especially increased after realizing the utility of phospholipids. The natural bio-friendly molecules which in collaboration with water can form diverse type of polymolecular/ super molecular structure with retardant release in sustained release formulation 25 .
The topical delivery has been attempted and made successful using a number of lipid based systems viz., vesicular systems 26 , lipid microsphere, lipid nanoparticles 1 , lipid emulsion 4 , polymeric gels 27 . In a recent development, phospholipids in conjunction with some other additives have been shown to provide a Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 17 very promising topical drug delivery vehicle i.e., lecithin organogel. Lecithin organogels (LOs) are thermodynamically stable, clear, viscoelastic, biocompatible and isotropic gels composed of phospholipids (lecithin), appropriate organic solvent and a polar solvent 28 . Lecithin organogel, the jelly like phase consists of three dimensional network of entangled reverse cylindrical (polymer like) micelle, which immobilize the continuous or macroscopic external organic phase, thus turning liquid into a gel 25 . The formation of three-dimensional network in the organogel is the result of transition at the micellar level in a low viscous network liquid consisting of lecithin cause micelles in non-polar organic liquid 29 . This spherical reverse micellar state of lipid aggregates, twins on to form elongated tubular micelles with the addition of water, and subsequently entangle to form a temporal three dimensional network in the solution bulk. The latter serves to immobilize the external organic phase, thus producing a gel form or the jelly like state of the initial non-viscous solution. However, the transparency and optical isotropy of the organogel remain as before. For this reason, these systems are often called as polymer like micelles and are also termed as living or equilibrium polymer, worm like or thread like micelles 25 .
1.10.1 Advantages of Organogels 28,30,31 : Template vehicle: Lecithin organogels provide opportunities for incorporation of wide range of substances with diverse physicochemical characters viz., chemical nature, solubility, molecular weight, and size etc.
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 18 Process Benefits: Spontaneity of organogel formation by virtue of self-assembled supramolecular arrangement of surfactant molecules, makes the process very simple and easy to handle.
Structural/ Physical Stability: Being thermodynamically stable, the structural integrity of lecithin oranogels is maintained for longer time periods.
Chemical Stability: Lecithin organogels are moisture insensitive and being organic in character also resist microbial contamination.
Topical Delivery Potential: Being well balanced in hydrophilic and lipophilic character, they can efficiently partition with the skin and therefore enhance the skin penetration and transport of the molecules. Lecithin organogels also provide the desired hydration of skin in a lipid-enriched environment so as to maintain the bioactive state of skin. Lecithin might influence the structure of the skin by disorganizing the lipid layer in the stratum.
Safety: Use of biocompatible, biodegradable and non-immunogenic materials makes them safe for long-term applications.
1.10.2 Limitations of Organogels: In the lecithin organogels, the lecithin should be pure otherwise no gelling will occur. Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 19 Lecithin is most costly. Lecithin is not available on large scale. Should be stored in a proper condition. The organogel has greasy property. Less stable to temperature.
1.11 LECITHIN ORGANOGEL AN OVERVIEW: The first description of lecithin organogels was given in an article published by Scartazzini and Luisi in the year 1988 28 . In this study, water was added to various organic solutions of purified soyabean lecithin. It was observed that addition of soy- lecithin caused an abrupt rise in the viscosity (1010 4 times) 30 , producing a transition of the initial non-viscous solution into gel of jelly like state. The amount of water required to produce the gel was found to be critical. The phenomenon was observed with various non-polar media and the list includes more than fifty solvents 28 .
By now, lecithin organogels have been studied extensively in many laboratories worldwide with regard to their varied aspects such as formulation component, formation and gelling mechanism, physicochemical properties, etc. and have also been proposed as a matrix for topical drug delivery.
1.11.1 Organogelling Composition: The organogel matrix chiefly consists of surfactant (lecithin) as gelation molecules, a non-polar organic solvent as external or continuous phase and polar agent, usually water. Lecithin is a trivial name for 1,2-diacyl-Sn-3-phosphocholine. Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 20 It belongs to a biological essential class of substance termed phosphoglycerides or phospholipids. The latter form the lipid matrix of biological membrane and also play a key role in the cellular metabolism.
As a biocompatible surfactant, it is widely used in every day life including human and animal food, medicine, cosmetics and manifold industrial applications 32 . Synthetic lecithin containing residues of saturated fatty acids failed to form organogel 28,30,33 . However, it has been established that unsaturation in phospholipid molecules is a desired property for the formation of lecithin organogels.
Besides lecithin as gelation molecules, the role of organic solvent in providing the desired solvent action into the gelatin molecules is much emphasized. A large variety of organic solvent are able to form gel in the presence of lecithin. Among them are linear, branched and cyclic alkenes, ethers and esters, fatty acids and amines. Specific examples includes ethyl laurates, ethyl myristate, isopropyl myristate (IPM), isopropyl palmitate (IPP), cyclopentane, cycloclane, trans-decalin, trans-pinane, n-pentane, n-hexane, n-hexadecane nd tripropylamine 28 .
Amongst the above, the fatty acid esters i.e., application of lecithin organogels. This has been attributed to their skin penetration enhancing property besides their biocompatible and biodegradable nature 32,34 .
The third component of polar agent acts as a structure forming and stabilizing agent and has a very crucial role to play in the process of gelling. Water is the most commonly employed polar agent although some other polar solvents such as glycerol, Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 21 ethylene glycol and formamide have also been found to possess the capability of transferring an initial non-viscous lecithin solution into jelly like state on organogel 25 .
As described earlier, the major limitation in formation of lecithin organogels is the requirement of high purity lecithin, the high purity grade lecithin is not only expensive but also difficult to obtain in large quantities. However, recent reports indicates the incorporation of synthetic polymers i.e., pluoronic in lecithin organogels, for their usefulness as cosurfactant and stabilizer 35 . It has been shown that the inclusion of pluronic as cosurfactant makes the organogelling feasible with lecithin of relatively lesser purity 36 . The term pluronic refers to series of non-ionic closely related block copolymers of ethylene oxide and propylene oxide 32 . Also known as poloxamers, poloxamer polyols or Lutrol
. These are primarily used in
pharmaceutical formulations as co-surfactants, emulsifier, solubilizers, suspending agents and stabilizers. These pluronic containing lecithin organogels have been termed as pluronic lecithin organogels, poloxamer organogels, pluronic organogels, PLO gel or simply PLOs.
1.11.2 Phase-behavior of organogels: Sameles containing different weight ratios (k m ) of lecithin/IPM (20:80) (40:60) (60:40) (80:20) were prepared, phase studies were carried out by adding water while stirring. After each addition of 1 liter of aqueous phase of pure water to the lecithin solutions, the resulting systems were examined for clarity and viscosity. The course of each addition was monitored through cross polaroids in order to determine the boundaries of any organogel and briefringent liquid crystalline Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 22 domains. The endpoint of the organogel domain at a given k m was determined when the system became turbid after the addition of a specific amount of water. The phase behavior of the systems was mapped on phase diagrams with the top apex representing the lecithin and the other apices representing IPM and water solution. The transparent, homogeneous, nonbirefringent area enclosed by the line connecting the endpoints was considered as microemulsion based organogel 37 .
1.11.3 Organogel structure and mechanism of organogelling 37 : The initially spherical reverse micelles that are formed by lecithin molecules in a nonpolar organic solvent transform into cylindrical micelles, once water is added. This was established with the help of light scattering and small angle neutron scattering techniques. This one dimensional growth of micelles is caused by the formation of hydrogen bonds between water molecules and phosphate groups of lecithin molecules so that two adjacent lecithin molecules are bridged together by one water molecule IR and NMR spectroscopic methods have revealed that water molecules could interact simultaneously with phosphate groups of neighboring lipid molecules via hydrogen bonding, acting as a bridge between them. In this case solvent molecules and lecithin phosphate groups can arrange in such a way that a hydrogen-bonded network will be formed. The increase in the amount of water results in the formation of long tubular and flexible micelles. These micelles can be entangled and therefore build up a transient three-dimensional network, that is responsible for the viscoelastic properties of the lecithin organogels. At a critical concentration of water, network shrinks and phase separation occurs. At still higher Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 23 contents of water a transformation to a solid, nontransparent precipitate can be observed. This diluted solution is composed of rod-like micelles, which their length is not enough to overlap and form a three-dimensional network. The existence of microdomains of different polarity within the same single-phase solution enables water-soluble and oil-soluble drugs to be incorporated. Results have shown that for a given system, as k m increases, the incorporation capacity increases. This could be attributed either to the increase in the number of cylindrical micelles or to the further growth of the cylindrical micelles or both, leading to the increase in the solubilizing capacity.
1.12 METHOD OF PREPARATION: The oil-surfactant mixture was heated at 60C to obtain a clear solution which on cooling forms organogels 38 . Based on the phase diagrams constructed, lecithin solutions were prepared by first dissolving lecithins in an organic solvents with the aid of magnetic stirrer. Formation of organogels takes place on addition of water with the help of micropiopette syringe. Sometime heat is applied for complete solubilization of drug 29 .
The oil phase is prepared by mixing lecithin and organic solvent, the mixture is allowed to stand overnight to ensure complete dissolution. The aqueous (polar) phase is prepared by adding pluronic to ice cold water, the mixture is agitated to ensure complete dissolution. The prepare PLO, the oil phase is mixed with aqueous phase of pluronic using a high shear mixing method by magnetic stirrer 35 .
