Development and Anti-Microbial Study of Herbal Hand Wash Using Nimba, Tulsi, Sourabhanimba, Kumari

Abstract

Hands are the main route of transmission of germs in healthcare, so hand hygiene is the most important measure to prevent the spread of harmful germs and prevent healthcare-related infections. Many infectious diseases can be transmitted from one person to another through contaminated hands. These diseases include gastrointestinal infections, such as salmonella, and respiratory infections, such as the flu. Proper hand washing can help prevent the spread of germs (such as bacteria and viruses) that cause these diseases. So an attempt is made to prepare herbal hand wash using drugs like nimba, girinimba, tulsi and kumari which are having antibacterial, antiviral, anti-fungal activities and skin hydration properties. Herbal hand wash was prepared as per standard method using herbal ingredients and swab samples streaked on Blood and Mac-conkey agar, incubated at 37°C for 24 hrs in aerobic condition for microbial load assessment study. The results suggest that hand-washing herbs can create excellent suppression zones to protect against skin pathogens. This could be the reason for using herbs in preparation for hand washing and using these compounds in the production of disinfectant lotions or soaps instead of chemicals. INTRODUCTION Food and water-borne diseases combined with nosocomial infections kill millions of people each year. A discussion of hospital acquisition and control of food-related infections and poisoning would be incomplete without including hand hygiene. They describe the means by which surgical procedures are most likely to be free from human pathogens and contact surface contaminants in food handling and consumption, without being complicated by hand-carried contaminants [1]. Hands are the main route of transmission of germs in healthcare, so hand hygiene is the most important measure to prevent the spread of harmful germs and prevent healthcare-related infections. Many infectious diseases can be transmitted from one person to another through contaminated hands. These diseases include gastrointestinal infections, such as salmonella, and respiratory infections, such as the flu. Proper hand washing can help prevent the spread of germs (such as bacteria and viruses) that cause these diseases. Some forms of gastrointestinal and respiratory infections can cause serious complications, especially in children, the elderly, or people with weakened immune systems. Therefore, simple hand washing is still an effective way to prevent infection [2]. Frequent hand washing with soap and water can dry out and crack your skin. This can be uncomfortable and increases the risk of skin infections. Frequent use of chemical handwashing causes obsessive-compulsive disorder (OCD) and dry skin, in extreme cases eczema and psoriasis. There is need of herbal hand wash which will protect ourselves from infection and also avoid the harmful effects of chemical hand wash available in market. So, an attempt is made to prepare herbal hand wash using drugs like nimba, girinimba, tulsi and kumari which are having antibacterial, antiviral, anti-fungal activities and skin hydration properties [3,4].

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BEPLS Vol 12 [3] February 2023 167 | P a g e ©2023 AELS, INDIA
Bulletin of Environment, Pharmacology and Life Sciences
Bull. Env. Pharmacol. Life Sci., Vol 12 [3] February 2023 : 167-170
©2023 Academy for Environment and Life Sciences, India
Online ISSN 2277-1808
Journal’s URL:http://www.bepls.com
CODEN: BEPLAD
ORIGINAL ARTICLE OPEN ACCESS
Development and Anti-Microbial Study of Herbal Hand Wash Using
Nimba, Tulsi, Sourabhanimba, Kumari
Nameera H. A., Priyanshi Goyal, Maanya. M. Lingam, Vinaykumar R Kadibagil, Govinda Sharma.K
Sri Dharmasthala Manjunateshwara College of Ayurveda and Hospital, Hassan-573201
Correspondence Email: drvinaykadibagil@gmail.com
ABSTRACT
Hands are the main route of transmission of germs in healthcare, so hand hygiene is the most important
measure to prevent the spread of harmful germs and prevent healthcare -related infections. Many infectious
diseases can be transmitted from one person to another through contaminated hands. These diseases include
gastrointestinal infections, such as salmonella, and respiratory infections, such as the flu. Proper hand
washing can help prevent the spread of germs (such as bacteria and viruses) that cause these diseases. So an
attempt is made to prepare herbal hand wash using drugs like nimba, girinimba, tulsi and kumari which are
having antibacterial, antiviral, anti-fungal activities and skin hydration properties. Herbal hand wash was
prepared as per standard method using herbal ingredients and swab samples streaked on Blood and Mac-
conkey agar, incubated at 37°C for 24 hrs in aerobic condition for microbial load assessment study. The
results suggest that hand-washing herbs can create excellent suppression zones to protect against skin
pathogens. This could be the reason for using herbs in preparation for hand washing and using these
compounds in the production of disinfectant lotions or soaps instead of chemicals.
