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ABSTRACT 1. INTRODUCTION
An ever increasing number of customers might want The Hands are the chief mode of transmission of
Good hygieneene is a minister to health, comfort, and microbial infections and diseases [1]. Hygiene of
social interactions. Current society is well aware of hands is hence the most vital thing to avoid the
hygiene. Washing hands ds is one of the simplest, most transmission of harmful germs and prevent the
effective ways to get rid of germs and avoid infection. infections. Hand hygiene is the simplest and least
Transient microorganisms can be of any type, from expensive means of preventing nosocomial infections
any source may contact the body. As hhands carry out [2]. Hand-washing is significant in food production,
the majority of functions of the human’s body and are food service and also chief in healthcare setting,
exposed to a variety of substances,, hands hygiene homes and day care preparations.
preparations Though good &
plays a chief conscientiousness. Herbal plants are the simple hygiene technique is single most important,
importan
unique resources of every country with potent easy and least expensive means of preventing health
medicinal properties pave ways towards sustainable care-associated
associated infections and the spread of
health. In this present study, the combination of antimicrobial multidrug resistance; but, unfortunately
Trachyspermum copticum, Coleus aromaticus
aromaticus, Acorus poor hand-hygiene
hygiene practices are still observed due to
calamus, Mentha piperita, Piper nigramnigram, Elatteria lack of scientific knowledge, unawareness of risks
ri and
cardamomum and Aloe barbadensis were used to unavailability of hand hygiene facilities[3].
facilities
formulate herbal hand sanitizer. The hand wash
samples were subjected to microbial analysis. The Plant extracts and products have been used for
isolated strains (Yersinia, Proteus, Escherichia coli
coli, centuries in traditional medicine, functional food,
Klebsiella, Salmonella, and Pseudomonas
Pseudomonas) were natural dyes, cosmetics, and in the treatment of
characterized by their cultural characterization and diseases [4,5,6]. The lead advantage of using natural
confirmed using selective media. Herbal sanitizer source is that they are easily available, cheap and
possesses vitamin C (.4%), vitamin A (0.062) and harmless without side effects compared to chemical
considerable microbial load. The antioxidant activity products. Plants have been used for long time by
was found to be 49% and the susceptibility test traditional
raditional healers to prevent or cure infectious
showed more activity found in Yersinia sp. > Proteus conditions. As plants
lants are rich in wide variety of
sp > Salmonella sp. > Klebseilla sp. > Pseudomonas secondary
ndary metabolites such as tannins, terpenoids,
sp. > E.coli. alkaloids,
loids, and flavonoids etc., they have been found to
possess antimicrobial properties.Thus
properties. research has
Keywords: Hygiene, Hand sanitizer, Anti microbial been increased enormously focusing on the
and Anti oxidant production of natural products with improved quality.
quality
The emergence
ergence of bacterial resistance to the presently
2.3 In Vitro Evaluation of Transient Bacterial on 2.6 In vivo efficacy of Herbal hand sanitizer
human palm flora:
Total of five volunteers with different age groups
Collection of palm washes samples by Wash/rinse were involved to evaluate in vivo efficacy of the
Procedure [7] (Hand washing). sample. Total eight palm washes were taken from the
volunteers. Control palm washes were taken by use of
sterile water only.
@ IJTSRD | Available Online @ www.ijtsrd.com | Volume – 2 | Issue – 4 | May-Jun 2018 Page: 326
International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470
2.6 a. Bacteriological study [10] organisms, Escherichia coli, Klebsiella sp. Salmonella
sp. were found to be the most predominant pathogen.
Point inoculation method was used for viable count of Our skin contains large numbers of microorganisms,
different bacterial flora. Aseptically 10 µl palm wash mainly Gram-positive. Most of the gram positive
samples were transferred to 90 µl sterile saline (85%) bacteria are natural flora, which are commonly found
solution. Appropriate serial dilutions were made and on the hands, face and in deep layers of the skin [12,
aseptically 5 µl droplets were point inoculated on 13]. The goal of hygiene is a sufficient reduction of
selective media plates followed by incubation at 37°C microbial counts on the skin to prevent cross-
for 6 hrs. Typical colonies showing characteristic transmission of pathogens among humans.
coloration were counted and representative colonies
were picked up randomly and counter using colony 3.2 Antibacterial Susceptibility Testing
counter.
