TO PREPARE AND SUBMIT ANTI-FUNGAL CREAM

Project Report
Submitted in partial fulfillment of the for
the Degree of

BACHELOR OF PHARMACY

Submitted By
SANGAM KUMAR
Reg. No.190730050

Under the supervision of

Dr. Tanushka
Assistant Professor

Co-Supervisor
Mr. Shiv Kumar
Assistant Professor

Department of Pharmaceutical Sciences and Technology

Maharaja Ranjit Singh Punjab Technical University, Bathinda
June 2023
 DEPARTMENT OF PHARMACEUTICAL SCIENCES & TECHNOLOGY
MAHARAJA RANJIT SINGH PUNJAB TECHNICAL UNIVERSITY
Badal Road, Bathinda -151001
(Established by Govt. of Punjab vide Punjab Act No. 5 of 2015)

Ref. No. – MRSPTU / PHARM / 2023 / Date….....................

CERTIFICATE

This is to certify that the work reported in the project report entitled “To Prepare and submit
Anti-Fungal Cream has been carried out by Mr. Sangam Kumar (190730050) under my
supervision at Department of Pharmaceutical Sciences and Technology, Maharaja Ranjit
Singh Punjab Technical University, Bathinda, Punjab, India during the academic session
2022-2023. The work reported in this report has not been submitted for the award of any other
degree. He has attended stipulated number of days as per the prevailing rules of Maharaja Ranjit
Singh Punjab Technical University.

Dr. Tanushka Mr. Shiv Kumar

Supervisor Co-supervisor
Assistant Professor Assistant Professor
Department of Pharmaceutical Sciences and Department of Pharmaceutical Sciences
Technology and Technology
Maharaja Ranjit Singh Punjab Technical University, Maharaja Ranjit Singh Punjab Technical
Bathinda University, Bathinda

Prof. (Dr.) Amit Bhatia

Head of Department
Department of Pharmaceutical Sciences
and Technology
Maharaja Ranjit Singh Punjab Technical University,
Bathinda
 DECLARATION

I hereby declare that the work reported in the project report entitled “To Prepare and
submit Anti-Fungal Cream” has been carried out by me under the Co-supervision of Dr.
Tanushka at Department of Pharmaceutical Sciences and Technology, Maharaja Ranjit
Singh Punjab Technical University, Bathinda, Punjab, India during the academic session
2022-2023. The work reported in the project report has not been submitted for the award of
any other degree.

Date: …............ Sangam Kumar

Place: Bathinda Reg. No- 190730050
 DECLARATION

I hereby declare that the work reported in the project report entitled “To Prepare and
submit Anti-Fungal Cream” has been carried out by me under the Co-supervision of Mr.
Shiv Kumar at Department of Pharmaceutical Sciences and Technology, Maharaja Ranjit
Singh Punjab Technical University, Bathinda, Punjab, India during the academic session
2022-2023. The work reported in the project report has not been submitted for the award of
any other degree.

Date:............. Sangam Kumar

Place: Bathinda Reg. No- 190730050
 Acknowledgement

First and foremost, I would like to thank God, the Almighty, who has granted countless
blessings, knowledge, and opportunity to me, so that I have been finally able to accomplish
the Project Report. I am also thankful to the people who inspired and guided me all way
through.
I would like to pay my special regards to my esteemed Supervisor Dr. Tanushka,
Department of Pharmaceutical Sciences and Technology, Maharaja Ranjit Singh Punjab
Technical University, Bathinda, for his invaluable supervision and support throughout my
research project. The achievements and successes that I have managed to accumulate were
made possible by the patient guidance, encouragement and invaluable advice that my guide
provided me throughout my work. It has been my proud privilege to work under the guidance
of such a dynamic and inspiring personality. I am thankful to God for providing me with an
ideal mentor who deserves a special honor.
I would like to express my gratitude and appreciation to Prof. (Dr.) Amit Bhatia, Head of
Department of Pharmaceutical Sciences & Technology, Maharaja Ranjit Singh Punjab
Technical University, Bathinda, Punjab, for proving me constant motivation and excellent
infrastructure to accomplish this research endeavor successfully.
My sincere thanks to Prof. (Dr.) Ashish Baldi and Prof. (Dr.) Rahul Deshmukh for their
helpful suggestion and inspiring advice.
Words fail me to express my appreciation to Mr. Shiv Kumar, Co-Supervisor, at MRSPTU,
Bathinda for his illuminating guidance and support in this project.
My deep and sincere gratitude to my family for their continuous and unparallel love, help and
support. I am grateful to my father, Mr. Dhananjay Kumar for giving me the life I ever
dreamed and my mother Mrs. Babita Devi for always making me her priority. I am forever
indebted to my parents for giving me the opportunities and experiences that have made me
who I am. I am so lucky to have a family like you.
I would like to thanks my respected teachers Mr. Subham Singh, Mr. Yogesh Garg, Mr.
Mohit Kumar, Mr. Ram Kumar, Ms. Nitasha, Ms. Divya.
I would like to thanks Mr. Jagdev Singh for helping me in my office work during my B.
Pharm.
I would like to thanks my friends Mr. Dilip Kumar, Mr. Harsh Raj, Mr. Rahul Yadav,
Mr. Mithun Minhas, Mr. Angad Gupta, Ms. Sneha, and my classmates for their support
and encouragement throughout the tenure my project report.
I also thankful non-teaching staff Mr. Sukhpreet, Mr. Param, Mr. Mukesh and Mr.
Gurmail for helping me and making my working environment comfortable.
Last but not least, I would like to thanks the entire person who helped and support me to
fulfillment of an objective. Thanks for all your encouragement.