Keywords: Herbal hand wash, nimba, tulasi, saurabhanimba, antimicrobial study
Received 02.11.2022 Revised 22.01.2023 Accepted 27.01.2023
INTRODUCTION
Food and water-borne diseases combined with nosocomial infections kill millions of people each year. A
discussion of hospital acquisition and control of food-related infections and poisoning would be
incomplete without including hand hygiene. They describe the means by which surgical procedures are
most likely to be free from human pathogens and contact surface contaminants in food handling and
consumption, without being complicated by hand-carried contaminants [1]. Hands are the main route of
transmission of germs in healthcare, so hand hygiene is the most important measure to prevent the
spread of harmful germs and prevent healthcare-related infections. Many infectious diseases can be
transmitted from one person to another through contaminated hands. These diseases include
gastrointestinal infections, such as salmonella, and respiratory infections, such as the flu. Proper hand
washing can help prevent the spread of germs (such as bacteria and viruses) that cause these diseases.
Some forms of gastrointestinal and respiratory infections can cause serious complications, especially in
children, the elderly, or people with weakened immune systems. Therefore, simple hand washing is still
an effective way to prevent infection [2].
Frequent hand washing with soap and water can dry out and crack your skin. This can be uncomfortable
and increases the risk of skin infections. Frequent use of chemical handwashing causes obsessive-
compulsive disorder (OCD) and dry skin, in extreme cases eczema and psoriasis. There is need of herbal
hand wash which will protect ourselves from infection and also avoid the harmful effects of chemical
hand wash available in market. So, an attempt is made to prepare herbal hand wash using drugs like
nimba, girinimba, tulsi and kumari which are having antibacterial, antiviral, anti-fungal activities and skin
hydration properties [3,4].

Nameera H. A. et al.
BEPLS Vol 12 [3] February 2023 168 | P a g e © 2023 AELS, INDIA
METHODOLOGY
Pharmaceutical study
Preparation of herbal hand wash
Table 1 Ingredients of herbal hand wash
Sr.no.
Ingredients
Latin/English Name
Quantity
1.
NimbaArka
Azadirachta indica
183 ml
2.
TulasiArka
Ocimum sanctum
183 ml
3.
SaurabhanimbaArka
Murraya koenigii
183 ml
4.
Kumari swarasa (Aleo vera)
Aloe barbadensis
100 ml
5.
Lavana
Sodium chloride (Salt)
50 g
6.
Lemon grass essential oil
4 ml
7.
Methyl Paraben
2 g
8.
Sodium Lauryl Ether Sulfate (SLES)
300
First nimba, tulasi and saurabhanimbaarka was prepared. Kumari swarasa was extracted from freshly
collected drugs. To arka and swarasa, other ingredients i.elavana, essential oil, Methyl Paraben and
Sodium Lauryl Ether Sulfate (SLES) were added one by one slowly. Mixture was sti rred continuously for
30 minutes for homogenous mixing of ingredients.
Analytical study
Appearance:
The prepared formulation of hand wash appears as greenish brown.
1) pH:
The pH of formulations was measured by digital pH meter.
2) Viscosity:
The viscosity of hand wash was determined by using Brookfield viscometer. 50ml of herbal hand wash is
taken into 100ml of beaker and the tip of viscometer was dipped into the beaker containing hand wash
formulation and its viscosity was measured.
3) Foam Height:
0.5gm of sample of Herbal handwash was taken and dispersed in 25 ml distilled water. Then, transferred
it into 500 ml stoppered measuring cylinder; volume was making up to 50 ml with water. 25 strokes were
given & stand till aqueous volume measured up to 50ml& measured the foam height; above the aqueous
volume.
4) Foam Retention:
50 ml of the Herbal handwash was taken into a 200ml graduated cylinder & shaken 10 times. The volume
of foam at 1-minute intervals for 4 minutes was recorded [2].
Table 2 Summary of analytical results of herbal hand wash
Sr. no
Test name
Result
1
pH
6.24
2
Viscosity
62 CPS
3
Foam height
300 ml
4
Foam Retention
20 ml
Assessment of Antimicrobial Study
Collection of Samples for evaluation
a. Wet Swab method.
