*Corresponding Author: Dr. Kalyani Kakad, Department of Pharmacognosy, P. E. Society’s Modern
College of Pharmacy, Nigdi, Pune Maharashtra- 411044. Email: [email protected]. 1
International Journal of Zoology and Applied Biosciences ISSN: 2455-9571
Volume 10, Issue 5, pp: 1-8, 2025 http://www.ijzab.com
https://doi.org/10.55126/ijzab.2025.v10.i05.001
Research Article
FORMULATION AND EVALUATION OF HERBAL ORAL GEL
CONTAINING EXTRACTS OF POWDERED VITEX NEGUNDO LEAVES
WITH ANACYCLUS PYRETHRUM TO TREAT MOUTH ULCER
*1Kalyani Kakad, 1Vikas Gore, 2Sneha Nawale
*1Department of Pharmacognosy, P. E. Society’s Modern College of Pharmacy, Nigdi, Pune Maharashtra- 411044.
2Department of Pharmacognosy, Gokaraju Rangaraju College of Pharmacy, Bachupally,
Hyderabad-500090 (Osmania University).
Article History: Received 2nd July 2025; Accepted 29th August 2025; Published 30th September 2025
ABSTRACT
Mouth ulcers, known as canker sores or aphthous ulcers, are common painful lesions within the oral cavity, often resulting
from factors like nutritional deficiencies, stress, hormonal changes, or physical trauma. This study investigates the
development of a topical gel incorporating natural therapeutic agents extracted from Vitex negundo and Anacyclus
pyrethrum to alleviate symptoms associated with mouth ulcers. These botanical extracts exhibit anti-inflammatory,
analgesic, and antimicrobial properties that can reduce pain and promote oral ulcer healing. The gel formulations,
formulated with appropriate gelling agents and excipients, underwent evaluation based on appearance and consistency to
identify the optimal formulation. The selected formulation will undergo further antimicrobial studies to assess its
effectiveness in managing mouth ulcers. In conclusion, the development of a topical gel containing bioactive compounds
from Vitex negundo and Anacyclus pyrethrum represents a promising approach for treating mouth ulcers. This research
contributes to the understanding of natural remedies for oral health conditions and highlights the potential of botanical
extracts in topical formulations for managing oral ulcers.
Keywords: Anti-ulcer activity, Wound healing, Gel formulation, Oral health, Evaluation.
INTRODUCTION
Gels are mostly semi-solid combinations including a liquid
phase that has been augmented with additional components
to harden it. Topical gel forms are placed locally on
particular mucosal surfaces when trying to spread the drug
or generate an immediate effect (Singh, 2014). Small sores
or scratching that develop inside the mouth or at the base of
the teeth are known as mouth ulcers. Apthopus ulcers as
canker sores are various names to mouth ulcers. A mouth
ulcer may also be referred to as a break or breach in the
membrane that lines inside of the mouth. typically, it starts
out as a white-colored or pale depression in your mouth.
Mouth ulcers may occur by nutritional shortages, poor oral
hygiene, infections, stress, indigestion, mechanical injury,
food allergies, hormonal imbalance, skin disease etc.
Mouth ulcers, also known as aphthous ulcers, can be
painful while eating, drinking or brushing teeth
(Indhumathi, 2020).
Topical gel formulations are consistent, semisolid
preparations which are simple to remove from the
epidermis and are less oily compared to other drug delivery
strategies. They are comprised of solutions or dispersions
of one or more medication in acceptable hydrophilic or
hydrophobic bases. They are applied externally to the skin
or certain mucous membranes for beneficial prevention, or
defensive purposes (Mhaske S. D., 2022). Several
excipients are utilized in the gel-making technique:
propylparaben, triethanolamine, methylparaben, carbopol
934, disodium edeate, and propylene glycol. The gelling
element carbopol 934 was utilized in three distinct levels to
make the gel formulation. One batch is picked according to
its appearance that displays the majority of the gel-like
qualities (Mhaske S. D., 2022).
