Formulation and Evaluation of Ketoconazole Microemulsion Antidandruff Shampoo

“FORMULATION AND EVALUATION OF KETOCONAZOLE MICROEMULSION ANTIDANDRUFF SHAMPOO”

MASTER OF PHARMACY DISSERTATION PROTOCOL

SUBMITTED TO THE

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

BANGALORE, KARNATAKA.

BY

SHILPA SENAN

B.pharm

Under The Guidance of

Dr. A.R. SHABARAYA

M.Pharm, MBA,Ph.D.

DEPARTMENT OF PHARMACEUTICS.

SRINIVAS COLLEGE OF PHARMACY, VALACHIL, MANGALORE – 574143

2013-2015

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

BANGALORE, KARNATAKA

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. / Name of the Candidate and Address: / MISS. SHILPA SENAN
1stYEARM.PHARM,
DEPT. OF PHARMACEUTICS,
SRINIVAS COLLEGE OF PHARMACY,
VALACHIL, MANGALORE-574143.
2. / Name of the Institution: / SRINIVAS COLLEGE OF PHARMACY,
VALACHIL, FARANGIPETE POST, MANGALORE-574143.
3. / Course of Study and Subject: / MASTER OF PHARMACY
(PHARMACEUTICS)
4. / Date of Admission: / 25-07-2013
5. / Title of the Project:
“FORMULATION AND EVALUATION OF KETOCONAZOLE MICROEMULSION ANTIDANDRUFF SHAMPOO”
6.
7. / Brief Resume of the intended work:
6.1 - Need of the study:

• Fungal infections are extremely common and some of them are serious and even fatal. With the control of most bacterial infections in the developed countries, fungal infections have assumed greater importance.

Hair is one of the vital parts of the body derived from the ectoderm of the skin and is protective appendages on the body and considered accessory structure of the integument along with sebaceous gland sweat glands and nails1,2.

• Dandruff is a common embarrassing scalp disorder affecting a large chunk of population. It affects the aesthetic value and often causes itching. It occurs when dead skin is shed,producing irritating white flakes and possibly an itchy scalp. Dandruff results from excessive drying of skin and over-activity of the oil glands, known as ‘seborrhea’. Although dandruff is associated with the scalp, flakes may also appear on face, nose and eyebrows, as well as on the skin behind the ear, and the skin of trunk, particularly increases dandruff, which is visible desquamation of scalp is the mildest manifestation of Seborrheic dermatitis and caused by Plasmodium ovale combined with multiple host factors1,2.
• The commonly associated symptoms of dandruff are itching with scalp soreness. The differential diagnosis of dandruff includes conditions such as eczema, atopic dermatitis, candidiasis, contact dermatitis, dermatomyositis, drug eruptions, lichen simplex, tinea capitis, xerotic eczema, and vitamin B and/or zinc deficiency2.
• Therapeutic shampoos offer a convenient option for treating scaling and scalp. These products have the advantage of being more cosmetically elegant than other topical formulations, such as solutions, ointments, and foams.The shampoos simultaneously clean the hair and scalp by emulsifying oily secretions while treating the underlying disease3.
• There are several classes of antifungal drugs which destroys or prevents

the growth of fungi. Polyenes, which causes an increase in fungal cell wall permeability leading to its death. Examples: amphotericin B, natamycin, nystatin. Azoles like clotrimazole, econazole, fluconazole, itraconazole, ketoconazole, miconazole1,2