One swab will be collected before any application from both the palms from the same volunteer which
will be marked as before treatment sample. Both palm will be treated with 10ml of herbal hand wash by
Scholar for 30 seconds and then washed with water. One swab will be collected from each palm following
exposure to herbal hand wash in sterile swab cover itself and labeled appropriately.
Preparation of Media
Culturing method: Streak culture method
Requirements:
MacConkey agar plate, blood agar plates, nichrome loop, gas burner, incubator (37oC).
For culturing the bacteria in sputum sample, the culture media was needed to be prepared. Culture media
was prepared by the following methods:
MacConkey Agar preparation
For preparation of 500 ml of MacConkey agar solid media: 17.5 gm of MacConkey agar was weighed and
mixed with 7.5 gm of Agar. Further the powder was dissolved in 500 ml of distilled water and transferred
to a 1000ml conical flask. Mouth of the conical flask was sealed tightly with cotton plug and was
autoclaved at 121oC for 20 minutes.

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BEPLS Vol 12 [3] February 2023 169 | P a g e © 2023 AELS, INDIA
Procedure for culturing of bacteria
The zone of inoculation were marked over the outer lower surface of the sterile MacConkey and blood
agar plates by using a glass marking pencil. The nichrome loop is heated red and was allowed to cool, then
a loop full of bacteria was transferred aseptically to each of the previously marked zone of inoculation on
the MacConkey and blood Agar plates and was streaked perpendicular to the zone of inoculation. The
plates which were streaked in this manner were kept in the incubator at 37°C and observed the results
after 24-48 hours. Further microscopic examination was done for the identification and characterisation
if the bacteria.
Microbial Load Assessment
1ml of each dilution will be freshly pipetted onto a sterile, appropriately labeled Petri dish. Warm media
will be poured over the 1 ml sample and shaken in all directions on the planar surface to ensure uniform
spread and distribution of media. Vacuum shunting will be ensured to minimalize cross-contamination
from external sources. After proper incubation, standardized to 24 to 36 hours in the incubation chamber,
the samples will be taken out. Distinct Colony Forming Units (CFU) will be subjected to photo
documentation as well as counting by identifying unique colonies.
Table 3 Aerobic culture report after 24 hrs of incubation
Sr No
Surface sampling Site
Before Hand wash
After Hand wash
1
Swab 01
40 – 45 Colonies
25 – 30 Colonies
2
Swab 02
25 – 30 Colonies
10-15 Colonies
3
Swab 03
35 – 40 Colonies
15 – 20 Colonies
4
Swab 04
45 – 50 Colonies
30-35 colonies
5
Swab 05
25 – 30 Colonies
20-25 colonies
Five swab samples were collected from health workers of SDM College of Ayurveda and Hospital. Swab
samples streaked on Blood and Mac-conkey agar, incubated at 370 c for 24 hrs in aerobic condition.
Results shows that number of colonies was reduced after washing with herbal hand wash.
DISCUSSION
Neem (Azadirachta indica) plants parts shows antimicrobial role through inhibitory effect on microbial
growth/potentiality of cell wall breakdown. Azadirachtin, a complex tetranortriterpenoid limonoid
present in seeds, is the key constituent responsible for both antifeedant and toxic effects in insects.
Leaves contain ingredients such as nimbin, nimbanene, 6-desacetylnimbinene, nimbandiol, nimbolide,
ascorbic acid, n-hexacosanol and amino acid, 7-desacetyl-7- benzoylazadiradione, 7-desacetyl-7-
benzoylgedunin, 17-hydroxyazadiradione, and nimbiol. Quercetin and ßsitosterol, polyphenolic
flavonoids, were purified from neem fresh leaves and were known to have antibacterial and antifungal
properties and seeds hold valuable constituents including gedunin and azadirachtin[3.4,5].
Methanolic fraction and aqueous fraction of Ocimum sanctum showed anti-fungal activity against
dermatophytic fungus i.e. T. rubrum etc. Aqueous fraction showed better anti dermatophytic activity as
compared to methanolic fraction. Antibacterial activity of the aqueous, alcoholic, chloroform extract and
oil obtained from leaves of Ocimum sanctum were studied against E. coli, P.aeruginosa, S. typhimurium and
S.aureus. Extract obtained from Ocimum sanctum were observed equally effective against pathogenic
gram positive and gram-negative bacteria [6,7].
