Indo American Journal of Pharmace Pharmaceutical utical Research, 2015
ISSN NO: 2231-68 2231-6876 76
FORMULATION AND EVALUATION OF NOVEL HERBAL HAND SANITIZER *
Mithun A. Thombare , Babaso V. Udugade, Tushar P. Hol, Manoj B. Mulik, Dnyaneshwar A. Pawade Satara College of Pharmacy, Satara - 415004. (M. S.) India. ARTICLE INFO Article history Received 20/01/2015 Available online 30/01/2015
Keywords Hand Hygiene, Extraction, Antimicrobial Activity, Herbal Sanitizer.
ABSTRACT Herbal hand sanitizer was prepared using leaves extracts of Ocimum sanctum, sanctum, Eugenia caryophyllus and caryophyllus and Cymbopogon flexuous. flexuous. The antibiotic sensitivity test of the prepared herbal hand sanitizer against skin pathogens was checked using disc diffusion method and results were compared with commercially available synthetic hand sanitizer. Results depicted that the herbal hand sanitizer gives larger inhibition zone than the commercially available synthetic hand sanitizer against Staphylococcus aureus aureus and Pseudomonas aeruginosa. aeruginosa. The efficacy of herbal hand sanitizer was evaluated using microorganism suspensions; which revealed that the herbal hand sanitizer is efficient in reducing higher number of microorganism from the hands as compared to commercial synthetic hand sanitizer. Thus, owing to higher antimicrobial activity and efficacy these herbal extracts can be used in the preparation of herbal herbal hand sanitizers on on commercial scale. scale.
Corresponding Corres ponding auth or
Mithun A. Thombare Satara College of Pharmacy, New Additional Additional MIDC, A/P- Degaon, SataraSatara- 415004, (M. S.) S.) India.
[email protected] +91-9423616476.
Please cite this article in press as M ithun A. Thombare et al. Formulation and Evaluation of Novel Herbal Hand Sanitizer. Indo Thombare American Journal of Pharm Research.2015:5(01). Research.2015:5(01).
3 8 Copy right © 2015 This is an Open Access article distributed under the terms of the Indo American journal of Pharmaceutical 4 e Research, which permits unrestricted unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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INTRODUCTION Hygiene is defined as maintenance of cleanliness practice which carries utmost importance in maintenance of well-being. Keeping bodily hygiene and usage of cleanser are requisite of healthy living. These concepts highlight the need of maintaining hygiene in prevention of diseases. [1] Although good and simple hygiene technique is single most important, easy and less expensive means of preventing health care associated infections and the spread of antimicrobial multidrug resistance; but, unfortunately poor hand hygiene practice are still observed due to lack of scientific awareness, lack of knowledge of risk and unavailability of hand hygiene facilities. Nosocomial infections are those which acquired or originated in a hospital or health care setting and are prevalence of compromised hosts, efficient mechanisms of transmission from patient to patient. Thus, occurrence of nosocomial infections is alarmingly increasing and has emerged as a serious concern in hospital care outcome; resulting in prolonged hospitalization, ample disease and mortality, and excessive costs. Escherichia coli, many Pseudomonas spp. and Staphylococcus aureus are commonly involved opportunistic microorganisms that primarily cause nosocomial infections. Generally infectious sites ar e urinary trac t, surgical wounds, respiratory tract, skin, blood, gastrointestinal tract and central nervous system. These pathogens also tend to become incorporated in to normal flora of health care workers. Pseudomonas aeruginosa is the most commonly detected microorganism in hospitalized patients and immunosuppressed people. Opportunistic fungal infections have become very important, especially in HIV patients. The highest frequencies of opportunistic fungal infections documented are candidacies, aspergillosis and cryptococcosis. Usually, microbes residing on hands are divided into resident and transient flora. Resident flora (e.g. Corynebactarium diphtheriae, Staphylococcus epidermidis, Streptococcus viridians) colonizing deeper skin layers are more resistant to mechanical removal, has lower pathogenic potential. While transient flora (e.g. Staphylococcus aureus, Gram-negative bacilli, Candida spp.) colonizes the superficial skin layer for shorter time periods, is usually acquired by contact with a patient or contaminated environment and are responsible for most health care a ssociated infection and the spread of antimicrobial resistant, but