Rasool Bazigha Kadhim et al / IJRAP 2011, 2 (6) 1811-1813

International Journal of Research in Ayurveda & Pharmacy

ISSN 2229-3566 Research Article www.ijrap.net

FORMULATION AND EVALUATION OF HERBAL HAND WASH FROM

MATRICARIA CHAMOMILLA FLOWERS EXTRACTS Ali Heyam Saad1, Shehab Naglaa Gamil2, Rasool Bazigha Kadhim1* and Rana Samour1

1Department of Pharmaceutics and Pharmacy Practice, Dubai Pharmacy College, Dubai, UAE 2Department of Pharmacognosy, Faculty of Pharmacy, Cairo University, Egypt

Received on: 19/10/11 Revised on: 04/11/11 Accepted on: 20/04/11

*Corresponding author Bazigha K. Abdul Rasool, PhD, Associate Prof. ,Department of Pharmaceutics and Pharmacy Practice, Dubai Pharmacy College, Dubai, UAE Email: bazigha_rasool@hotmail.com, bazigharasool@yahoo.com ABSTRACT Nasocomial infection has emerged as a critical issue in hospital care outcome, resulting in substantial morbidity and mortality. The hands of health care workers are the primary routes of transmission of infection to patients. Hence, it brings up the use of antiseptic for hand washing purposes. Many of the antiseptic available in market are alcohol based sanitizers which have some shortcomings or adverse effects. Their frequent use can lead to skin irritation. Chamomile is one of the most widely used and well-documented medicinal plants in the world. This study aimed to formulate effective herbal hand wash using Matricaria chamomilla (German chamomile) flowers with emphasis on safety and efficacy and to avoid the risk posed by synthetic antimicrobials. Evaluation of the antimicrobial activity against skin pathogens of the prepared herbal hand wash was performed using disc diffusion method. Its efficacy was checked and compared with the commercial ones. Results revealed that chamomile soap formulation was more efficient in reducing the number of organisms from hands than the commercial antiseptic soaps thus it can be used as an antiseptic soap with less or no side effects. Keywords: Herbal hand wash, Formulation, Evaluation, Chamomile Extracts, Skin pathogens, Staphylococcus aureus, Pseudomonas aeruginosa. INTRODUCTION Skin being the most exposed part of our body requires protection from skin pathogens. Nasocomial infection has emerged as a critical issue in hospital care outcome, resulting in extended hospitalization and substantial morbidity and mortality1. The hands of Health Care Workers (HCWs) are the primary routes of transmission of multidrug resistant pathogens and infection to patients. Hence, it brings up the use of antiseptic for hand washing purposes2. Many of the chemicals antiseptic are now available in market as alcohol based sanitizers, cholorohexidine products, etc. These soaps or solutions help reduce health care associated transmission of contagious disease more effectively3 but they have some shortcomings or adverse effects. Their frequent use can lead to skin irritation and also resistance among pathogens4. Organisms such as Staphylococcus aureus and Pseudomonas aeruginosa are some of the causative agents of skin infections5. Some studies been conducted had showed that resistance to chemical antiseptic have led to outbreaks6, 7. Historically, plants have provided a good source of antiinfective agents. Plant based antimicrobials represent a vast untapped source for medicines. They are effective in the treatment of infectious diseases while simultaneously mitigating many of the side effects that are often associated with synthetic antimicrobials. Plants containing flavonoids and polypeptides used in traditional medicine have been found to be active against a wide variety of microorganisms8. Matricaria chamomilla (German chamomile) is from the Asteraceae family and is the major source of blue oil and flavonoids9, 10. It has been widely used in folk and traditional medicines for its multi-therapeutic, cosmetic and nutritional values11,

12. Chamomile flowers contain essential oil 0.4-2% v/w13 which has antibacterial and fungicidal action14, 15. In addition to that, flowers contain more than 120 chemical constituents and have been identified as secondary metabolites16 including 28 terpenoids, 36 flavonoids, and 52 additional compounds with potential pharmacological activity17. Components, such as α-bisabolol and cyclic ethers are fungistatic, whereas chamazulene and α-bisabolol are antiseptics18. Chamazulene, α -bisabolol, flavonoids and umbelliferone displayed antifungal properties against trichophyton mentagrophytes and Candida albicans, all of them have the

