THE FORMULATION AND EVALUATION OF HERBAL SOAP

AGAINST BACTERIAL AND FUNGAL SKIN INFECTION USING

CURCUMA LONGA, OCIMUM TENUIFLORUM AND AZADIRACHTA
INDICA AS THE MAIN ACTIVE INGREDIENT

By
BIKOBO DESTAING VALERY
HS23BPT018
A Research Project Submitted to The Department of Pharmaceutical
Studies, School of Health Sciences of Biaka University Institute of Buea
For the Partial Fulfilment of The Requirements for An Award of The
Bachelor in Pharmacy Technology.

Supervisor:
DR. SIMON AKO
 CHAPTER ONE
1.1 Introduction
The skin is the largest organ in the body that act as the first line of defense against pathogenic

agents with underlying tissue (Dryden, 2009). Skin and soft tissue infections are very common

withing the population due to constant exposure of the skin to pathogenic agents (Dryden, 2009).

Studies have shown that skin infection cause by bacterial, viral, and fungal ranges from 42 to

65% of the overall skin illness (Enfeksiyonlari., et al 2015)). The most bacterial skin infections

is cause by Staphylococcus aureus which is commonly found on the skin particularly in moist

areas such as the anterior nares (nose), axilla and groin and mucous membrane and causes skin

infections such as cellulitis, folliculitis, boils and abscesses (Debra, louisa et al., 2023). Tinea

versicolor is a dermatophyte that is cause by fungus known as Pityrosporum ovale causes skin

infection resulting in the change of the pigmentation of skin (Sanuth and Efuntoye, 2010).

Trichophyton rubrum is another dermatophytic fungus in the phylum Ascomycota that causes

athletes-foot, fungal infection of the nail, jock itch and ring worm (Zaugg et al., 2009)

Since ancient civilization soap has been one of most important and the oldest chemical product

of removing dirt and eliminating pathogenic agents on the skin. Soap is a chemical product made

by the chemical process of saponification of an alkali salt such as wood ash or strong alkali

solution with a long chain fatty acid (Grace X. F, Sowmya K et al, 2015). The most common

used fat or oils for production of soap through saponification reactions are animal tallow,

coconut oil, palm oil, kernel oil and linseed oil. (Kubmarawa, 2000). Similarly, potassium and

sodium hydroxides are widely used as the caustic alkaline for the purpose. (Eromosele, 1997).

Herbal soap preparation is a medicine or drugs containing phytochemical from different

botanical sources and has antibacterial and antifungal agents which involve the uses of plants
parts such as like leaves, stem, roots and fruits for the treatment of injuries or disease or to

achieve good health (Kareru, et al., 2010). This preparation possess antimicrobial property are

administered topically and available to apply in various forms like creams, lotion, gel, soap,

solvent extract or ointment the variety of creams and soap properties have been used to treat

various skin disorder (Bandyopadhyay et al., 2004). Herbal skin care formulation can be made

from collection of various herbal plant part such as leaves, roots, bark, fruits seed and flowers

which are topically applied as antimicrobial and anti-inflammatory agents in treating skin disease

such as eczemas, ringworm and pruritus (Sharma, et al., 2014).

Antibacterial agents and antifungal agents play crucial roles in combating infection caused by

bacterial and fungi, these agents are used to target specific pathogens and prevent their growth or

eliminate them (Andrea et al., 2019). The incorporation of antibacterial and antifungal agent into

soap products for the treatment of skin infection has been a common practice. However, several

challenges have emerged over time such as the development of microbial resistance by bacterial

and fungi to these agents. This resistance reduces the effectiveness of treatment (Deborah A et

al., 2017, Paloma et al., 2023). The distinctive characteristics of soap production for both

industrial and domestic uses depend specifically on the type of oil which give the product

specific criterial like aroma, clarity, color, low moisture content, absence of fat and rancid odor

