WORLD JOURNAL OF PHARMACY

Desai et al.
AND PHARMACEUTICAL SCIENCES
World Journal of Pharmacy and Pharmaceutical Sciences
SJIF Impact Factor 7.632

Volume 10, Issue 6, 1552-1563 Research Article ISSN 2278 – 4357

FORMULATION AND EVALUATION OF HERBAL ANTI-

MICROBIAL CREAM CONTAINING HIBISCUS ABELMOSCHUS
LINN EXTRACT

Anita R. Desai1*, Laxman S. Vijapur2, Chetankumar A. Teli3 and Sachin Terdal4

1
Professor and HOD Department of Pharmaceutics, H.S.K. College of Pharmacy Bagalkot-
587101.
2
Assistant Professor Department of Pharmaceutics, H.S.K. College of Pharmacy Bagalkot-
587101.
3
Assistant Professor Department of Pharmaceutics, BLDE’S College of Pharmacy
Basavanbagewadi-586203.
4
PG research scholar Department of Pharmaceutics, H.S.K. College of Pharmacy
Bagalkot-587101.

Article Received on
ABSTRACT
05 April 2021, Hibiscus abelmoschus fractions has previously demonstrated good
Revised on 26 April 2021,
Accepted on 16 May 2021 antimicrobial & antifungal property, to exploit the above property we
DOI: 10.20959/wjpps20216-19095 have conducted a study by formulating & evaluating the cream
containing Hibiscus abelmoschus oil extract and to study of anti-

*Corresponding Author
microbial activity, as creams can be easily applied topically & increase
Dr. Anita R. Desai contact time and minimize side effects as compare to oral
Professor and HOD administration. Oil was extracted from the Hibiscus abelmoschus seeds
Department of
using petroleum ether. Cream was formulated using various excipients
Pharmaceutics, H.S.K.
like borax, beeswax, white soft paraffin and other additives like methyl
College of Pharmacy
Bagalkot-587101.
paraben, propyl paraben, and distilled water. All the formulations were
evaluated for pH, viscosity, extrudability, spreadability, ATR and anti-
microbial activity. The ATR spectral analysis suggested that there was no interaction between
the drug and formulation additives, pH of formulations were fairly constant about 7.4–7.9
within the range of skin pH. All the cream formulations showed good viscosity, all
formulation were spread by applying low share which shows the formulations were having
good spreadability. All the formulation was showing good & F2 showed excellent

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Desai et al. World Journal of Pharmacy and Pharmaceutical Sciences

extrudaibility. All the formulation showed good antimicrobial activity, where as F1 & F2
showed good antifungal activity.

KEYWORDS: Hibiscus abelmoschus L, Cream, antimicrobial, antifungal, formulation.

1. INTRODUCTION
An antimicrobial is an agent that inhibits the microbes or stops their growth. Antimicrobial
drugs are the greatest contribution of the 20th century to therapeutics. Their advent changed
the outlook of the physician about the power drugs on diseases. They are one of the few
curative drugs. Their importance is magnified in the developing countries, where infective
diseases predominate. As a class they are one of the most frequently used as well as misused
drugs.[1]

Creams are semisolid preparation containing one or more medicinal agents dissolved or
dispersed in ether a water-in-oil emulsion or an oil-in-water emulsion, as these semisolid
preparation are used for beautifying, cleansing, for improved appearance & has topical
protective property with local effect for the delivery of drug to the skin disorders. Many
patients and physicians prefer creams to ointments because they are easier to spread and
remove. Pharmaceutical manufacturers frequently manufacture topical preparations of a drug
in both cream and ointment bases to satisfy the preference of the patient and physicians. As
creams contain both aqueous phase & water phase it becomes easy for formulator to
incorporate all type of herbal extracts,