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 24 1.13 CHARACTERIZATION OF ORGANOGELS: In contrast to the ease of preparation, characterization of LOs is relatively complicated on account of their interior structural design build up on the self- associated supramolecules. These microstructures, the resultant of varied polar non- polar interactions, are highly sensitive and pose difficulties in the investigative studies. However, different characterization studies are extremely useful while investigating the potential applications of organogel systems as a topical vehicle. For instance, it has been reported that many of the physicochemical properties of Los viz. Rheological behavior, physical and mechanical stability, and drug release behavior are dependent upon how do molecules arrange themselves to provide the specific structural network within the organogel system 25,38 .
1.13.1 Rheological behavior For any vehicle to be used for topical drug delivery applications, it is essential to study its rheological behavior. The latter is important for it efficacy in delivering the molecules onto or across the skin site. The critical parameters like spreadibility, adhesiveness (property related to bioadhesion on skin site), cohesiveness (which indicates structural reformation following application of shear stress, and consistency need to be modified in a favorable manner. Lecithin organogels (LOs) have been studied extensively for their rheological attributes and determined to be viscoelastic in nature 25 .
At higher lecithin concentrations, there is more extensive entanglement of long cylindrical micelles with each other, forming a network-like structure with a Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 25 very high viscosity. The entrapment of the drug within this network lowers the amount of free drug available for release, causing a decrease in the release across the membrane 29 .
1.13.2 Determination of gelation temperature 39 : Formulations were enclosed in glass tubes (2 mm inside diameter) and observed over a temperature range of 4-5C. The change from solution to gel ov vice-versa was determined by inverting the tube. The temperature was changed at a rate of 5C h and the temperature at which the physical state of the formulation was changed was regarded as the gelation temperature. In all cases the gelation temperature was reproducible to within 0.1C. The gel melted completely within a 0.2 0.3 C range.
1.13.3 Gelation Kinetics 40 : The gelation properties of organogels were investigated in the presence of various solvents. Gel-sol and sol-gel transitions were evaluated by the inverse method and gelation kinetics were determined by turbidimetry.
1.14 IN VITRO DRUG RELEASE 29,41 : The permeation apparatus designed as described by Chowdary et al was employed to study the release profile of drugs from the semisolid formulations. Phosphate buffer 6.4 used as receptor fluid.
The release/ permeation of drugs from lecithin gels through various membrnes was determined using Franz diffusion cell. Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 26
1.15 TOPICAL DRUG DELIVERY APPLICATIONS OF LECITHIN ORGANOGEL-BASED SYSTEMS: Organogel formulation Major findings Lecithin (200mM) IPP gel of broxaterol and scopolamine 42 . Transdermal delivery of compounds Phosphatidylcholine (PC) gel in isopropyl palmitate (IPP) or cyclooctane 30 . Investigated for transdermal transport of various drugs along with aminoacids and peptides IPP-lecithin gel of diclofenac and indomethacin 43
Enhanced efficacy of NSAIDs administered through topical route Phytosphingosine or sphingosine lecithin organogel comprising soy PC, IPP, ethanol and water 44
Treatment of scars Soya lecithin-isopropyl myristate (IPM) organogel containing ketamine hydrochloride and amitryptiline hydrochloride 45
Enhance skin penetration and partitioning of the drugs into the skin layers Nicardipine lecithin-IPM organogel 46 Enhanced skin permeation across guinea pig and human skin Methimazole in LO gel 31 Significant percutaneous absorption of methimazole LO gel of cardiac glycoside digoxin 47 Topial administration of the compound in LO gel was found to be effective for the treatment of muscle spasm Cyclobenzaprin in lecithin organogel 48
(lecithin 10-30%, IPM 10-30%, water 30- 60%) Topical formulation for bauxism.
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 27 1.15.2 Topical delivery of therapeutic substances in pluronic lecithin organogels: PLO gel formulation Applications Ketoprofen PLO gel 30 Administration of ketoprofen in PLO gel offered convenience, produced less side effects and alleviated pain in a specific location PLO gel of Diclofenac, Ibuprofen, Ketamine 48
Randomized, placebo controlled study on lateral epicondylites employing diclofenac in PLO gel reduced pain and increased functional status Preparation also found to be effective treatment for osteoarthritis Lecithin organogjel in combination of Pluronic F-127 (poloxamer 407) solution/ Cyclobenzaprin 36
Effective formulation for topical treatment of carpal tunnel syndrome Lecithin (20-40% v/v) in isopropyl palmitate or isopropyl myristate containing suitable amount of pluronic and water with or without short chain alcohol 44 . The components of PLO gel provide desired hydration state to the skin, thus effective in the treatement of eczema or psoriasis
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 28 1.16 SAFETY AND SKIN COMPATABILITY STUDIES: Lecithin-based organogel system i.e., LO or PLO gels are composed of pharmaceutically approved (non-immunogenic and biocompatible) excipient. However, the level of surfactant and organic solvents in lecithin organogels is faily high. Therefore, it is important to consider the safety and irritancy of the formulation on prolong use. In this context, skin (human skin) compatibility of the gels have been evaluated employing various techniques before and after applications with either IPP alone or with 200 mM-IPP gel. No significant alteration in the skin were apparent after three days and stratum corneum was still intact. The irritation potential of LOs has been assessed by Dreher et al by carrying out human skin irritation study 50 . Result indicated a very low cumulative skin irritation potential of LOs. That supports the stability of LO based gels as topical vehicle for long-term application.
1.17 FUTURE PROSPECTS: In the field of topical drug delivery, LOs have emerged as one of the most potential carrier systems. In contrast to other lipid-based system such as vesicular system (liposomes and niosomes) lecithin-organogel systems may prove to have an edge in term of efficacy, stability and most importantly, the technological feasibility. Morevoer, the topical drug delivery of new biotech generated proteinaceous molecules in the protective non-polar microenvironment of these systems may help protect these sensitive macromolecules from and degradation, while their transport to the desired site 48 .
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 29 Thus, amidst the increasing opportunities and challenges, the LOs may prove to be highly promising system in realizing the drug delivery objectives while scientists are desperately trying for more viable alternative viz-a-viz existing carrier system.
PLO is probably due to financial constrains as well as the industry focusing on area such as biotechnology and genomics. However, the great interest in PLO in the US has led to formulation of a second generation lecithin organogel premium, lecithin organogel base by Xenex Labs and Max Pharmaceuticals, USA 35 .
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 30 CHAPTER2 OBJECTIVES
2.1 NEED FOR THE STUDY: A gel can be described as the cross-linked material that retains a large amount of solvent inside its medium and if the solvent retained in organic one, such material is known as organogel. Traditionally organogel type systems are applied topically when the active agent is oil soluble one or if we need the sustained release of the drug into the deep skin layers 51 . A polar organic solvents, soya bean lecithin can form thermo-reversible, isotropic, non-birefrigerant gel like system so called micro- emulsion based organogels, characterized by high viscosity and optical transparency 13 .
In the present study, various polymers such as soya bean lecithin, isopropyl myristate, poly sorbate 80, mineral oil (liquid paraffin), pluronic F-127, have been employed for organogels 29,52,53 .
Miconazole nitrate is a synthetic imidazole derivative with molecular formula C 18 H 14 C 14 N 2 CHNO 3 and practically insoluble in water has the antifungal activity with a broad spectrum activity against pathogenic fungi (including yeast and dermatophytes and gram positive Candida albicans, Staphylococcus and Streptococcus) 54 . It may act by interfering with permeability by inhibiting the fungal cytochrome P-450 enzyme responsible for the synthesis of ergosterol the main sterol in the fungal cell membrane 55 . Miconazole is topically active drug and only rarely Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 31 administered parenterally due to its extensive first pass metabolism and severe toxicity 56 . Miconazole on oral administration causes nausea, vomiting and diarrhea 57 . Miconazole readily penetrate the stratium corneum of the skin 58 , but less than 1% in blood. Irritation and burning are rare after cutaneous application. It is also used in the treatment of several systemic fungal infections including Candidiasis.
Organogels may be formulated to enhance the release and to provide more sustained topical antifungal effect of miconazole nitrate.
2.2 OBJECTIVES OF THE STUDY: 1. Formulation of lecithin based organogels of miconazole nitrate. 2. Evaluation of in vitro antifungal activity of optimized miconazole nitrate organogels. 3. To evaluate the influence of formulation variables on the release rate of miconazole nitrate.
2.3 SCHEME OF WORK: Phase-I: 1. Extensive literature survey. 2. Procurement of materials.
Phase-II: Standardization of Materials: a) Standardization of miconazole nitrate b) Standardization of polymers Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 32 1. Lecithin 2. Tween-80 3. Isopropyl myristate 4. Pluoronic F-127 5. Liquid paraffin.
Phase-III: 1. Formulation of lecithin based organogels 2. Preliminary evaluation of organogels. 3. Selection of best composite based on preliminary evaluation.
Phase-IV: 1. Incorporation of drug 2. Preparation of standard calibration curve of miconazole nitrate 3. Evaluation of Organogels. a) pH b) Spreadibility c) Viscosity d) Gelation kinetics e) Gel life (stability) Phase-V: 1. Drug content uniformity 2. In vitro drug diffusion 3. In vitro antifungal activity
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 33 CHAPTER3 REVIEW OF LITERATURE
3.1 REVIEW OF LITERATURE Rajani V and Verma PRP (1993) determine the diffusion studies of ibuprofen from ointment bases using cellophase membrane. Data reveals that percent drug release were concentration dependent. Similar observations have been made with respect to the release of benzocaine, sorbic acid, salicyclic acid and benzocaine acid. The diffusion rate was higher in the first hour and therefore decline. The general rank of order of the drug release was found to be water soluble >water miscible > hydrophilic >oleaginous. The slowest release was found with a water-oil emulsion. Release was generally dependent on the drug concentration 59 .