(Okoye et al., 1999, Manji et al., 2013). The quality soap is achieved by reacting different

concentration of oil or fat with lye and the quality of the soap depends on the hardness, the

ability of the soap to penetrate and remove dirt, conditional lathering potential and the antiseptic

nature of the soap (Manji et al., 2013). Majority of the commercial soap today sold in the market

are highly incorporated with harmful chemical agent such as aluminum, barium, bis phenols,

plastics formaldehyde, triclosan, sodium lauryl sulfate, dioxane, fragrance, cocamidopropyl

betaine, butylated hydroxy anisole and butylated hydroxytoluene, triethanolamine and

methyllisothiazoline and methylchloroisothiaoline which are absorbed into the body via the

internal lungs from vaporization of the chemicals as well as skin absorption with negative side

effects on the health of individuals (Aiello et al ., 2007). Synthetic antiseptic soap is expensive

and unaffordable especially in developing countries, herbal soap or herbal cosmetics provide an

affordable and cheap with comparative health and safety benefits (Joshi and Pawal, 2015;

Sharma et al., 2008).

Herbal cosmetic has many potential health benefits such as antioxidant, anticancer and

antimicrobial properties that manages various skin and hair conditions. The presence of

phytochemicals such as vitamins, proteins, terpenoids and other bioactive ingredients rejuvenate,

freshen and protect the hair and skin from various conditions such as psoriasis, eczema, skin

dryness, skin cancers, sun burn, skin dryness, boil, candidiasis, athletes’ foot, chicken pox solar

keratosis, dermatitis, impetigo and others (Fathima et al., 20011, Kapoor, 205; Joshi and Pawal,

2005). Neem, turmeric and tulsi are all-natural plant ingredients in herbal soap, and this

combination has antibacterial, antifungal, and anti-inflammatory properties (Sankholkar et al.,

2009).

The aim of this work is to conduct an experimental study on the herbal analysis and evaluation of

Curcuma longa, Ocimum tenuiflorum and Azadirachta indica against S. aureus, Trichophyton

rubrum and Tinea versicolor and the herbal formulation of medicated soap against skin infection

cause by fungi and bacterial.

1.2 Problem statement

Synthetic antiseptic soap is more expensive making it unaffordable for consumers seeking

effective skincare solution and the microbial resistance of chemicals substances used in soap,

along with the potential use of harmful chemical, posses significant risk to public health and the

environment. This issue not only impacts the effectiveness of soap against resistance

microorganisms but also raises concern about the negative consequences of harmful chemicals

on human health and the ecosystem. Addressing these challenges requires innovative approaches

to develop soap formulations that are both effective against microbial resistance and free from

harmful chemical, ensuring the well being of consumers and the environment while maintaining

cost effectiveness.

1.3 RATIONAL

The high cost of medicinal soap used for specific skin conditions, cosmetic or therapeutic

purposes has act as barrier for low income individuals from accessing essential skincare products

and the scarcity of affordable alternatives of medicinal soap for specific skin conditions has

worsen skin conditions and an increase in health care expenses. challenges have emerged over

time such as the development of microbial resistance by bacterial and fungi to these agent, this

resistance reduces the effectiveness of treatment and many commercial medical soap contain a

high amount of synthetic chemical substance and additives such parabens, sulfate and artificial

fragrance causes harm to human health and the aquatic ecosystems thus use of herbal soap as an

alternative could be a potential benefit in the treatment of skin infection since it is relatively

cheap, has little or no adverse effect and ecofriendly and more effective against skin infections.
 1.4 Hypothesis

I. Extract from Curcuma longa, Ocimum tenuiflorum and Azadirachta indica will produce

antifungal and antibacterial effects that meets both the Encyclopedia of Industrial Chemical

standards and KEBs standards respectively.

II. The antibacterial and antifungal soap produced from Curcuma longa, Ocimum tenuiflorum

and Azadirachta indica leaf extract will cure diseases such as ring worm, boils, wounds,

pityriasis and even pimples.