these microorganisms can be easil y removed by mechanical means such as hand washing. In the scenario mechanized life style; a consumer will always prefer readyma de formulation of alcohol hand rub rather than hand washing (application of a non-antimicrobial or antimicrobial soap; and mechanical friction is generated by rubbing the hands together for 1 minute, followed by rinsing with water and then drying thoroughly with a disposable towel). Hand sanitizer or hand antiseptic is a supplement or alternative to hand washing with soap and water. Various preparations are available in cluding gel, foam and liquid solution. The active ingredient in hand sanitizer is mostly alcoh ol, inactive ingredients includes a thickening agent such a polyacrylic acid for alcohol gel, humectants such as glycerin for liquid rubs, polypropylene glycol and essential oils of plants. Alcohol based hand sanitizer are more effective that kills most microorganism compared to soap and do not dry out hands as much. The level of alcohol varies between 60 -85% (most commonly used level is 62%). Alcohol rub sanitizers kill most bacteria, fungi and stop some viruses too. Alcohol rub sanitizers containing at least 70% alcohol (mainly ethyl alcohol) kill 99.9% of the bacteria on h and 30 seconds after application and 99.999% in 1 minute. [2] Traditional healers have long used plant to prevent or cure infectious conditions. Plants are rich in wide variety of secondary metabolites such as tannins, terpenoids, alkaloids, and flavonoids etc. which have been found to possess in-vitro antimicrobial properties. Considering this ultimatum; an attempt has been made to screen classical literature for the herbs with antimicrobial properties and it has been found that Ocimum sanctum (tulsi leaves), Eugenia caryophyllus (clove), Cymbopogon flexuous (lemon grass), and Aloe barbadensis (aloe) holds that antimicrobial potency.[3] Thus, we aimed to formulate and evaluate herbal hand sanitizer comprising of alcoholic extracts combination of these astonishing herbs using other suitable excipients; which can be used as ready to use herbal hand sanitizer. MATERIALS Leaves of tulsi (Ocimum sanctum), lemon grass (Cymbopogon flexuous), aloe ( Aloe barbadensis) and flowering buds of clove ( Eugenia caryophyllus) were collected from the local area of Satara region, Maharashtra, India and authenticated by expert botanist from Department of Botany of Yashvantrao Chavan Institute of Science, Satara. Nutrient agar (for bacterial cultivation), marketed preparation (Dettol Hand Sanitizer) and other excipients were purchased from market. Escherichia coli (Gram – ve), Pseudomonas aeruginosa (Gram – ve), Staphylococcus aureus (Gram +ve) and Basilus subtilis (Gram +ve) pathogens were selected for evaluation of antimicrobial activity of the herbal hand sanitizer. METHODS [4] Crude Drug Extraction 10 gm of well dried crude drug (tulsi, clove and lemon grass) was taken.
Grinded into fine powder.
4 8 4 e The powder of each plant material was added separately to 100 ml of methanol.
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The mixture was heated on water bath at 60⁰C for 1 h, then filtered and plant extract filtrate was collected. Formulation of Herbal Hand Sanitizer The compositions of alcohol based and non-alcohol herbal hand sanitizers prepared were as given in Table 1 and 2 respectively. Carbopol 940 was added to deionized water with constant stirring. After uniform mixing, triethanolamine (TEA) was added with slow stirring to avoid formation of possible air bubbles in the product and the blend was kept aside for 24 h. All the extracts were added to denatured al cohol along with glycerin, polysorbate 20 and were then mixed with aqueous phase. Finally 0.25% each of methyl and propyl paraben was added as preservative and 0.5% of perfume was mixed with slow stirring to obtain uniform product. Prepared product was stored in air tight containers. [5] Table 1: Composition of alcohol based herbal hand sanitizer formulation.
Sr. No. 1 2 3 4 5 6 7 8 9 10 11
Ingredients and Excipients Deionized water Alcohol (denatured) Tulsi leaves extract Lemon grass oil Clove extract Carbopol 940 Triethanol amine Glycerin Polysorbate 20 Preservative Perfume
Quantity Given (%) 30.00 62.00 1.00 1.00 1.00 0.50 0.70 2.30 0.50 0.50 0.50
Quantity Taken (gm/ml) 9.0 18.6 0.3 0.3 0.3 0.15 0.21 0.69 0.15 0.15 0.15
Uses Vehicle Antibacterial Antibacterial Antibacterial Antibacterial Thickening agent Solubilizing agent Emollient Emulsifier Preservative Fragrance
Table 2: Composition of non-alcohol herbal hand sanitizer formulation.
