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Zeeshan Afsar & Salma Khanam. Int. Res. J. Pharm. 2016, 7 (8)

INTERNATIONAL RESEARCH JOURNAL OF PHARMACY

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ISSN 2230 – 8407

Research Articles
FORMULATION AND EVALUATION OF POLY HERBAL SOAP AND HAND SANITIZER
2
Zeeshan Afsar * 1 & Salma Khanam
1Department of Pharmacognosy, Farooqia College of Pharmacy, Mysore, India
2Department of Pharmacognosy, Al-Ameen College of Pharmacy, Bangalore, India
*Corresponding Author Email: [email protected]

Article Received on: 02/07/16 Revised on: 7/29/16 Approved for publication: 04/08/16

DOI: 10.7897/2230-8407.07896

ABSTRACT

A poly herbal soap and hand sanitizer was formatted using the leaf and bark extracts of Cassia fistula, Ficus religiosa and Milletia pinnata. The
antimicrobial efficacy of the prepared formulations was tested using agar well diffusion method against the organisms E coli (MTCC-1698), S aureus
(MTCC-1143) and P aeruginosa (MTCC-2453). The prepared poly herbal formulations demonstrated good antimicrobial effect. The prepared
formulations were evaluated for various physicochemical parameters for which good characteristics were observed. Hence owing to good antimicrobial
effect and acceptable parameters, the prepared formulations can be further standardized and used as effective antiseptics and disinfectants.

Keywords: Cassia fistula, Ficus religiosa, Milletia pinnata, Hand sanitizer, Zone of inhibition.

INTRODUCTION different parts of Cassia fistula has been reported. It was found to
contain Flavonoids, phenolic compounds, and proanthocyanidins.
5
Opportunistic excellence causes nosocomial infections; and multidrug- Literature survey of this plant shows that it is
6 7
resistant pathogens are commonly involved in nosocomial infections. reported to pose' good antimicrobial properties. , The
Multidrug resistant infectious diseases of bacterial and fungal origin are chemical composition of Milletia pinnata was studied. It was reported to
leading killers and account for approximately 25% of global deaths and contain saponins, tannins, carbohydrates, alkaloids, sterols and
8
are difficult The alarming rate at which the human pathogens like flavonoidal glycosides. The pharmacological
1 to treat. properties of Milletia pinnata reported in literature also suggest it has
9
Staphylococcus aureus (Methicillin & Multiantibiotic resistant), Escherichia good antimicrobial properties. The phytochemical
coli, Pesudomonas aeruginosa, Candida albicans and Cryptococcus screening of the bark extracts of Ficus religiosa showed the presence
neoformans are evolving themselves as multidrug resistant “Superbugs” tannins, saponins, flavonoids, steroids, terpenoids and cardiac glycosides.
ten
towards the newly generated classes of antibiotics, demands for The pharmacological properties of Ficus
exploration of new chemical sources from biodiversity and develop religiosa showed that the methanolic extracts showed to poses good
11
therapeutic regimes to combat infectious superbugs. antimicrobial properties.
2
Hand hygiene is a vital principle and
exercise in the prevention, control, and reduction of healthcare acquired The aim of the present study was to prepare hand sanitizer and soap
infections. Right hand washing and drying methods stop the chain of formulations using the extracts of Cassia fistula, Milletia pinnata and
transmission of deadly pathogens (from the contaminated surface/site) Ficus religiosa and to investigate the antimicrobial activity of the extracts
form hands to other parts of the body. Hand sanitization is the preeminent against the common organisms which cause nasocomial infections.
aid in preventing nosocomial infections caused by different opportunistic Furthermore to evaluate the stability and phycochemical parameters of
microorganisms. the prepared formulations so that they can be further standardized and
3
The hands of health care workers are the primary used commercially.
mode of transmission of these multidrug-resistant pathogens and
infections to patients. Skin being the most exposed part of our body
requires protection from skin pathogens. Hence it brings up the use of MATERIALS AND METHODS
hand sanitizers and soaps for hand wash purposes.
3
Many of chemical Collection of samples
antiseptics available in the market are alcohol based sanitizers.
These formulations including soaps and solutions reduce health care The plants Cassia fistula, Milletia pinnata and Ficus religiosa
associated transmission of contagious diseases but they have some were collected from Mysore district, the specimen were authenticated at
short comings or adverse effects, their frequent use RRL, Bangalore.
3
can lead to skin irritation and also resistance among pathogens.
Plants are rich in a wide variety of secondary metabolites, such as Preparation of extracts
phenolic compounds, tannins, terpenoids, alkaloids, and flavonoids,
which have been found to have antimicrobial properties. Hence there is The leaves and bark of Cassia fistula, Milletia pinnata and Ficus religiosa
an upsurge of developing herbal disinfectants and evaluate its were dried in hot air oven at 35oC for three days, powdered to a mesh
4
effectiveness. The chemical analysis of size of # 40 and stored in air tight