1.5 Research question

i. What are the physicochemical (PH, moisture content, odor, shape, dirt dispersion, wetting

time, foaming forming ability, foaming stability and color) properties of the formulated soap?

ii. What are the phytochemicals present in Curcuma longa, Ocimum tenuiflorum and

Azadirachta indica?

iii. How stable is the herbal soap under varying storage conditions of two weeks (temperature,

humidity)?

iv. How effective is the herbal soap formulation in inhibiting the growth of S. aureus,

Trichophyton rubrum and Tinea versicolor compare to conventional chemical soap?

v. What are the minimum inhibitory concentrations (MICS) of the herbal ingredients against S.

aureus, Trichophyton rubrum and Tinea versicolor?

vi. Does the herbal soap cause any adverse reaction or allergies in human subject during a

prolong use of two weeks?

1.6 General objective

To formulate herbal soap and evaluate the effect of Curcuma longa, Ocimum tenuiflorum and

Azadirachta indica against S. aureus, Trichophyton rubrum and Tinea versicolor.

1.7 Specific objectives

I. To evaluate the physicochemical properties of the soap formulation.

II. To evaluate the phytochemicals constituent in Curcuma longa, Ocimum tenuiflorum and

Azadirachta indica.

III. To monitor changes during storage under different condition within a period of two

weeks.

IV. To conduct in vitro antifungal and antibacterial assay to determine the inhibitory effect of

the herbal soap on S. aureus, Trichophyton rubrum and Tinea versicolor and to compare

the results with chemical soaps.

V. To observe any adverse reaction or allergies in human subject during a prolong use of

two week.

1.8 Si gnificant of the study

 This research contributes to the development of natural and effective alternatives to synthetic

antimicrobial soaps.

 The findings may lead to the production of safe and eco – friendly herbal soap for health skin

which are more affordable and easily assessable in nature.

 This finding will greatly impact the public health sector by decreasing the cost of health care

and reliance on chemical - based products which will potential prevent skin infections.
1.9 Limitation of the study

 This research work focuses on in vitro experiment and it may not fully represent real world

conditions.

 The herbal soaps effectiveness may differs based on the skin type of individuals and usage

patterns

 The study did not explore long term effects or chronic use of the soap.

1.10 Definition of terms

Dirty t dispersion:

Wetting time:

Foaming stability:

Moisture content

Ph:

Optimal inhibitory concentration:

Phytochemicals:

Bactericidal:
 CHAPTER TWO
LITERATURE REVIEW

2.1 A review of S. aureus, R. rubrum and T. versicolor

2.1.1 Overview of Staphylococcus aureus.

Staphylococcus aureus is a member of Bacillota gram - positive facultative anaerobe bacteria

which is spherical in shape and is one of the normal body flora often found on the skin, nares and

mucous membrane of healthy individuals (Masalha et al., 2001, Wertheim et al., 2005 ) and

releases alpha cytotoxin hemolysin also known Hla or alpha toxin that bind to metalloprotease

ADAM10 which initiate the breakdown of the epidermis cells of the skin (86) . Staphylococcus

aureus skin infection can be transmitted from one person to another by coming with a

contaminated skin individual and touching contaminated object (Felix, 2022). Staphylococcus

aureus causes serious skin infection ranging from mild to server such folliculitis is the

inflammation of the follicles leading to pustules or red bumps (Laureano et al., 2014), impetigo

which is highly contagious superficial infection impacting the keratin cells of the epidermis (56)

more common in warm and humid climates (64) and it is characterized by honey-colored crusts

and small blister (56), cellulitis is another skin infection cause by staphylococcus aureus

affecting deeper layers of the skin causing redness, warmth and swelling (39), boils a painful

pus-filled lump caused by infected hair follicles (55)

Classification of S. aureus

Kingdom: Bacteria

Phylum: Firmicutes

Class: Bacilli

Order: Bacillales

Family: Staphylococcaceae

Genus: Staphylococcus

Species: Aureus

Scientific name: Staphylococcus aureus

2.1.1 Overview of Trichophyton rubrum

Trichophyton rubrum is an anthropophilic saprotroph which are white and cottony like on their

surfaces although some colony may appear red, yellowish or brownish underside (Kane, Julius,

1997). Trichophyton rubrum colonizes the upper layers of death skin and it is the most common

cause of athletes-foot, fungal infection of the nail, jock itch and ring worm (Zaugg et al., 2009).

