THE FORMULATION AND EVALUATION HERBAL SOAP

AGAINST BACTERIAL AND FUNGI SKIN INFECTION USING

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 word cosmetic originated from the Greek word meaning the power, arrange, skills in

decorating (Hughe et al., 1959). But cosmetics, according to the Drugs and Cosmetics Act is

defined as articles intended to be rubbed, poured, sprinkled or sprayed on, introduced into or

otherwise applied to the human body or any part there for cleansing, beautifying, promoting

attractiveness or altering the appearance (Samiksha et al., 2021). Cosmetic do not fall under the

under the review of and approval of process required for drug or medicine, which general need a

drug license. Herbal cosmetics are the preparations containing only plant based chemical

substances from a variety of plant botanical sources, which effect the physiology of the skin and

provide essential substances to nourish the skin or hairs of the body. Herbal cosmetic is the

aromatic used of natural herb and their products in cosmetic (Kapoor., Natural Product

Radiance, p 306-314, Sankholkar, 2009). According he Drug and Cosmetics Act specify that

herbs and essential oils that are used in cosmetics should not penetrate beyond the surface layers

of the skin nor should have any therapeutic effect (Sankholkar, 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

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). Most skin infection are caused by fungi,

staphylococcus aureus and streptococcus species (Kareru, et al., 2010). 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.

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?

vii. Is there any difference in skin compatibility between the herbal soap and commercial

chemical soap?
1.6 General objective

Is 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.

V. To compare the results with chemical soaps.

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

two week.

VII. To observe any difference in skin compatibility between the herbal soap and commercial

chemical soap within two weeks of use.

1.8 Significant 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.

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 Prevalence of skin infection

Skin infection has imposed a significant health threat worldwide and it is characterize by

attacking skin layers and underlying soft tissues (Eposito, Noviello., et al 2016). 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 burden of bacterial skin infection world

wild is of considerable importance, studies done in 2019 show that the standardized age

incidence of bacterial infection was approxiametly 169.72 million cases world wild with the

highest bacterial skin infection recorded in sub – Saharan Africa and the lowest burden recorded

in high – middle sociodemographic index aera (Zhou, Bao et al., 20222). 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). Dermatophytes are among the most prevalent infection-causatives in the world

(Bakheshwain et al., 2011) and millions of dollars are spend annually in their treatment (Brooks

et al., 2004). They can be persistent and worrisome though not debilitating or life-threatening.

Skin diseases however, constitute a significant problem all over the world (Van Hees and Naafs,

2001). Studies have shown that skin infection is a global phenomenon (Enemuor and Amedu,

2009). Tinea versicolor is a relatively common skin infection caused by the fungus

Pityrosporum ovale and associated with pigmentary changes on the skin. Epidemophyton

flocussum and Tinea corporis infect the skin and nails (Sanuth and Efuntoye, 2010).
2.2 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 commonly found in the

respiratory tract and on the skin (Masalha et al., 2001).

2.2.1 Classification of S. aureus

Kingdom: Bacteria

Phylum: Firmicutes

Class: Bacilli

Order: Bacillales

Family: Staphylococcaceae

Genus: Staphylococcus

Species: Aureus

Scientific name: Staphylococcus aureus

2.2.2 Localize S. aureus skin infections

The manifestation most skin infection is associate with S. aureus which produces four main toxin

that interact with the skin which are Panton Valentine leucocidin exfoliatins, enterotoxin and

toxic shock syndrome toxin 1 which causes either primary and secondary localize skin

infections. Cutaneous skin infection occur without any previous appearance of a lesion examples

of such infections are impetigo, folliculitis, furuncles and primary abscesses while secondary

skin infection occur as result of an already existing cutaneous lesion which include
impetiginization, secondary abscesses, lymphangistis, cellulitis and secondary wound infection

(Pascal, 2020).

Impetigo is a highly is highly contagious superficial infection cause by Staphylococcus aureus

or Streptococcal pyogenes common in warm and humid climates affecting keratin cells of the

epidermis which is characterized by honey-colored crusts and small blister (Stanley et al., 2006).

Staphylococcus aureus causes impetigo by locally producing exfoliatins A and B toxins which

target a protein called desmoglein , this protein play a crucial role in keeping the skin cell

together. When these toxins interfere with desmoglien 1, they disrupt the cohesion between

keratinocytes, causing the layers of the skin to separate and form blisters known as bullae

(Stanley et al., 2006, Hanakawa et al., 2002, Amagai et al., 2002)

Folliculitis is the inflammation of hair follicles mostly cause by Staphylococcus aureus affecting area of
the body with high hair density

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

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