• Microemulsion is defined as a dispersion consisting of “oil, surfactant, co-surfactant, and an aqueous phase. It is a single optically isotropic and thermodynamically stable liquid solution with a droplet diameter usually within the range of 10-100nm. Microemulsions have number of special properties such as enhanced drug solubility, good thermodynamic stability, ease of manufacturing and permeation enhancement ability. Microemulsions are promising delivery systems that allow sustained or controlled drug release for percutaneous, topical, transdermal, ocular, and parenteral administration. Antifungal agents e.g. miconazole, ketoconazole anditraconazole being lipophilic in nature have been formulated as microemulsions to impart to them the advantages like ease of preparation due to spontaneous formation, thermodynamic stability, transparent and elegant appearance, increased drug loading, enhanced penetration through the biological membranes, increased bioavailability compared to conventiontal dosage forms. Microemulsion of poorly water soluble antifungal drugs miconazole, ketoconazole, and itraconazole were designed and developed using either mineral oil or microemulsions of poorly water soluble antifungal agents were successfully developed with in vitro release rates comparable to that of the gel formulation4.
• Ketoconazole (KTZ) is a broad-spectrum imidazole derivative of antifungal agent developed for treatment of human mycotic infections and plays an essential role in the antifungal chemotherapy.It is weak base with poor water solubility. Ketoconazole which arean imidazoleantifungal agents commonly applied topically (to the skin) or mucus membranes to cure fungal infections. These works by inhibiting the synthesis of ergosterol, a critical component of fungal cell membranes. It can also be used against certain species of Leishmaniaprotozoa (which are a type of unicellular parasite) as these also contain ergosterol in their