It colonizes the skin by secretion more than 20 specific proteases that including exopeptidases

and exopeptidases which allow for the digestion of human keratin, collagen and elastine (kwong-

Chung, Bennett et al., 1992).

Classification of Trichophyton rubrum

Kingdom: Fungi
Division: Ascomycota
Class: Eurotiomycetes
Order: Onygenalles
Family: Arthrodermataceae
Genus: Trichophyton
Species: T. rubrum

2.1.2 Overview of Tinea versicolor

Tinea versicolor is a dimorphic lipophilic fungus that form one of the body normal flora

(Diongue et al., 2018) and interferes with the skin normal pigmentation leading to the formation

of small, discolored patches which are either lighter or darker in color than the surrounding skin

and mostly affect the trunk and shoulder (Mayor clinic staff, 2022).

2.2 Overview of the physicochemical properties of herbal soap

Herbal soap, derived from natural plant based ingredients, have gained popularity due to their

perceived health benefits and environmental friendliness. Understanding the physicochemical

properties of herbal soaps is essential for product development, quality assurance and consumer

satisfaction in this review, key aspect related to physicochemical properties of herbal soaps will

be discuss

2.2.1 PH value of herbal soap formulation

The PH of a soap formulation significantly influences it cleansing efficacy and skin

compatibility. Herbal soap often maintain a slightly acidic PH of around 5.5 which is closer to

the skins natural PH. This Acidity helps preserve the skins protective barrier and prevents

excessive dryness or irritation. Research have investigated the PH levels of herbal soap

formulations using PH meters and found them to be within the desired range of 5.5 to 10 ( Smith,

2019).
2.2.2 Solubility of herbal soap formulation
Solubility affect how well a soap dissolves in water during use. Herbal soap, containing plant

extracts and essential oils, may exhibit varying solubility properties. Studies have shown that

certain herbal ingredients enhance solubility, leading to better lathering and cleansing. Solubility

test involve measuring the weight of soap dissolved in a given volume of water over time (Gupta

and Kumar, 2007). The solubility range of a soap will depend on the it unique composition and

specific active ingredient it contains (Layne, 2019).

2.2.3 Wetting ability of herbal soap formulation

Is the ability of soap molecule to spread evenly across a surface and penetrate into material being

cleaned. Surface tension affect the wetting ability of soap on the skin. Soap with lower surface

tension allows better spreadability and coverage. Herbal soap containing emollients and

surfactant exhibit favorable tension properties(Singh, 2013). Wetting ability of soap sample is

determined by sucking a piece of cotton fabric in given volume of soap solution over a given

period of time (Shreya et al., 2023).

2.2.4 Moisture content of herbal soap formulation

Moisture content is parameter that is used in assessing the self – life of a product. High moisture

content in soap would lead to reaction of excess water with un – saponified fat to give free fatty

acid and glycerol in a process known as hydrolysis of soap upon storage. The moisture content of

soap is determined by weighing a specific mass of the soap sample and drying at a temperature

of 101 degree Celsius in an oven for two hours and repeated untill a constant weight was

reached. The % moisture was callculated using the formular: %moisture = (Cs – Cl /Cs -Cw) x

100. Where Cw is weight of crucible, Cs is the weight of crusible plus sam b ple, Cl is the

weight of the crucible plus sample after floating (Mak mensah and Firempong, 2011).
2.2.5 Determination of Total Alkali
Total alkalinity is the total amount of alkaline present in the soap. They include alkaline

component like hydroxides, sodium (II) oxide, carbonate and bicarbonate. This is a very

important parameter which determine the efficacy of soap in cleaning. Monitoring the total

alkali in soap ensure that the soap is neither too harsh nor too mild on the skin, making it suitable

for it intended use. The standard range for total alkalinity in should be 0.2% to 1.2%.