Antimicrobial creams destroy bacteria or suppresses their growth or their ability to reproduce.
Heat, chemicals such as chlorine and antibiotic drugs all have antimicrobial properties. They
can also be classified according to their function. Agents that kill microbes are microcidal,
while those that merely inhibit their growth are called biostatic. The use of antimicrobial
medicines to treat infection is known as antimicrobial chemotherapy, while the use
antimicrobial medicines to prevent infection is known as antimicrobial prophylaxis.[2]

Hibiscus abelmoschus generally known as okra, used from ancient time for the treatment of
various diseases like diarrhea, syphilis, gonorrhea, urinary tract infection & inflammation &
also used to treat itchiness & skin moisturizer. Leaves extract of hexane, ethyl acetate,
methanol, and aqueous has demonstrated antimicrobial activity against a number of
pathogens such as Staphylococcus aureus, Bacillus megaterium, Shigella flexneri, Proteus

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mirabilis, Proteus vulgaris, and Corynebacterium diphtheriae. The hexane fraction

containing essential oil showed strong antibacterial activity against Corynebacterium
diphtheriae. Aqueous extract of Abelmoschus moschatus seeds exhibited antimicrobial
activity against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Pseudomonas
aeruginosa, Proteus vulgaris, and Salmonella enterica paratyphi. Hydroalcoholic extract of
leaves of plant also showed antimicrobial effect against Candida albicans.[3,4] Hence the
present study was designed to formulate the herbal cream of with different concentrations of
Hibiscus abelmoschus L seeds extract & to evaluate for their antimicrobial & antifungal
activity against various microorganisms.

2. MATERIALS AND METHODS

2.1 Materials
Hibiscus abelmoschus were purchased from Handral seeds and fertilizers, Bagalkot
Karnataka. Borax (Thermo fisher Sci Ind Pvt Ltd. Mumbai), Besswax (Genuine chemical co.
Mumbai), Petroleum ether (SDFCL. Mumbai), Petroleum jelly (LOBA Chem Mumbai),
Methyl Parabben (NR CHEM Mumbai) and propyl paraben (Genuine chemical co. Mumbai)
used in the formulation of cream were analytical grade.

2.2 Drug Profile

Drug: Hibiscus abelmoschus Linn.
Family: Malvaceae
Synonyms: Abelmoschus moschatus
Habitat: In India-dry tropical and semi evergreen forest, and found in most tropical regions.
it is also found hilly regions of deccan and karnataka and foothills of himalayas.

Vernacular names Given below

In Kannada : Kaadu-Kastoori, Kasturi bende.
In English. : Musk mallow.( Ladies finger)
In Sanskrit : Latakasturika.
In Hindi : Maskdana.
In Tamil : Vettrilaikkasturi, Kattukasturi.
In Telugu : Kasturibendavittu.
Parts used : Seeds.
Habitat : A herbaceous annual grown in many hotter parts in India (common in
Bengal) and found in most tropicals regions.

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2.3 Plant Botanical Description

Hibiscus abelmoschus (Abelmoschus moschatus) is annual, herb which grows up to 1.6m in
height. Leaves are polymorphous, more or less cordite shown in fig.1, the lower ovate, acute
or roundish- angled with the upper palmate 3-7 lobed divided nearly to the base. Lobed are
narrow-acute or oblong-ovate, crenate serrate or irregularly toothed, and hairy on both
surfaces. Flowers are large, bright yellow with dark purple base in color and usually appear
solitary axillary. Capsules are 6.5-7.5cm long, ovate, acute, and hispid. Seeds are
subreniform, black or grayish brown in color, concentrically ribbed, and scented shown in
fig.1.[5]

2.4 Preparation of Plant extract: Petroleum ether extract of Hibiscus Abelmoschus L.

The dry seeds of Hibiscus abelmoschus were purchased from Handral seeds and fertilizers,
Bagalkot Karnataka; seeds were identified and authenticated by Prof. M.K. Ganachari,
Botanist. Department of Botany, Basaveshwar Science College, Bagalkot Karnataka, were
deposited and stored in air tight container, the seeds of Hibiscus abelmoschus were cleaned
and air dried then subjected to coarse powdering and passed through sieve #44 to get uniform
size powder. About 250 g of crushed seed powder was taken in a round bottom flask and
soaked with 500 ml of petroleum ether. The sealed round bottom flask was kept for 2 days
with occasional shaking and stirring. The extracts were filtered through a two layered muslin
cloth. The filtrate was concentrated by evaporating excess solvent by using hot air oven at
temperature 450c.[6]