Sangam Kumar
 Table of Content
Page No.
Sr. No. Topic

1. List of Figure I
2. List of Table I
3. Abstract II
4. Introduction 1-4
5. Literature Review 5-11
6. Aim and Objective 12
7. Plan of Work 13
8. Materials and Methods 14-16
9. Result 17-18
10. Conclusion 19
11. Bibliography 20-21
 List of Figure

Sr. No. Figure Page No.

1. Fungal infection 1

2. Trial of anti-fungal cream 14

List of Table
Sr. No. Table Page No.
1. Composition of Antifungal cream 14

I
Abstract
Fungal infections of the skin are one of the often faced with dermatological diseases in
worldwide. Topical therapy is an attractive choice for the treatment of the cutaneous
infections due to its advantageous such as targeting of drugs to the site of infection and
reduction of the risk of systemic side effects. Currently, antifungal drugs are generally
used as conventional cream and gel preparations in topical treatment. The efficiency of
that treatment depends on the penetration of drugs through the target layers of the skin at
the effective concentrations. However, stratum corneum, the outermost layer of the skin,
is an effective barrier for penetration of drugs into deeper layers of the skin. The
physicochemical characteristics of drug molecules and the types of the formulations are
effective factors in topical drug delivery. Therefore, a number of formulation strategies
have been investigated for delivering antifungal compounds through targeted site of the
skin.

II
 INTRODUCTION 2023

1. Introduction

The concept of beauty and cosmetics is as ancient as mankind and civilization. Indian herbs
and its significance are popular worldwide. An herbal cosmetic has growing demandin the
world market and is an invaluable gift of nature. Herbal formulations always have attracted
considerable attention because of their good activity and comparatively lesser or nil side.
Fungal infections are infections caused by a fungus, a type of microorganism. Two
commoncauses of fungal infections are a fungus called tinea and yeast infections caused by
the fungusCandida albicans.

Fig.1: Fungal infection

Herbal cosmetics are defined as the beauty products which possess desirable physiological
activity such as healing, smoothing appearance, enhancing and conditioning properties
because of herbal ingredient. Now-a-days the usefulness of herbs in the cosmeceutical
production has been extensively increased in personal care system and there is a great
demand for the herbal cosmetics.
Cosmetics are the substances intended to be applied to the human body for cleansing,
beautifying, promoting attractiveness, and altering the appearance without affecting the
body's structure or functions. But the usage of synthetic products becomes very harmful from
long time for the youth as well as our environment. Various synthetic compounds, chemicals,
dye, and their derivative proved to cause various skin diseases having numerousside effects.
Thus, we are using herbal cosmetics as much as possible.
The basic idea of skin care cosmetic lies deep in the Rig-Veda, Yajurveda, Ayurveda, Unani
and Homeopathic system of medicine. These are the products in which herbs are used in
crude or extract form.

