Introduction
The skin is one of the most uncovered part of the body and it must be protected from pathogenic microorganisms like bacteria, viruses, fungus etc. Hand washing is essential for protecting the skin layer from pathogenic microorganisms and preventing the spreading of communicable diseases. Hands are generally primary mode of transmission of microbes and infections. So, hand cleanliness is maintained by using hand wash.1
Andrographis paniculata which is also popularly known as green chiretta and it belongs from the family Acanthaceae. As andrographolide is chief active constituent of the plant, it could be considered as excellent candidate of antimicrobial drug development. It is one of the highly used potential medicinal plants in the world, currently. Traditionally, it has been believed to be a treatment for bacterial infections and some diseases. It exhibits antibacterial, antioxidant, anti-inflammatory, anticancer, and immune stimulating properties.2
Nowadays, herbal plant bio actives are preferred over chemicals for formulation due to the fewer side effects. There are large numbers of medicinal plants available which are extensively used in different treatments of skin infections and possess pharmacological activity. The therapeutic activity of any plant depends upon their active constituents. Secondary metabolites found in plants include glycosides, alkaloids, tannins, saponins, terpenoids and flavonoids among others which are responsible for various pharmacological activity. According to the literature study, Andrographis paniculata leaves extract possess antimicrobial and antibacterial activity. Thus, we aimed to formulate and evaluate herbal handwash gel using most effective excipients in the preparation of herbal handwash.3
Materials and Methods
Preparation of ethanolic extract of Kalmegh
The leaves of Andrographis paniculata (Green chirretta) were collected from Raigarh, Chhattisgarh, India during August-September2021 and its botanical identity was confirmed by Dr. Saeargaonkar SL, Department of Plant Breeding, College of Agriculture & Research Station Raigarh, Chhattisgarh, India. Using the soxhlet apparatus, one kg shade dried powdered leaves were extracted with ethanol. A concentrate was prepared by distilling off the solvent from the total extract and the concentrate was placed on a water bath and converted toasyrupymass and then completely dried by evaporating (31gram). 4
Qualitative analysis
The preliminary phytochemical screening was carried out for testing the different phytoconstituents present in alcoholic extract of Andrographis paniculata (Kalmegh). The phytoconstituents tests were carried out and the results are represented in following-
Identification of phytoconstituents by chemical tests5, 6, 7, 8, 9, 10
Test for alkaloid
Wagner’s test: 1 ml of concentrate added in 2 ml of Wagner's reagent. The presence of alkaloids indicated by the precipitates reddish brown color.
Test for glycosides
Fehling’s test: A small amount of extract is hydrolyzed with 5 ml of hydrochloric acid for several hours in a water bath and the hydrolyzate is subjected to Fehling's test. Add 2 ml of Fehling's solution (1 ml of Fehling's solution A and 1 ml of Fehling's solution B), add 2 ml of the extract, mix and bring to a boil. The presence of reducing sugars indicated by the precipitates yellow or red color.
Test for saponins
In a water bath, 2 g of powdered sample was boiled in 20 ml of distilled water and filtered. 10 mL of the filtrate was mixed with 5 mL of distilled water and vigorously shaken to form a stable persistent froth emulsion.
Test for phytosterol
Salkowski test: Phytosterols were detected using the Salkowski technique. 1 ml concentrated sulphuric acid was applied to 1g plant extract and left to sit for 5 minutes in this test. The presence of phytosterols is shown by the creation of a golden yellow tint in the lower layer after shaking.
Test for flavonoids
Concentrated sulphuric acid was used to treat the extract. The presence of anthocyanins is indicated by a yellowish orange appearance. Flavones are found in yellow to orange colors, while flavonones are found in orange to crimson colors.