Kalyani Kakad et al. Int. J. Zool. Appl. Biosci., 10(5), 1-8, 2025
www.ijzab.com 2
Oral ulcers
An ulcer is a defect in the integrity of the epithelium
resulting from molecular necrosis. Ulcers are frequently
noticed near the cavities in the mouth, and patients
typically seek neither dental or medical therapy for such
conditions. Redness, suffering, and/or a burning feeling
all typical warning signs. They can arise anywhere in the
oral cavity, but they can lead to difficulty if they do so in
the area that's flexible (Mhaske S. D., 2022). A mouth
ulcer, frequently referred to as an oral ulcer or a mucosal
ulcer, is an injury of the oral cavity's membrane. Usually,
those are painful round or oval sores that arise on the inside
of the cheeks or lips behind the mouth. Mouth ulcers are
fairly prevalent and may develop from a number of diseases
and their therapies. despite the fact these usually have not
any significant underlying causes underlying it. Mouth
ulcers tend to be triggered by nutritional deficiencies such
as iron deficiency, vitamin a lack (particularly B12 and C),
infections, stress, indigestion, mechanical harm, food
allergies, hormonal imbalances, issues with the skin, etc.
Mouth ulcers, additionally known as aphthous ulcers, might
discomfort when you eat, drink, or brushing your teeth
(Mhaske S. D., 2022).
Mouth ulcers can be described as moderate, at deeper, or
herpetiform depending on the severity and amount. Mouth
ulcer forms that are typical are: Moderate ulcers: These
typically go entirely in 10 days to 2 weeks with a diameter
for 2 to 8 mm. Deeper ulcers: These usually have an
unequal or irregular border. This type of ulcer may leave a
scar in the mouth and take a long time to heal. Herpetiform
ulcers: these kinds of ulcers are the result by multiple small
pinhead-sized infections packed close (Raghvendr K. Y.,
2021).
Vitex negundo is a verbenaceae family tree which climbs
to a height if 4.5 feet and is native to India. Anti-
inflammatory, antibacterial, anti-fertility, antispasmodic,
analgesic, hepato-protective, estrogenic, anticonvulsant,
antiarthritic, diuretic, antimicrobial, anti-parkinsonian,
antipsychotic, antidepressant, antihistamine-releasing
activity, mosquito repellent activity, anti-feedant, anti-
filarial, juvenomimetic, and anti-androgenic are only some
among its numerous medicinal uses. The leaves can be
helpful in rheumatism, arthritis, catarrhal fever, cephalagia,
sprains, orchitis, syphilis, inflammations, and ulcers. It also
serves as warm, tonic, and vermifuge (Sharma P.C., 2005).
Anacyclus pyrethrum (L) is commonly known as African
pyrethrum, akarkara, tigendesste, and igendess. It is a
species belonging to the family Asteraceae, which is
indigenous to Morocco, Algeria, and Spain. This species
includes the two varieties Anacyclus pyrethrum var.
pyrethrum (L) and Anacyclus pyrethrum var. depressus
(Ball) Maire. The roots of A. pyrethrum (L) are used to
cure toothaches, salivary production, angina, digestive
issues, lethargy, female infertility, and even tongue and
limb paralysis in traditional medicine. They are used to
treat gout and sciatica and to keep disease at bay in the
form of cream-based animal fats. Anesthetic, anti-
inflammatory, anticonvulsant, antioxidant, antidiabetic, and
memory enhancer effects of Anacyclus pyrethrum (L) roots
have been documented in the literature (Neha R. Jumde.,
2022).
MATERIAL AND METHODS
Collection and authentication of plant materials
The leaves of plant powder Vitex negundo and rhizomes of
Anacyclus pyrethrum we procured from the local area of
PCMC, Pune, India in month of December 2023 and the
plant specimens are authenticated by “Manakarnika
Aushadhayala”. Chinchwad -33 respectively.
Chemicals
Ethanol, carbopol 940, methyl paraben, propyl paraben,
propylene glycol 400, disodium edetate, triethanolamine,
distilled water.
Equipment’s
Digital balance, pH meter, Magnetic stirrer, water bath,
Ultra sonicator, Brookfield viscometer, spredability
apparatus.
Preparation of plant extract
A total of 20 g of powder of leaves parts of Vitex negundo
and Anacyclus pyrethrum was extracted by maceration with
200 mL of 90% ethanol at room temperature for 48 h. Then
mixtures were filtered and evaporated at 40 °C, and the
residue obtained was then kept a until further use. Then
extraction yield for leaves powder was 16% w/w and 10%
w/w, respectively. (Sharma, P. K., 2018 and Sharma, P. K.,
2012).
