Article WJPR 1467435569

World Journal of Pharmaceutical Research
Bahuguna et al. SJIF Impact

World Journal of Pharmaceutical Factor 6.805
Research
Volume 5, Issue 7, 1559-1577. Research Article ISSN 2277– 7105
FORMULATION AND EVALUATION OF HAND WASH

1
Megha Bahuguna and 2Shilpi Kashyap*
1
Himachal Institute of Pharmacy.
2
Assistant Professor Himachal Institute of Pharmacy Poanta Sahib.
INTRODUCTION
Article Received on
19 May 2016, Hygiene
Revised on 08 June 2016, It is basically defined as branch of science which is involved in
Accepted on 29 June 2016
knowledge and practices related to promotion of health.[1] Spread of
DOI: 10.20959/wjpr20167-6663
infections (bacterial and viral) can be prevented by following hygiene
practices. Since cleaning processes (e.g hand washing, taking bath)
*Corresponding Author
removes dirt and soil as well as infectious microbes they are
Shilpi Kashyap
commonly used to achieve hygiene.[2] Hygiene is defined as
Assistant Professor Himachal
Institute of Pharmacy Poanta maintenance of cleanliness practices which carries utmost importance
Sahib. in maintenance of health. Keeping bodily hygiene and usage of
cleansers are requisites of healthy living. These concepts highlight the
need of maintaining hygiene in prevention of diseases.[3]
Hand Hygiene
The act of cleaning hands with water or soap or liquid is generally referred to as hand
hygiene.[4] The importance of washing hands is that it cleanses the hands of harmful
chemicals and pathogens (bacteria and virus).[5] Hand hygiene is especially important for
people working in medical field and restaurants or who cook and serve food to general
public.[6] Proper hand hygiene can decrease the transmission of cold viruses and other germ is
an established fact. Washing hands is considered the best way to maintain personal hygiene
and protect oneself from diseases.[7]
Promotion and implementation of hand washing program in the schools, brought a reduction
of 42% in school absenteeism[8,9] and reduced the incidence of gastrointestinal and respiratory
illness among children.[10]
www.wjpr.net Vol 5, Issue 7, 2016. 1559

Bahuguna et al. World Journal of Pharmaceutical Research
Hand Washing
It refers to washing hands with plain or antimicrobial soap or water. [11] In actual practice, it
can vary considerably from a brief rinse of hands to extensive scrubbing. The purpose of
hand washing in the health care setting is to remove pathogenic microorganisms ("germs")
and avoid transmitting them. It has been reported that lack of hand washing remains at g
unacceptable levels in most medical environments, with large numbers of doctors and nurses
routinely forgetting to wash their hands before touching patients.[12] One study showed that
proper hand washing and other simple procedures can decrease the rate of catheter-related
Bloodstream infections by 66 percent.[13]
Skin is one of the most exposed part of the body requires protection from the pathogens. To
protect the skin from harmful micro organisms and to prevent spreading of many contagious
diseases hand washing is absolutely an important precaution. Correct use of a fingernail brush
to wash hands and fingertips is the best way to assure removal of transient
microorganisms.[14]
Hand washing removes visible dirt from hands and reduce the number of harmful
microorganisms. Harmful bacteria and viruses such as, E. Coli and Salmonella can be carried
by people, animals or equipment and transmitted to food.[15]
Six steps to effective hand washing[16]

Step 1
Wet hands and apply soap. Rub palms together until soap is bubbly.
Fig-1
Step 2
Rub each palm over the back of the other hand.

Step 3
Rub between your fingers on each hand.
Step 4
Rub your hands with the fingers together
Step 5
Rub around each of your thumbs.
Step 6
Rub in circles on your palms. Then rinse and dry your hands

Hands perform the majority of functions of the human‟s body and are exposed to a variety of
substances which include soil during farming, food during cooking, touching raw and
contaminated food material, during personal hygiene. Clean hands stop the spread of germs;
therefore hand washing is often emphasized as the single most important measure in any
infection control programme for preventing cross transmission of microorganisms between
patients.
Plant extracts and products have been used for centuries in traditional medicine, functional
food, natural dyes, cosmetics, as a detergent and in the treatment of diseases.[17,18,19]
In present study we formulate herbal hand wash using different plant extracts with potential
antibacterial activity and thereby establishing them as a potent antimicrobial agent in the
formulation of herbal hand wash.
COMPOSITION OF HERBAL HAND WASH