Shoba Rani R Hiremath, et al 60 had carried out the permeation studies of marketed clotrimazole creams. The drug release was less than 20% with all the formulations tested at the end of 8 hours and hence isopropyl myristate was chosen next as a medium because of its bipolar properties.
Gondaliya DP and Pundarika Kshudu K 61 had carried the investigation- examined preparation and evaluation of nimesulide clear aqueous gels and emulgel using acrypol 940p. A 3 factorial design was adopted for the optimization of aqeous gel formulation. Propylene glycol and polyethylene glycol 400 were chosen as independent variable to study their effects as cosolvents. The clear aqueous gel formulation containing 15% w/w ethanol, 20% w/w propylene glycol and 30% w/w Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 34 PEG-400 showed maximum drug penetration (18.68%) in 5 hours in in-vitro diffusion study. Drug diffusion was increased by addition of chromophore EL, a lipophilic penetration enhancer.
Ilango R et al 62 developed transdermal preparation of nimesulide gel. The effect of polymer-concentration on in vitro nimesulide release from carbpol 940 HPMC gel and effect of permeation enhancer like Tween-80 and SLS at different concentration on drug release were studied.
Magdy C Mohammed 63 have studied optimization of chlorphenesin emulgel formulation. He prepared emulgel using different polymers and evaluate emulgel for various parameter like rheological study, in vitro study release, antifungal activity and stability studies.
Miconazole nitrate used in the treatment of severe systemic fungal infection, chronic muco cutaneous candidasis, fungal meningitis, vulvo vaginal candidiasis, tinea infection, otomicosis, cutaneous candidiasis and rarely given as i.v for system mycosis 58 .
Mucoadhesive buccal patches of miconazole nitrate was prepared and its in vitro / in vivo performance were evaluated by Wafee and Ismail 64 . The patches were prepared with ionic polymers sodium corboxy methylcellulose and chitosan and non ionic like hydroxy ethyl cellulose and hydroxy propyl methyl cellulose. Convenient bio adhesion, acceptable elasticity, swelling and surface pH were obtained patches exhibited sustained release over more than 5 generally enhanced the release rate. Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 35
Dibiase MD and Rhodes CT 65 had carried out the work on pluronic F-27 gel and evaluated as a potential topical vehicle for epidermal growth factor delivery. The chemical stability of the polypeptide within the gel matrix was investigated using HPLC. Thermal stability studies was performed on the base gel formulation. Modifications to the formulation was made to improve physical characteristics and chemical stability. Humectants and antioxidants was investigated as potential formulation additives and the microbial status of the product was also evaluated.
Minghetti Patel 66 reported dermal patches of miconazole were evaluated for their technological characters by Minghetti et al for the treatment of tinea argium infection. Artificial silk used as backing layer. Eudragit and plastoid were used which provided release of at least 24 hours.
Khurana and Ahuja 67 prepared and evaluated mucoadhesive tablets of miconazole nitrate for the treatment of oral candidiasis. It produced satisfactory drug release.
David H et al 68 investigation of two hundred eighty (280) patients with symptomatic vulvovaginal candidiasis were randomly assigned to treatment with either miconazole nitrate 4% vaginal cream for 3 days, followed by placebo for 4 days, or Monistat-7 (miconazole nitrate 2%) vaginal cream for 7 days in this double- blind, parallel-group, outpatient study. Sixteen US centers participated. Patients were seen on admission and then at 8 to 10 days and 30 to 35 days after completion of treatment. Clinical, microbiologic, and therapeutic efficacy was assessed. This Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 36 study compared the safety and efficacy of a new cream formulation of 5g of miconazole nitrate 4% administered once daily for 3 days with that of 5g of miconazole nitrate 2% vaginal cream, the currently marketed product, administered for 7 days. Although not significantly clinically different, cure rates were slightly higher with the 3-day treatment. Relapse rates were low in both treatment groups and symptom relief was also comparable. The most frequent adverse experiences were genital (itching, burning, irritation, and discharge), as well as headache and respiratory congestion; reports of adverse experiences were similar in the two treatment groups. Miconazole nitrate 4% vaginal cream administered for 3 days was found to be promising new candidate for over-the-counter treatment of vulvovaginal candidiasis.
Chandia Valenta et al 34 studied the soya-lecithin aggregates prepared by a technique using compressed gas to formulate new dermal preparations. Ketoprofen (KP) a Non-steroidal anti-inflammatory drug (NSAID) is included as a model drug. The novel soya-lecithin aggregates are promising candidates for new drug delivery system in dermatology and cosmotology. Lecithin aggregates loaded with drugs are multifunctional causes that also act as penetration enhancers. The improvement in skin permeation is related to both the solubilizing effect of lecithin matrix and penetration enhancing effect of lecithin itself.
Murda S 35 reported the great interest in pluronic lecithin organogel in the US has led to the formulation of a second generation lecithin organogel, premium lecithin Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 37 organogel base by Xenex laboratories and has non-greasy, non-tacky and improved stability to temperature as compared with original pluronic lecithin organogels.
Reza Aboofazeli et al 69 stated that the partial phase diagrams were constructed with soybean lecithin water, sodium salicylate, alcohol and isopropyl myristate. Phase diagrams showed the area of existence of a stable isotropic region along the surfactant / oil axis (i.e., reverse microemulsion area).
Shilpa K et al 75 showed that the microemulsion based organogel are useful in iontophoretic drug delivery vehicles include their potential to increase the maximum loading of a water soluble agent and the drug ability may be improved especially in comparison to the use of hydrogels where the presence of an aqueous continuous phase may allow degrative processes to occurs.
Singh R et al 70 investigation reveals that the antifungal activity measured as zone of inhibition of lecithin organogel ketoconazole as a model drug was better as compared to the activity recorded for hydrogel base.
William N et al 30 concludes that lecithin gels may be efficient vehicles for the transdermal transport of various drugs. The presence of lecithin in organic solvent result with increase in drug solubility as compared with heat solvent the transport rate is 10 times higher that with the aqueous solution of drug. Lecithin organogel can be prepared easily and rapidly and can be obtained with biocompatible components. They are stable for a long time, can incorporate sizeable amount of different chemical as guest molecules and fulfill the cosmetics and pharmacological application. Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 38
Shelke VB et al 38 research result shows that the organogels can be used for all type of drug molecule, controlled release increased resistance to microbial contamination and reduced risk of toxicity.
Meiying Ning et al 24 reveals vulvovaginal candidiasis is the infection with Candida albicans. Approximately 75% of women have a vaginal infection with candida strain during their life and about 40 to 50% of them suffer a second one and small percentage shows a chronic cause. The entrapment of drug in vesicles may help in the localized delivery of the drug and an improved solubility and availability of the drug at the site may reduce the dose and systemic side effects.
Hoffmann G et al 31 showed that lecithin organogels were developed for transdermal transport of drugs and consists of a spaghetti like network of lecithin micelles, which can host various guest molecules by solubilization in their transdermal methimazole treatment research work to treat cat hyperthyroidism.
Marco Antonio M et al 71 evaluate the system containing water lecithin/ polysorbate 80/ isopropyl myristate the results showed high stability, very low toxicity for the parentral use.
Vitonial M, Bentley M et al 39 studies showed that in vitro permeation of a model drug triamcinolone acetonide was decreased when the lecithin concentration was increased. The presence of lecithin in the poloxamer (pluronic) gel improved the characteristics for topical drug delivery. Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 39
Gelatin containing microemulsion based organogels (MBGs) have been formulated using pharmaceutical acceptable surfactant and oils such as tween-85 and isopropyl myristate. MBGs are clinically conducting employed for ionotphoretic delivery of a model drug. MGBs also appear to offer improved microbial resistance in comparison to aqueous solution or hydrogels. This work is done by Kantaria S et al 72 .
Scartazzini R, Luisi R 28 showed that the interest in lecithins as basic components for gel materials lies in biomimetic chemistry. This is vague term and one that should be used sparingly. The point can be made that lecithin gels may be related more closely than others to gel like lipidic aggregates that exist in vivo.
Luisi PL et al 30 suggested organogels can be used to solubilize a variety of drug candidates therefore it is widely applied in chemical, pharmaceutical, cosmetic applications.
Dreher F et al 43 in investigation to estimate the function of the gel as a potential transdermal penetration enhancer system performed the interaction of lecithin, isopropyl palmitate with the human stratum corneum using DSC and FTIR and found more significant irritancy. Lecithins and isopropyl palmitate affects the stratium corneum lipid organization.
Murdan S et al 77 determined the sorbitan monostearate organogels can be used as delivery vehicles for hydrophilic and hydrophobic drug and vaccines. The gels Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 40 may also provide sustained release of appropriate active entity after intramuscular and subcutaneous administration.
In short communication, Murdan S et al 26 showed that sorbitan monostearate organogels are opaque, thermo reversible, semisolid whose microstructure consists of surfactant tubules dispersed in organic continuous phase. Inverse toroidal vesicles are the precursors of the surfactant tubules. The toroids are thought to be analogues to other well known vesicles, liposomes and niosomes except for their toroidal (rather than spherical) shape and their inverse nature.
Murdan S et al 76 suggested in their studies that the sorbitan stearate and isopropyl pulmitate organogels may have potential applications as delivery vehicles for drugs and antigens.