Preparation of herbal gel
Carbopol 934 was dissolved slowly with stirring in 60 mL
of demineralized water for 1 hrs to avoid agglomeration
Then disodium edetate and triethanolamine were dissolved
in 10 mL of demineralized water separately and stirred for
10 min. Mixed 4.83 mL of propylene glycol in 12 mL of
demineralized water with stirring for 10 min. Disodium
edetate and triethanolamine solution were added to
carbopol solution and the pH was then adjusted to 7.4 by
stirring the solution for 10 min. Then propylene glycol
solution was added with stirring for 10 min until a clear
consistent gel base was obtained (Nappinnai., 2006).
Further required quantity of extract was added to the above
mixture and this solution was mixed properly to the
Carbopol 940 gel with continuous stirring (Das S., 2011).
www.ijzab.com 2
Oral ulcers
An ulcer is a defect in the integrity of the epithelium
resulting from molecular necrosis. Ulcers are frequently
noticed near the cavities in the mouth, and patients
typically seek neither dental or medical therapy for such
conditions. Redness, suffering, and/or a burning feeling
all typical warning signs. They can arise anywhere in the
oral cavity, but they can lead to difficulty if they do so in
the area that's flexible (Mhaske S. D., 2022). A mouth
ulcer, frequently referred to as an oral ulcer or a mucosal
ulcer, is an injury of the oral cavity's membrane. Usually,
those are painful round or oval sores that arise on the inside
of the cheeks or lips behind the mouth. Mouth ulcers are
fairly prevalent and may develop from a number of diseases
and their therapies. despite the fact these usually have not
any significant underlying causes underlying it. Mouth
ulcers tend to be triggered by nutritional deficiencies such
as iron deficiency, vitamin a lack (particularly B12 and C),
infections, stress, indigestion, mechanical harm, food
allergies, hormonal imbalances, issues with the skin, etc.
Mouth ulcers, additionally known as aphthous ulcers, might
discomfort when you eat, drink, or brushing your teeth
(Mhaske S. D., 2022).
Mouth ulcers can be described as moderate, at deeper, or
herpetiform depending on the severity and amount. Mouth
ulcer forms that are typical are: Moderate ulcers: These
typically go entirely in 10 days to 2 weeks with a diameter
for 2 to 8 mm. Deeper ulcers: These usually have an
unequal or irregular border. This type of ulcer may leave a
scar in the mouth and take a long time to heal. Herpetiform
ulcers: these kinds of ulcers are the result by multiple small
pinhead-sized infections packed close (Raghvendr K. Y.,
2021).
Vitex negundo is a verbenaceae family tree which climbs
to a height if 4.5 feet and is native to India. Anti-
inflammatory, antibacterial, anti-fertility, antispasmodic,
analgesic, hepato-protective, estrogenic, anticonvulsant,
antiarthritic, diuretic, antimicrobial, anti-parkinsonian,
antipsychotic, antidepressant, antihistamine-releasing
activity, mosquito repellent activity, anti-feedant, anti-
filarial, juvenomimetic, and anti-androgenic are only some
among its numerous medicinal uses. The leaves can be
helpful in rheumatism, arthritis, catarrhal fever, cephalagia,
sprains, orchitis, syphilis, inflammations, and ulcers. It also
serves as warm, tonic, and vermifuge (Sharma P.C., 2005).
Anacyclus pyrethrum (L) is commonly known as African
pyrethrum, akarkara, tigendesste, and igendess. It is a
species belonging to the family Asteraceae, which is
indigenous to Morocco, Algeria, and Spain. This species
includes the two varieties Anacyclus pyrethrum var.
pyrethrum (L) and Anacyclus pyrethrum var. depressus
(Ball) Maire. The roots of A. pyrethrum (L) are used to
cure toothaches, salivary production, angina, digestive
issues, lethargy, female infertility, and even tongue and
limb paralysis in traditional medicine. They are used to
treat gout and sciatica and to keep disease at bay in the
form of cream-based animal fats. Anesthetic, anti-
inflammatory, anticonvulsant, antioxidant, antidiabetic, and
memory enhancer effects of Anacyclus pyrethrum (L) roots
have been documented in the literature (Neha R. Jumde.,
2022).
MATERIAL AND METHODS
Collection and authentication of plant materials
The leaves of plant powder Vitex negundo and rhizomes of
Anacyclus pyrethrum we procured from the local area of
PCMC, Pune, India in month of December 2023 and the
plant specimens are authenticated by “Manakarnika
Aushadhayala”. Chinchwad -33 respectively.