2.2.6 Determination of Free Caustic Alkali

2.2.7 Determination of Total Fat Matter

2.3 Overview of the phytochemical constituent of C. longa, O. tenuiflorum and

A. indica

2.1Phytochemical constituent of C. longa

Curcuma longa also known as turmeric, has been extensively used for their medicinal properties.

In the context of skin infections, turmeric stands out due to its diverse phytochemical constituent,

mechanism of action and therapeutic applications (Prasad et al., 2011). Turmeric contain several

bioactive compound, with curcumin being the most prominent. Curcumin is a polyphenol with

potential anti-inflammatory, antioxidant and antimicrobial properties, other constituents include

demethoxycurcumin and bisdethoxycurcumin this compound collectively contribute to turmeric

therapeutic effects on the skin health and infections (Aggarwal and Harikumar, 2009). The

phytochemical analysis ethanolic extract of turmeric shows important preliminary phytochemical

constituent such as tannin, saponins, phenolic compound, phytosterols, terpenoids, alkaloids and

flavonoids which has very important pharmacological activity such as antimicrobial,

antimutagenic, anti-inflammatory and antioxidant (Rajesh, Rao et al., 2013).

 FIGURE 1: chemical structure of Curcumin

2.2Overview of Azadirachta indica

Neem is a tree belonging to the family Meliaceae. Is one of the two species found in the genus

Azadirachta indica and it is native to india and Burma, growing in tropic and sub tropic regions.

Which can grow up to 15 to 20mheight. Neem leaves are evergreen but under drought they may

shield their leaves. Taxonomical classification of neem: Rutales (Order), Rutinea (suborder),

Melioideae (Sub family), Meliacaee (Family), Azadirachta (Genus) and indica (Species). (Alok

et al., 201) The most important active ingredient is azadirachtin, which is followed by quercetin,

salannin, sodium nimbinate, nimbolinin, nimbin, nimbidin, and nimbidol. In addition to ascorbic

acid, n-hexacosanol, 6-desacetyl-7-benzoylazadiradione, 17-hydroxyazadiradione, and nimbiol,

leaves also contain the following substances: nimbin, nimbanene, 6-desacetylnimbinene,

nimbandiol, nimbolide, and nimbandiol. The polyphenolic flavonoids quercetin and beta

sitosterol, which are known to have antibacterial and antifungal properties, were extracted from

fresh neem leave[18,19]. Neem is useful against specific fungi that infect humans since the dawn

of time. Several important fungi have been found to be resistant to neem treatments, including

ringworm, which assaults the skin and nails, athlete's foot fungus, which impacts the skin, nails,

feet, fungi that develop in the bronchi, lungs, and mucous membranes. Neem leaf and oil seed

extracts are effective against a variety of fungi, including Candida, Epidermophyton, Microspor,

Trichosporon, and others. Antioxidant, Antiviral activity, Antibacterial activity [20,21,22].

Figure 2

2.3Phytochemical constituent of Ocimum tenuiflorum

Ocimum tenuiflorum also known as tulsi is an erect, much branched sub – shrub 30 – 60 cm tall,

with simple opposite green or purple leaves that are strongly scented and hairy stems. Leaves

have petiole and ovate, up to 5cm long, usually somewhat toothed flowers are purplish in

elongate racemes in close whorls. The scientific name is Ocimum tenuiflorum and it belong to

the (family) Lamiaceae, (genius) Ocimum.The primary chemical components of tulsi are

oleanolic acid, ursolic acid, rosmarinic acid, eugenol, carvacrol, estragole, linalol, and

caryophyllene. These compounds have been used for many years in food goods, cosmetics,

dental treatments, and other products for the mouth and teeth. which is made up of roughly 71%

eugenol and 20% methyl eugenol. Ocimum sanctum fresh leaves and stem extract produced

significant amounts of the phenolic compounds (antioxidants) cirsilineol, circimaritin, Leaves of