Murraya koenigii has broad types of characteristics, such as antibacterial activity, activity of antifungal,
activity of antiprotozoal. The essential oil from M. koenigii leaves showed antibacterial effect against B.
subtilis, Staph. aureus, C. pyogenes, P. vulgaris and Pasteurella multicida. The pure oil was active against
the first three organisms even at a dilution of 1: 50027. The acetone extract of the fresh leaves of M.
koenigii on fractionation gives three bioactive carbazole alkaloids named as mahanimbine, murrayanol
and mahanine, which has shown mosquitocidal, antimicrobial and topisomerase I and II inhibition
activities. Acetone extract of M. koenigii is active against Aspergillus niger, benzene extract is most active
against Alternaria solani and Helminthosporium solani and ethanol extract is active against Penicillum
notatum. The leaves of this plant also are wealthy in diverse compounds inclusive of flavonoids,
polyphenols, alkaloids, etc. The primary constituent is tannins. Tannins are water soluble polyphenols
which can be typically located in classes hydrolysable and condensed tannins. Tannins are found in many
foods. Tannins were stated to be bacteriostatic or bactericidal towards Staphylococcus aureus. They act at
the membranes of the organisms.
In the current situation, the plants studied are rich in these various compounds and are therefore more
effective against skin pathogens. The main ideology behind the combination of plant materials is the
observation of the additive effect of the active ingredients of various plants. This combination has proven
to be beneficial and is used to hand wash herbs. The results clearly prove that the herbal soap thus
produced is much more active than the commercial antiseptic soap. The active ingredients in

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BEPLS Vol 12 [3] February 2023 170 | P a g e © 2023 AELS, INDIA
manufactured soap may be more effective at killing or eliminating organisms than the chemicals used in
soaps. Thus, these compounds can be extracted and incorporated into soap bases to produce a superior
antiseptic soap with little or no side effects. Thus, a new way can be found to combat antibiotic resistance
of pathogenic organisms and enable a safer and healthier life through germ-free hands. Although the
elimination is not 100%, a significant number can be reduced[8,9].
Mucopolyssacarides aid in the skin's moisture retention. It was claimed that products containing Aloe
vera increased skin hydration through a humectant mechanism. Several approaches have been used to
demonstrate the activity of Aloe vera inner gel against Gram-positive and Gram-negative bacteria. Aloe
vera gel has been shown to suppress the bacteria Streptococcus pyogenes and Streptococcus faecalis. In a
monolayer culture, aloe vera gel proved bactericidal against Pseudomonas aeruginosa, whereas
acemannan inhibited it from attaching to human lung epithelial cells. A processed Aloe vera gel
preparation reportedly inhibited the growth of Candida albicans.Aloe extracts' antiviral properties could
be attributed to indirect or direct actions. They have these effects both indirectly and directly by boosting
the immune system. Various enveloped viruses, such as Herpes simplex, Varicella zoster, and Influenza,
are inactivated by the anthraquinone aloin [10]. The inclusion of six antiseptic agents, namely Lupeol,
salicylic acid, urea nitrogen, cinnamonic acid, phenols, and sulphur, contributes to Aloe vera's
antibacterial activity. They all have antifungal, antibacterial, and antiviral properties [11,12].
CONCLUSION
The drugs used for developing herbal hand wash i.e. nimba, tulsi, saurabhanimba and kumari are having
antibacterial and antifungal properties. Kumari will help in skin hydration possibly by means of a
humectant mechanism. The analytical study shows that values are under normal range and suitable for
hand wash. The microbial load study results suggest that hand-washing herbs reduces the microbial load
to protect against skin pathogens. This could be the reason for using herbs in preparation for hand
washing and using these compounds in the production of disinfectant lotions or soaps instead of
chemicals.
Acknowledgement
Rajiv Gandhi University of Health Sciences, Bengaluru, Karnataka. Short term research grant for UG
students with project name Development and anti-microbial study of herbal hand wash and project code-
UG21AYU042.
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CITATION OF THIS ARTICLE
Nameera H. A. Priyanshi Goyal, Maanya. M. Lingam, Vinaykumar R Kadibagil, Govinda Sharma.K.
Development and Anti-Microbial Study of Herbal Hand Wash Using Nimba, Tulsi, Sourabhanimba,
Kumari.Bull. Env. Pharmacol. Life Sci., Vol 12[3] Feb 2023 : 167-170.