Chemicals
Ethanol, carbopol 940, methyl paraben, propyl paraben,
propylene glycol 400, disodium edetate, triethanolamine,
distilled water.
Equipment’s
Digital balance, pH meter, Magnetic stirrer, water bath,
Ultra sonicator, Brookfield viscometer, spredability
apparatus.
Preparation of plant extract
A total of 20 g of powder of leaves parts of Vitex negundo
and Anacyclus pyrethrum was extracted by maceration with
200 mL of 90% ethanol at room temperature for 48 h. Then
mixtures were filtered and evaporated at 40 °C, and the
residue obtained was then kept a until further use. Then
extraction yield for leaves powder was 16% w/w and 10%
w/w, respectively. (Sharma, P. K., 2018 and Sharma, P. K.,
2012).
Preparation of herbal gel
Carbopol 934 was dissolved slowly with stirring in 60 mL
of demineralized water for 1 hrs to avoid agglomeration
Then disodium edetate and triethanolamine were dissolved
in 10 mL of demineralized water separately and stirred for
10 min. Mixed 4.83 mL of propylene glycol in 12 mL of
demineralized water with stirring for 10 min. Disodium
edetate and triethanolamine solution were added to
carbopol solution and the pH was then adjusted to 7.4 by
stirring the solution for 10 min. Then propylene glycol
solution was added with stirring for 10 min until a clear
consistent gel base was obtained (Nappinnai., 2006).
Further required quantity of extract was added to the above
mixture and this solution was mixed properly to the
Carbopol 940 gel with continuous stirring (Das S., 2011).
Kalyani Kakad et al. Int. J. Zool. Appl. Biosci., 10(5), 1-8, 2025
www.ijzab.com 3
Table 1. Ingredients of herbal mucosal gel.
Ingredients Quantity (gm) Role %w/w
Basil oil 1 Reduce mouth Ulcer,
antibacterial activity.
0.3
Nirgundi 0.5 Antibacterial, Laxative,
Antioxidant, Anti-inflammatory.
0.15
Akarkara 0.5 Anti-inflammatory,
Antibacterial used
0.15
Apple cider 0.5 Destroy bacteria in mouth 0.15
Carbopol 934 1.5 Mucoadhesive polymer used as gelling agent 0.45
Methyl paraben 0.00015 Preservative 0.000045
Propyl paraben 0.01 Preservative 0.03
Propylene glycol 5 Co-solvent 1.5
Triethanolamine 1.5 For adjust pH 0.45
Disodium edeate 0.005 Chelating agent 0.0015
Distilled water q. s - Up to 30ml
Evaluation parameters
Physical evaluation
Physical parameters such as color, odour and consistency
were checked visually. The color of the formulations was
checked by visual inspection. The consistency of
formulations was checked by applying on skin. The odour
of the formulations was checked by mixing the gel in water
and observing the smell (Table 2).
Percentage yield
Weigh the empty container in which the gel formulation
was stored then again weigh the container with gel
formulation. To obtain the practical yield subtract the
weight of empty container with the container with gel
formulation. Then the percentage yield was calculated by
the formula given below: Percentage yield = (practical
yield/theoretical yield) × 100 (Table 3).
Measurement of pH
The pH of gel formulations was determined by using digital
pH meter. Take 1 gm of gel and dissolved in 10 ml of
distilled water and keep apart for two hours (Table 4). Then
the measurement of pH of formulations was done by
dipping the glass electrode completely into the gel system
three times and the average values are reported (Mokashi
M., 2015).
Homogeneity
All prepared gel formulations were tested for homogeneity
by visual inspection after the gels have been set in to the
container. They were tested for their presence and
appearance of any aggregates (Table 5).
Viscosity
The measurement of viscosity of the formulated gel was
determined by Brookfield Viscometer with spindle no. 1 at
25°C. The gels were rotated at speed 0.3, 0.6 and 1.5
rotations per minute and at each speed, the corresponding
dial reading was noted. Then viscosity of the prepared gels
was obtained by multiplication of the dial reading with
factor given in the Brookfield Viscometer catalogues (11).
Viscosity of gel formulation was reported in Table 6.
