INTRODUCTION:

Hand washing is also known as hand hygiene it is the act of cleaning hands to remove soil, dirt, and microorganisms. Instead, if water and soap are not available, hands can be washed with ash. Hospital hand hygiene refers to medical procedure-related hygiene procedures.¹ before providing medication or medical treatment, hand washing may avoid or mitigate the spread of illness.

The primary medical purpose of hand washing is to cleanse the hands of bacteria and chemicals that may cause harm or illness (such as bacteria or viruses). This is particularly relevant for people in the medical profession who deal with food or work.² Hand washing with soap reliably prevents the spread of diseases such as diarrhea and cholera that are transmitted via fecal-oral routes at crucial moments during the day. For instance, if people do not wash their hands and they touch their eyes, nose, or mouth they will be infected with respiratory infections such as influenza or common cold.³ The key mode of transmission of microbes and pathogens is hands. Hand hygiene is also the most effective measure to decrease harmful germ transmission and to prevent infection. The most effective, easiest and least costly means of preventing nosocomial infections is hand hygiene. The infected hands will serve as microorganism transmission vectors.⁴ Hand washing is a necessary measure to protect the skin from infectious microorganisms and to prevent many infectious diseases from spreading. In readiness for jobs, food processing staff and food service employees need to be taught how to use proper hand and fingertip washing by management. Any health care provider, caregiver or person involved in direct or indirect patient care must be concerned with hand hygiene and should be able to do so at the right time and properly.⁵ Hand washing extracts visible dirt from the hands and lowers the amount of destructive microorganisms such as E. coli and Salmonella may be transferred from humans, livestock or machines to food. Fecally polluted hands are by far the most prevalent manner in which the tainted poultry handler transmits pathogens to food. Hand washing with soap and water has been considered a personal hygiene practice for decades. Originally, the practice of washing hands with an antiseptic agent began in the early 19th century.⁶ Murraya koenigii is known as ‘curry patta’ in hindi and widely used as spice and condiment in India and other tropical countries. It belongs to the family Rutaceae. Anti-oxidant, anti-tumour, anti-microbial, anti-inflammatory, anti-trypanocidal and mosquitocidal activities have been indicated for some of these alkaloids.⁷ Syzygium aromaticum, also known as clove, is a dried flower bud belonging to the Myrtaceae family. Several reports have documented the antibacterial, antiviral, anticarcinogenic, and antifungal activities of some aromatic herbs including cinnamon, oregano, clove, thyme, and mint. However, clove has gained much attention among other spices due to its potent antimicrobial and antioxidant activities.⁸ The evaluation tests carried out are as follows (i) Physical evaluation, (ii) Chemical evaluation, and (iii) Antimicrobial activity.

MATERIALS AND METHODS:

Chemicals and Reagents:

Methanol (HPLC grade), Distilled water, Broth (Nutrient agar), Brookfield Viscometer (DV-1 Prime).

Procurement of Plant Material:

Leaves of Murraya koenigii and Buds of Eugenia aromaticam plants were collected from the grocery store kasegaon, Dist- Sangli, Maharashtra, India.

Extraction:

The Leaves of cultivated plants Murraya koenigii and buds of Eugenia aromaticam are picked and grind coarsely. 50grams of coarsely grinded plant leaves and buds are soaked separately in 200ml of methanol and held for 3-4 days for maceration. The extract is purified during maceration and the filtrate was collected and used to prepare hand wash.^9-15

Preparations of Herbal Hand Hash Formulation:

The formulation of herbal hand wash was prepared from the extract of leaves of cultivated plants Murraya koenigii and buds of Eugenia aromaticam using Sodium hydroxide, Potassium hydroxide, Stearic acid, Triethanolamine, Lanolin, Glycerin, Lauryl sulphate, Rose merry oil. 10gm of each plant material extract filtrate was used to prepare hand wash. Add 1gm of sodium hydroxide to a 3ml of Glycerin, 0.5gm of stearic acid, 1gm of potassium hydroxide continuously stirring in methanolic extract solution. Finally 2gm triethanolamine, 2gm lanoline, 2gm SLS, and 5ml of rose merry oil was added and adjusted to make 100ml of final volume.

Evaluation of Foam Herbal Hand Wash:

Antimicrobial activity evaluation:

Screening of the anti-microbial efficacy of the formulated herbal hand wash was performed on different microorganisms using the agar plate method as a standard procedure. The anti-microbial activity was tested against four different microorganisms, Bacillus subtillis, Staphylococcus aereus, Pseudomonas aeruginosa and Escherichia coli, using four sterile petri plates. A solution of the nutrient agar was filled with the plates and allowed for solidification. Upon solidification, the subculture microorganisms were inoculated into the agar media of the nutrient and three cavities were made therein. The first cavity is filled with usual amoxicillin antibiotics, the second with herbal hand wash without lime water, and the third cavity with herbal hand wash is filled with lime water. Note was taken for the sample to be placed at the level of the cavity. To test the activity the plates are placed in the incubator at 37⁰C. After 24 hours the plates for inhibition zone formation were observed. The anti -microbial formulation activity is calculated from the inhibition region.^16-17

Physical Assessment:

Colour: Visually determined.