2.5 Formulation of Hibiscus abelmoshus Antimicrobial Cream

Hibiscus abelmoschus cream were prepared by w/o emulsion method. The cream were
formulated by using five different concentration of oil such as 1%, 2%, 3%, 4% and 5%.
Borax was used as emulsifying agent. Bees wax was used to create a barrier which helps to
seal moisture into the skin. White soft paraffin is used as a moisturizer. Methyl paraben and
propyl paraben was used as preservatives in cream. The composition all ingredients are
shown in Table.1. The formulation was prepared by adding two phases which are mentioned
as follows: Beeswax and white soft paraffin were melted together in a china dish at 700C.
Borax was dissolved in water and the temperature of the solution was brought to 70 0C. The
extracts and preservatives were dissolved in the borax solution. The water phase containing
the extract and borax was then added to the oil phase with stirring. It was then agitated slowly

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and cooled; this procedure was carried out for all the formulations. The cream was filled into
jars on cooling, prepared formulation were labeled and stored.[7,8]

2.6 EVALUATION OF HERBAL CREAM

2.6.1 ATR studies
The calibration sample were prepared and analyzed using a fiber optic ATR probe with an
IRE made of zirconium dioxide connected to the Fourier transformer IR spectrometer. The
ZrO2 IRE was designed as a pyramid with a 900 angle on the top to enable 2 internal
reflections at the angle of 450 to the surface. The ATR element was cleaned with acetone
after each measurement; the emersion part of the probe was made of Hastelloy C-22 alloy
well suited for field applications. For the measurements, the probe was mounted on a tripod.
The probe was immersed 1 to 1.5 cm deep into a samples & ATIR was recorded.[9]

2.6.2 Charecterization of Cream

a. pH determination
The pH of various formulations was determined by using digital pH meter. About 1 g of the
cream was weighed and dissolved in 100 ml of distilled water and stored for two hours. The
measurement of pH of each formulation was done in triplicate and average values were
calculated.[10]

b. Viscosity
Viscosity of the prepared formulation were evaluated by using Brookfeild Viscometer by
placing the formulation in beaker were spindle no.64 was rotated at 50 rpm, at each speed
corresponding dial reading was noted.[11]

c. Spreadability
Spreadability of the formulation was done by using two sets of glass slides of standard
dimensions. The herbal cream formulation was placed over one of the slides. The other slide
was placed on the top of the formulation, such that the cream was sandwiched between the
two slides weight was placed upon the upper slides so that the cream between the two slides
was pressed uniformly to form a thin layer. The weight was removed and the excess of
formulation adhering to the slides was scrapped off. The upper slide allowed slipping off
freely by the force of weight tied to it12. The time taken for the upper slide was noted &
calculated by using the formula Spreadability = m ×l /t

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m = weight tied to the upper slide (30g) l =length of glass slide (5cm) t = time taken in
seconds

d. Extrudability
Extrudability of the prepared formulation under study was filled in clean, lacquered
aluminum collapsible tube with nozzle tube of 5mm opening and pressure was applied on
tube by the help of finger. Tube extrudability was then determined by measuring the amount
of cream extruded through the tip when the pressure was applied on the tube.[13]

e. Antimicrobial Activity
Antibacterial activity of formulated cream was performed by modified Agar well diffusion
method using Staphylococcus aureus (MTCC 912), Bacillus subtilis (MTCC 7424),
Pseudomonas aeruginosa (MTCC 424), Klebsiella pneumonia(MTCC 3384), Candida
tropicalis (ATCC 1369) & Candida albicans (ATCC 14053). Agar plates were prepared by
Muller-Hilton agar medium poured in a petri dish & microorganism were inoculated with test
microorganism’s culture & agar plates were at 37°C for 24h for bacteria and at 28°C for 24 h
for fungal strains. After 24h wells of 6mm diameter was made with sterile cork borer & wells
were loaded with the formulation F1 to F5. After 24 h of incubation, formulations efficacy
was determined in terms of zone of inhibition. The antibacterial activity was evaluated by
measuring the diameter of the resulting zone of inhibition against the tested microorganisms
in millimeters.[14]