Desai A and Kadam V et al 73 noted that, mother nature has gifted India with great variety of flora and fauna. Since centuries man has made an effective use of materials from natural origins in the medical and pharmaceutical field, natural materials have been used in controlled release drug delivery system. The natural release retardants include phospholipids (lecithins).
Kavitha K et al 74 reported, a topical drug delivery system localizing the drug at skin will be much favourable for the treatment of skin infection.
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 41 Rajiv Kumar and Om Prakash Katare 25 determined enhanced skin penetration and site specific delivery of bioactive into the deeper layer of skin has been achieved employing organogels as a topical vehicles.
Sudaxshina Murdan 35 showed that, in man the in vivo studies in man, where pluronic lecithin organogels was applied using diclofenac as a model drug may be beneficial as a delivery vehicle for local action during treatment. Patient feels less pain.
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 42 3.2 DRUG PROFILE Miconazole Nitrate is included in National List of Essential Medicines, 2003 78 .
Miconazole Nitrate: Azoles: Azoles are group of synthetic fungistatic agents with a brand spectrum activity.
Mechanism of action: Miconazole nitrate inhibit fungal cytochrome P-450 enzyme responsible for the synthesis of ergosterol the main sterol in the fungal cell membrane.
The resulting depletion of ergosterol alter the fluidity of the membrane and this interfers with the action of membrane associated enzymes the overall effect is inhibition of replication. A further repercussion is the inhibition of the transformation of candidal yeast cells into hyphae-the invasive pathogenic form of the parasite 23 .
The requirement for IV administration and toxicity of the older antifungal agents created a need for antifungal agents with a better therapeutic profile. The relatively non toxic oral azole medications represents the 1 st major advance in this direction since their introduction in 1980s played important role in systemic therapy of fungal diseases.
According to number of nitrogen atom azoles are classified as imidazoles and triazoles. Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 43
Imidazoles: Ketoconazole, Miconazole and Clotrimazole.
Molecular weight Miconazole nitrate 479.15 Properties: Colour White to pale cream Form Crystalline or microcrystalline powder Description Odourless or almost odourless Solubility Practically insoluble in water 1 in 9.5 of alcohol, 1 in 15 of ether, 1 in 4 of isopropylalchol 1 in 5.3 of methyl alcohol, 1 in 9 of propylene glycol Cl OCH 2 Cl CH CH 2 N N Cl Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 44 Melting point - 179 180C Shelf life - at least 5 years Storage condition - Store in a well closed container protect from light
Indications: Administered by intravenous infusion in the treatment of severe systemic fungal infections including candidiasis, coccidioidomycosis, crypto-coccossis, para coccidomycosis and infections due to pseudeliescheria boydii, has been given prophylactically to patients at high risk of opportunistic fungal infections.
It is administered by intravenous infusion in the treatment of severe fungal infections including Candidiasis, Cryptococcosis 58 .
Used for vulvovaginal candidiasis.
Used for dermatophytic infections including tinea corpis (ring worm), tinea pendis (atheletes foot) and tinea crucis.
Therapeutic doses: Adult Oral Oral and intestinal candioliasis. 125 to 250 mg as tablets or gel 4 times a day
Parenteral: I.V. dose 02. To 1.2 g 3 times daily
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 45 In fungal meningitis intravenous treatment may be supplemented by a single daily dose of 20mg given by intrathecal injection every 3 to 7 days.
Skin: Once or twice a day as 2% gel, cream lotion or powder. Vaginal: 5-10 g of a 2% cream once daily for 7-14 days or tampons containing 100mg twice a day for 5 days. Children: Oral: Oral and intestinal candidiasis 125 mg 4 times dialy. Contraindications: Should not be used in patients known to be hypertensive to this drug. First trimester of pregnancy.
Routes of Entry: Oral for oral and intestinal candidiasis. Dermal: For treatment of fungal infections of the skin and nails. Eye: Solution of miconazole as topical. Parenteral: As intravenous infusion in the treatment of severe systemic fungal infections 79 .
Adverse Reactions 79 : Cardiac arrhythmias (IV administration), tachycardia, haematological alterations aggregation of erythrocytes, gastrointestinal disturbancesanorexia, nausea, vomiting, diarrhea, local irritation and clinical effects. Vulvovaginal burning, itching, irritation, pelvic cramps, cutaneous pruritis, flushes. Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 46
Kinetics Absorption by route of exposure Oral: After oral administration, miconozole is incompletely absorbed from the gastrointestinal tract; peak plasma concentrations of about 1 mcg/mL have been achieved 4 hours after a dose of 1g 79 .
Parenteral: By intravenous infusion, doses above 9 mg/kg body-weight usually produce plasma concentration above 1mcg per mL.
Topical: There is little absorption through skin or mucous membranes when miconazole nitrate is applied topically 79 .
Following intravaginal administration of miconazole nitrate, small amounts are absorbed 81 .
Administration of a single dose of miconazole nitrate suppositories (100 mg) to healthy subjects resulted in a total recovery from the urine and faeces of 0.85% (+0.43%) of the administered dose 82 .
Distribution by route of exposure: Data available indicates that over 90% of miconazole is bound to plasma proteins. Penetration into the cerebrospinal fluid and sputum is poor but miconazole diffuses well into infected joints. Animal studies indicate that the drug crossed the placenta but it is unknown whether micronazole nitrate is excreted in breast milk 79 .
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 47 Biological half-life by route of exposure: Oral: The information available indicates that miconazole orally administered has a half-life of about 24 hours.
Parenteral: The decrease of plasma concentrations after an intravenous perfusion is biphasic, with half-life of 30 minutes (t 1/2a) and 24 hours (t 1/2b).
Metabolism: Micronazole is metabolised in the liver to inactive metabolites. Miconazole has an important first pass effect. Hepatic metabolism of miconazole is by O-dealkylation and oxidative N-dealkylation.
Elimination by route of exposure: Oral: Approximately 50% of an oral dose may be excreted mainly unchanged in the faeces. From 10 to 20% of an oral dose is excreted in the urine, mainly an metabolites, within 6 days.
Parenteral: From 10 to 20% of an intravenous dose is excreted in the urine, mainly as metabolites, within 6 days. After an intravenous dose of miconazole, 40% of the radioactivity is found in faeces and 18% in urine. About 14 to 22% of the dose is excreted via the kidneys, mainly as inactive metabolites.
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 48 Pharmacology and toxicology: Mode of action: Toxicodynamics: Since composition of fungal and mammalian cells membrane is different, miconazole does not affect the human cells. Many adverse effects have been associated with the injection vehicle, which contains Cremophor EL.
Pharmacodynamics: The pharmacological mode of action of miconazole is unknown. In-vitro studies suggest that imidazoles impair the synthesis of ergosterol, which is a vital component of the fungi cell membranes. In-vitro miconazole inhibited the ability of neutrophils to reach the site of infection proptly (chemotaxis) and it demonstrates marked immunosuppressive properties. The clinical significance of these data is unknown.
Interactions 81 : Miconazole given systemically may enhance the activity of anticoagulant or suphonylurea hypoglycaemic drugs. The combination of amphotericin and miconazole appeared to be less effective when either drug used alone. Miconazole enhanced the activity of clomipramine, carbamazepine and phenytoin.
The base contained in certain suppository formulations may interact with some latex products, such as that used in vaginal contraceptive diaphragms.
Since concomitant administration of rifampin and ketoconazole, reduces the blood levels of the latter, the concurrent administration of miconazole intravenously and rifampicin should be avoided. Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 49
Ketoconazole increases the blood level of cyclosporin a, therefore, there is the possibility of a drug interaction involving cyclosporin a and intravenous miconazole; blood levels of cyclosporin a should be monitored if the two drugs are given concomitantly.
Preliminary studies in-vitro and in a few patients indicate that systemic use of both amphotericin B and miconazole may result in antagonistic effect rather than additive antifungal effect.
Several patients have developed an enhanced hypoprothrombinemic response to warferin following miconazole therapy.
Management: One should be alert for an altered hypoprothrombinemic response to oral anticoagulants if miconazole therapy is initiated or discontinued, adjust oral anticoagulant dose as necessary 80,83 .
-phosphatidycholine where R 1 and R 2 are fatty acids which may be different or identical. Lecithin is a complex mixture of materials. The structure above shows phosphatidylcholine, the principal component of egg lecithin, in its -form. In the - form the phosphorus containing group and the R 2 group exchange positions.
Applications in Pharmaceutical Formulation or Technology: Lecithins are used in a wide variety of pharmaceutical applications. They are also used in cosmetics and food products. Lecithins are mainly used in pharmaceutical products as dispersing, emulsifying and stabilizing agents and are included in intramuscular and intravenous injections, parenteral nutrition formulations and topical products, such as creams and ointments. Lecithins are also CH 2 O C R 1 CH 2 O P OCH 2 CH 2 N + (CH 3 ) 3 O O O CH O C R 2 O Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 51 used in suppository bases, to reduce the brittleness of suppositories and have been investigated for their absorption enhancing properties in an intranasal insulin component of enteral and parenteral nutrition formulations. Liposomes in which lecithin is included as a component of the bilayer have been used to encapsulate drug substances and their potential as novel delivery systems has been investigated.
Lecithin as a naturally biocompatible surfactant is capable of producing balance microemulsions in the presence of short chain alcohols, as cosurfactants alcohol can decrease the polarity of the polar medium or incorporate into the ligand layer and change the critical packing parameters (CPP) of lecithin molecule. This effect in turn promote micellization of the microemulsion. Lecithin also deorganize the skin organelles and helps in drug permeation.