Spread ability
Spread ability is expressed in terms of time in seconds
taken by two slides to slip off from gel that is placed in
between the slides under the direction of certain load. If the
time taken for separation of two slides is less then better the
spread ability (Kaur LP., 2010) 12). Spread ability is
calculated by using the formula:
S = M × L / T
Where M = weight tied to upper slide
L = length of glass slides
T = time taken to separate the slides
Spread ability of gel formulations were reported in Table 7.
Extrudability
The formulated gel was filled in standard capped
collapsible aluminum tubes and sealed by crimping to the
end. The weight of filled tubes was recorded and the tubes
were sandwiched between two glass slides and were
clamped. 500gm weight was placed over the slides and then
the cap was removed to extrude (Table 8). The amount of
extruded gel was collected and weighed. Extrudability was
www.ijzab.com 3
Table 1. Ingredients of herbal mucosal gel.
Ingredients Quantity (gm) Role %w/w
Basil oil 1 Reduce mouth Ulcer,
antibacterial activity.
0.3
Nirgundi 0.5 Antibacterial, Laxative,
Antioxidant, Anti-inflammatory.
0.15
Akarkara 0.5 Anti-inflammatory,
Antibacterial used
0.15
Apple cider 0.5 Destroy bacteria in mouth 0.15
Carbopol 934 1.5 Mucoadhesive polymer used as gelling agent 0.45
Methyl paraben 0.00015 Preservative 0.000045
Propyl paraben 0.01 Preservative 0.03
Propylene glycol 5 Co-solvent 1.5
Triethanolamine 1.5 For adjust pH 0.45
Disodium edeate 0.005 Chelating agent 0.0015
Distilled water q. s - Up to 30ml
Evaluation parameters
Physical evaluation
Physical parameters such as color, odour and consistency
were checked visually. The color of the formulations was
checked by visual inspection. The consistency of
formulations was checked by applying on skin. The odour
of the formulations was checked by mixing the gel in water
and observing the smell (Table 2).
Percentage yield
Weigh the empty container in which the gel formulation
was stored then again weigh the container with gel
formulation. To obtain the practical yield subtract the
weight of empty container with the container with gel
formulation. Then the percentage yield was calculated by
the formula given below: Percentage yield = (practical
yield/theoretical yield) × 100 (Table 3).
Measurement of pH
The pH of gel formulations was determined by using digital
pH meter. Take 1 gm of gel and dissolved in 10 ml of
distilled water and keep apart for two hours (Table 4). Then
the measurement of pH of formulations was done by
dipping the glass electrode completely into the gel system
three times and the average values are reported (Mokashi
M., 2015).
Homogeneity
All prepared gel formulations were tested for homogeneity
by visual inspection after the gels have been set in to the
container. They were tested for their presence and
appearance of any aggregates (Table 5).
Viscosity
The measurement of viscosity of the formulated gel was
determined by Brookfield Viscometer with spindle no. 1 at
25°C. The gels were rotated at speed 0.3, 0.6 and 1.5
rotations per minute and at each speed, the corresponding
dial reading was noted. Then viscosity of the prepared gels
was obtained by multiplication of the dial reading with
factor given in the Brookfield Viscometer catalogues (11).
Viscosity of gel formulation was reported in Table 6.
Spread ability
Spread ability is expressed in terms of time in seconds
taken by two slides to slip off from gel that is placed in
between the slides under the direction of certain load. If the
time taken for separation of two slides is less then better the
spread ability (Kaur LP., 2010) 12). Spread ability is
calculated by using the formula:
S = M × L / T
Where M = weight tied to upper slide
L = length of glass slides
T = time taken to separate the slides
Spread ability of gel formulations were reported in Table 7.
Extrudability
The formulated gel was filled in standard capped
collapsible aluminum tubes and sealed by crimping to the
end. The weight of filled tubes was recorded and the tubes
were sandwiched between two glass slides and were
clamped. 500gm weight was placed over the slides and then
the cap was removed to extrude (Table 8). The amount of
extruded gel was collected and weighed. Extrudability was
Kalyani Kakad et al. Int. J. Zool. Appl. Biosci., 10(5), 1-8, 2025
www.ijzab.com 4
determined by calculating the percentage of extruded gel
(Mendhekar SY., 2017).
Clarity
The clarity of herbal batch was determined by visual
inspection (Pandey N., 2011).