strongest activity against Gram-positive and Gram-negative bacteria19. Despite its widespread use as health care remedy and its in vitro antibacterial and antifungal effectiveness against Staphylococcus and Candida, no trial has been documented about its use as liquid hand wash soap. In the present context, Matricaria chamomilla had been selected in preparing herbal hand wash. Chamomile flowers were extracted with alcohol to assure maximum taking out of all the phytochemicals which act on skin pathogens and ensure maximum antimicrobial efficiency. Chamomile extract hand wash soap was then prepared, screened against skin pathogens and checked for its efficacy. MATERIALS AND METHODS Plant collection Samples of Matricaria chamomilla flowers were collected during March-April (2011) from plants cultivated in Haj Saed Campus garden, Muhaisnah-1, Dubai, United Arab Emirates and verified in the Department of Pharmacognosy at Dubai Pharmacy College, UAE. Preparation of plant extracts Fresh flowers of the investigated plant (200 g) were mixed and homogenized with ethanol 70%in a mixer, followed by exhaustive maceration at room temperature with the 70% alcohol (2 liters x2) then filtered. The combined alcoholic extract was evaporated under reduced pressure. The residue free solvent (12.5 g) was suspended in distilled water (40 ml) to be used for further study. Preparation of chamomile hand wash soap The herbal hand wash was prepared by adding 4ml of the suspended water extract (1.25g /4 ml w/v) to 3 g of sodium lauryl sulphate (SLS)20. The solution was made homogenous under room temperature and used for the antibacterial screening. Blank soap was prepared as per the previous procedure without herbal extract. The candidate formula was compared to standard soaps commercially available in UAE pharma market including: Savlon® (Novartis, UK) and Huiji® (Huiji Biotechnology, China). The commercial herbal soap contains green tea, aloe, wild chryeanthemum and lemon. Antimicrobial assay The pathogens were recovered from the hands of HCWs. Amongst the isolated pathogens Staphylococcus aureus and Pseudomonas aeruginosa were selected and identified and used for this study. The

Rasool Bazigha Kadhim et al / IJRAP 2011, 2 (6) 1811-1813

International Journal of Research in Ayurveda & Pharmacy

anti-microbial activity of the hand wash soaps was investigated by the disc diffusion method21. Tested culture organisms used were S.aureus, Ps.aeruginosa, Bacillus subtilis, Escherikia coli and Candida albicans. Nutrient agar was used as a culture medium. The herbal hand wash soap was vigorously shaken for homogeneity. One mL portion of the soap solution was added to 20 mL of sterile molten culture media in Petri-dishes and allowed to set. Then a 0.1 ml of standard inoculum of each test organism was streaked on the plates which were then incubated under the standard conditions. Following incubation, plates were observed for the presence or absence of microbial growth. In similar ways, discs of commercial antiseptic soaps and blank soap were prepared. In vivo efficacy study Experimental design Four volunteer groups (n=4) were involved for in vivo efficacy study of four different soaps (chamomile hand soap formulation, blank soap, commercial standard soap, commercial herbal soap). Volunteers with damaged skin and those who were on antimicrobial therapy were excluded. Study was done after approval of ethical committee of the College. The protocol had been explained and the informed consents were obtained from the volunteers. Procedure A rinse wash method was adopted for this study. Total 40 palm washes were taken for a single volunteer for a single soap. Control palm washes were taken by use of sterile water only. Rinse palm for 10 seconds with small amount of tap water. Palm is completely lathered with the desired soap for a specified time (1 min). Hand wash samples were collected in the sterile beakers under sterile conditions. Temperature of sterile water was near to 40 °C for 30 seconds and 75 mL water was 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 37 °C for 18-24 hr22. Bacteriological study Point inoculation method as mentioned by Miles and Mishra23, was used for viable count of different bacterial flora. RESULTS AND DISCUSSION Preliminary antimicrobial sensitivity screening tests findings shown in Table 1. Chamomile soap formulation proved to be beneficial and had an excellent activity against all the tested organisms. A significant (p<0.05) result was obtained against S. aureus. Hence it was encouraging to be used in the preparation of herbal hand wash. Many of these organisms are natural flora of the skin and mucous membranes infections of man24. Data for in vitro antimicrobial evaluation of chamomile soap is presented in Table 2. Chamomile soap formulation had greater antimicrobial activity against the test organisms than the commercially available hand soaps. No significant effect obtained by the blank soap which indicates that the antibacterial activity of herbal hand wash is solely due to the activity of active components. The activity against Staphylococcus aureus was of significant interest (p<0.05). The skin carries large numbers of bacteria flora, mainly Gram-positive picked up from the various objects with which it comes in contact. Staphylococcus aureus is one of these natural flora commonly found on the hands, face and in deep layers of the skin. Staphylococcus aureus is ubiquitous and is not easily eliminated by routine washing and scrubbing even with some antiseptic soap26. Thus, the potency of the chamomile herbal soap against Staphylococcus aureus was very remarkable. Similar results were reported by Fuerst using ethanol extract against Staphylococcus aureus isolates from boils, carbuncles, breast abscess, and infantile-impetigo25. The herbal soap was not tested against Gram-negative bacteria, since they are not auchthchonous skin flora. However, they are only encountered when the normal Gram-positive bacteria are depleted by antibiotic application, other non-physiological conditions such as