Description: Lecithin vary greatly in their physical form, from viscous semiliquids to powders, depending upon the free fatty acid content. They may also vary in color from brown to light yellow, depending upon whether they are bleached or unbleached. Lecithins have practically no odor. Those derived from vegetable sources have a bland or nut like taste, similar to soybean oil.
Method of Manufacture: Lecithins are essential components of cell membranes and may thus in principle be obtained from a wide variety of living matter. In practice however, lecithins are usually obtained from vegetable products such as soybean, peanut, cottonseed, sunflower, rapeseed, corn or groundnut oil, Soybean lecithin is the most Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 52 commercially important vegetable lecithin. Lecithin obtained from eggs is also commercially important and was the first lecithin to be discovered.
Vegetable lecithins are obtained as a byproduct in the vegetable oil refining process. Polar lipids are extracted with hexane and after removal of the solvent a crude vegetable oil obtained. Lecithin is then removed from the crude oil by water extraction. Following drying the lecithin may then be further purified.
With egg lecithin, a different manufacturing process must be used since the lecithin in egg yolks is more tightly bound to proteins than in vegetable sources. Egg lecithin is thus obtained by solvent extraction from liquid egg yolks using acetone or from freeze dried egg yolks using ethanol. Synthetic lecithins may also be produced.
Safety: Lecithin is a component of cell membranes and is therefore consumed as a normal part of the diet. Although excessive consumption may be harmful, oral doses of up to 80 g daily have been used therapeutically in the treatment of tardive dyskinesia. When used in topical formulations lecithin is generally regarded as a non-irritant and non-sensitizing materials.
3.3.2 Polyoxyethylene Sorbitan Fatty Acid Esters 32,71 : Nonproprietry Names: BP: Polysorbates 80 Ph Eur: Polysorbatum 80 USPNF: Polysorbates 80 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 53
Applications in Pharmaceutical Formulation or Technology: Polysorbates are hydrophilic nonionic surfactants used widely as emulsifying agents in the preparation of stable oil-in-water pharmaceutical emulsions. They may also be used as solubilizing agents for a variety of substnces including essential oils and oil soluble vitamins, and as wetting agents in the formulation of oral and parenteral suspensions.
Description: The color is yellow, oil liquid at 25C.
Stability and Storage: Polysorbates are stable to electrolytes and weak acids and bases; gradual saponification occurs with strong acids and bases. The oleic acid esters are sensitive to oxidation. Polysorbates should be stored in a well-closed container, protected from light, in a cool, dry place.
Applications in Pharmaceutical Formulation or Technology: Isopropyl myristate is a non-oleaginous emollient that is absorbed readily by the skin. It is used as a component of semisolid bases and as a solvent for many substances applied topically. Applications in topical pharmaceutical and cosmetic formulations include; bath oils; make-up; hair and nail care products; creams; lotions; lip products; shaving products; suntan preparations; skin lubricants; deodorants; otic suspensions and vaginal creams.
Description: Isorpopyl myristate is a clear, colorless, practically odorless mobile liquid with a bland taste. It consists of esters of propan-2-ol and saturated high molecular weight fatty acids, principally myristic acid.
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 55 Stability and Storage Conditions: Isopropyl myristate is resistant to oxidation and hydrolysis and does not become rancid. It should be stored in a well closed container in a cool, dry, place and protected from light.
Incompatibilities: When isopropyl myristate comes into contact with rubber, there is a drop in viscosity with concomitant swelling and partial dissolution of the rubber; contact with plastics, e.g., nylon and polyethylene, results in swelling. Isopropyl myristate is incompatible with hard paraffin, producing a granular mixture. Also incompatible with strong oxidizing agents.
Regulatory Status: Included in the FDA inactive ingredients guide (otic, topical and vaginal preparations). Used in non-parenteral medicines licensed in the UK.
3.3.4 Pluronic 32 : Nonproprietary Names: USPNF: Poloxamer Synonyms: Monolan, poloxalkol; polyethylene-propylene,glycol, copolymer, polyoxyethylene-polyoxypropyle, cepolymer, supronic; Symperonic. Chemical Name and CAS Registry Number: -hydro--hydroxypoly(oxyethylene) poly (oxypropylene) poly (oxyethylene) block copolymer. Functional Category: Emulsifying agent; solubilizing agent; wetting agent.
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 56 Applications in Pharmaceutical Formulation or Technology: Poloxamers are nonionic polyoxyethylene-polyoxypropylene copolymers used primarily in pharmaceutical formulations as emulsifying or solubiling agents. The polyoxyethylene segment is hydrophilic whilst the polyoxypropylene segment is hydrophobic. Poloxamers are used as emulsifying agents in intravenous fat emulsions, and as solubilizing and stabilizing agents to maintain the clarity of elixiers and syrups. Poloxamers may also be used as wetting agents, in ointments, suppository bases, gels and as tablet binders and coatings.
Poloxamer 188 has also been used as an emulsifying agent for fluorocarbons used as artificially blood substitutes.
Description: Poloamers generally occurs as white-colored, waxy, free flowing prilled granules or as cast solids. They are practically odorless and tasteless. At room temperature, poloxamer 124 occurs as a colorless liquid.
Stability and Storage Conditions: Poloxamers are stable materials. Aqueous solutions are stable in the presence of acids, alkalis and metal ions. However, aqueous solutions do support mold growth. The bulk material should be stored in a well-closed container in a cool dry, place.
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 57 3.3.5 Liquid Paraffin 32 : Nonproprietary Names: BP: Liquid paraffin Ph Eur: Parrafinum liquidum USP: Mineral oil.
Synonyms: 905 (mineral hydrocarbons), Avatech; Citation; heavy liquid petrolatum; heavy mineral oil, liquid petrolatum; paraffin oil; white mineral oil. Functional Category: Emmolient, solvent, tablet and capsule lubricant; therapeutic agent; oleaginous vehicle.
Applications is Pharmaceutical Formulation or Technology: Mineral oil is used primarily as an excipient in topical pharmaceutical formulations where its emollient properties are exploited as an ingredient in ointment bases. It is additionally used in oil-in-water emulsions, as a solvent.
Description: Mineral oil is a transparent, colorless, viscous liquid, free from fluorescence in daylight. It is practically tasteless and odorless when cold, and has a faint odor when heated.
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 58 Stability and Storage Conditions: Mineral oil undergoes oxidation when exposed to heat and light. Oxidation begins with the formation of peroxides, exhibiting an induction period. Under ordinary conditions, the induction period may take months or years. Mineral oil should be stored in an airtight container, protected from light, in a cool, dry place.
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 59 CHAPTER4 METHODOLOGY
MATERIALS AND METHODS: A] Materials and Source: Sl. No. Material Source 1. Miconazole nitrate Gift Sample Adelphi A/C Sigma Lab, Tivium, Goa Date 05.03.2005 Batch No. 2004-05/ GRL/SPR/00013/ MZN/ 16/03/04 2. Isopropyl myristate Loba Chemicals Pvt. Ltd., Mumbai Batch No. 3. Soya bean lecithin Hi-media, Mumbai Batch No. RM-637 4. Pluronic (F-127) 5. Tween-80 SD Fine Chem Ltd., Mumbai 6. Paraffin Liquid SD Fine Chem Ltd., Mumbai 7. Cellophane Membrane Hi-media, Mumbai 8. Sodium dihydrogen phosphate SD Fine Chem Ltd., Mumbai 9. Disodium hydrogen phosphate SD Fine Chem Ltd., Mumbai 10. Sodium hydroxide SD Fine Chem Ltd., Mumbai 11. Sodium chloride SD Fine Chem Ltd., Mumbai
All other chemicals and reagents used were of analytical grade.
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 60 Equipment: Sl. No. Equipment Source 1. UV-visible spectrophotometer Shimadzu Corporation, Japan Model 1700 2. Brookfield DV-II+programmable viscometer Sanjay Technologies, Mumbai Model M-97/164-E-0102 3. Digital Balance Shimadzu Corporation, Japan Model BL-220II 4. Thermostatic Hot plate with magnetic stirrer Remi Udyog, Mumbai Model DW M51102 5. Digital pH meter (pen type) Hanna Instruments, Italy 6. Keshary-Chein type diffusion cell Fabricated by Sai Enterprises, Pune 7. Autoclave Oswal Companies 8. Hot air oven Sheetal Scientific Industries, Mumbai 9. Incubator Rotex Instrument 10. Micropipette Scientex Pvt Ltd. 11. Digital Stirrer Remi Motors, Mumbai Model RQ-121/D
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 61 4.1 RAW MATERIAL CHARACTERIZATION: 4.1.1 Standardization of Drug 84 : Tests were carried out on the sample of miconazole nitrate to establish its identify and purity as per IP 1996.
4.1.2 Standardization of Polymers: The polymers used in the study were tested as per pharmacopoeial or official manufacturers standards.
4.1.2a Lecithin was characterized as per the USP XXV limits 85 . 4.1.2b Tween 80 was tested as per the USP XXV limits 85 . 4.1.2c Pluronic was tested for the compliance with USP XXV specification 85 . 4.1.2d Isopropyl myristate as per the USP XXV limits 85 . 4.1.2e Liquid paraffin was tested for compliance with BP limits 86 .
4.2 PREPARATION OF MICONAZOLE NITRATE LOADED LECITHIN MICROEMULSION BASED ORGANOGEL: 4.2.1. Preparation of Organogels loaded with Miconazole Nitrate 29 : Lecithin organogels based systems were prepared by lecithin and tween-80 in different composition were dissolved in isopropyl myristate (IPM). After incorporation of drug, gelation was achieved on addition of the aqueous phase.