Preparation of herbal hand-wash
Herbal handwashgelwaspreparedwiththehelpofCarbopol-934as a gel forming agent in the concentration of 1% w/w for a period of one day. Then thein fledged was agitated with the help of amechanical agitator to confirm even distribution of the gel. Then adjust the pHto7.0 by adding small quantities of TEA with nonstop stirring. Then, the base was used to integrate extract of Andrographis paniculata to develop a formulation of herbal handwash. To this Sodium lauryl sulfoacetate (0.4g) as surfactant and Rosewater (15ml) was also assimilated. 11, 12
Antibacterial activity
The antibacterial activity of herbal handwash was investigated using the agar well diffusion method. The test organisms were dispersed with bacterial suspension of optical density of 0.8 at 800 nanometers. Wells of 8-millimeter diameter were then made on plate. Each well was filled with 100ml of extract and subjected to incubation at 37°Cfor the duration of 24 hours. After 24 hours of incubation antagonist action is determined using zone of inhibition (centimeter). The antibacterial activity of Herbal handwash was tested against microorganism in triplicates. Ciprofloxacin is used as standard. The organism used are S. aureus, E. coli, S. auroginosa and B. subtilis. 13
Table 1
The phytochemical evaluation of Andrographis paniculata leaves extract.
Physico-chemical evaluation of formulation: 14, 15, 16, 17
Physico-chemical evaluation of the herbal hand wash gel was determined by appearance, pH, viscosity, foam height and retention.
Homogeneity and appearance
Morphological or physical evaluation of hand wash gel for color, odor and texture were carried out by visual inspection.
pH
200 ml distilled water was taken and 2 gm of herbal handwash was dissolved in it. A previously standardized digital pH meter was used to determine the pH of the solution.
Determination of viscosity
Digital Brookfield viscometer was used to determine the viscosity of herbal handwash.
Determination of foam height
Herbal gel-based hand wash sample of about 1gm was taken and it was mixed with 50 ml of water. It is after transferred to 250 ml stoppered beaker. With distilled water, the capacity was increased to 100 mL. After 25 motions, the solution was allowed to stand until the aqueous volume reached 100 ml and the foam height was measured above the solution.
Results and Discussion
Phytochemical Screening- The qualitative analysis of alcoholic Andrographis paniculata leaves extract was performed for phytochemical screening. The results were shown in Table 1.
The results show that saponins, flavonoids, terpenoids, alkaloids, and glycosides are present in the alcoholic extract of Andrographis paniculata leaves. The extract was further formulated in gel dosage form and then evaluated for its anti-microbial activity.
Antibacterial activity-18
The antibacterial efficacy of the formulation was tested on S. aureus, E. coli, S. auroginosa and B. subtilis. Herbal hand wash has shown significant antimicrobial activity against B. subtilis (Table 2).
The results show that the herbal hand wash formulations are used in different concentrations was shown significant effect against B. subtilis. The different concentrations of 10, 20, 50 mg/ml of formulation the zone of inhibition against B. subtilis was found to be 6.00 ± 1.00, 9.26 ± 1.63 and 13.23 ± 1.12 mm, respectively. On the other side, ciprofloxacin (5 μg/ml) was used as standard drug for positive control and it shows 18.12± 1.42 mm zone of inhibition against B. subtilis. In previous studies, Lai Xiaoping et al. (2014), received patent grant (CN103301049A) entitled “Andrographis paniculata hand sanitizer and preparation method thereof”. So, results confirmed that Andrographis paniculata leaves alcoholic extract containing formulation has good potential as herbal handwash.
Evaluation of formulation
Herbal handwash was also evaluated by physical parameters (Table 3) physical and chemical standards like odor, color, consistency, pH, viscosity, foam retention and foam height were performed and results were found to be in the acceptable limits.
Previously, Patel et al. (2017), formulated and evaluated herbal hand wash containing ethanolic extract of Glycirrhiza glabara root extract. On comparison with that study, our study shows concordant significant antibacterial activity as well and used in future after clinical trials.
Conclusions
An Andrographis paniculata leaves extract containing herbal hand wash was developed with potential antibacterial and enhanced quality using Andrographis paniculata leaves extract as antibacterial agent, Carbopol-934 as gelling agent, TEA as neutralizer and Sodium lauryl sulfoacetate (SLSA) as surfactant. The antibacterial efficacy results confirmed its effectiveness of hand wash against B. subtilis micro-organism. The finding also revealed that herbal hand wash are promising formulation preparation which protect us from many daily encounter bacteria and beneficial to eliminate various skin disorders. As a future scope, these preclinical findings must be extrapolated to human subjects in order to confirm their clinical value. Finally, it is concluded that this herbal gel hand wash provided an effective and safe alternative to existing marketed hand wash gels.
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