PLANT EXTRACT
SOLVENT
HUMECTANT
FLAVOURING AGENT
PRESERVATIVE
THICKENING AGENT
EMULSIFIERS
SURFACTANTS
DETERGENTS
AYURVEDIC APPROACH IN HAND HYGIENE

Ayurveda is the traditional medicinal system of India and believed to have originated over
6000 years ago. It describes the ways to remain healthy as well as methods to treat disease.
The name itself means “Knowledge (Veda) of life (Ayu)”
Ayurveda is not only a curative medical science but also is a comprehensive way of healthy
living. Guidelines related to healthy or „swastha‟ individual are termed as „Swastha
vritta‟.[20,21]
Though proper hygiene and in particular hand hygiene is the single most important, simplest,
and least expensive means of preventing health care-associated infections and the spread of

Antimicrobial resistance; but, unfortunately poor hand-hygiene practices are still observed
due to lack of scientific knowledge, unawareness of risks and unavailability of hand-hygiene
facilities.[22,23,24]
Considering ayurvedic approach; an attempt has been made to screen classical literature for
the herbs with cleansing properties and found that, Pancha Valkala [25] (the barks of five
plants) has activities like
[26]
[27]
[28]
[29]
[30]
Pancha Valkala is the combination of barks of five different plants- Vata (Ficus bengalensis
Linn.)
iosa Linn.)
Ex Correa.)
HISTORY OF HERBAL HAND WASH

We cannot ignore the fact that Austrian-Hungarian physician Ignaz semmelweis discovered
the importance of hand hygiene in 1858, opening the era of infection control. Semmelweis
demonstrated that puerperal fever (also known as childbed fever) was contagious and that this
incidence could drastical He made this discovery in 1847 while working in the Maternity
Department of the Vienna Lying-in Hospital. His failure to convince his fellow doctors led to
a tragic conclusion, however, he was ultimately vindicated and cleared of blame. While
employed as assistant to the professor of the maternity clinic at the Vienna General Hospital
in Austria in 1847, Semmelweis introduced hand washing with chlorinated lime solution for
interns who had performed autopsies. This immediately reduced the incidence of fatal
puerperal fever from about 10 percent (range 5–30 percent) to about 1–2 percent. At the time,
diseases were attributed to many different and unrelated causes. Each case was considered
unique, just as aly be reduced by appropriate hand washing by medical care-givers. Human
person is unique. Semmelweis‟s hypothesis, that there was only one cause, that all that
mattered was cleanliness, was extreme at the time, and was largely ignored, rejected or

ridiculed. He was dismissed from the hospital for political reasons and harassed by the
medical community in Vienna, being eventually forced to move to Pest.
Semmelweis was outraged by the indifference of the medical profession and began writing
open and increasingly angry letters to prominent European obstetricians, at times denouncing
them as irresponsible murderers. His contemporaries, including his wife, believed he was
losing his mind, and in 1865 he was committed to an asylum. In an ironic twist of fate, he
died there of septicaemia only 14 days later, possibly as the result of being severely beaten by
guards. Semmelweis‟s practice earned widespread acceptance only years after his death,
when louis pasteur developed the germ theory of disease offering a theoretical explanation
for Semmelweis‟s findings. He is considered a pioneer of antiseptic procedures.
No wonder, 150 years later, it was the semmelweis university(SU) in Budapest that first
adapted a digital tool to effectively teach proper hand disinfection technique. The Department
of Surgical Research and Techniques at SU began to employ a UV dye-based, computer-
imaging empowered device after it received the 1st ICPIC Innovation Academy Awar.[63]
BENEFITS OF USING HERBAL HAND WASH

Ease of availability
herbs are easily availaible in rural as well as urban areas, so they can be easily used by
anyone.
Cheap
Cost of herbal plants is less as compared to chemical ingredients used in synthetic hand
washes.
Increased efficiency
Herbal hand washes are more efficient in promoting hand hygiene.
Less side effects