The antimicrobial activity of herbal hand sanitizer
2.7 Antioxidant Activity by Power Reducing Assay against different pathogens was determined by Agar
[11] Well diffusion method. The sample hand sanitizer
possessed more antibacterial effective against isolated
1 ml of Plant extract was mixed with phosphate buffer microorganisms with the inhibition zone ranging from
(2.5 ml 0.2 M, pH 6.6) and potassium ferric cyanide 15–30.8 mm. More activity was found in Yersinia
(2.5 ml). The mixture was incubated at 50°C for 20 sp.> Proteus sp> Salmonella sp.> Klebseilla sp.>
minutes. A portion (2.5 ml) of trichloroacetic acid Pseudomonas sp.> E.coli. Aloe vera used for its
(10%) was added to the mixture, which was then moisturizing activity in many cosmetics [6]. Plants are
centrifuged at 3000 rpm for 10 min. The upper layer rich in potential secondary metabolites, such as
of solution (2.5 ml) was mixed with distilled water flavonoids, tannins alkaloids polyphenols ext. These
(2.5 ml) and Ferric chloride (0.5 ml, 0.1%) and compounds have diverse biological activity including
absorbance measured at 700 nm. Increased antibacterial activity [14]. Most of the polyphenols are
absorbance of the reaction mixture indicates stronger extracted in the methanol [15]. The results suggested
reducing power. The activity was compared with that herbal extracts in mixture giving higher activity
ascorbic acid as standard. than the individual extracts. The combination of the
Calculation: antibacterial compounds from different plant extracts
may show synergistic effect enhancing their
Percentage scavenging activity = AcontrolAtest antimicrobial activity.
×100 Mithun A. Thombare [16] et al. 2015 observed that
the alcohol based herbal hand sanitizer (at
Acontrol concentration 400 μg/ml) showed greater inhibition
zones against bacterial species Escherichia coli,
Where Acontrol is the absorbance of the control. Atest is Pseudomonas aeruginosa, Bacillus subtilis and
the absorbance in the presence of the sample. Staphylococcus aureus in comparison to
3.RESULTS AND DISCUSSION commercially available hand sanitizer.
The herbal hand sanitizer was prepared and it was
used for evaluate the antimicrobial and antioxidant Microbial Load
activity.
Microbial load was analysed before applying sanitizer
3.1 Identification of Human bacterial flora and after applying of herbal based sanitizer. The result
showed that microbial load was considerably low
The bacterial flora like Yersinia sp. Proteus sp. after applying herbal hand sanitizer. Highest reduction
Escherichia coli, Klebsiella sp. Salmonella sp. and was observed in college student and slum boy with 75
Pseudomonas sp. were isolated from human palm and and 50% respectively after the application of herbal
identified based on Bergy’s manual. In this study, hand sanitizer. Many studies have been done earlier in
prevalence of gram positive and gram negative view of evaluation of total transient microflora of
bacteria organisms were 66.7% and 33.3% hands. Staphylococci especially coagulase –negative
respectively (fig: 1). Among Gram negative S.aureus were found to be the dominant part of
@ IJTSRD | Available Online @ www.ijtsrd.com | Volume – 2 | Issue – 4 | May-Jun 2018 Page: 327
International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470
2456
transient hand microflora [17]. Colonization of health Table 2: List of selective medium and test
care worker hands with S.aureus has been described organisms
to range between 10 to 76.3% and transient flora
varies considerably from person to person [18]. It was S. No. Test Organism Medium used
concluded that the herbal based sanitizer had potential
1 Yersinia sp. Yersenia agar
to control and prevent the over population of bacterial
flora in the palm. 2 Proteus sp. Macconkey agar
Kavatheker et. al. (2004)[19],, reported that pure 3 Escherichia coli Brilliant green agar