Ocimum sanctum contain 0.7% volatile oil. isothymusin, and rosameric acid from

Eugene[33,34]. Ocimum sanctum contains antifungal properties, and the extracts from the leaves

may be helpful for treating dermatophytic infections. Tulsi exibit antifungal action against the

certain pathogenic fungus. The ancient medical practices of Ayurveda, Greek, Roman, Siddha,
and Unani all make extensive use of tulsi leaves. The herb tulsi has also been used to treat

respiratory conditions. Tulsi leaves are frequently used to make Ayurvedic medicine, which is

used to treat a variety of illnesses and ailments.It is quite helpful for chronic fever [35,36].The

whole plant used has; Hypotension, 4 Raw materials used in Soap making

Figure three;

2.4 In-vitro antifungal and antibacterial assay and materials used in soap
making
Studies has shown that herbal soap formulations containing Curcuma longa, Ocimum

tenuiflorum, and Azadirachta indica demonstrate promising antimicrobial effects against S.

aureus, T. rubrum, and Tinea versicolor. The observed zones of inhibition indicate their potential

in managing skin infections.

2.4.1 Antibacterial Activity

A study by Sharma et al. (2018) evaluated a herbal soap containing turmeric, holy basil, and

neem extracts against S. aureus. The soap showed significant inhibition, with a zone of inhibition

of 18 mm. Another study by Patel et al. (2019) investigated a similar herbal soap against S.

aureus and reported a zone of inhibition of 20 mm.

2.4.2 Antifungal Activity

In a study by Gupta et al. (2017), a herbal soap containing these active ingredients was tested

against T. rubrum. The soap exhibited a zone of inhibition of 16 mm. For Tinea versicolor, a

study by Singh et al. (2016) found that the herbal soap formulation showed a zone of inhibition

of 14mm

2.4.3 The raw materials to use in soap making include;

Fats and Oils; All animal fats and vegetable oils intended for soap making should be as free as

possible from unsaponifiable matter of a good color appearance and in sweet, fresh condition

(Simmons and Appleton, 2007). In the past, animal fat was obtained directly from a slaughter

house but nowadays soap makers use fat that has been processed into fatty acid. This eliminates

many impurities and it produces water as a by-product instead of glycerin. Many vegetables fats

including olive oil, coconut oil, palm kernel oil can also be used in the soap making. Each offer

quite different fatty acid content and hence results in soaps of distinct feel. Soap can be made

using only one type of fat or oil by blending animals and vegetable or blending more than one

vegetable oil. Soap made only from vegetable oils lathers easily but does not harden properly

while soap made from animal fats are hard, tends to be grainy and lathers poorly. A mixture of

the two or more types of fats or oils brings out the best qualities of both. Coconut and palm oils

are very good for soap making (Francion and callings, 2002). Some vegetable oils such as palm

kernel oil, jojoba seed oil, cocoa butter, beeswax, animal and others are rich in minerals and

vitamins and are very good when used in bathing soap, for it nourishes the skin and cures many

skin diseases and infections.

The type of alkali metal determines the kind of soap product. The alkali most commonly used is

sodium hydroxide or potassium hydroxide. Potassium-based soap creates a more water-soluble

product than sodium –based and hence it is known as soft soap, which is often liquid while

sodium-based soaps are firm (Shoge, 2011). The soft soap alone or in combination with sodium-

based soap is commonly used in shaving products. Lithium metal can also be used as an alkali to

form lithium soaps which also tend to be hard and are exclusively used in greases. The alkalis to

use in soap making can be from two different sources. These sources are; Lye, caustic soda or

potash purchased from the market. Lye obtained by leaching or washing water through the ashes

of plant.