Herbal hand washes have fewer side effects than other hand washes.
EVALUATION OF HERBAL HAND WASHES

PHYSICAL PARAMETERS
ph

CHEMICAL PARAMETERS
[31,32,33-35]
EVALUATION TECHNIQUES OF HAND WASH ACTIVITY AFTER USE

ng[36]-involves applying an adhesive tape to the skin surface then removing the
tape with an adhering layer of skin containing colonies of bacteria. This method allows one to
determine the distribution of bacterial colonies in relation to area and depth of the epidermal
layers.
[37]
-is performed with cotton or calcium alginate swabs. The swabs are
placed in water or hexametaphosphate solution, the suspension is shaken thoroughly to
distribute the organisms, and an aliquot is plated bacteriologically. Variations in bacterial
counts can occur because of non reproducibility in the pressure used in applying the swabs to
the skin and because the number of bacteria removed from the swabs may vary with the
efficiency of the shaking.
-are made by adding liquefied nutrient agar to a Rodac plate or small

container, allowing the agar to harden, then pressing the plate with uniform pressure to the
skin area to be tested. Plates are incubated at 37˚C for 48 hours, where upon colonies are
counted.
TECHNIQUES USED FOR EVALUATION OF TRANSIENT HAND MICROFLORA

- finger impressions are taken on the separate media plates for
both leftand right fingers and incubating the plates for 18-24 hr at 370C followed by colony
counting.
od- hand wash samples are collected in the sterile beakers/bags under
sterile conditions. Temperature of sterile water should be near to 400˚C and 75 ml water is

used for a single hand wash. Further enumeration of bacterial flora is carried out by serially
diluting and plating on media plates and incubating them at 370C for 18-24 hr.
ANTIMICROBIAL SCREENING METHODS

-Disc/Well diffusion and Agar/Broth dilution methods.[38]
-
O Carrier tests: In these tests, a carrier such as a silk or catgut thread or a little stick is
contaminated by submersion in a liquid culture of the test organism. The carrier is then dried
and is brought in contact with the disinfectant for a specified exposure time. After the
exposure, it is cultured in a nutrient broth; no growth indicates activity of the disinfectant
tested whereas growth indicates no activity. By multiplying the number of test concentrations
of the disinfectant and the contact times, a potentially active concentration-time relationships
of the disinfectant is obtained.[39]
O Suspension tests: In these tests, a sample of the bacterial culture is suspended into the
disinfectant solution and after exposure it is verified by subculture whether this inoculum is
killed or not. Suspension tests are preferred to carrier tests as the bacteria are uniformly
exposed to the activity of disinfectant. There are different kinds of suspension tests such as
determination of phenol coefficient by Rideal Walker method; Chick Martin test and
Disinfectant kill time tests.[39] In a study Rideal Walker method was used to test the activity
of disinfectants the activity was compared with phenol and the Rideal Walker coefficient was
evaluated.[40]
O Capacity tests:In these tests the disinfectant is challenged repeatedly by successive

additions of bacterial suspension until its capacity to kill has been exhausted. Each time a
soiled instrument is placed into a container with disinfectant, a certain quantity of dirt and
bacteria is added to the solution. The ability to retain activity in the presence of an increasing
load is the capacity of the disinfectant. The best known capacity test is the Kelsey-Sykes test.
Kelsey-Sykes test is a triple challenge test, designed to determine concentrations of
disinfectant that will be effective in clean and dirty conditions.[39]

Evaluation of Herbal Hand Wash

PHYSICAL PARAMETERS
Ph
Before every single ph measurement or before any series of uses ph meter must be calibrated.
After calibration rinse electrode and submerge it in the tested solution. Read the result and
write it down in table.
Viscosity
The viscosity of hand wash was determined by using digital Brookfield viscometer.
Measured quantity of herbal hand wash was taken into a beaker and the tip of viscometer was
immersed into the hand wash gel and the viscosity was measured in triplicate.
Stability
The stability studies were carried out by storing at different temperature conditions like 40ᵒC,
25ᵒC & 37ᵒC for 1 week
Color
The colour was inspected visually.
Odor
The odor was percepted by sensing the formulation.
Media
• Nutrient Agar Media
INGREDIENT QUANTITY (gm/l)