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 62 Table-4.2.1: Preparation of Organogels Loaded with Miconazole Nitrate Sl. No. Formulation Code Lecithin % w/w Tween- 80 % w/w IPM % w/w Water % w/w Drug % w/w 1. AM 1 7.54 3.77 30.16 56.56 2 2. AM 2 20.63 10.31 30.95 36.11 2 3. AM 3 35.43 1.77 11.81 21.26 2 4. AM 4 56.02 28.01 14.00 28.01 2
4.2.2 Preparation of Pluronic Lecithin Organogel 35 : The oil phase is prepared by mixing lecithin and isopropyl myristate and allowing the mixture to stand overnight to ensure complete dissolution. The aqueous phase is prepared by adding pluoronic F-127 to ice cold water, placing the mixture in refrigerator and agitating periodically to ensure complete dissolution.
To prepare PLO, the oil phase containing 2% drug miconazole nitrate mixed with the aqueous phase using a high shear mixing method by magnetic stirrer/ digital stirrer. It is important that the aqueous phase is cool before mixing because an aqueous solution of pluoronic F-127 is in the liquid state at a low temperature and gels at a higher temperature.
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 63 Table-4.2.2: Preparation of Pluronic Lecithin Organogels (PLO) Sl. No. Formulation code Lecithin % w/w IPM % w/w 30% Pluronic in water % w/w Drug % w/w 1. FM 1 7.54 30.16 60.33 2.0 2. FM 2 17.85 26.73 53.47 2.0 3. FM 3 36.76 24.50 36.76 2.0 4. FM 4 56.02 14.00 28.01 2.0
4.2.3 Preparation of Miconazole Nitrate Loaded Organogels 29 : Based on the different composition ration, the lecithin solutions were prepared by first dissolving lecithin in the organic solvent isopropyl myristate (IPM) with the aid of a magnetic stirrer and then while still stirring the required 2% miconazole nitrate is added followed by necessary amount of water (double distilled) into the mixture to obtain a clear gel. Formation of clear, homogenous and non-birefringent gels, after addition of water by a micropipette syringe.
Table-4.2.3: Preparation of Miconazole Nitrate Loaded Organogels Sl. No. Formulation Code Lecithin % w/w IPM % w/w Water % w/w Drug % w/w 1. SR 1 8.90 35.70 53.50 2.0 2. SR 2 20.22 30.33 47.50 2.0 3. SR 3 43.2 28.8 25.95 2.0 4. SR 4 63.33 15.08 22.63 2.0
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 64 4.2.4 Hot Melt Type Organogel 87 : Lecithin in various concentrations was used as a gelator and paraffin as the oil medium, the material was heated with stirring on hot plate stirrer and water is added with micropipette. Upon cooling of the melted disperse system, gellation was achieved. Table-4.2.4: Preparation by Hot Melt Type Organogels Sl. No. Formulation Code Lecithin % w/w Paraffin % w/w Water % w/w Drug % w/w 1. MN 1 13.43 53.72 30.82 2.0 2. MN 2 31.60 47.40 18.97 2.0 3. MN 3 49.00 34.67 16.33 2.0 4. MN 4 67.61 16.90 13.52 2.0
4.3 CONSTRUCTION OF CALIBRATION CURVE OF MICONAZOLE NITRATE 88 : 40 mg Of miconazole nitrate accurately weighed & added in a mixture of 10ml of 0.1M Hcl & 50 ml of iso propanol (propan-2ol). The volume is made 100ml with iso propranol. The UV absorption maximum is 272 nm. The Beer-Lamberts range used is 4g-32g. Previous studies have shown that miconazole has relatively low absorption in the UV range and are difficult to analyze in the low concentration. Therefore, the method adopted by Agarwal and Katare was followed to second the results.
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 65 4.4 EVALUATION OF LECITHIN-MICROEMULSION BASED ORGANOGELS: 4.4.1 pH 74 : Digital pen pH meter was used to measure the pH of gels at constant temperature. Prior to this pH meter was calibrated using buffer solution of pH 7.0 and 9.2 then the electrode was wasted with demineralized water. The electrode was inserted in to the sample 10 min prior to taking the readings.
4.4.2 Spreadability 17,41 : Introduction: One of the criteria for gels to meet the ideal qualities is that it should possess good spreadability. Spreadability is a term express to denote the extent of area to which the gel readily spreads on application to skin or the affected parts. The therapeutic efficiency of formulation also depends upon its spreading value. Hence, determination of spreadability is very important in evaluating gel characteristics.
Spreadability of the formulations was determined by an apparatus suggested by Mutimer et al, which was suitably modified in the laboratory and used for the study. It consists of a wooden block, which was provided by a pulley at one end. A rectangular ground glass plate was fixed on the block. An excess of gels (about 2 gm) under study was placed on this ground plate. The gel was then sandwiched between this plate and another glass plate having the dimensions of the fixed ground plate and provided with the hook. A 300 gm weight was placed on the tip of two plates for five minutes to expel air and to provide a uniform film of the gel between the plate. Excess of the gel was scrapped off from the edges. The top plate was then Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 66 subjected to pull of 300 gm. With the help of a string attached to the hook and the time (in seconds) required by the top plate to cover a distance of 10 cms be noted. A shorter time interval indicates better spreadability.
The spreadability was determined by special apparatus and it was calculated using the formula: S = t ml
Where, S = spreadability m = weight tied to the upper slide l = length of the glass slide t = time taken in seconds.
4.4.3 Determination of Viscosity 74 : The viscosity of formulated organogels were determined. The viscosity was determined using a Brookfield viscometer. The sample holder taken for the viscosity measurement was filled with the samples and then inserted into a flow jacket mounted on the viscometer. The samples adaptor (spindle), rotated at an optimum speed was used to measure the viscosity of the preparation, the samples was allowed to settle for five minutes prior to taking the readings.
4.4.4 Stability studies 38,71,89 : The stability of the formulations kept in the containers was assessed at different temperatures. Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 67
Samples were kept at constant temperature 25C for their physical stability, organoleptic properties and macroscopically appearance of the microemulsion.
4.4.5 Gelation Kinetics 40 : Gelation kinetic was determined by turbidimetric method using Nepheloturbidometer. In this method, the sample cell containing formulation components without water was placed in the cuvette chamber and meter is set to 0 NTU. Then water is added drop by drop using micropipette. Until the meter shows maximum NTU i.e., 100 NTU and the water quantity is measured of which turbidity is maximum.
4.4.6 Determination of Miconazole Nitrate Content 74 : An accurately weighed amount of each preparation was dissolved in phosphate buffer pH 6.4. The content of miconazole nitrate was determined spectro- photometrically at 272 nm using Shimadzu UV-visible spectrophotometer.
4.4.7 In vitro Diffusion Study 74 : The apparatus consists of a cylindrical glass tube (with 22 mm internal diameter and 76 mm height), which was opened at both the ends. One gram of (1 gm) of gel formulation containing 20 mg of miconazole was spread uniformly in the surface of cellophane membrane (previously soaked in receptor medium for overnight 24 hours) and was fixed to the one end of tube such that the preparation occupies inner circumference of the tube. The whole assembly was fixed in such a way that the lower end of tube containing gel was just touched (1-2 mm deep) the surface of Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 68 the diffusion medium i.e., 45 ml pH 6.4 phosphate buffer present in 50 ml beaker, which was placed on thermostatic hot plate with magnetic stirrer and maintained at 372C. The cellophane membrane act as barrier between the gel and pH 6.4 phosphate buffer. The contents were stirred using magnetic bar at 50 rpm. A quantity of 1 ml samples were withdrawn at an interval of hourly and the same amount is replaced with pH 6.4 phosphate buffer each time.
The released drug was estimated by using Shimadzu UV-visible spectrophotometer at 272 nm.
4.4.8 In Vitro Antifungal Activity 63,70,74 : Ditch plate technique: For evaluation of fungistatic mainly for semisolids. Agar plate were prepared and sterilized as per standard procedure. A ditch is made in the centre of agar plates (2.5 x 0.50 cm) and the formulation (0.5 gm) were filled in the ditch. The prepared culture loops (culture fungi) were streaked across the agar at a right angle from the ditch to the edge of the plate. Control plate of marketed formulation was also prepared. After incubation of 72 hours at 25C the fungal growth was observed and the percentage inhibition was measured as follows: Percentage inhibition = 1 2 L L x 100 Where L 1 = total length of the streaked fungal culture L 2 = length of inhibition.
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 69 4.4.9 Drug-Polymer Interaction Studies (Infrared Spectroscopy): The infrared spectra (IR) of miconazole nitrate, IPM, pluronic, lecithin, tween 80, liquid paraffin and some selected formulation was obtained using FTIR (Perkin- Elenmeyer 1600 Series).