Sr. No. 1 2 3 4
Ingredients and Excipients Aloe Vera gel Tulsi leaves extract Lemon grass oil Clove extract
Quantity (ml) q. s. 1 1 1
Antibacterial Activity The screening of antibacterial activity of the extracts against pathogens was performed using disc diffusion method. N utrient agar media was prepared (Table 3), sterilized and aseptically spreaded on four sets of Petri plates (each set containing three plates) which were previously marked as AHS, ANHS and CHS (formulation coding). Microorganisms used were Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Pseudomonas aeruginosa. The plates were inoculated with microorganism suspension and incubated at 37°C for 24 h . Next day filter paper discs loaded with a lcohol based herbal hand sanitizer, non -alcohol hand sanitizer and synthetic hand sanitizer were placed in the respectively marked plates. It was taken care that the sterile discs completely absorbs the formulation. After 24 h test results were observed to determine the efficacy of formulations in terms of zone of inhibition of microorganism. Higher the zone of inhibition, the more effective is the test formulation. [6] Table 3: Composition of nutrient agar culture media.
Ingredients Peptic digest of animal tissue Sodium chloride Beef extract Yeast extract Agar Final pH at 25 C
Quantity (gm/lit) 5.0 5.0 1.5 1.5 15.0 7.4±0.2
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RESULTS AND DISCUSSION Total four bacterial species were selected in the present study to evaluate the antimicrobial activity of the prepared formulations. The antimicrobial potency of prepared formulations was evaluated using disc diffusion method (as mentioned before). After incubation the results observed in terms of zone of inhibition against microorganisms are quoted in Table 4. Alcohol based herbal hand sanitizer (at concentration 400 μg/ml) showed greater inhibition zones against bacterial species Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis and Staphylococcus aureus in comparison to commercially available hand sanitizer. Table 4: Antimicrobial study results.
Name of the Microorganism Escherichia coli Pseudomonas aeruginosa Bacillus subtilis Staphylococcus aureus
Zone of Inhibition (diameter in mm) AHS ANHS CHS Std.(SLS) 36 27 32 32 30 19 25 37 32 25 30 31 32 25 29 30
Fig No. 1: Zone of inhibition in diameter on P. aeruginosa.
Fig. No. 2: Zone of inhibition in diameter on E. coli.
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Fig. No. 3: Zone of inhibition in diameter on B. subtilis.
Fig. No. 4: Zone of inhibition in diameter on S. aureus.
Fig. No. 5: Result of antimicrobial study of Hand-sanitizer against various pathogens. DISCUSSION The prepared formulation of herbal hand sanitizer showed significant results against th ree bacterial species. The significance was found to be more in comparison to the commercially available hand sanitizer or hand disinfectant. The composition ( Ocimum sanctum Eugenia caryophyllus and cymbopogon flexuous) has been attributing with properties like anthelmintic, antimicrobial, antiinflammatory and analgesic etc. The alcohols have excellent, rapid (within few seconds) germicidal activity against vegetative bacteria, fungi, and many viruses and antimicrobial activity is based on protein denaturation of microorganisms. Alcohol based hand. sanitizers are highly effective against mycobacteria (the bacteria most resistant to the disinfection process) and multidrug-resistant pathogens. Alcohol based herbal hand sanitizer are approximately 100 times more effective against microorganism than commercially available hand sanitizer. Herbal hand sanitizer offer numerous advantages over commercially available hand sanitizer onto both hands,
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usually requires only 15 to 30 seconds. Whereas vigorous friction, rinsing with water, and drying with a towel are not needed like hand disinfectants or soaps. CONCLUSION Hands are the most common mode of transmission of pathogens to patients and proper hand hygiene can prevent health careassociated infections and the spread of antimicrobial resistance. Scientific evidence and ease of use support of alcohol-based hand sanitizers during patient care. It may be concluded that Herbal Hand Sanitizer has a significant anti-microbial effect on the specified microorganisms except P. aeruginosa. Thus, there is immense potential in establishing the use of antimicrobial herbal products as a measure to control the multidrug resistant microbes as well as to check their spread through hands from one geographical region to another. REFERENCES 1. Vyaset P., Antimicrobial Activity of Ayurvedic Hand Sanitizer, International Journal of Pharmaceutical and Biological Archives 2011; 2:2: 766. 2. http://en.wikipedia.org/wiki/Hand_sanitizer. 3. Kokate CK, Purohit AP, GokhaleSB. Text book of Pharmacognosy. 50 th ed. Pune: Nirali Prakashan; 2014.p. 14.6714.68,14.85,14.52. 4. Joshi MG., Kamat DV., Kamat SD., Evaluation of Herbal Hand Wash Formulation, Natural Product Radiance 2008; 7:5: 413415. 5. Wani NS., Bhaleroa AK., Ranaware VP., Zanje R., Formulation and Evaluation of Hand Sanitizer, International Journal of PharmTech Research 2013; 5:1: 40-43. 6. Kokare CR. Pharmaceutical microbiology experiments and techniques. 2nd ed. Nashik: Career Publication; 2008.p. 201-202.
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