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 INTRODUCTION 2023

These herbs should have varieties of properties like antioxidant, anti- inflammatory,
antiseptic, emollient, anti - seborrheic, antikerolytic activity and antibacterial etc. The
medicinal plant find application in pharmaceutical, cosmetic, agricultural and food
industry. The use of medicinal herb for curing disease has been documented in historyof all
civilization. Man in the prehistoric era was probably not aware about the health hazard
associated with the irrational therapy. With the beginning of medical research, it was
discovered that plants contain active ingredients that are accountable. Herbs are used for
their medicinal properties. Cosmetics are developed to reduce wrinkles, fight acne and to
control oil secretion. For various types of skin ailments formulations like skin protective,
sunscreen, antiacne, antiwrinkle and antiaging are designed using varieties of materials,
either natural or synthetic. Cream is a polyherbal formulation that consists of Tulsi oil. That
herbs have beenselected on the basis of a traditional system and scientific justification with
modern uses. An herbal cream that can give effective protection to skin and free from any
toxicity or toxic residue or any irritation when regularly used and should also be
cosmetically acceptable. Herbal medicine is one of the oldest and most universal system of
health care system. The advancement in the field of herbal drug delivery started recently
with the aim tomanage human diseases efficiently. World Health Organization (WHO)
estimates that 80%of the world populations presently use herbal medicine for primary health
care. Every nation is seeking health care beyond the traditional boundaries of modern
medicine; turning to self- medication in the form of herbal remedies. Modern herbal
medicine is based upon the combination of traditional knowledge, clinical experience,
understanding of medicinal science and scientific evidence of herbal medicine. People are
slowly and gradually switching to alternative forms of medicine.
Advantages of herbal system of medicines

• Lower risk of side effects

• Widespread availability

• Effectives with chronic medicine

• Low cost effectiveness makes them all the more alluring

• Natural detoxification process of the body is effectively enhanced by herbal

medicine

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 INTRODUCTION 2023
Disadvantages of herbal system of medicines
• Bulk dosing.
• Poor stability in higher acidic pH, liver metabolism etc.
• Large molecular size limiting the absorption via passive diffusion.
• High amount of raw material is required for processing the medicine.
• Isolation and purification of individual components from whole herbal extract lead
to partial or total loss of therapeutic activity.

Some very common types of fungal infections caused by tinea include

Athlete’s foot

Jock itch

Ringworm

Symptoms

Itching of the feet

Scaling and flaking of the skin of the feet

Itching of the groin area

Red, scaly rash in the groin area

Red, itchy area on the scalp, often in the shape of a ring

Hair loss in the affected area

Lesions or sores that are raised, are yellow-white in color, and appear in patches in
the mouth or throat and/or on the tongue

Sore, bleeding gums

Patches or lesions that become sore, raw and painful, making it difficult to eat or
swallow
Creams are emulsions of either the O/W or W/O type.
Creams are defined as “viscous liquidor semi-solid emulsions of either the oil-in-water or
water-in-oil type. They are semisolids usually consisting of solutions or dispersions of one
or more medicaments in suitable bases. They are formulated using hydrophilic or
hydrophobic bases to provide preparations that are essentially miscible with the skin
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 INTRODUCTION 2023
secretion.
Cream
Pharmaceutical Creams (topical preparation) are homogeneous, semi-solid or viscous
preparations that possess a relatively fluid consistency and are intended for external
application to the skin or certain mucous membranes for protective, therapeutic or
prophylactic purposes especially where an occlusive effect is not necessary.
Ingredients used in skin creams
1. Water
2. Oil, fats and waxes
3. Mineral oil

4. Glyceride oil

5. Waxes
6. Fats
7. Lanolin.
8. Emollients
9. Humectants
A typical manufacturing process breaks down into four individual operations
1. Preparation of the oil phase: - Flake/powder ingredients, sometimes dry blended
in advance, are dispersed into mineral oil or silicone oil. Heating maybe required to
melt someingredients.
2. Hydration of aqueous phase ingredients: - Emulsifiers, thickeners and stabilizers
are dispersed into water in a separate vessel. Heating may be required to accelerate
hydration.
3. Forming the emulsion: -The two phases are blended under vigorous agitation toform
theemulsion.

4. Dispersion of the active ingredient: -The active ingredient often makes up onlya
small proportion of the formulation; this must be efficiently dispersed to maximize
yield and product effectiveness.

