Research Article

Formulation of Herbal Soap against

Acne Causing Bacteria
Anjum Attaullah1,*, Aruna Govindarajulu1, Mohana Priya K1, Mubeena KF1, Sreenithi R1, Summera
Rafiq2, Shaik Jasmine Shahina2
1
Department of Biochemistry, Justice Basheer Ahmed Sayeed College for Women, Chennai, Tamil Nadu, INDIA.
2
Department of Microbiology, Justice Basheer Ahmed Sayeed College for Women, Chennai, Tamil Nadu, INDIA.

Submission Date: 07-10-2021; Revision Date: 15-11-2021; Accepted Date: 03-12-2021

Correspondence:
ABSTRACT Dr. Anjum Attaullah,
Department of
The use of herbs on skin disorders has been done for thousands of years. Herbal remedies, Biochemistry, Justice
including those for many kind of disorder especially skin disorders, are currently gaining popularity Basheer Ahmed Sayeed
among patients. In Asia, especially in south east Asian countries, herbal treatments that have College for Women
(Autonomous),
been used from ages are now being studied scientifically. In India, records of Ayurvedic medicine, Chennai-600 018,
a method of medicine with natural roots date back to about 3000 BC. The system of Ayurvedic Tamil Nadu, INDIA.
medicine combines physiological and holistic principles. Propionibacterium acnes, Staphylococcus Phone no:
epidermidis and Staphylococcus aureus in the follicular canal and sebum production are the +919500137319
reason for leading to Acne vulgaris. Inflammation in acne is caused by Staphylococcus aureus. In
Email: anjum.a@jbascol-
the present study, aqueous extract of herbs like Guava leaves, Turmeric, Aloe Vera, Soap lege.edu.in
nuts and Rose petals were used to formulate a herbal soap to combat acne. The preliminary
phytochemical analysis of the aqueous extract of these herbs showed the presence of various
secondary metabolites such as saponins, phenols, tannins, terpenoids, glycosides and quinones.
In vitro antibacterial activity was performed against Staphylococcus aureus and E. coli using agar
well diffusion method. The zone of inhibition with guava leaves was observed to be 17mm, while
11 mm was observed with turmeric for Staphylococcus aureus. While with E. coli, Guava leaves
showed a zone size of 9mm and no zones were observed with Turmeric and Aloe vera extracts.
Although, Aloe vera is known for its antibacterial activity, in our study Aloe vera did not exhibit any
antibacterial activity.
Key words: Herbal soaps, Antibacterial activity, S. aureus, Guava leaves, Turmeric.

INTRODUCTION excess oil and dead sticky cells become trapped in the
pore, creating a blockage called comedo (blackhead). If
The major organ in the body is the human skin, which
the follicle ruptures, the comedo might turn into a more
is the outer layer of the body which is the first line
severe outbreak. Acne vulgaris has been associated
of defense. Skin contains many specialized cells and
with Staphylococcus aureus and Propionibacterium acnes.
structures. Acne vulgaris also known as acne is a chronic Substantial in vitro research shows that S. aureus may
skin disease involving blockage or inflammation in the have a pathophysiological involvement in acne vulgaris.
pilosebaceous gland that occurs when hair follicles are In other research, both S. epidermidis and P. acnes were
clogged with dead skin cells and oil. Worldwide, this identified as bacteria likely to cause acne vulgaris.[1]
has been the most common dermatological conditions Natural plant-based medications are increasing in
affecting approximately about 650 million people. These prominence due to a number of benefits, including
minimal adverse reactions improved patient endurance,
SCAN QR CODE TO VIEW ONLINE
lower cost, and acceptance owing to a historical use. Aside
www.ajbls.com from providing reasonable means for the treatment of
many diseases that are intractable and incurable in other
DOI: 10.5530/ajbls.2021.10.80
systems of medicine, herbal remedies give a reasonable
method for the cure of several ailments which are
difficult and incurable in other medicinal systems. As
608 Asian Journal of Biological and Life Sciences, Vol 10, Issue 3, Sep-Dec, 2021
 Anjum, et al.: Formulation of Herbal Soap against Acne Causing Bacteria