Gel strength
Gel strength was determined by the time in seconds
required by the weight to penetrate in the gel. A 3.5 gm
weight was placed on the surface of 5 gm formulated gel
(Tables 9). Gel strength was determined by reporting the
time in seconds required by the weight to penetrate 0.5 cm
in the gel (Thombre KP., 2018).
Anti-fungal activity
The antifungal and antibacterial activity of formulation and
blank formulation were carried out by Cup-plate method in
comparison with marketed antifungal formulation (Hiora sg
oral gel). The antifungal and antibacterial activity test was
performed by using Candida albicans and Staphylococcus
aureus. Prepared nutrient brought and poured in to sterile
petri plates and kept aside for drying and cooling. After that
Candida albican culture were spread by micron wire loop.
A sterile cork borer 6 mm diameter was used to drill holes
4 mm deep. Then place 0.5 gm of gel from each
formulation in to these holes. Plates were then incubated at
27°C for 48 hr. Then the zone of inhibition (diameter in
mm) was measured (Koland M., 2011, Satish Kumar A,
2016) (Table 10).
Stability study
Stability studies were done with open and close container.
Here, by subjecting the product to room temperature for 1
month. (Table 11). national Journal of Drug
RESULTS AND DISCUSSION
From the above results it is clearly shows that all the
prepared gel formulations were yellowish green in color
and having good homogeneity and gelling property. The
pH of all gel formulations was in the range compatible with
normal pH range of oral cavity. The rheological behaviors
of gel formulations were studied with Brookfield
viscometer which indicated that the viscosity of gel
formulation was consistent neither too thick nor too thin.
The study of spreadability shows that with increasing the
viscosity of formulation spreadabilty decrease and its vice-
versa. The gelling strength and extrudability is found in the
suitable range. Thus overall, the gel formulation complies
with all parameters of an marketed ideal gel. Accelerated
stability studies indicated that the physical appearance,
rheological properties, extrudability, spreadability in the
optimized formulation remain unchanged upon storage for
3 months. This optimized herbal mucosal gel showed good
antifungal and antibacterial activity against Candida
albicans and Staphylococcus aureus that is main
microorganism responsible for mouth ulcer.
Table 2. Physical evaluation of gel formulations and Marketed gel (Hiora-Sg oral gel).
Formulation Herbal gel Marketed gel
Color Yellowish green Dark brown
Consistency Good Good
Odor Characteristic Odorless
Table 3. Percentage yield of gel formulations and Marketed gel (Hiora-Sg oral gel).
Formulation Herbal gel Marketed gel
Percentage yield (%) 93.32 98.4
Table 4. pH of gel formulations and Marketed gel (Hiora-Sg oral gel).
Formulation Herbal gel Marketed gel
pH Day
1 7.0 7.2
2 6.8 6.9
3 6.4 6.7
Avg = 6.7 6.9
www.ijzab.com 4
determined by calculating the percentage of extruded gel
(Mendhekar SY., 2017).
Clarity
The clarity of herbal batch was determined by visual
inspection (Pandey N., 2011).
Gel strength
Gel strength was determined by the time in seconds
required by the weight to penetrate in the gel. A 3.5 gm
weight was placed on the surface of 5 gm formulated gel
(Tables 9). Gel strength was determined by reporting the
time in seconds required by the weight to penetrate 0.5 cm
in the gel (Thombre KP., 2018).
Anti-fungal activity
The antifungal and antibacterial activity of formulation and
blank formulation were carried out by Cup-plate method in
comparison with marketed antifungal formulation (Hiora sg
oral gel). The antifungal and antibacterial activity test was
performed by using Candida albicans and Staphylococcus
aureus. Prepared nutrient brought and poured in to sterile
petri plates and kept aside for drying and cooling. After that
Candida albican culture were spread by micron wire loop.
A sterile cork borer 6 mm diameter was used to drill holes
4 mm deep. Then place 0.5 gm of gel from each
formulation in to these holes. Plates were then incubated at
27°C for 48 hr. Then the zone of inhibition (diameter in
mm) was measured (Koland M., 2011, Satish Kumar A,
2016) (Table 10).
Stability study
Stability studies were done with open and close container.
Here, by subjecting the product to room temperature for 1
month. (Table 11). national Journal of Drug
RESULTS AND DISCUSSION
From the above results it is clearly shows that all the
prepared gel formulations were yellowish green in color
and having good homogeneity and gelling property. The
pH of all gel formulations was in the range compatible with
normal pH range of oral cavity. The rheological behaviors
of gel formulations were studied with Brookfield
viscometer which indicated that the viscosity of gel
formulation was consistent neither too thick nor too thin.