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2. Literature Review
Bhaskaran et al. formulated a fluconazole cream for the treatment of Candida albicans.
The optimized cream formulation was prepared using stearic acid, oleic acid, beeswax and
borax.The uniform distribution of the active ingredient fluconazole could be confirmed in
all formulated creams. The FC-C-C formulation showed satisfactory spreadability and
extrudability. FC-C-C delivered (95.07 ± 15.85) % in only 36 h, and the formulation
released the drug by an anomalous diffusion mechanism. The viscosity of FC-C-C was
found to be (63.20 ± 0.83) cP. The antifungal study and animal studies confirmed that the
prepared formulation is non-irritant and has an enhanced antifungal activity that reduces the
side effectsof fluconazole. The studies confirm that the prepared formulation may be useful
for the treatment of Candida albicans [1].
Da Silva Gündel et al. evaluated the antifungal activity of eucalyptus (Eucalyptus
globulus) and lemongrass (Cymbopogon flexuosus) essential oils in their free and
nanoemulsion forms in the murine model of vulvovaginal candidiasis (VVC). The nano-
emulsions were developed by the homogenization method under high agitation and
characterized according to the mean droplet size, polydispersity index, zeta potential, and
pH. The murine VVC model was inducedin BALB/c mice by the administration of estradiol
valerate preceding the Candida albicans challenge. Nanoemulsions containing eucalyptus
or lemongrass oils showed average droplet size of less than 100 nm, a polydispersity index
about 0.2, negative zeta potential, and acidic pH. For in vivo treatment of VVC, the
essential oils in their free form did not show antifungal activity, while the two
nanoemulsions were able to reduce the fungal load similarly or better than the control
animal group treated with miconazole cream. The results obtained in this studydemonstrated
that the use of nanoemulsions containing eucalyptus or lemongrass essential oilscould be a
promising alternative to be used as a proof of concept to develop a new therapy forthe
treatment of VVC [2].
Lamie et al. proposed creams containing non-formulated ITZ or encapsulated in
aspasomes (0.1% or 0.5%) were topically applied in patients with diagnosed diaper
dermatitis complicated by candidiasis, tinea corporis (TC), and tinea versicolor (TVC).
Placebos (void aspasomal cream and cream base) were also utilized. The obtained results
for diaper rash revealed that aspasomal cream (0.5% ITZ) was eminent with respect to
complete cure and negative candida culture after 10-day therapy relative to counterparts
containing 0.1% ITZ aspasomes or non-formulated ITZ (0.1% and 0.5%). For tinea, the
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same trend was manifestedin terms of ‘cleared’ clinical response in 90% of patients and
absence of fungal elements after4-week treatment. Relative to non-formulated ITZ, ITZ
aspasomal cream was endorsed to be auspicious especially when ITZ concentration was
lowered to half commercially available cream concentration (1%), pushing further
exploitation in other dermal fungal infections [3].
Tuchiu, Stefan-van Staden and van Staden studied that antifungal agents are essential
drugs used to treat fungal infections caused by various types of fungi. Due to their
mechanism of action, these drugs bear serious adverse reactions, interact with a wide range
of other drugs, and negatively impact the environment. Therefore, there is a need for
accurate, sensitive, andreliable detection methods to minimize and possibly avoid their
potentially negative effects. Even though so far classical methods have proven to be
effective in detecting these drugs, some of their disadvantages have led the scientific
community to focus its efforts on electrochemical methods, as they are simpler to use, more
sensitive, and require a smaller quantity of sample and minimal sample pretreatment. This
mini-review focuses on electrochemical sensors developed between 2017 and 2022 to
detect and quantify antifungal azoles, highlighting their response characteristics, sensitivity,
and applicability in real samplesanalysis [4].
Bouchand et al. prepared topical voriconazole cream. Systemic use of voriconazole (VCZ)
might be restricted by adverse events, such as hepatotoxicity and neurotoxicity, or drug-
druginteractions. Topical VCZ application to skin may help to treat local infection more
effectivelyand limit unwanted whole-body exposure. Topical VCZ cream was stable for
90days when refrigerated. A patient with cutaneous Fusarium solani infection on his right
forearm was successfully treated with topical 1% VCZ cream after failure of oral VCZ
treatment [5].
Gupta, et al., studied that a variety of oral and topical antifungal agents are available for
thetreatment of superficial fungal infections caused by dermatophytes. This paper builds
onthe antifungal therapy update published in this journal for the first special issue on
Dermatophytosis. Since 2008, there have not been additions to the oral antifungal
armamentarium, with terbinafine, itraconazole, and fluconazole still in widespread use,
albeitfor generally more severe or recalcitrant infections. Griseofulvin is used in the
treatment of tinea capitis. Oral ketoconazole has fallen out of favor in many jurisdictions
due to risks of hepatotoxicity. Topical antifungals, applied once or twice daily, are the
primary treatment for tinea pedis, tinea corporis/tinea cruris, and mild cases of tinea