a result, a variety of herbs are being studied for the Soap making using Rose petals
therapy of skin conditions varying from itchiness to The Rose flowers were freshly picked from the flower
skin cancer. bed. Fresh rose petals were detached, spread in a thin
layer and were shade dried for four to five days at the
METHODOLOGY room temperature (25°- 35°C). The dried material
Sample collection was crushed and ground and stored in a tightly closed
container.
Herbal formulation was made using the following herbs
like Guava leaves, Turmeric, Aloe Vera, Soap nuts and Qualitative Phytochemical analysis
Rose petals. Preliminary qualitative phytochemical analysis was
carried out to identify the secondary metabolites present
Preparation of Guava leaves Extract
in the aqueous extracts of Guava leaves, Turmeric, Aloe
1 g of dried guava leaf was extracted with 50 ml of Vera, Soap nuts and Rose petals as per the standard
boiling water at 100°C for 20 min. The obtained guava methods.[5]
leaf extract was filtered using a vacuum pump with
Antibacterial activity
Whatman filter paper and evaporated using rotary
evaporator.[2] The extracts were freshly prepared in sterile distilled
water to a final concentration of 100 µg/ml respectively
Preparation of Turmeric Extract
for agar disc diffusion test.[6] Isolated bacterial species
12 g of turmeric was dissolved in 60 ml of distilled water of Staphylococcus aureus and E. coli were obtained from
and was kept in rotary shaker for about 10 hr and 30 min. the Department of Microbiology, JBAS College for
This extract was concentrated using rotary evaporator Women, Chennai. The antibacterial activity of the
and the extract was stored at 4°C in a refrigerator.[3] extracts of guava leaves, turmeric and Aloe vera was
Preparation of Aloe Vera Extract
carried out using the agar disc diffusion method.

The Aloe Vera leaves were freshly collected and were Preparation of Inoculum
cleaned individually by distilled water. The spikes and Cultures for experiments was prepared by transforming
the margins were removed before slicing the leaves and a loopful of bacterial cell from the stock cultures to test
the cortex was carefully separated from the parenchyma tube containing Muller-Hinton broth and was incubated
using knife. Filets were washed thoroughly with distilled for 2-6 hr at 37°C to match the turbidity with 0.5 Mac
water to remove the exudate from the surfaces. The Farland standard.
fresh filets were then stored in a refrigerator.
Agar Well Diffusion Method
Preparation of Soap Nut Extract Antimicrobial activity was evaluated by using agar well
About 50 g of soap nut powder was taken and was diffusion method. The inoculum was spread evenly on
boiled with 200 ml of distilled water with continuous the Muller-Hinton agar’s (MHA) surface using a sterile
stirring and was filtered. The filtrate was then placed cotton swab and was allowed to dry for 10-15 min.
in the hot air oven at 60°C for drying and was used for Wells of 6mm were cut with the help of sterile well
further analysis.[4] cutter. 100µl of different samples were added in the
cut wells of MHA and the plate was incubated at 37°C
Soap making using soap nuts
for 24hr. After incubation, a clear zone around the
Whole soap nuts were taken and were crushed. well determines the antibacterial activity of the herbs.
Required amount of water was added and was boiled Diameter of the zones of inhibition was measured in
with continuous stirring for 30 min. The extract was millimeters.
allowed to cool and was strained off and stored in a
Double Boiling Method for Soap formulation
bottle. The extract appeared light brown in colour due
to the presence of saponins. The double boiler was placed on the hot plate. The
soap bases were cut into cubes, and were filled up to
Preparation of Rose water Extract 75% volume in container. 1-2% of water was added to
The rose petals were freshly picked and they were boiled compensate the soap base. The top was covered and
in distilled water. It was boiled until the petals have lost allowed to melt with stirring lightly in between. The
their entire colour and the water has gained pink colour. melting temperature was around 75-80°C. Once the
The water was strained and stored in a container. base completely melted, different extracts were added
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 Anjum, et al.: Formulation of Herbal Soap against Acne Causing Bacteria

which included- 10gm of Aloe Vera gel, 20 gm of respect to the human body in preventing diseases and
Guava leaf extract, 10ml of Soap nut extract and 5gm skin infections. The antibacterial activity of the aqueous
of Rose pellet extract. It was then poured into a mould. extract of components namely guava leaves, turmeric
It was allowed to cool at room temperature and then and aloe vera were compared using agar well diffusion
demoulded. method against Staphylococcus aureus and E. coli. The
zone of inhibition with guava leaves was observed to
RESULTS be 17mm, while 11mm was observed with turmeric
for Staphylococcus aureus. While with E. coli, Guava leaves
Qualitative Phytochemical analysis
showed a zone size of 9mm and no zones were observed
Preliminary qualitative phytochemical analysis was with Turmeric and Aloe Vera extracts. Although, Aloe
carried out to identify the secondary metabolites present Vera is known for its antibacterial activity, in our study
in the aqueous extracts of Guava leaves, Turmeric, Aloe Aloe Vera did not exhibit any antibacterial activity
Vera, Soap nuts and Rose petals as per the standard (Table 1).
methods. The results of this study indicate that the aqueous extract
of two different components namely guava leaves and
turmeric possesses anti-bacterial activity. The turmeric
Guava Leaves

Rose Petals
Secondary
metabolite

Turmeric

Soapnut
showed less inhibition when compared to guava leaves.
Alovera
S. No.