3. RESULTS AND DISCUSSION

The present investigation attempted to develop the herbal antimicrobial cream using extract
of Hibiscus abelmoschus seeds oil. Formulated creams were subjected for visual observations
& all formulation was found to be homogeneous white colored semisolid with characteristic
odor of the raw materials, then formulations were subjected for evaluation studies such as pH,
viscosity, extrudability, spreadability and antimicrobial activity.

3.1 ATR spectroscopy studies

ATR spectra of the Hibiscus abelmoschus cream, without drug cream, Hibiscus abelmoschus
oil and combination of them were represented in Figure.2, the characteristic peak found in
pure oil were also found in cream which indicates there were no changes in these main peaks
in IR spectra of oil and cream, which shows that there were no physical & chemical
interactions. The peaks obtained in the spectra’s of each sample correlates with the peaks of

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oil spectrum. This indicates that the oil was compatible with the formulation component &
drug exists in original form and available for the biological action.

3.2 pH determination of Hibiscus abelmoschus cream

The various Hibiscus abelmoschus cream formulations were subjected to pH determination
studies and data shown in the table 2. Skin pH is normally acidic, ranging in pH values of 4-
6, while the body’s internal environment maintains a near neutral pH 7-9. This creates a steep
pH gradient of 2 to 3 units between the stratum carenum and underlying epidermis and
dermis. The pH of all formulated creams were in the range of 7.4 to 7.9, which lies in the
normal pH range of theskin.[13]

3.3 Viscosity of cream

Viscosity plays a important role in effective formulation of creams as it helps the ingredients
of cream to be in intimate contact at the applied area of the skin & also effect the
spreadibility, peraped formulation were subjected to to viscosity studies and data is shown in
the table 2. All the cream formulations showed good viscosity and they were capable to
remain in the site of application for prolonged time. In topical formulations, consistency is a
substantial factor affecting viscosity and overall acceptance of the formulations. The apparent
viscosity of all the formulation are having low shear rate which is an indicator of viscosity
upon topical application.[14]

3.4 Spreadability of creams

The Spreadability was expressed in terms of time in seconds taken by two slides to slip off
from the cream, placed in between the slides, under certain load. Lesser the time taken for
separation of the two slides, better the Spread ability. Spreadability plays a considerable role
in patient compliance and ensures uniform application of cream to a larger area of the skin.
The various Hibiscus abelmoschus cream formulations were subjected to spreadability
studies and data is shown in the table 2. The low value of spreadability coefficient of the
cream was sufficient suggesting easy spreading and no signs of grittiness. The values of
spreadability indicate that the cream was easily spreadable by small amount of shear. The
lower value of spreadability indicates the lesser work required to spread the cream over the
skin, which means all the formulated creams was easily spreadable by applying small amount
of shear. Above all the formulation F1 and F2 were having good spredability.[15]

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3.5 Extrudability of cream

It is usual empirical test to measure the force required to extrude the material from tube.
More quantity extruded better was extrudability, formulations were subjected to extrudability
studies and data shown in the table 2. The measure of the force required to extrude the
material from a collapsible tube when certain amount of force has been applied on it. The
formulation F1, F3, F4 and F5 showed good extrudability and the F2 showed excellent
extrudability. All the formulations were in the official range ofextrudability.[16]

3.6Antimicrobial activity
The topical formulations were tested against selected pathogenic strains of bacteria and fungi.
The antimicrobial activity of Hibiscus abelmoschus cream was assayed by agar disc diffusion
method against the selected microbes. The various Hibiscus abelmoschus cream formulations
were active against all the pathogenic bacteria and fungi under study as revealed by their
respective zones of growth inhibition. However, the formulations displayed a variable degree
of antimicrobial activity against the tested strains. As shown in table 3. Formulation F2
containing hibiscus abelmoschus oil (2%) was found to be most effective against all the
selected microorganisms accept pseudomonas aeruginosa.[3,4]

Table.1 Preparation of Hibiscus abelmoshus Antimicrobial Cream.