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unguium. Newer topical antifungal agents introduced include the azoles, efinaconazole,
luliconazole, and sertaconazole, and the oxaborole, tavaborole [6].
Bseiso et al. prepared oral antifungal agents. Fungal infections are amongst the most
commonly encountered diseases affecting the skin. Treatment approaches include both
topical and oral antifungal agents. The topical route is generally preferred due to the possible
side effects of oral medication. Advances in the field of formulation may soon render
outdated conventional products such as creams, ointments and gels. Several carrier systems
loaded with antifungal drugs have demonstrated promising results in the treatment of skin
fungal infections. Examples of these newer carriers include micelles, lipidic systems such
assolid lipid nanoparticles and nanostructured lipid carriers, microemulsions and vesicular
systems such as liposomes, niosomes, transfersomes, ethosomes, and penetration enhancer
vesicles [7].
Marriott et al. studied a new imidazole antifungal agent with broad-spectrum activity. It’s
invitro activity against common dermal pathogens is generally better than miconazole by a
factor of 2–8. This activity is paralleled by good topical efficacy in a guinea pig
dermatomycosis model. Pharmacokinetic studies in animals have demonstrated minimal
systemic exposure following dermal application. Acute general pharmacology studies have
shown that the compound is well tolerated in animals and unlikely to produce side-effects
in man [8].
Spiekermann and Young studied the efficacy and safety of the broad spectrum, topically
applied antifungal agent clotrimazole and evaluated in two double-blind, multicentric
trials. Ten investigators reported on a total of 1,361 cases in which a 1% solution or a 1%
cream formulation was compared with its respective vehicle. Clotrimazole was
therapeutically effective, as confirmed by mycological cure (negative microscopy and
culture) and clinical improvement, in tinea pedis, tinea cruris, tinea corporis, pityriasis
versicolor, and cutaneouscandidiasis. Furthermore, species identification established the
efficacy of clotrimazole against Trichophyton rubrum, T mentagrophytes, Epidermophyton
floccosum, Microsporumcanis, Malassezia furfur (Pityrosporum orbiculare), and Candida
albicans. Safety was demonstrated by the low incidence of possibly drug-related adverse
experiences,namely, 19(2.7%) of 699 patients who were treated with clotrimazole, of whom
four (0.6%)discontinuedtreatment [9].

Espino et al. reported that candida albicans is an opportunistic pathogenic yeast commonly
found in mouth, gastrointestinal tract and vagina. Under certain conditions, it causes skin,
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mucosal and systemic infections. With growing concern over the emergence of resistant
strains to conventional antifungals, the development of novel antifungal agents for the
management of this pathogen is an urgent need. In the present work, novel bioextracts from
folk medicinal plants were directly used as active ingredient in a topical formulation for
dermal candidiasis. With the aim to replace hazardous traditional reagents, a natural solvent
composed by lactic acid: glucose: water (LGH) was used as vehicle for bioactive
compoundextraction. Furthermore, phenolic and alkaloid composition were determined by
HPLC and their individual antifungal effect was evaluated. LGH extracts of Larrea spices
demonstrateasignificant antimicrobial activity against C. albicans being higher than their
individual bioactive constituents. Notably, the mixture of Larrea cuneifolia and L
divaricata extracts intopical formulations reveal a synergistic antifungal effect highlighting
their potential for candidiasis treatment [10].