The study of spreadability shows that with increasing the
viscosity of formulation spreadabilty decrease and its vice-
versa. The gelling strength and extrudability is found in the
suitable range. Thus overall, the gel formulation complies
with all parameters of an marketed ideal gel. Accelerated
stability studies indicated that the physical appearance,
rheological properties, extrudability, spreadability in the
optimized formulation remain unchanged upon storage for
3 months. This optimized herbal mucosal gel showed good
antifungal and antibacterial activity against Candida
albicans and Staphylococcus aureus that is main
microorganism responsible for mouth ulcer.
Table 2. Physical evaluation of gel formulations and Marketed gel (Hiora-Sg oral gel).
Formulation Herbal gel Marketed gel
Color Yellowish green Dark brown
Consistency Good Good
Odor Characteristic Odorless
Table 3. Percentage yield of gel formulations and Marketed gel (Hiora-Sg oral gel).
Formulation Herbal gel Marketed gel
Percentage yield (%) 93.32 98.4
Table 4. pH of gel formulations and Marketed gel (Hiora-Sg oral gel).
Formulation Herbal gel Marketed gel
pH Day
1 7.0 7.2
2 6.8 6.9
3 6.4 6.7
Avg = 6.7 6.9
Kalyani Kakad et al. Int. J. Zool. Appl. Biosci., 10(5), 1-8, 2025
www.ijzab.com 5
Table 5. Homogeneity of gel formulations and Marketed gel (Hiora-Sg oral gel).
Formulation Herbal gel Marketed gel
Homogeneity Good Good
0975-9
Table 6. Viscosity of gel formulations and Marketed gel (Hiora-Sg oral gel).
Formulation Herbal gel Marketed gel
Viscosity(cps) Rpm 10 4100 5200
Rpm 20 5300 6400
Rpm 30 6200 6900
Table 7. Spreadability of gel formulations and Marketed gel (Hiora-Sg oral gel).
Formulation Herbal gel Marketed gel
Spreadabilty (gm.cm/sec)
Wt.(gm)
5 9.19 14.16
10 12.31 11.17
Table 8. Extrudability of gel formulations and Marketed gel (Hiora-Sg oral gel).
Formulation Herbal gel Marketed gel
Extrudability (%) 86.3 89.5
Table 9. Gelling strength of gel formulations and Marketed gel (Hiora-Sg oral gel).
Formulation Herbal gel Marketed gel
Gelling strength
Wt.(gm)
5 39.11 37.16
10 28.87 27.19
Table 10. Antifungal studies of gel formulations and Marketed gel (Hiora-Sg oral gel).
Formulation Zone of inhibition (mm) Zone of inhibition (mm)
Candida albicans Staphylococcus aureus
Standard drug ciprofloxacin 47 ciprofloxacin 45
Herbal gel 0.25 gm = 23 0.25 gm = 25
0.5 gm = 21 0.5 gm 30
Marketed (Hiora-Sg oral gel) 0.25 gm = 25 0.25 gm = 15
0.5 gm = 28 0.5 gm = 26
www.ijzab.com 5
Table 5. Homogeneity of gel formulations and Marketed gel (Hiora-Sg oral gel).
Formulation Herbal gel Marketed gel
Homogeneity Good Good
0975-9
Table 6. Viscosity of gel formulations and Marketed gel (Hiora-Sg oral gel).
Formulation Herbal gel Marketed gel
Viscosity(cps) Rpm 10 4100 5200
Rpm 20 5300 6400
Rpm 30 6200 6900
Table 7. Spreadability of gel formulations and Marketed gel (Hiora-Sg oral gel).
Formulation Herbal gel Marketed gel
Spreadabilty (gm.cm/sec)
Wt.(gm)
5 9.19 14.16
10 12.31 11.17
Table 8. Extrudability of gel formulations and Marketed gel (Hiora-Sg oral gel).
Formulation Herbal gel Marketed gel
Extrudability (%) 86.3 89.5
Table 9. Gelling strength of gel formulations and Marketed gel (Hiora-Sg oral gel).