Peptic Digest of Animal Tissue 5.000
Sodium Chloride 5.000
Beef Extract 1.500
Yeast Extract 1.500
Agar 15.000
CHEMICAL PARAMETERS
Antimicrobial Studies
The screening of anti-microbial efficacy of the extract was performed on microorganism by
using dip well method as per standard procedure. Petri plates were taken for testing the

antimicrobial activity of extract against Staphylococcus aureus. The plates were filled with
Nutrient agar solution and allowed for solidification. After solidification the microorganisms
from the subculture were inoculated into the nutrient agar media and three discs were
inoculated with extract, antibiotic drug and formulation sample respectively. The plates were
incubated at 37ºc for overnight. After 24 hours of incubation, the plates were
Observed for the zone of inhibition.

Sample
Gram positive bacteria i.e. Staphylococcus aureus was collected from department of
microbiology, Himachal Ins Determination of MIC (MINIMUM INHIBITORY
CONCENTRATION)
The MIC is defined as the lowest concentration that completely inhibits the growth of
microorganisms for 24 hrs incubation. Determination of minimum inhibitory concentration of
calotropis gigantea was determined by adding 0.5 ml of extract into the wells of nutrient agar
plates that were already seeded with the standardized inoculums of the test bacteria. The agar
plates were incubated at 37ºc for 24 hrs. The lowest concentration of oil showing a clear zone
of inhibition was considered as MIC.
Foam Height
1gm of sample of Poly Herbal Gel Based hand wash was taken and dispersed in 50 ml
distilled water. Then, transferred it into 500 ml stoppered measuring cylinder; volume was
make up to 100 ml with water. 25 strokes was given & stand till aqueous volume measured
upto100 ml & measured the foam height; above the aqueous volume.
Foam Retention
50 ml of the Poly Herbal Gel Based hand wash was taken into a 250ml graduated cylinder
and shaken 10 times. The volume of foam at 1-minute intervals for 4 minutes was Recorded.
Foam retention should remain stable for at least 5 minutes.
Spreading Coefficient
Two glass slides were taken of known dimensions and an excess of gel (about 2g) under
study was placed on the ground slide. The gel was then sandwiched between.
This slide and another glass slide having the dimension of fixed ground slide. A 500g
weighted was placed on the top of the two slides for 5 minutes to expel air and to provide a

uniform film of the gel between the slides. Excess of the gel was scrapped off from the edges.
The top slide was then separated with friction and the time (in seconds) required by the top
slide to cover a distance of 7.5 cm be noted. A shorter interval indicates better
Spreadability
Spreadability was calculated using the following formula: S=ML/T
Where, S = Spreadability,
M = Weight in the pan (tied to the upper slide),
L = Length moved by the glass slide
T= Time (in sec.) Taken to separate the slide completely each other
Skin Irritation
The formulation was applied on skin and left for 30 min
RESULTS AND DISCUSSIONS

The present study was carried out to formulate Calotropis gigantea based hand wash using gel
base as carriers. The formulation was prepared by using generally approved excipients that
are compatible with any similar hand cleansing formulations. It was organoleptically
evaluated to ensure product stability and performed in-vitro antimicrobial test to prove its
efficacy to act against infectious bacteria collected from volunteers.
PHYSICAL PARAMETERS
Color
Green color of the formulation was inspected visually.
Odor
The odor of the formulation was like that of a lemon.
Viscosity (dyne/cm)
The viscosity was carried out by Brookfield viscometer. The results of the observation are
shown in table 4. The viscosity of Formulation 1st is 50 P, Formulation 2nd is 55 P and of
Formulation 3rd is 58 P. The viscosity of Formulation 1st is found to be the best for herbal
hand wash.