Other soap ingredients can be used to enhance the color, texture and scent of the soap.

Fragrances and perfumes are added to the soap mixture to cover the odor of dirt and leave behind

the fresh smelling scent. Either essential oils or artificial perfumes can be used abrasives are also

used to enhance the texture of the soap and they include; talc, silica and marble pumice (volcanic

ash). Modern manufacturers color soap to make it more enticing to the consumers as compared

to the past where soap was made without dye hence a dull grey or brown color (Francion and

Callings,2002).

2.4.4 Soap making processes

The cold process method is one of the most common ways to produce soap even at home. This

process gets its name from the general low temperature used to mill the type of the soap. The low

temperature or sufficient temperature is to ensure the liquefaction of the fat used. This soap

making process requires exact measurements of alkali and fat amounts and computing their ratio,

using saponification charts to ensure that the finished product mild and skin friendly.(

Donkor,1997)
In the hot process method, alkali and fat are boiled together at 80-100 degrees until

saponification occurs, which the soap maker can determine by taste or by eye. After

saponification has occurred, the soap is precipitated from the solution by adding salt and the

excess liquid drained off. The hot-soft soap is then spooned into a mold (Mittelbachet al,, 1996)

Hughes, G.R., J.Soc. Cosmet. Chem., 1959, X, 159.

2. Encyclopaedia. Britannica, 14th Edn; 1929.

 3. Kapoor. V.P., Herbal Cosmetics for Skin and Hair Care, Natural Product Radiance, p 306-
314.

4. Harry R.G, In: Modern Cosmeticology, Vol 1(Revision Eds), Wilkinson J.B., Clark.R., Green
E.,Mclaughlin T.P., 1962, Leonard Hill (Books) Ltd, London.

5. Sankholkar. D.S, Current Regulations and Suggested Way Forward, The Pharma Times,
Vol.41, No.8,2009, p 30-31

6. Kareru, P. G., Keriko, J. M., Kenji, G. M., Thiong’o, G. T., Gachanja, A. N., and Mukiira, H. N.
(2010). Antimicrobial activities of skincare preparations from plant extracts. African Journal of
Traditional, Complementary and Alternative Medicines, 7(3).

7. Bandyopadhyay, U., Biswas, K., Sengupta, A., Moitra, P., Dutta, P., Sarkar, D., ... and
Banerjee, R. K. (2004). Clinical studies on the effect of Neem (Azadirachta indica) bark extract on gastric
secretion and gastroduodenal ulcer. Life sciences, 75(24), 2867-2878.

8. Sharma, J., Gairola, S., Sharma, Y. P., and Gaur, R. D. (2014). Ethnomedicinal plants used to
treat skin diseases by Tharu community of district Udham Singh Nagar, Uttarakhand, India. Journal of
ethnopharmacology, 158, 140-206.

9. Kapoor, V. P. (2005). Herbal cosmetics for skin and hair care.4(4). 306-315.

10. Charaka Samhita, Handbook on Ayurveda, Editor, Gabriel Van Loon, 2002-2003 Vol 1.

11. Prashant, L., Kole et al, Cosmetics potential of herbal Extracts, natural Product Radiance,
Vol 4(4), 2005, p 315-321.

12. The Wealth of India: A Dictionary of Indian raw Materials and Industrial products- Raw
materials Series, Publication and Information Directorate, CSIR, New Delhi, Vols I-XI, 1948-1976; Revised
Series IA, 1985; 2B, 1988; 3 Ca-Ci, 1992.

13. Chopra R.N., Nayar S.I., Chopra I.C., Glossary of Indian Medicinal Plants, Publications and
Information Directorate, CSIR, New Delhi, 1956.