the spillage from the gastrointestinal tract (GIT) onto the skin around the anal and genital regions as well as on soles of the feet 26. Table 3 and Fig.1 (a-d) represents growth reduction in bacterial flora obtained after the use of chamomile herbal soap formulation, Savlon, commercial herbal soap and blank soap. Highest reduction was observed by chamomile herbal soap followed by commercial herbal soap, Savlon and blank soap. The % of reduction shown by chamomile herbal soap was 80.57%, while it was found to be to be least in case of control blank soap (67.74%). The commercial herbal antiseptic soaps showed more reduction than Savlon (68.29%). A percentage reduction in bacterial flora was calculated using the formula. % Reduction= Control (cfu/ml) – Test (cfu/ml) X 100 ……. (eq. 1) Control (cfu/ml)

The significant (p<0.05) antimicrobial activity of the prepared formula can be attributed to the presence of tannins in the water extract. Tannins are water soluble polyphenols27 and have been reported to be bacteriostatic or bactericidal against Staphylococcus aureus by acting on the membranes of the organisms28. The results clearly showed that the natural antimicrobials in the prepared herbal soap were far more active than the synthetic antimicrobials used in commercial antiseptic soap. CONCLUSION In the present study, natural active constituents in chamomile extract manifested superior inhibition against skin pathogens than synthetic antimicrobials present in the commercially available antiseptic soaps. Therefore, these compounds were extracted and incorporated in soap bases in order to prepare superior antiseptic soap with less or no side effects. Hence, a new way can be found to combat antibiotic resistance of pathogenic organisms and provide safe and healthier living through germfree hands, although the removal is not 100%, but a major number can be reduced with natural economic and safe chamomile soap. ACKNOWLEDGMENT Authors are grateful for and would like to express gratitude to the Dean of Dubai Pharmacy College and Dubai Pharmacy College undergraduate students, Hamda J. Obaid and Nourhan A. Baroud for their support in lab work. REFERENCES 1. Boyce JM and Pittet D. Guideline for Hand Hygiene in Health-Care Settings.

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13. Das M, Mallavarapu GR and Kumar S. Isolation of a genotype bearing fascinated capitula in chamomile (Chamomilla recutita). J Med Aromat Plant Sci 1999; 21: 17-22.

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Table 1 Preliminary Antimicrobial Activity of Matricaria chamomile

Test microorganisms Antimicrobial activity IZD (mm)± SD*

Staphylococcus aureus 19.5± 0.55** Pseudomonas aeruginosa 12.2± 0.33

Bacillus subtilis 13.4± 1.00 Escherikia coli 10.2± 0.23

Candida albicans 8.20± 0.66

*: Values calculated as means ± SD (n=3); **: Results are significant (p<0.05) and

IZD: Inhibition zone diameter.

Table 2: In Vitro Antimicrobial Evaluation of Chamomile Hand Wash Soap

Test organisms Zone of Inhibition (mm)

Chamomile soap Standard soap Blank

soap Staphylococcus aureus + + -

Pseudomonas aeruginosa

+ + -

-: indicates No inhibition zone or less than 7mm; +: presence of

microbial growth. Values are means of triplicates.

Table 3: Reduction in Hands’ Bacterial Flora

Formulation Bacterial flora reduction

Control (cfu/ml)

Test (cfu/ml)

Reduction (%)

Chamomile soap formulation 28.05Í103 5.35Í103 81 Blank soap 22.45Í103 7.5Í103 68

Commercial Herbal soap 18.4Í103 5.32Í103 72 Savlon 22.75Í103 7.3Í103 68

Fig 1: Effect on general transient bacterial microflora after use of: a) chamomile herbal soap; b) blank soap; c) Savlon; d) commercial herbal soap