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Zeeshan Afsar & Salma Khanam. Int. Res. J. Pharm. 2016, 7 (8)

containers. The powder was then successively by refluxation for Determination of clarity, color and odor: Clarity and color was
eight hours using five different solvents with increasing polarity. checked by naked eyes against white background, the odor was
Viz: - Petroleum ether, Chloroform, Ethyl acetate, Methanol, and smelled.
40% methanol.
pH: The pH of all the prepared formulations was determined by
Preliminary antimicrobial screening of the extracts using Digital pH Meter. The formulations were dissolved in 100
ml of distilled water and stored for two hours. The measurement
All the extracts of leaf and bark were subjected to preliminary of pH of formulation was done in previously calibrated pH
antimicrobial screening by agar well diffusion method against the meter.
organisms E coli (MTCC-1698), S aureus (MTCC-1143)
and P aeruginosa (MTCC-2453). The extracts which demonstrated Determination of percentage free alkali: About 5 gm of sample
maximum activity were selected for the formulation. was taken in a conical flask and added to it into 50 ml of
neutralized alcohol. It was boiled under reflux on a water bath for
Preparation of formulations 30 minutes, cooled and 1 ml of phenolphthalein solution was
added. It was then titrated immediately with 0.1N HCL.
Three extracts that showed maximum antimicrobial activity were
prepared in combinations in two different concentrations ie 250 Foam Height: 0.5gm of sample of soap was taken, dispersed in
mg each (750 mg) and 500 mg each (1500 mg) and these 25 ml distilled water. Then, transferred it into 100 ml measuring
combinations of extracts were incorporated in the prepared cylinder; volume was make up to 50 ml with water. 25 strokes
formulations. were given and stand till aqueous volume measured upto50 ml
and measured the foam height, above the aqueous volume.
Herbal soap
Foam Retention: 25 ml of the 1% soap solution was taken into a
Solidified basic glycerine soap was broken down to smaller pieces 100ml graduated measuring cylinder. The cylinder was covered
and melted on water bath. 1.5grams of the extract combinations with hand and shaken 10 times. The volume of foam at 1-minute
were added to the melted soap along with 5ml of ethanol. 0.033 g intervals for 4 minutes was recorded.
of stearic acid, 1ml each of cinnamon oil and citronella oil was
added to the melted soap. The melted soap was gently mixed for Alcohol Insoluble Matter: 5 gm of sample was taken in a conical
about 30 minutes and molded in circular moulds. The soap was flask. Added it to 50 ml of warm ethanol and shaken vigorously to
allowed to solidify at room temperature until set and kept under dissolve The solution was filtered through a tarred filter paper with
physical observation for any characteristic changes. 20 ml warm ethanol and dried it at 105ºC for 1 hour. The weight
of dried paper was taken.

Preparation of Hand sanitizer Formula

% alcohol insoluble matter = Wt. of residue × 100 / Wt. of
The hand sanitizer was prepared using the following formula. sample

Ingredients Quantity taken (10ml) High Temperature Stability: Liquid soap was allowed to stand
Extract combination 0.75 g at 50ºC for one week. The stability of liquid soap was observed
Citronella oil 1.0 ml during this period. The sample which was homogeneous and
Cinnamon oil 1.0 ml stable liquid after standing was indicated as stable and the sample
Carbopol 0.1 g in which the crystals were roughened and the sample in which
Triethanolamine 0.1 g precipitation was caused; then liquid was said to be as unstable.
glycerine 0.5 ml
Polysorbate-20 0.1 ml
Perfume Qs
Antimicrobial testing of the prepared formulations
Methyl Paraben 0.1 mg
Alcohol 4.0 ml The prepared soap and hand sanitizer were subjected to
Water 2.0 ml antimicrobial screening by agar well diffusion method.
Organisms used were E coli (MTCC-1698), S aureus (MTCC
Procedure 1143) and P aeruginosa (MTCC-2453). One gram of soap was
mixed with 5 ml of sterile water; 1ml of sanitizer was mixed with 5
The extract combinations were added in water and all the ml DMSO and used for evaluating the antimicrobial activities. The
ingredients were added and stirred well except triethanolamine, plates were incubated at 370C for 24 hours and the zones of
inhibition were recorded.
alcohol and perfume. Citronella oil and cinnamon oil were added
to triethanolamine along with perfume stir well and both the
RESULTS
contents were mixed together thoroughly and the volume was
made up using alcohol.
Preliminary antimicrobial screening of the extracts
Evaluation of physicochemical parameters of the prepared The initial susceptibility testing of the different extracts of leaves
formulations and stem bark of Cassia fistula, Milletia pinnata and Ficus religiosa
was done by using agar diffusion method. The stem bark extracts
Various physicochemical parameters which are mentioned below of Cassia fistula showed considerable antimicrobial activity in
were performed to establish quality of the prepared formulations. terms of zones of inhibition. The extracts of leaves showed no
inhibition zones. The leaves and bark of Milletia pinnata and Ficus
religiosa also showed significant zones of inhibition. The
observations are recorded in
Table 1.