Ph
The ph was determined by using digital ph meter. The ph of formulation 1st is 6.87 and the
ph of of formulation 2nd is 7.0 and the ph of formulation 3rd is 7.5. The observations are
listed in the table 4 according to which ph of formulation 1 was found to be the best for
herbal hand wash.
Sr. Evaluation
F1 F2 F3
No. Parameter
1 Color Green Green Green
2 Odor Lemon like Lemon like Lemon like
3 Viscosity(poise) 50 55 58
4 Ph 6.87 7.0 7.5
Stability
The study was carried out for 7 days and no change was noticed. The observations are listed
in table no.7.
MEASUREMENT OF STABILITY
S.No. Appearance Ph Viscosity
F 1 NO CHANGE 6.87 50
CHEMICAL PARAMETERS
Determination of MIC (Minimum Inhibitory Concentration)

According to the zone of inhibition formed resulting from the plant extract and formulation
prepared against bacteria isolated, showed that hand wash prepared with Calotropis gigantea
has great activity. The zone of inhibition with extract, formulation and the antibiotic drug
ampicillin are listed below:
Sample
Zone of Inhibition
Plant Exract
32mm
Herbal Hand Wash
18.5mm
Ampicillin
Showed resistance
Foam Height
Stand till aqueous volume measured upto 100 ml after 25 strokes & measured the foam
height, above the aqueous volume.
Foam height of the formulation was found to be:
Sr. No. F1 F2 F3
1 11cm 9cm 7.3cm
Since foam height of formulation 1 was almost stable for 5 min thus it was best to be used as
herbal hand wash.
Spreading Coefficient
The top slide when separated with friction the time (in seconds) required by the top slide to
cover a distance of 7.5 cm was noted. A shorter interval indicates better spreadability.

Skin Irritation
No irritation was felt on skin and no redness was observed.
CONCLUSIONS
Like cosmetics, cosmeceuticals are topically applied but they contain ingredients that
influence the biological functions of skin.
Hand hygiene can also be a problem in between the people. Prevention and control of
infectious activities are designed to limit the spread of infection and provide a safe
environment for all people, regardless of the setting. In light of the emergence of antibiotic
resistant organisms, effective infection control measures, such as handwashing, are essential
to prevention.
Hand wash gels are used for the purpose of cleaning hands. Its composition is prepared
according to delicateness of skin so that it can not cause any type of irritation.
The tests were performed on Calotropis gigantean which is used as an active ingredient in
antimicrobial activity through well diffusion method.
Other tests were also performed
18.5mm, which was found to be good for a herbal hand wash.
ph: ph of formulation 1st was 6.87 which was nearer to ph of skin.
1st.
foam retention: Foam formed was almost stable in formulation 1stfor 5

min thus it was best to be used as herbal hand wash.
From the results it can be concluded that above preparation using Calotropis gigantea as plant
extract and gelling agent HPMC in 1% w/v conc. Can be used as an antimicrobial herbal
hand wash as it shows good zone of inhibition against the tested bacteria i.e. S. Aureus. Other
evaluation tests also proved to be efficient for the usage.

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World Journal of Pharmaceutical Research

Bahuguna et al. SJIF Impact

FORMULATION AND EVALUATION OF HAND WASH

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Six steps to effective hand washing[16]

Rub each palm over the back of the other hand.

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Rub between your fingers on each hand.

Rub your hands with the fingers together

Rub around each of your thumbs.

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COMPOSITION OF HERBAL HAND WASH

AYURVEDIC APPROACH IN HAND HYGIENE

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HISTORY OF HERBAL HAND WASH

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BENEFITS OF USING HERBAL HAND WASH

Less side effects

EVALUATION OF HERBAL HAND WASHES

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EVALUATION TECHNIQUES OF HAND WASH ACTIVITY AFTER USE

-are made by adding liquefied nutrient agar to a Rodac plate or small

TECHNIQUES USED FOR EVALUATION OF TRANSIENT HAND MICROFLORA

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ANTIMICROBIAL SCREENING METHODS

O Capacity tests:In these tests the disinfectant is challenged repeatedly by successive

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Evaluation of Herbal Hand Wash

INGREDIENT QUANTITY (gm/l)

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Observed for the zone of inhibition.

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RESULTS AND DISCUSSIONS

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Other tests were also performed

18.5mm, which was found to be good for a herbal hand wash.

ph: ph of formulation 1st was 6.87 which was nearer to ph of skin.

foam retention: Foam formed was almost stable in formulation 1stfor 5

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