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RAW MATERIAL CHARACTERIZATION: Table-5.1.1: Standardization of Miconazole Nitrate Sl. No. Characteristic IP Limit Observation 1. Description White or almost white, crystalline or microcrystalline powder White micro crystalline powder 2. Solubility Freely soluble m method slightly soluble in ethonol and m chloroprm, very slightly soluble m water and in ether Meets the specification 3. Identification A IR spectra of sample must be identified to that of standard Complies 4. Identification E Melts between 178 to 184 C 180C 5. Loss on drying Not more than 0.5 determined on 1 g by drying in in an oven at 105 for two hrs. 0.3% 6. Assay 98.5% to 101.5% w.r.t. dried substance 99.6% w/w 7. Clarity and coloured solution 1% w/v solution in methanol is clear Clear
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Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 72 Table-5.1.2a: Standardization of Lecithin Sl. No. Characteristic USP Limit Observation 1. Description Lecithins from viscous semiliquid to powders depending upon the freely fatty acid content, colour from brown to light yellow. Light yellow powder 2. Solubility Lecithins are soluble in aliphatic and aromatic hydrocarbons, halogenated hydrocarbons, mineral oil, and fatty acid. Meets the limits
Table-5.1.2b: Standardization of tween 80 Sl. No. Characteristic USP Limit Observation 1. Description Polysorbates have a characterization odor and a worm, some what bitter taste the colors. Yellow 2. Solubility Soluble methanol and water insoluble in solver mineral oil and vegetable oil. Meets the limit 3. PH 6-8 7 4. Viscosity -- 425 (mPas) Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 73 Figure-5.1.2a: IR Spectra of Lecithin
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Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 75 Table-5.1.2c: Standardization of pluronic (poloxames) Sl. No. Characteristic USP Limit Observation 1. Description White, almost odourless laste less waxy flakes White flakes 2. Solubility Freely soluble in water allows, chloroform pratically insoluble in propylane glycol Meets the limits 3. Melting point -- 52C 4. pH 5.0-7.5 7
Table-5.1.2d: Standardization of Isopropyl myristate Sl. No. Characteristic USP Limit Observation 1. Description Isopropyl myristate is a clear, colorless, practically odourless mobile liquid with a bland taste. Meets the limits 2. Solubility Miscible with acetone chloroform, ethanol ethylacetate, fats, fatty alcohol, fixed oils, liquid hydrocarbons. Practicatly in soluble in propylene glycol and water Meets the limits 3. pH Between 6-7 6.5 4. Residue on ignition - 0.1%
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Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 78 Table-5.1.2e: Standardization liquid paraffin Sl. No. Characteristic BP Limit Observation 1. Description Mineral oil is transparent colorless. Viscus liquid, odorless. Transparent viscous liquid 2. Solubility Practically insoluble in ethanol, and water soluble in acetone, benzone, ether and missalbe with volatile oil and fixed oils. Meet the limits
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Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 80 5.2 PREPARATION OF MICONAZOLE NITRATE ORGANOGELS Organogels were prepared as described in methodology (section-4.2)
5.3 CALIBRATION CURVE OF MICONAZOLE NITRATE: Date 12 th Aug. 2005 Drug Miconazole Nitrate Mobile Phase SPB 6.4 Wavelength 272 nm Unit of concentration mcg/ml Slope of Calibration curve 77.4256 Constant of calibration curve 1.2403 R of calibration curve 0.9998
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 83 Effect of addition of water on turbidity of microemulsion based organogel of formulations (Gelation kinetics)
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 92 Figure-5.4.2: Effect of addition of water on turbidity of microemulsion based organogel of formulations FM 1 , FM 2 , FM 3 & FM 4
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 93 Table-5.4.3a: Gelation Kinetics SR 1
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 97 Figure-5.4.3: Effect of addition of water on turbidity of microemulsion based organogel of formulations SR 1 , SR 2 , SR 3 & SR 4
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 98 Table-5.4.4a: Gelation Kinetics MN 1
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 100 Figure-5.4.4: Effect of addition of water on turbidity of microemulsion based organogel of formulations MN 1 , MN 2 , MN 3 & MN 4
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 101 Table-5.4.5: Gelation Kinetic Data Sl. No. Formulation Code Water gm% at maximum NTU 1. AM 1 26.60 2. AM 2 23.07 3. AM 3 19.75 4. AM 4 18.60 5. FM 1 23.07 6. FM 2 16.60 7. FM 3 15.25 8. FM 4 9.09 9. SR 1 28.50 10. SR 2 24.20 11. SR 3 16.60 12. SR 4 16.60 13. MN 1 9.09 14. MN 2 9.09 15. MN 3 9.09 16. MN 4 9.09
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 103 5.4.7a In Vitro Percentage Drug Release Name of the drug Miconazole Nitrate Loading dose (mg) 20 Total No. of readings including zero time reading 13 Diffusion medium SPB 6.4 RPM 50 Volume of diffusion media (ml) 45 Volume of sample removed (ml) 1 Dilution factor 5 Slope of calibration curve 77.4256 Constant of calibration curve 1.2403 R of calibration curve 0.9998
Table-5.4.7a: In vitro percentage drug release of formulations AM 1 , AM 2 , AM 3 and AM 4
Average % Drug Release Sl. No. Time (Hours) AM 1 AM 2 AM 3 AM 4
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Figure-5.4.7a: In vitro percentage drug release of formulations AM 1 , AM 2 , AM 3 and AM 4
0 20 40 60 80 100 120 0 2 4 6 8 10 12 Time (Hrs) P e r c e n t
D r u g
R e l e a s e d AM1 AM2 AM3 AM4 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 105 5.4.7b In Vitro Percentage Drug Release Name of the drug Miconazole Nitrate Loading dose (mg) 20 Total No. of readings including zero time reading 13 Diffusion medium SPB 6.4 RPM 50 Volume of diffusion media (ml) 45 Volume of sample removed (ml) 1 Dilution factor 5 Slope of calibration curve 77.4256 Constant of calibration curve 1.2403 R of calibration curve 0.9998
Table-5.4.7b: In vitro percentage drug release of formulations FM 1 , FM 2 , FM 3 and FM 4
Average % Drug Release Sl. No. Time (Hours) FM 1 FM 2 FM 3 FM 4
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Figure-5.4.7b: In vitro percentage release of formulations FM 1 , FM 2 , FM 3 and FM 4
0 10 20 30 40 50 60 70 80 90 100 0 2 4 6 8 10 12 Time (Hrs) P e r c e n t
D r u g
R e l e a s e d FM1 FM2 FM3 FM4 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 107 5.4.7c In Vitro Percentage Drug Release Name of the drug Miconazole Nitrate Loading dose (mg) 20 Total No. of readings including zero time reading 13 Diffusion medium SPB 6.4 RPM 50 Volume of diffusion media (ml) 45 Volume of sample removed (ml) 1 Dilution factor 5 Slope of calibration curve 77.4256 Constant of calibration curve 1.2403 R of calibration curve 0.9998
Table-5.4.7c: In vitro percentage drug release of formulations SR 1 , SR 2 , SR 3 and SR 4
Average % Drug Release Sl. No. Time (Hours) SR 1 SR 2 SR 3 SR 4
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Figure-5.4.7c: In vitro percentage drug release of formulations SR 1 , SR 2 , SR 3 and SR 4
0 10 20 30 40 50 60 70 0 2 4 6 8 10 12 Time (Hrs) P e r c e n t
D r u g
R e l e a s e d SR1 SR2 SR3 SR4 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 109 5.4.7d In Vitro Percentage Drug Release Name of the drug Miconazole Nitrate Loading dose (mg) 20 Total No. of readings including zero time reading 13 Diffusion medium SPB 6.4 RPM 50 Volume of diffusion media (ml) 45 Volume of sample removed (ml) 1 Dilution factor 5 Slope of calibration curve 77.4256 Constant of calibration curve 1.2403 R of calibration curve 0.9998
Table-5.4.7d: In vitro percentage drug release of formulations MN 1 , MN 2 , MN 3 and MN 4
Average % Drug Release Sl. No. Time (Hours) MN 1 MN 2 MN 3 MN 4
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Figure-5.4.7d: In vitro percentage drug release of formulations MN 1 , MN 2 , MN 3 and MN 4
0 5 10 15 20 25 30 35 0 2 4 6 8 10 12 Time (Hrs) P e r c e n t
D r u g
R e l e a s e d MN1 MN2 MN3 MN4 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 111
Table-5.4.8: In Vitro Antifungal Activity of Miconazole Nitrate Organogels Zone of Inhibition (mm) Formulation Code Candida Albicans P Chysogenum A Niger Marketed 9.00 11.00 14.00 AM 1 11.00 13.00 15.00 FM 1 10.00 12.00 14.00 SR 1 9.00 11.00 12.00 MN 1 6.00 8.00 10.00 * mean of three readings.
Figure-5.4.8: Comparative Antifungal Activity of Selected Miconazole Nitrate Organogel Formulations with Control
5.4.9 Drug Polymer Interactions: The IR spectra of formulations AM 1 , FM 1 , SR 1 , MN 1 are depicted in figure 5.4.9. a,b,c,d. Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 112
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 114 Figure-5.4.9a: IR Spectra of AM 1 Organogel
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Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 116 Figure-5.4.9c: IR Spectra of SR 1 Organogel
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 117 Figure-5.4.9d: IR Spectra of MN 1 Organogel
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 118 CHAPTER6 DISCUSSION
Lecithin, a biocompatible material has recently gained wide popularity for development of better drug delivery system. In present study, lecithin along with Tween-80, pluronic, isopropyl myristate and liquid paraffin were used for the development of lecithin-microemulsion based organogels for the topical delivery of miconazole nitrate. Miconazole nitrate is a synthetic imidazole derivative with molecular formula C 18 H 14 N 2 CH NO 3 and practically insoluble in water. Miconazole nitrate was obtained from Adelphi A/c Sigma Lab., Goa and its standards complied with IP limits (table-5.1.1). Lecithin, pluronic, tween-80 and isopropyl myristate complied with USP XXV standards (table-5.1.2a, b, c and d). Liquid paraffin complied with the BP standards (table-5.1.2.e).