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Zeeshan Afsar & Salma Khanam. Int. Res. J. Pharm. 2016, 7 (8)

Table 1: Zones of inhibition (mm) of leaf and bark extracts of Cassia fistula, Milletia pinnata and Ficus religiosa

Extracts E coli S aureus P aeruginosa

CB-PET 8.0 6.0 16.0
CB-CHL 14.0 __ 16.0
CB-ETH 18.0 18.0 16.0
CB-MOH 20.0 18.0 20.0
CB-40MOH 12.0 8.0 6.0
CL-PET __ __ __
CL-CHL __ __ __
CL-ETH __ __ __
CL-MOH __ __ __
CL-40MOH __ __ __
MB-PET 6.0 10.0 12.0
MB-CHL 12.0 8.0 16.0
MB-ETH 16.0 12.0 16.0
MB-MOH 18.0 14.0 18.0
MB-40MOH 12.0 10.0 16.0
ML-PET 12.0 __ 8.0
ML-CLOSE 4.0 __ 6.0
ML-ETH 8.0 __ 8.0
ML-MOH 6.0 __ 10.0
ML-40MOH 6.0 __ 12.0
FL-PET __ __ __
FL-CHL __ __ __
FL-ETH __ __ __
FL-MEOH 8.0 14.0 10.0
FL-40MOH 15.0 __ 17.0
FB-PET __ __ __
FB-CHL __ __ __
FB-ETH 18.0 16.0 16.0
FB-MEOH 10.0 __ 12.0
FB-40MOH __ 8.0 __
Note: CB= Cassia fistula bark, CL= Cassia fistula leaf, MB= Milletia pinnata bark, ML= Milletia pinnata leaf. FL= Ficus religiosa leaf, FB= Ficus
religiosa bark, PET= Petroleum ether, CHL= Chloroform, ETH= Ethyl acetate, MOH= Methanol, 40MOH= 40% Methanol. __ indicate no
activity.

Preparation and evaluation of physicochemical parameters appearance characteristics as well as the pH was found in the
of formulations range of 6.5 to 7.5 which is the desired pH. Other such
parameters as percentage free alkalie, foam height, foam
The physicochemical parameters of the prepared soap and hand retention, alcohol insoluble matter, test for chlorides and high
sanitizer were determined. Parameters such as color, odor, temperature stability were determined; the results are tabulated in Table 2.
appearance, pH were tested. The formulations performed well

Table 2: Physicochemical parameters of hand sanitizer and soap formulations

Formula Color Odour Appearance pH % free alkalie Foam Foam Alcohol High temperature
height retention insoluble stability
(cm) (min) matter
Hand Cream Aromatic Good 7.5 0.15 ten 2.5 4.5 Good
sanitizer brown

Soap Light Fragrant Good 7.0 0.27 27 6.0 18.0 Soap melts above
brown 600C

Antimicrobial screening of the prepared formulations three extracts were prepared in combinations and incorporated
in formulations in two different concentrations ie 250 mg each
The extracts that exhibit maximum antimicrobial activity were (750 mg) and 500 mg each (1500 mg). Both the formulations
ethyl acetate bark extracts of Cassia fistula, Ficus religiosa and demonstrated good zones of inhibition from 18 to 26 mm.
methanolic bark extracts of Cassia fistula and Milletia pinnata The results are tabulated in Table 3.
with zones of inhibition ranging from 14 to20 mm. Hence these

Table 3: Antimicrobial screening of the prepared formulations

Zones of inhibition in mm
No 01 Formulation E coli S aureus P aeruginosa
02 HS-1 18.0 20.0 22.0
03 HS-2 26.0 22.0 24.0
04 SP-1 20.0 18.0 20.0
SP-2 22.0 24.0 22.0
Note: HS-1 = Hand sanitizer with 750 mg concentration of extracts, HS-2 = Hand sanitizer with 1500 mg concentration of extracts, SP-1 = Soap with 750 mg
concentration, SP- = Soap with 1500 mg concentration.

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Zeeshan Afsar & Salma Khanam. Int. Res. J. Pharm. 2016, 7 (8)

DISCUSSION AND CONCLUSION 8. Simin SK, Usmanghani M, Shaiq A, Viqaruddin A.

Chemical constituents from the seeds of Pongamia pinnata
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that most of the extracts exhibits good antimicrobial effect among antibacterial properties of leaves of Pongamia pinnata Linn.
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