Ingredients F1 F2 F3 F4 F5
Oil Extract 1ml 2ml 3ml 4ml 5ml
White soft paraffin 55gm 55gm 55gm 55gm 55gm
Bees wax 13.17gm 12.17gm 11.17gm 10.17gm 9.17gm
Borax 0.83gm 0.83gm 0.83gm 0.83gm 0.83gm
Methyl paraben 0.16gm 0.16gm 0.16gm 0.16gm 0.16gm
Propyl paraben 0.16gm 0.16gm 0.16gm 0.16gm 0.16gm
Water 30ml 30ml 30ml 30ml 30ml

Table 2: Evaluation of Hibiscus abelmoshus Antimicrobial Cream.

Formulation pH Viscosity in Spreadability Extrudability
F1 7.6 ±0.14 27020±20 10.83±0.16 88.02±0.60
F2 7.5 ±0.0 27230±30 13.00±0.18 91.92±0.81
F3 7.7 ±0.12 27110±10 14.04±0.17 88.82±0.85
F4 7.9 ±0.04 27050±20 13.00±0.20 80.58±0.73
F5 7.4 ±0.16 27110±10 16.25±0.15 89.42±0.76
Values are expressed as mean±SEM (n=3)

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Table 3: Antimicrobial activity of Hibiscus abelmoshus Creams.

Serial Zone of inhibition (mm)
Organism
No. F1 F2 F3 F4 F5
1 Staphylococcus aureus (MTCC 912) 10.1±1.1 10.1±1.3 12.1±1 11.2±1.1 R
2 Bacillus subtilis (MTCC 7424) 13.2±1.1 13.3±1.2 12.2±1.1 10.1±1.2 8.1±1.1
3 Pseudomonas aeruginosa (MTCC 424) R R R R R
4 Klebsiella pneumonia (MTCC 3384) 13.1±1.2 12.1±1.3 10.1±1.2 8.1±1.1 8.2±1.2
5 Candida tropicalis (ATCC 1369) 12.2±1.1 13.2±1.2 R R R
6 Candida albicans (ATCC 14053) 12.2±1.1 13.1±1.1 R R R
all the results are mean±SD (n=3), R=Resistant

(a) (b)
Fig 1. (a) Branch bearing capsule of Hibiscus abelmoschus (b) Seeds of Hibiscus
Abelmoschus.

Fig 2 ATR spectra (1) Hibiscus abelmoschus oil (2) Cream without oil (3) Cream with
Hibiscus abelmoschus oil.

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(a) (b) (c)

(d) (e) (f)

Fig. 3: Agar disk diffusion method of Hibiscus abelmoschus formulations against the (a) Staphylococcus aureus (b) Bacillus subtilis (c)
Pseudomonas aeruginosa (d) Klebsiella pneumonia (e) Candida tropicalis (f) Candida albicans to evaluate the antimicrobial activity.

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4. CONCLUSION
The concept of above formulation was to incorporate the oil of Hibiscus abelmoschus seeds
in the cream, as creams are widely accepted & better absorbed by skin with its moisturizing
& emollient effect. Hence in the present investigation we prepared Hibiscus abelmoschus
cream by using conveniently excipients. Results of evaluation demonstrated the pH of the
creams were in normal range of the skin with good viscosity, spreadability & extrudability
which indicated creams were capable to remain in the site of application for prolonged time.
Thus we concluded that Hibiscus abelmoschus cream would provide safe & healthy germ
free skin.

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