Test

Formulation of Soap
Soaps act as emulsifiers or surfactants, softening the
1 Phenols Ferric + _ + + + horny layer of the epidermis and acts as a germicide
and tannins chloride test by enhancing the permeability of microbial envelope
2 Alkaloids Wagner’s ++ + _ _ + thereby disrupting the integrity of microbial cells.
test Antimicrobial activity of soaps makes them useful
3 Saponins Foam test + _ + + ++ agent for bathing, laundry, washing, and cleansing of
surfaces.[7,8] The cleansing and germicidal properties
4 Flavonoids Sodium + + + + _ of the soapy-plants are comparable to those of the
hydroxide
test
standard soaps, which are salts of higher fatty acids.
Crude preparations of soapy plants are able to soften
5 Terpenoids Salkowski + + + + +
test the skin epidermis, enhance greater penetration and
6 Cardiac Keller killani + + + + + cleansing of sores and acne and thereby promote rapid
glycoside test healing and resolution of blemishes.
7 Test for Action of + + + + +
quinones ammonia
Table 1: Antibacterial activity of the herbs.
8 Test for Biuret test + + + _ +
proteins Herbs Staphylococcus aureus E.coli

9 Test for Libermann _ _ + _ _ Guava leaves 17mm 9mm

steroids Burchard Turmeric 11mm No zone
test
Aloe vera No zone No zone

The secondary metabolites were found in abundance in

Guava leaves which showed the presence of phenols,
tannins, alkaloids, saponins, flavonoids, terpenoids,
cardiac glycoside, quinones and proteins. This
supported the antibacterial activity of Guava leaves
when compared to other herbs used in this study. Other
herbs also showed considerable amounts of secondary
metabolites when tested.
Antibacterial Activity
Antimicrobial activity of any substance is defined as
its ability to either kill bacteria or inhibit the growth
of bacteria. Antimicrobial activity is significant with
610 Asian Journal of Biological and Life Sciences, Vol 10, Issue 3, Sep-Dec, 2021
 Anjum, et al.: Formulation of Herbal Soap against Acne Causing Bacteria