cell membranes. In addition to its antifungal and antiparasitic actions, it
also has some limited antibacterial properties5. In the present study an attempt will be made to design and evaluate ketoconazole antidandruff microemulsion shampoo.
Advantages of medicated antidandruff shampoo:
Effective against skin problems with bacteria, yeast, mold. Fungi and viruses.
Broad-spectrum activity protects against difficult to identify opportunistic microorganisms.
Safe for shampooing animals with skin injuries including cuts. Tears. And abrasions.
Speeds healing.
Economic.
Ease of transport.
Ease of applying and removing.
Good patient compliance.
Maximum rate of skin permeation of drug.
Termination of therapy is possible if toxicity is observed.
6.2 - Review of literature:
Design of antidandruff shampoo is a new concept to study hence a thorough literature survey will be done on the topic, through primary, secondary and tertiary sources of drug information at Srinivas College of Pharmacy.
•Anshul Jain, Goswami RB, Neelima Goswami, Nishi Prakash Jain2 has formulated and evaluated antidandruff shampoo of fluconazole and ketoconazole. The antidandruff shampoo were prepared in ten different batches which contain fluconazole and ketoconazole along with excipients in different concentrations in each batches. By performing evaluation test they concluded that all the evaluation tests of ten batches are according to
standard limits. It also produce good foam and penetration.
•Jayaraj Kumar, JayachandranE, GridharB, Rahul Nair, Jayakandan M, KathiravanM et al6.,has developed and evaluated antidandruff shampoo of povidone iodine liquid. Povidone iodine is an iodophore complex of iodine is continually delivered. The different concentrations of dioctyl sodium sulfo succinate(20%), sodium lauryl sulphate(9%), along with (4%) of povidone iodine formulation were prepared and evaluated for foam, foam stability, cleaning action, wetting action, conditioning action, viscosity, microbial test, eye irritancy test. All the evaluation test showed better result than the marketed product on comparison study.
•Donald L7 Greer had performed successful treatment of tinea capitis with 2% ketoconazole shampoo. A total of 16 black children, aged 3-6 all with proven tinea capitis caused by Trichophyton tonsurans, were treated daily for 8 weeks with 2% ketoconazole shampoo. This study shpwed that ketoconazole 2% shampoo alone reduces the number of viable anthroconidia in children with tinea capitis thus reducing the transmissibility and contagious nature of thte disease.
•PrapapornBoonme, Nathida Pakpayat, Kanokwan Yotmanaee, Sarinnart Kunlawijitrungsee, Duangkhae Maneenuan8, performed a study to evaluate silicone quaternary microemulsion. This study aimed to characterise properties and to evaluate conditioning performance of silicone quaternary microemulsion. Addition of silicone quaternary microemulsion in the investigated formulation did not markedly affect the characteristics of the obtained shampoos. Samples provided stable foam, surface tension reduction and low viscosity with Newtonian flow.
•Alia A Badavi9,performed the evaluation of microemulsion systems containing salicylic acid. Microemulsions are used to solubilise drug and to improve topical drug availability. Salicylic acid is keratolytic agent
used in topical product with antimicrobial action. Different concentrations of salicylic acid were incorporated in an microemulsion base composed of isopropyl myristate, water, and tween 80: propylene glycol in the ratio of 15:1. Evaluation by examination under cross polarising microscope, measuring of percent transmittance, determination of specific gravity, assessment of rheological properties and accelerated stability test were carried out. The data showed that addition of salicylic acid affect physical properties of base.
•Adnanazeem10,performed a study of emerging role of microemulsion in cosmetics. Microemulsion represent a promising carrier system for cosmetic active ingredients due to their numerous advantages over the existing conventional formulations. They are capable of solubilising both hydrophilic and lipophilic ingredients with relatively higher encapsulation. They have also found application in after shave formulations which upon application to the skin provide reduced stinging and irritation and a comforting effect without tackiness. These newer formulations elicit very good cosmetic attributes and hogh hydration properties with rapid cutaneous penetration which may accentuate their role in topical products.
•Saboji JK, Manvi FV, Gadad AP,11 has formulated and evaluated ketoconazole microsponge gel for prolonged topical administration. Microsponge containing ketoconazole drug with six different proportions of Eudragit RS100 as polymer were studied successfully using quasi-emulsion solvent diffusion method. These formulations were studied for particle size and physical characterisation. The physical characterisation showed that microsponge formulation MS IV (containing 1gm of ketoconazole and 0.2gm of Eudragit RS 100) and MS VI (containing 2gm of ketoconazole and 0.2gm of Eudragit RS 100) showed a better loading efficiency and production yield. These two microsponges formulation were prepared as gel in 0.35%w/w carbopol and studied for pH,viscosity,spreadability, drug content, in vitro release, antimicrobial activity and in vivo antifungal activity studied on guinea pig.
The microsponge formulation gel,MKG 1(contains 1.0%w/w of ketoconazole microsponge equivalent to ketoconazole 1.0%w/w) showed viscosity 4390cps, spreadability of 19.27gcm/s and drug content of 85.2%. The antimicrobial studies showed azone of inhibition with 13.5mm and 12.0mm for microsponge formulation gel MKG 1 and MKG 2(contains 1.0%w/w of ketoconazole microsponge equivalent to ketoconazole 1.0%w/w) respectively when compared with the pure drug, zone of inhibition 18.2mm. These formulations also showed better antifungal activity on fungal induced guinea pig when compared with control group without of drug.
•Praveen S Patil, Vinod M Reddy, Karnakumar V Biradar, Chandrashekhar B Patil, Sreenivasa Rao K12, have developed and evaluated clotrimazole anti-dandruff hair gel. The different formulations were developed using polymers such as Carbopol 940, Carbopol 934, PEG 200 etc. These polymers selected based on their use in gel. All the formulations were evaluated Active Content, Physical appearance, pH, Viscosity, Extrudability, Antifungal activity, Drug release profile, Compatibility and Stability study. In stability study, formulation F7 (containg clotrimazole 1.5%) was shows no appreciable changes as compared to other formulation during the period of three months. Formulation F7 was shows maximum zone of inhibition to an anti-fungal activity during in vitro study. The viscosity of formulation F6 (containing clotrimazole 1%) was found to be highest and viscosity of formulation F1 (containing clotrimazole 0.5%) was found to least. The result of the extrudability study indicate that the F5 to F8 had better extrudability than F1 to F4. Therefore F7 could be used as an effective formulation for anti-dandruff hair gel of clotrimazole as compared to other formulation.
•Shireesh R, Chandra Shekar B, Vishnu P, Prasad MVV13 have formulated and evaluated the ketoconazole nail lacquer as ungual drug delivery system for the treatment of onychomycosis. It includes penetration enhancers of urea hydrogen peroxide, thioglycolic acid and an
antifungal agent (ketoconazole).From the experiment they concluded that urea shows slow release than thioglycolic acid this demonstrates the potential use of such penetration enhancer systems with ketoconazole in the topical treatment of onychomycosis.
•Van Cutsem3, comparing the in vitro antifungal activity of ketoconazole 2%, selenium sulphide 2.5%, and zinc pyrithione 1% and 2% against M. Furfur in guinea pig, Ketoconazole was found to be most effective for reducing M. furfur counts but results with selenium and 1% and 2% zinc pyrithione was comparable.The antidandruff effects ofketoconazole were superior to those of selenium sulphide.
•Leyden3, studied the combination of 2% sulphur and 2% salicylic acid in a shampoo base (Sebulex etc.) in double- blind, controlled trial using both clinical assessment of scaling and corneocyte counts. They observed significantly greater and earlier reductions in both the degree of scaling and in corneocyte counts in subjects using the 2% sulphur/ 2% salicylic acid combination than in those using either the active ingredient alone or the shampoo vehicle.
6.3 - Objectives of the study:
To evaluate with respect to drug excipient interaction studies (FTIR)
To carry out the preformulation studies of ketoconazole.
To design and develop ketoconazole antidandruff microemulsion shampoo by microemulsion method.
To carry out evaluation of formulated ketoconazole antidandruff microemulsion shampoo.
To carry out stability studies as per ICH guidelines.
Materials and Methods:
Materials:

1. Drug :Ketoconazole14
2. Surfactants : Anionic surfactants like ammonium lauryl sulphate,

Sodium lauryl sulphate,lauric, stearic acid and their
salts etc14

1. Foam builders : Betaines or alkanolamides, sacrosinate etc14.

1. Conditioning agent : Lanolin, dimethicon etc14.
2. Thickners : Methylcellulose, Sodium chloride, Ammonium

chlorideetc14

1. Sequestering agent : EDTA etc14.
2. Opacifying agent : Propylene glycol, stearic acid etc14.
3. Clarifying agent : Ethanol. Isopropanol etc14.
4. Preservatives : Methyl paraben and propyl paraben etc14.
5. Perfumes : Fruity or floral etc14.
6. Solvent : Water.

All other chemicals used will be of analytical grades.
Method:
Medicated antidandruff shampoo will be prepared by microemulsion method16.
In this method formulation of ketoconazole microemulsion will be done by incoperating oil, surfactant,and cosurfactant. This ketoconazole microemulsion will be mixed with other ingredients of shampoo in different concentrations.
7.1-Source of data:
Review of literature from
a)Journals such as :-
International Journal of Pharmacy and Pharmaceutical Sciences.
International Journal of Research in Pharmaceutical Sciences.
International Journal of PharmTech Research.
International Journal of CurrentPharmaceutical Research.
International Journal of Universal Pharmacy and Life Sciences.
International Journal of Applied Pharmaceutics.
Indian Journal of Dental Research.
International Journal of Dermatology.
International Journal of Pharma Professional’s Research.
Scholar research library.
b)Internet Browsing.
c)Laboratory based studies.
7.2 - Method of Collection of Data:

1. An overview of Medicated Antidandruff Shampoo.
2. Formulation of Medicated Antidandruff Shampoo.
3. Evaluation of Medicated Antidandruff shampoo

Drug and excipients interaction: By F. T. I. R Spectroscopy.
Physical appearance / visual inspection6,15
Determination of globule size4 .
Transmission electron microscopy4 .
Determination of pH15.
Foam and foam stability6,15.
Determination of solid content15.
Wetting action6.
Dirt dispersion15.
Surface tension measurement15.
Detergency ability15.
Microbiological assay15.
Stability studies as per ICH guidelines2.
7.3 - Does the study require any investigations or interventions to be
conducted on patients or other humans or animals? If so, please
describe briefly.
- NA
7.4 -Has ethical clearance been obtained from your institution in case of 7.3?
-NA
List of references:

1. Purushottam RK,Ashok K, Aamer Q,Prashant S. Formulation and preclinical evaluation of ketoconazole gel prepared by hydrotropic phenomenon. Int J Uni Pharm Life Sci 2011;1(1):72-84.