herbal hand sanitizers were found to be more effective
in reducing the total microbial load. Alcohol rubs are 4 Klebsiella sp. EMB agar
approximately 100 times more effective against
viruses than any form of hand washing [20]
[20].Sanitizers 5 Salmonella sp. SS agar
offer numerous advantages over non-alcoholic
alcoholic hand 6 Pseudomonas sp. Macconkey agar
disinfectants as, rubbing sanitizers onto both hands
&until it completely evaporate, usually requires only
15 to 30 seconds. Whereas vigorous friction, rinsing
with water, and drying with a towel are not needed Table: 3. Microbial load before and after applying
like hand disinfectants or soaps [21]. herbal hand sanitizer:
sanitiz
Load Load
In vitro Antioxidant activity of Herbal hand Percentage
before after
sanitizer Volunteers of
applying applying
reduction
sanitizer sanitizer
A concentration dependent scavenging activity was
observed in herbal hand sanitizer with maximum College
activity of 49.4% % achieved in the highest tested 4×103 3×10
3 3
75
student
concentration at 10mg/ml (fig 3). The sample had
relatively higher reducing power due to the presence Software
of high bioactive compounds which was present in working 6×103 2×10
2 3
33
selected medicinal ingredients and it has been lady
attributed with properties like free radical scaven
scavenging,
anthelmintic, antimicrobial, antiinflammatory and Home
5×103 2×10
2 3
40
analgesic etc. which are widely used in cosmetic, and maker
pharmaceutical industries, natural antioxidants
Servant
[22,23,24].Hence,, it could have promising skin care 7×103 3×10
3 3
43
women
properties, can able to protect the human body from
free radicals. Slum boy 8×103 4×10
4 3
50
SELECTIVE PLATING
The test organisms were confirmed by plating them in
selective medium and the results were observed and
showed in table.2
5. Acknowledgment
6. Reference
Hand hygiene is a vital principle exercise in the 6) Raskin I, Ribnicky DM, Komarnytsky S, Ilic N,
prevention, control, and reduction of healthcare Poulev A, Borisjuk N, Brinker A, Moreno DA, DA
acquired infections. Proper hand washing and drying Ripoll C, Yakoby N, O’Neal JM, Cornwell T,
methods prevent the chain of transmission of deadly Pastor I, Fridlender B. Plants and human health in
pathogens from the contaminated site to other parts of the twenty-first
first century. Trends Biotechnol.
the body. Hand sanitization is the paramount aid in 2002;20(12):522-532.
preventing nosocomial infections caused by different 7) Monique Courtenay, Lina Ramirez,
Ramirez Beth Cox,
opportunistic microorganisms .InIn this present study, Inyee Han, Xiuping Jiang,
Jiang Paul Dawson (2005).
results showed that the herbal sanitizer has Effects of various hand hygiene regimes on
antimicrobial property, antioxidant
dant activity, and removal and/or destruction of Escherichia coli on
considerable microbial load. When compare to hands. Volume5, Issue2‐4.
4. Pages 77-84.
alcohol based hand sanitizers, herbal hand sanitizer is
8) Ekrakene T and Igeleke CL(2007).
CL Microorganis
more effective against microbes, non toxic, formulate
ms associated with public mobile phones along be
hand soft and makes the hand hygienic. The further
nin‐ sapele express way. J. Appl. Sci.
study will be conducted for evaluation
luation of bioactive
Res. ;3(12):2009‐12.
compounds in herbal hand sanitizer responsible for
medicinal properties and its shelf life time
time. Thus, 9) Perez, C.; Pauli, M. and Bazerque, P. (1990). An
there is immense potential in establishing the use of antibacterial assay by agar well diffusion method.
antimicrobial herbal products as a measure to control Acta Bio Et Med Exp. 15, 113-115.
113
the multidrug resistant microbes as well as to check
@ IJTSRD | Available Online @ www.ijtsrd.com | Volume – 2 | Issue – 4 | May-Jun 2018 Page: 330