Wannissorn B. extracted lemon grass oil by steam distillation of wilted leaves of lemon
grass. Lemon grass oil was extracted by steam distillation of wilted leaves of lemon grass
(Cymbopogon citratus (DC.) Stapf.) cultivated in Thailand. The minimum inhibitory
concentration (MIC) and minimum lethal concentration (MLC) of this oil and citral against
35 clinical isolates of 4 dermatophytes (Trichophyton mentagrophytes, T. rubrum,
Epidermophyton floccosum, and Microsporum gypseum) were determined by agar dilution
method. It was found that the MIC and MLC of lemon grass oil were higher than those of
citral. The most resistant strain was M. gypseum followed by T. rubrum, T.
mentagrophytes and E. floccosum, respectively. The mode of action of lemon grass oil and
citral were proven to be fungicidal. The comparative study of the efficacy of cream
containing four different concentrations (1.5%, 2%, 2.5% and 3%) of lemon grass oil was
performed in vitro by hole diffusion assay. The 2.5% lemon grass oil was demonstrated to
be the minimum concentration for preparation of an antifungal cream for subsequent
clinical study [11].
Thomas S P et al. described the use of combination aminoglycoside-steroid-antifungal
creams or ointments. We present the case of a 60-year-old man with a perforated tympanic
membrane who suffered a total hearing loss after the instillation of cream containing
triamcinolone, neomycin, gramicidin and nystatin (Tri-Adcortyl™ cream) into his ear
canal. On balance, we believe that a number of potentially ototoxic constituents in this
cream were responsible. Other possible causes of sensorineural hearing loss and the
possible mechanisms of ototoxicity of this cream are discussed. The reasons why such
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creams may be particularly ototoxic, compared with drops, are also considered. The authors
caution against the use of such creams or ointments in the ear if there is any suspicion of a
tympanic membrane perforation [12].
Tayah Dalal Y. & Eid Ahmad M. studied about Miconazole is a synthetic derivative
of imidazole, a medication with a broad-spectrum antifungal agent that is used to treat
localized vaginal, skin, and nail infections. The aim of the study was to develop an
innovative technique to improve the permeability and efficacy of topical miconazole
nitrate. A nanoemulgel of miconazole nitrate was formulated by the incorporation of
a nanoemulsion and a hydrogel. The nanoemulsion was first optimized using a self-
emulsifying technique, and the drug was then loaded into the optimum formulation and
evaluated prior to mixing with the hydrogel. Miconazole nitrate nanoemulgel formulations
were evaluated for their physical characteristics and antifungal activity. Based on the
results, the formulation with 0.4 % Carbopol showed the highest release profile
(41.8 mg/ml after 2 h); thus, it was chosen as the optimum formulation. A cell diffusion
test was performed to examine the ability of the Miconazole nitrate nanoemulgel to
penetrate the skin and reach the bloodstream. Percentage cumulative drug releases of
29.67 % and 23.79 % after 6 h were achieved for the MNZ nanoemulgel and the
commercial cream, Daktazol, respectively. The antifungal activity of the novel MNZ
nanoemulgel formulation was tested against Candida albicans and compared to Daktazol
cream and almond oil; the results were: 40.9 ± 2.3 mm, 25.4 ± 2.7 mm and 18 ± 1.9 mm,
respectively. In conclusion, a novel MNZ nanoemulgel showing superior antifungal
activity compared to that of the commercial product has been developed. This
nanotechnology technique is a step toward making pharmaceutical dosage forms that has a
lot of promise [13].
Patel Kanu et al. studied that topical Microemulsion based gel drug administration is a
localized[14] drug delivery system anywhere in the body through ophthalmic, rectal,
vaginal and skin as topical routes. Many advantages of gels a major limitation is in the
delivery of hydrophobic drugs. So to overcome this limitation, Microemulsion based
approach is being used so that even a hydrophobic therapeutic moiety can enjoy the unique
properties of gels. Whenever, it is used for fungal disease for topical delivery system so it
is good for compare to oral delivery. When gels and Micro emulsions are used in combined
form the dosage form are referred as Microemulsion based gel. Skin is one of the most
extensive and readily accessible organs on human body for topical administration and is

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main route of topical drug delivery system. It is prepared by mixing an oil-in-water type or
water-in-oil type emulsion with a gelling agent. The use of Micro emulsion-based gels can
be extended in analgesics and antifungal drugs [14].

Spiekermann Paul H. et al. studied about efficacy and safety of the broadspectrum,
topically applied antifungal agent clotrimazole were evaluated in two double-blind,
multicentric trials. Ten investigators reported on a total of 1,361 cases in which a 1%
solution or a 1% cream formulation was compared with its respective vehicle. Clotrimazole
was therapeutically effective, as confirmed by mycological cure (negative microscopy and
culture) and clinical improvement, in tinea pedis, tinea cruris, tinea corporis, pityriasis
versicolor, and cutaneous candidiasis. Furthermore, species identification established the
efficacy of clotrimazole against Trichophyton rubrum, T mentagrophytes, Epidermophyton
floccosum, Microsporum canis, Malassezia furfur (Pityrosporum orbiculare),
and Candida albicans. Safety was demonstrated by the low incidence of possibly drug-
related adverse experiences, namely, 19 (2.7%) of 699 patients who were treated with
clotrimazole, of whom four (0.6%) discontinued treatment [15].