In the present study, herbs like Guava leaves, Turmeric,

Aloe Vera, Soap nuts and Rose petals were used to
formulate herbal soap to combat acne. The preliminary
phytochemical analysis of the aqueous extract of the
Guava leaves, Turmeric, Aloe vera, Rose and Soap nut
revealed the presence of various phytochemicals such
as saponins, phenols, tannins, terpenoids, glycosides
and quinones which was found to be in agreement with
Kokate et al., 2003[12] and Harborne, 1998.[13]
The family Myrtaceae includes the genus Psidium,
which is considered to have originated in tropical South
America. The leaves of Guava are 2 to 6 inches long
and 1 to 2 inches wide, appear dull-green with stiff but
coriaceous with pronounced veins and aromatic when
crushed.[14] Guava leaf has bioactive components which
DISCUSSION
can fight against pathogens, manage blood glucose
Acne is a common skin condition, caused by changes levels, and can also help in weight loss. The leaves of
in the pilosebaceous units, skin structures consisting Guava leaves contain essential oils which are rich in
of hair follicle and its associated sebaceous glands via cineol, tannins, triterpenes, flavonoids, resin, eugenol,
androgen stimulation. Basically, acne is a disease whose malic acid, and mineral salts.[15-17]
initial pathology is the microscopic microcomedo. The Turmeric is the processed rhizome of Curcuma longa.
microcomedo may involve into visible open comedones South Asian women have traditionally used turmeric
(Blackheads) or closed comedones (Whiteheads). roots for skin care. Scientists have used inventive
Inflammatory papules, pustules and nodules may technique to create a colourless derivative, tetrahydro
develop in due course of time.[9] The severity of
curcuminoids, which has similar health advantages as
acne varies greatly among individuals and a genetic
yellow turmeric extract. The UV protectant, protein
background plays an important role. It is estimated that
integrity support, tyrosinase inhibitory and antioxidant
as many as 90% of all teenagers are affected by acne,
properties of tetrahydro curcuminoids would work
with virtually 100% of teenagers experiencing some
together in an anti-acne formulation to provide
degree of comedones formation.[10]
multifaceted benefits.[18,19] A famous homemade
Herbs and their products have been highly esteemed
treatment utilized among people is using a mixture of
source of medicine throughout the human history.
turmeric and coconut oil, which is applied to the afflicted
Herbs are widely used today indicating that these are
region as well as to surrounding area and when left left
a growing part of modern high-tech medicine. Some
overnight the size of the acne is reduced. Because of its
of the herbs and herbal treatment which have their
beneficial benefits on the skin, it is a major component
role in the management of acne and other skin related
disorders are compiled here. Number of herbs with a in various cosmetic goods such as creams, soaps, and
history of use in traditional cultures has entered the cleansers.
growing cosmeceutical market. An herbal formulation Aloe Vera is a robust, short-stemmed shrub with water-
which contains many herbal extracts and have negligible storing leaves. It is broadly utilized in the cosmetic,
adverse effects compared with modern medicine, are pharmaceuticals, and food sectors, and its worldwide
commonly indicated for moderate and severe forms of annual market price is expected to be $13 billion.[20] Aloe
acne. Some of the herbs that possess promising role Vera has dense, pointy, succulent green leaves which can
against Acne vulgaris includes: Tephrosia purpurea, Saw grow to be 12-19 inches (30-50 cm) long. Every leaf
palmetto, Rubia cordifolia, Viola tricolor, Aloe vera, Asparagus contains a sticky substance which retains water, causing
officinalis, Calendulla officinalis, Betula alba, Simmondsia the leaves to be dense. The “gel” people connect with
chinensis, Vitex agnus-castus, Coriandrum sativum, Lavandula Aloe Vera products is this slimy, water-filled tissue. This
angustifolia, Citrus aurantium, Citrus limon, Rheum officinale, hydrogel includes majority of the plant’s beneficial
Urtica dioica, Curcuma longa, Saponaria officinalis, Juglans components, such as vitamins, minerals, amino acids,
nigra, Azadirachta indica, Melaleuca alternifolia, Populus and antioxidants. The aloe shrub includes a variety of
candicans, Arctium lappa, Amaranthus hypochondriacus, natural phytochemicals with wellness properties, including
Allium sativum.[11] anthraquinones, vitamins, minerals, polysaccharides,
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 Anjum, et al.: Formulation of Herbal Soap against Acne Causing Bacteria

sterols, amino acids, saponins, salicylic acids, and use different plant-based products for curing problems
others.[21,22] associated with skin. Compared with the conventional
Sapindus mukorossi is a deciduous tree in the sapindaceae allopathic drugs, ayurvedic medicines have relatively low
family which grows in the higher parts of the Indo- cost and can be of great benefit to the Indian people
Gangetic plains, Shivaliks, and sub-Himalayan pathways especially the poor people. Herbal medicines are a rich
at elevations ranging from 200m to 1500m. It is among source of active ingredients and can be safer and cost-
the main significant plants in Asia’s tropical and effective skin infection treatment ranging from rashes to
subtropical areas, and is also referred as soap-nut tree. dreadful skin cancer. Our study showed that the aqueous
The fruit is valued for the saponins (10.1%) present extracts of Guava leaves were found to be very effective
in the pericarp constitutes up to 56.5% of the drupe in killing Staphylococcus aureus and E. coli. Turmeric
known for inhibiting tumour cell growth.[23] Much of this extracts could only inhibit the growth of Staphylococcus
plant’s pharmacological properties have recently been aureus and did not have any effect on E. coli. Aloe Vera
investigated, including antibacterial,[24] cytotoxoic,[25] a well-known plant for its antibacterial activity did not
molluscicidal,[26] insecticidal,[27] and fungicidal properties.[23] show any inhibition with both the bacteria tested.
One of the most talked about activities of this plant
is the contraceptive of the saponins extracted from CONFLICT OF INTEREST
the pericarp of the fruit.[28] Sapindus mukorossi is widely
The authors declare that there are no conflicts of
recognised for its ancient therapeutic properties.[29]
interest.
The occurrence of alkaloids, phytosterols, phenolic
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Cite this article: Anjum A, Aruna G, Priya KM, Mubeena KF, Sreenithi R, Rafiq S, Shahina SKJ. Formulation of Herbal Soap
against Acne Causing Bacteria. Asian J Biol Life Sci. 2021;10(3):608-13.

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