1. Anshul J, Goswami RB, Neelima G, Nishi PJ . Formulation and evaluation of antidandruff shampoo of fluconazole and ketoconazole. World J Pharm Res 2012;2(1):182-202.

1. Angela S, Joseph C. An overview of medicated shampoos used in dandruff treatment. Int J dermatology 2006;31(7):396-400.

1. Tenjarla S. Microemulsion an overview and pharmaceutical application.

CritRe V Ther Drug Carrier System 1999;16(1):461-521.

1. Tripathi KD. Essentials of medical pharmacology. 6th ed. New Delhi: Jaypee Brothers Med Publishers 2008;(1):762.

1. Jayraj KK, Jayachandran E, Gridhar B, Rahul N, Jayakandan M. Formulation and evaluation of povidone iodine liquid antidandruff shampoo. Int J Pharm Sci and Res 2009;1(3):108-11.

1. Donald LG.Successful treatment tof tinea capitis with 2% ketoconazoleshampoo. Int J dermatology 2000;39(4):302-4.

1. Prapaporn B, Nathida P, Kanokwan Y, Sarinnart K, Duangkhae M. Formulation and evaluation of silicone quaternary microemulsion. Int J Chem Tech 2011;1(4):1442-8.

1. Alia AB. Formulation and evaluation of salicylic acid microemulsion shampoo. European Review Med Pharmacol Sci 1999;3(1):11-18.

1. Adnanazeem. Study on emerging role of microemulsion in cosmetics. Recent Patents on Drug Delivery and Formulation 2008;2(3):275.

1. Saboji JK, Manvi FV, Gadad AP, Patel BD. Formulation and evaluation of ketoconazole microsponge gel by quassi solvent diffusion. J Cell Tissue Res 2011;11(1):2691-6.

1. Praveen SP, Vinod MR, Karnakumar VB, Chandrashekhar B P. Development and evaluation of clotrimazole antidandruff hair gel. Int J Res Pharm Chem 2011;1(4):936-44.

1. Shireesh R, Chandra B, Vishnu P, Prasad MVV. Ungual drug deliverysystem of ketoconazole nail lacquer. Int J Appl Pharm 2010;2(4):17-19.

1. Perry R. How shampoos are made. J dermatology 2011;(1):17-19.

1. Ashok K, Rakesh RM . Evaluation of prepared shampoo formulations and to compare formulated with the marketed shampoos. Int J Pharm Sci Res 2010;3(3)120-26.

1. Adnan A, Mohammed R, Farhan J. A, Zeenath I. K , Roop K. K, Mohammad A. Emerging role of microemulsion in cosmetics. Recent patent on Drug Delivery and Formulations 2008;2(3):285-89.

9. / Signature of the candidate / (SHILPA SENAN)
10. / Remarks of the Guide / The work which is assigned to
SHILPA SENANis under my guidance.
11. / 11.1 Name and Designation of the
Guide / Dr. A. R. SHABARAYA M.Pharm., Ph.D.
Principal and Director,
Srinivas College of Pharmacy,
Valachil, Mangalore- 574143.
11.2 Signature
11.3 Name and Designation of the
Co-Guide / ------
11.4 Signature / ------
11.5 Head of the Department / Dr. A. R. SHABARAYA M.Pharm,MBA,PhD.
Principal and Director,
SrinivasCollege of Pharmacy,
Valachil, Mangalore- 574143.
11.6 Signature
12. / 12.1 Remarks of the Principal / Recommended and forwarded for favourable consideration.
12.2 Signature / Dr. A. R. SHABARAYA