Pai S.T. & Platt M.W. studied about Otomycosis due to saprophytic keratolytic fungi
represents a small percentage of clinical external otitis. Although there are certain
antibacterial and antifungal agents available, they usually are very caustic, potentially
ototoxic and cannot be used if the ear drum is perforated. Garlic is utilized as a folk
medicine in many countries for its antimicrobial and other beneficial properties. In
response to a lack of otic preparations, the authors studied the efficacy of garlic extracts
against the fungi belonging to the genus Aspergillus which are the most common cause of
this infection. Aqueous garlic extract (AGE) and concentrated garlic oil (CGO) along with
various commercial garlic supplements and pharmaceutical prescriptions were used in
an in-vitro study. AGE and especially CGO were found to have antifungal activity. These
agents showed similar or better inhibitory effects than the pharmaceutical preparations and
demonstrated similar minimum inhibitory concentrations [16].
Adam Konstantia et al. studied about the essential oils of Origanum
vulgare subsp. hirtum, Mentha spicata, Lavandula angustifolia, and Salvia fruticosa exhibited
antifungal properties against the human pathogens Malassezia furfur, Trichophyton rubrum,
and Trichosporon beigelii. Of the four oils, O. vulgare subsp. hirtum oil showed the highest
fungicidal activity and at a dilution of 1/50000 caused a 95% reduction in the number of
metabolically active cells within 6 h of exposure. Among the main components of the four oils,

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carvacrol and thymol exhibited the highest levels of antifungal activity. The therapeutic efficacy of
the O. vulgare subsp. hirtum essential oil was tested in rats experimentally infected with T.
rubrum and yielded promising results. Furthermore, the above essential oils were tested with the
Ames test and did not exhibit any mutagenic activity [17].
Kazi & Channa Tahseen studied about Candidiasis is a fungal infection caused by
Candida albicans. Allium sativum (garlic) and Curcuma longa (turmeric) have been used
as antifungal agents. The main aim of this study was to identify the effectiveness of these
natural products towards C. albicans and on their pharmacological and toxicity aspects.
Thus, agar disc diffusion method was used to study the antifungal activity of the ethanolic
extracts. Fluconazole served as the positive control while solvent (ethanol) served as the
negative control. Minimum inhibitory concentration (MIC) of the plant extracts were tested
by using two-fold agar dilution method at concentrations ranging from 0.390g/L to 100g/L.
As delivery agents, cream and gel formulations demonstrated good stability test results.
Furthermore, both plants showed synergistic effects. Active compounds of garlic and
turmeric which is allicin and curcumin were observed through Thin Layer Chromatography
(TLC). Moreover, all the formulation resulted in optimum MIC at pH 5.5 and temperature
25.5ºC. Toxicity test using Brine Shrimp Lethality Test (BSLT) showed that ethanolic
extracts of both plants displayed LC50 values at 77.93µg/mL and 31.97µg/mL. Whereas,
LC50 value of synergistic experiments of extract was 10.77µg/mL. Besides, synergistic
cream formulation was most potent against brine shrimp larvae compared to others with
LC50 value of 5.35µg/mL. Synergistic experiment using gel formulation also showed
potency against brine shrimp larvae with LC50 value of 3.58µg/mL. As a conclusion, both
plant extracts and preparations showed significant effectiveness against C. albicans and
potency on shrimps [18].

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 AIM AND OBJECTIVE 2023

3. Aim and Objective

3.1 AIM

To prepared and submit anti-fungal cream

3.2 Objectives

• To screen out the drugs and excipients

• To prepare anti-fungal cream

• To characterized and evaluate the prepare Anti-fungal cream

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 PLAN OF WORK 2023
4. Plan of work

4.1 Pre formulation studies

• Physio chemical property

• Partition coefficient
• pH
• Compatibility
• Solubility
• Morphology

4.2 Formulation of anti-fungal cream

• Levigation method
• Fusion method
• Cold fusion method
• Trituration method

4.3 Evaluation of anti-fungal cream

• Physical appearance
• Residence time
• Spread ability
• Rheology
• Homogeneity
• Dilution test
• Dissolubility test

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 MATERIALS AND METHODS 2023
5. Materials and methods
Formula

Table 1 - Composition of Antifungal cream (10gm)

Percentage Amount
S. No Ingredients Quantity taken
1. API ------- ------
2. White bees wax 20.0 % ------
3. Light Mineral oil 50.0 % ------
4. Borax 0.7 % ------
5. Distilled water 28.8 % ------
6. Perfume & Preservative 0.5 % ------

Methods of Preparation

• Melted white beeswax in a water bath (70°C).