14. D’Amelio F.S, Sr, In: Botanicals A Phytocosmetic Desk Reference (Ed. FS D’Amelio, Sr), 1999,
CRC Press, London.

15. Kumar S, Medicinal Plants in Skin Care Director, Central Institute of Medicinal and Aromatic
Plants, Lucknow, 1994.

16. Thakur R.S., Puri, H.S., Hussain, A, In: Major Medicinal Plants of India, 1989, CIMAP,
Lucknow.

17. The British herbal Pharmacopoeia, British Herbal Medicine Association, 1996.

18. Ceres A, The healing power of herbal teas. Thorsons Publishers, London, 1984.

19. Warra, A. A. (2013) Soap making in Nigeria using indigenous technology and raw materials,
African Journal of Pure and Applied Chemistry, 7(4): 139-145
 20. Okeke, S. U. N. (2009) Home economics for schools and colleges, Onitsha: Africana First
publishers Plc Nigeria

21. Adaku, U. and Melody, M. (2013) Soap Production Using Waste Materials of Cassava Peel
and Plantain Peel Ash as an Alternative Active Ingredient, Implication for Entrepreneurship, IOSR Journal
of VLSI and Signal Processing,3(3): 2319 – 4197

22. Antezana, W., Calve, S., Beccaccia, A., Ferrer, P., Blas, C. D., Rebollar, P. G. and Cerisuelo, A.
(2015) Effects of nutrition on digestion efficiency and gaseous emissions from slurry in growing pigs: III.
Influence of varying the dietary level of calcium soap of palm fatty acids distillate with or without orange
pulp supplementation, Animal Feed Science and Technology, 209: 128-136

23. Phanseil, O. N., Dueno, E. and Xianghong, W. Q. (1998) Synthesis of exotic soaps in the
chemistry laboratory, Journal of Chemistry Education, 75(5): 612

24. Kuntom, A., Siew, W. L. and Tan, V. A. (1994) Characterization of Palm acid oil, Journal of
American Oil and Chemical Society, 71: 525-528

25. C. P. Bryan, The Papyrus Ebers, London, 1930, 12, 139, etc.

26. J. R. Partington, Origine and Development of Applied Chemistry, London, 1933, 198.R. vun
Grot, Historische Studion aus dem Phannabologischen Institut, Hallo, 1889, i, 94

27. R. C. Thompson, Assyrian Herbal, London, 1924, 191, 270. Pliny, Natural History, lib. XXVIII,
emp. 51.

28. Panda, H. (2011). Herbal soaps and detergents handbook. NIIR Project Consultancy Services

29. Reddy, Y. R. R., Kumari, C. K., Lokanatha, O., Mamatha, S., and Reddy, C. D. (2013).
Antimicrobial activity of Azadirachta Indica (neem) leaf, bark and seed extracts. Int. J. Res. Phytochem.
Pharmacol, 3(1), 1-4.

30. Joshi, M. G., Kamat, D. V., and Kamat, S. D. (2008). Evaluation of herbal handwash
formulation.7 (5), 413-15.

31. Kumar, K. P., Bhowmik, D., Tripathi, K. K., and Chandira, M. (2010). Traditional Indian Herbal
Plants Tulsi and Its Medicinal Importance. Research Journal of Pharmacognosy and Phytochemistry, 2(2),
93-101

32. Afsar, Z., Khanam, S., and Aamir, S. (2018) Formulation and comparative evaluation of
polyherbal preparations for their disinfectant effects, 1 (1), 54-65.

33. Dhanasekaran, M. (2016) International research journal of pharmacy. 7(2), 31-35

Kane, Julius (1997). Laboratory handbook of dermatophyte: a clinical guide and llaboratory handbook of
dermatophytes and otherfilamentous fungi from sskin, hair, and nails. Belmont, CA: Star Pub. ISBN
9780898631679
Curcumba longa L . plants of th world online, kew science, Kew Gardens, Royal Botanic Gardenss. 2018
Retrived 26 march 2018