Lecithin organogels based systems were prepared by three different methods. In the first method lecithin and tween-80 in different compositions were dissolved in isopropyl myristate (IPM) after incorporation of drug gelation was achieved on addition of the aqueous phase (table-4.2.1).
In the second method, pluronic lecithin organogels (PLOs) were prepared employing lecithin, isopropyl myristate and pluronic in water (table-4.2.2).
The third method was based on preparation of lecithin solution by employing isopropyl myristate (IPM) followed by incorporation of drug. Addition of aqueous phase resulted in the lecithin organogel system (table-4.2.3). Finally hot melt type Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 119 organogels were prepared by using lecithin and liquid paraffin in various concentration. Water was added and gelation was achieved on cooling (table-4.2.4).
The parameters studied included pH, spreadability, viscosity and gel life (stability). The results indicated change in pH with change in composition of lecithin organogel. An increase in percent of lecithin resulted in increase in viscosity and decrease in the spreadability of lecithin organogels. An organogel (MN 4 ) prepared by hot melt technique was found to possess highest viscosity i.e., 22868 CPs and lowest spreadability of 0.57 gm-cm/sec, whereas AM 1 lecithin microemulsion based organogel containing tween-80 (3.77%) showed lowest viscosity 1572 CPs. Gel stability studies indicated that organogels with higher viscosity were more stable than less viscous gels. MN 4 hot melt type organogels containing 67.61% of lecithin produce the stable gel 192 hours at 25 (table-5.4).
The physical stability of an organogel to be used as drug delivery system is important as the gel should retain a structure during storage for uniform consistency during use and for reproducible release profile. Lower the amount of lecithin less stable the organogels were formed. Organogels containing lecithin and Tween-80 were found to be the most stable, tweens with polyoxy ethylene chain precipitate in tubular aggregates of sorbiton monostearate and thus stabilize the organogel further AM 4 showed highest gel-life of 220 hours at 25C. The general trend in gel life observed was AM 4 >MN 4 >FM 4 >SR 4 .
Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 120 Gelation kinetics studied using nephalo-turbidimeter produce interesting results the amount water required for gelation was found to be less as the amount of lecithin increased in the organogel system. SR 1 organogel containing lecithin and IPM showed maximum gelation on addition of 28.5 gm% of water. Organogels produced by hot melt method were an exceptional. The amount of water required for maximum gelation was observed to be a constant i.e., 9.09 gm% (table-5.4.5).
Percent drug content of organogels were determined using calibration curve of miconazole nitrate in phosphate buffer pH 6.4 at 272 nm. Percent drug content was found to be between 97.82 and 101% (table-5.4.6).
In vitro release of miconazole nitrate from the organogels was determined using the method suggested by Chowdary et al 41 . Phosphate buffer pH 6.4 was employed as diffusion medium in the receptor compartment the organogels containing lecithin and tween-80 showed retarded release with an increase in the amount of lecithin. The order of release may be given as AM 1 >AM 2 >AM 3 >AM 4
(table-5.4.7a). Highest release is shown by AM 1 formulation. Similar results were observed for organogels containing lecithin and pluronic FM 1 >FM 2 >FM 3 >FM 4 with FM 1 showing highest release 94.98% (Table-5.4.7b). Formulation containing only lecithin showed a comparatively low percent release of miconazole nitrate in the order SR 1 >SR 2 >SR 3 >SR 4 . SR 1 showing highest release 59.21% (Table-5.4.7c). The organogels prepared by hot melt method showed the least percent drug release in the order MN 1 >MN 2 >MN 3 >MN 4 with MN 1 showing highest release of 30.05% (Table- 5.4.7d). Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 121
It may be noted that the organogel formulations with higher viscosity showed less release of miconazole nitrate. Since the number and dimension of aqueous region available for diffusion of the drug are reduced with the possible increase in micro viscosity in the three dimensional structure of the gel.
In vitro antifungal activity of miconazole nitrate organogels were determined by Ditch plate method using best release formulation from amongst the four different types of organogels. A marked preparation was used as a control for comparison the maximum zone of inhibition studies indicated highest antifungal activity for lecithin, Tween-80 formulation. AM 1 followed by lecithin, pluronic formulation FM 1 , SR formulation containing only lecithin showed comparatively less antifungal activity whereas organogel prepared by hot melt method were found to have least antifungal activity. The antifungal activity against Candida albicans, Penicillin P.chrysogenum, Aspergillus niger was in the order AM 1 >FM 1 >Marketed>SR 1 >MN 1 . The in vitro antifungal activity significantly correlated with the in vitro release of miconazole nitrate from the mentioned formulation (table-5.4.8).
FTIR of miconazole nitrate showed principle peaks at wavelength 1089, 1313, 827, 812 cm 1 . FTIR studies were also done for lecithin, tween-80, pluronic, IPM, liquid paraffin and formulation AM 1 , FM 1 , SR 1 and MN 1 as shown in figure respectively. Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 122 CHAPTER7 CONCLUSION
Lecithin microemulsion based organogel have currently generated great interest as topical and transdermal drug delivery vehicle. In the present investigation lecithin-microemulsion based organogels were formulated using four different methods employing lecithin and tween-80, pluronic, and also preparing the organogels using hot melt method where mineral oil (liquid paraffin) was used. From the studies, the following conclusions can be drawn. 1. Lecithin organogels can be prepared easily and rapidly and can be obtained with biocompatible components. 2. Change in composition of the lecithin organogels resulted in a change in pH. 3. With an increase in amount of lecithin viscosity of the organogels were found to increase resulting in decrease spreadability MN 4 formulation showed maximum viscosity 22868 CPs, whereas AM 1 formulation containing tween- 80 (3.77%) was least viscous 15726 CPs. 4. Organogels with higher viscosity were found to be more stable. Organogel containing lecithin and tween 80 were the more stable AM 4 showed highest gel life of 220 hours at 25C. 5. 28.5 gm% of water was required to produce maximum gelation in SR 1
organogels containing lecithin and IPM. Organogels produced by hot melt method required least amount of water for maximum gelation 9.09 gm%. Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 123 6. In vitro release of miconazole nitrate from organogel showed that organogels containing lecithin and tween 80 in IPM (AM 1 ) gave highest release. The order of release for the best releasing formulation prepared by different methods was AM 1 >FM 1 >SR 1 >MN 1 . 7. In vitro antifungal activity of miconazole nitrate organogels against Candida albicans, P.chrysogenum, A. niger was in the order AM 1 >FM 1 > Mkt> SR 1 >MN 1 . 8. In vitro antifungal activity significantly correlated with in vitro release of miconazole nitrate.
Thus, finally it may be concluded that lecithin-microemulsion based organogel have good potential as carrier for topical delivery of antifungal agent such as miconazole nitrate.
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Lecithin organogels are readily obtained by adding a minimal amount of water to a solution of lecithin in organic solvents. Surprisingly high viscosities can be achieved in various organic solvent on addition of water. Such systems have currently generated great interest as topical and transdermal drug delivery carriers.
In the present study, lecithin based microemulsions were formulated as topical carriers for miconazole nitrate, an antifungal agent, a synthetic imidazole derivative with molecular formula C 18 H 14 C 14 CHNO 3 and practically insoluble in water.
The need for the study and objective of the present study have been discussed extensively in chapter-2. The scheme of work was divided into V-phases. Initially collection of theoretical and technical data by extensive literature survey, review of literature and drug profile is presented in chapter-3. This was followed by procurement of material. Miconazole nitrate was obtained rom Adelphi A/c Sigma Lab, Goa and standardized as per IP. Standardization of all other material were done and they met the pharmacopoeial and other established standards.
Lecithin based organogel systems were prepared by three different methods employing lecithin, tween-80, pluronic, isopropyl myristate was used as organic solvent whereas gelation was achieved on addition of aqueous phase. Organogels were also prepared by hot melt method using lecithin in liquid paraffin as non- aqeuous phase. Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 125
Various parameters were evaluated such as pH, spreadability, viscosity, gelation kinetics and gel life.
From the study, it was observed that: 1. Lecithin organogels can be prepared easily and rapidly and can be obtained with biocompatible components. 2. Change in composition of the lecithin organogels resulted in a change in pH. 3. With an increase in amount of lecithin viscosity of the organogels were found to increase resulting in decrease spreadability. MN 4 formulation shown maximum viscosity 22868 CPs whereas AM 1 formulation containing tween- 80 (3.77%) was least viscous 15726 CPs. 4. Organogels with higher viscosity were found to be more stable. Organogels containing lecithin and tween-80 were the more stable (AM 4 ) showed highest gel life of 220 hours at 25. 5. 28.5 gm% of water was required to produce maximum gelation in SR 1
organogels containing lecithin and IPM. Organogels produced by hot melt method required least amount of water for maximum gelation 9.09 gm%. 6. In vitro release of miconazole nitrate from organogel showed that organogels containing lecithin and tween-80 in IPM (AM 1 ) gave highest release. The order of release for the best releasing formulation prepared by different methods was AM 1 >FM 1 >SR 1 >MN 1 . Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com) 126 7. In vitro antifungal activity of miconazole nitrate organogels against Candida albicans, P. chrysogenum, A. niger was in the order AM 1 >FM 1 >Mkt> SR 1 >MN 1 . 8. In vitro antifungal activity significantly correlated with in vitro release of miconazole nitrate.
Thus, finally it may be concluded that lecithin microemulsion based organogel have good potential as carrier for topical delivery of antifungal agent such as miconazole nitrate.
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Assessment of Knowledge Attitudes Perception and Barriers Tow Ards Pharmacovigilance Activities Among Community Pharmacists and Final Year Phatmacy Students in Malaysia