• Added light mineral oil to it. In other heat water do about toe and dissolved in it.
• Mixed the aqueous Phase to the oily phase with constant stirring, till a creamy
emulsion Prepared.
• Added Perfume at about 40°c

Fig.2: Trial of anti-fungal cream

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 MATERIALS AND METHODS 2023
❖ Evaluation Parameters
• Physical appearance:The Physical appearance of the cream was judged by its color,
pearlscence, roughness, and graded.
• Irritancy test: Mark an area (1 sq. cm) on the left-hand dorsal surface. The cream was
applied to the specified area and time was noted. Irritancy, erythema, edema, was
checked if any for regular intervals up to 24 hrs and reported.
• Homogeneity: The formulation was tested for homogeneity by visual appearance and
by touch.
• Washability: Washability test was carried out by applying a small amount of cream on
the hand and then washing it with tap water. All three formulations were easily washable.
• pH measurement: The pH meter was calibrated using standard buffer solution. About
0.5 gm of cream was weighed and dissolved in 50ml of distilled water and its pH was
measured using digital pH meter.
• Determination of spread ability: Spread ability may be expressed by the extent of the area
to which the topical application spreads when applied to the affected parts on the skin. The
therapeutic efficiency of the formulation also depends upon its spreading value. Sample (about
2gm) was applied in between two glass slides and they were pressed together to obtain a film of
uniform thickness by placing 1000 gm weight for 5 minutes. There after a weight 10gm was
added to the panand the top plate was subjected to pull with the help of string attached to the
hook. The time in which the upper glass slide moves over the lower plate to cover a distance of
10cm is noted. The spread ability (S) can be calculated using the formula

S= m x L/T
Were,
S – Spreadability
m- weight tied to upper glass slide- length moved on a glass slide
t- time taken
The determination was carried out in triplicate and average of three readings was recorded.
• Viscosity: Viscosity of cream was done by using Brooke field viscometer at a
temperature of 25 ℃ using spindle No. 63 at 2.5 rpm. According to the results all the
three formulations showed adequate viscosity.

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 MATERIALS AND METHODS 2023
• Dye test: The scarlet red dye is mixed with the cream. Place a drop of the cream on a
microscopic slide then covers it with a cover slip, and examines it under a microscope.
If the disperse globules appear red the ground colorless. The cream is o/w type. The
reverse condition occurs in w/o type cream i.e. the disperse globules appear colorless.
• Test for microbial growth: Agar media was prepared then the formulated cream was
inoculated on the plate’s agar media by steak Plate method and a controlled is prepared
by omitting the cream. The plates were placed in the incubator and are incubated in 37
C for 24 hours. After the incubation period, the plates were taken out and the microbial
growth were checked and compared with the control.
• Stability testing: The stability study was planned to check the stability of the developed
formulation in 25 ºC and 75 % relative humidity. The formulation would be kept in the
stability chamber for 3 months and its pH, would be determined for one months.

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 RESULT 2023

5. RESULT
The prepared formulation was evaluated for physical appearance, homogeneity, stability, pH,
spreadability, viscosity, in vitro permeability, and skin irritation.

S. no Parameters Observation

1 Appearance White

2 PH 5.7
3 Homogeneity Uniform &Smooth

4 Speardability Easily spreadable

5 Irritancy No irritation

6 Washability Good

8 Viscosity Brookfield
Viscometer
9 Dye test Oil-in-Water

• Appearance: The cream prepared was found to be of a yellowish green color. Creams
showed consistent texture with good homogeneity and without lumps. Therefore, the
formulated creams showed good topical applicability.
• pH: The pH of cream was found to be 5.7, which is acidic value. which were acceptable
for skin formulations (pH ranging from 4 to 6).
• Homogeneity: It was found that the cream was homogeneous and smooth and consistent
in nature.
• Spreadibility: It was found that the cream was easily spreadable and moisturizes the skin
surface of human volunteer.
• Type of smear: It was found that the cream produced non-greasy film on the skin surface.
• Emollience: After observation, it was found that cream not left residue on skin surface
after application.
• Skin irritant: The formulated cold creams were non-irritant and did not cause adverse
effects on the applied area after 72 h of the study, indicating that they were safe for the
skin.
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 RESULT 2023

• Viscosity: Viscosity refers to the stickiness or thickness of a semisolid cream and was the
most important quality control parameter of the formulated creams. The tested cream
showed gradual changes in viscosity with an increase in rpm.
• Stability testing: Accelerated stability testing of prepared formulations was conducted for
2 most stable formulations at room temp, studied for 7 days. No change in colour and
phase separation was observed.

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 CONCLUSION 2023

Conclusion
The cold cream was to be prepared by simple methods and less equipment’s were required.
The prepared cold cream having the best emollient properties and passed all the evaluation
parameters of cream.

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 BIBLIOGRAPHY 2023

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