Odour: Manually determined.

Appearance: Visually determined

Homogeneity: Visually determined

Perfume test: For its acceptability, it was founded upon individual observation. Five people were asked for the scent to be appropriate and their opinions were taken and fragrance was assessed using the criteria described below.

A) The fragrance was good, just as good as the reference hand wash fragrance.

B) The scent wasn't as good as the reference hand wash but comparable.

Table 1: Composition of prepared formulation

Sr. No.	Ingredients	Quantity
1.	Sodium hydroxide	1 gm
2.	Potassium hydroxide	1 gm
3.	Stearic acid	0.5 gm
4.	Triethanolamine	2 gm
5.	Lanolin	2 gm
6.	Glycerin	3 gm
7.	Lauryl sulphate	1 gm
8.	Rose merry oil	5 ml
9.	Clove extract	10 gm
10.	Curry leaves extract	10 gm

Chemical Parameter:

pH Determination:

1gm of herbal hand wash sample was taken and dissolved into 100ml of distilled water. pH of Hand wash was balanced using NaOH solution of 40%.

Viscosity:

Herbal hand wash viscosity was measured by the use of a Brookfield optical viscometer.

Foam Height:

0.5gm of Herbal Hand wash sample was collected and dispersed in 25ml distilled water. Then it was transferred to a stopped measuring cylinder of 500ml, the volume was makeup to 50ml. Twenty-five strokes were given and stored until aqueous volume up to 50ml was measured; above the aqueous volume.

Foam Retention:

50ml Herbal hand wash was put into a 200ml graduated cylinder and 10 times shaken. The volume of foam was kept for 4 minutes at 1-minute intervals.

RESULT AND DISCUSSION:

Formulation and Evaluation of Foam Herbal Hand Wash from Leaves of Murraya koenigii and Buds of Eugenia aromaticam Extract:

Foam herbal hand wash formulations were prepared using Sodium hydroxide and Potassium hydroxide where used for the opacification and Stearic acid as emulsifying agent. Triethanolamine was used in formulations to adjust the pH neutralize the pH and Lauryl sulphate as Surface active agent.

Foam herbal hand wash formulations showed brown colour, Characteristic odour, and good homogeneity. The pH of formulations was 5.60 which lies in the normal pH range of the skin. The viscosity was found to be 29 cps, foam height 300ml, and foam retention was 20ml.

The prepared formulations of herbal hand wash were subjected for physical evaluation and antimicrobial efficacy.

Physical and Chemical Evaluation:

All the observation data for physical and chemical evaluations of hand wash presented as follows

Table 2: Physical evaluation tests

Sr. No.	Test	Observation
1.	Physical parameter
	Colour	Brown
	Odor	Characteristics
	Appearance	Translucent
	Homogeneity	Good
2.	Chemical Parameter
	pH	5.60
	Viscosity	29 cps
	Foam Height	300 ml
	Foam Retention	20ml

Antimicrobial Activity Screening Tests:

The anti-microbial activity of the prepared Herbal Hand Wash was tested by the cup plate technique on Candida albicans, Staphylococcus aureus and Bacillus subtillis. The analysis of the cup plate technique indicated that the hand wash prepared from the methanol extract of the combined plant materials was more efficient than the hand wash activity shown in the commonly available table^.18-21

Table 3: Zone of inhibition of formulations

Sr. No	Micro organism	Zone of inhibition of Std. hand wash(mm)	Zone of inhibition of Test hand wash(mm)
1.	Staphylococcus aureus	23	39
2.	Bacillus substillis	25	35
3.	Candida albicans	30	42

Figure 1: Comparative study of formulation for antimicrobial activity a. Staphylococcus aureus zone of inhibition; b. Bacillus substillis zone of inhibition; c. Candida albicans zone of inhibition; d. Standard anti-bacterial herbal hand wash zone of inhibition.

The formulation of herbal hand wash has proved beneficial with excellent activity against all the microorganisms tested as compared to standard hand wash.

CONCLUSION:

In the present study, active constituents were present in the methanolic extract of Murraya koenigii leaves and Eugenia aromaticam buds demonstrated superior inhibition of the synthetic antimicrobial agents present in the commercially available antiseptic spray hand wash against various gram positive, gram negative and fungal skin pathogens. Therefore, these compounds could be extracted and incorporated in hand wash bases in order to prepare superior antimicrobial hand wash. The formulated hand wash shows the better result of physical and chemical parameters as well as excellent activity against all the tested microorganisms as compare to standard hand wash. Formulation can also be used regularly to promote healthy adult and children's hygiene. Further pharmacological tests, toxicological research, and possible isolation of therapeutic antibacterial from this product are potential challenges.

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Received on 26.02.2021 Modified on 18.05.2021

Asian J. Res. Pharm. Sci. 2021; 11(3):195-198.

DOI: 10.52711/2231-5659.2021.00031