Formulation Herbal gel Marketed gel
Gelling strength
Wt.(gm)
5 39.11 37.16
10 28.87 27.19
Table 10. Antifungal studies of gel formulations and Marketed gel (Hiora-Sg oral gel).
Formulation Zone of inhibition (mm) Zone of inhibition (mm)
Candida albicans Staphylococcus aureus
Standard drug ciprofloxacin 47 ciprofloxacin 45
Herbal gel 0.25 gm = 23 0.25 gm = 25
0.5 gm = 21 0.5 gm 30
Marketed (Hiora-Sg oral gel) 0.25 gm = 25 0.25 gm = 15
0.5 gm = 28 0.5 gm = 26
Kalyani Kakad et al. Int. J. Zool. Appl. Biosci., 10(5), 1-8, 2025
www.ijzab.com 6
Figure 1. Zone of Inhibition Candida albicans.
Figure 2. Zone of Inhibition Staphylococcus aureus.
Table 11. Stability studies of gel formulations and Marketed gel (Hiora-Sg oral gel).
Formulation Herbal gel Marketed gel
Stability Open container Closed container Open container Closed container
Not stable Stable Not stable Stable
0975-93
www.ijzab.com 6
Figure 1. Zone of Inhibition Candida albicans.
Figure 2. Zone of Inhibition Staphylococcus aureus.
Table 11. Stability studies of gel formulations and Marketed gel (Hiora-Sg oral gel).
Formulation Herbal gel Marketed gel
Stability Open container Closed container Open container Closed container
Not stable Stable Not stable Stable
0975-93
Kalyani Kakad et al. Int. J. Zool. Appl. Biosci., 10(5), 1-8, 2025
www.ijzab.com 7
Optimization of batches
After analysis of formulations for their extrudability and gelling strength it is observed that the formulation containing
equal amount of Vitex negundo extract and Anacyclus pyrethrum extract showed good result. The batch F (mixed)
optimized with good pH, viscosity, spreadability, extrudability and gelling strength, hence this formulation is further used
for anti-fungal study.
Table 12. Optimization of batches.
Parameters Optimized batch F (Herbal gel) Optimized batch F (Marketed gel)
Color Yellowish green Dark brown
Odor Characteristics Odorless
pH 6.7 6.9
Viscosity 6200 7100
Spredability 12.31 11.17
Extrudability 86.3 89.5
Gelling strength 28.87 27.19
CONCLUSION
From the mentioned results, it was concluded that the
prepared gel formulations are in good appearance with
suitable pH range. Formulated gel have good homogeneity,
proper gel strength & spread ability. The gel is neither too
thick nor too thin. The all-formulated gels are found to be
stable in closed container as compared to open container.
The formulation showed the antibacterial and antifungal
activities against Staphylococcus aureus and Candida
albicans. It is very good attempt to establish herbal gel of
Vitex negundo extract and Anacyclus pyrethrum extract.
The data presented in this study, it was demonstrated that
the developed herbal gel formulation possess significant,
therapeutically efficacious, suitable vehicle for drug
delivery in low cost but definitely with high potential. The
result showed that due to combination dosage form
developed new herbal gel formulation having good activity
so it is safe, stable and good for mouth ulcer treatment.
ACKNOWLEDGMENT
The authors express sincere thanks to the Head of the
Department of Pharmacognosy, P. E. Society’s Modern
College of Pharmacy, Nigdi, Pune Maharashtra for the
facilities provided to carry out this research work.
CONFLICT OF INTERESTS
The authors declare no conflict of interest
ETHICS APPROVAL
Not applicable
FUNDING
This study received no specific funding from public,
commercial, or not-for-profit funding agencies.
AI TOOL DECLARATION
The authors declares that no AI and related tools are used to
write the scientific content of this manuscript.
DATA AVAILABILITY
Data will be available on request
REFERENCES
Singh, M., & Mittal, V. (2014). Formulation and evaluation
of herbal gel containing ethanolic extract of Ipomoea
fistulosa. International Journal of Scientific Research,
3(6), 1866.
Indhumathi, S., & Siva, K. K. (2020). A review on
medicated chewing gum and its role in mouth ulcers.
Research Journal of Pharmacy and Technology, 13(1),
2020-2023.
Mhaske, S. D., Jagdish, S. V., Holkar, P. U., Gulve, A. S.,
& Tarade, A. S. (2022). Formulation and evaluation of
oral antiulcer gel from liquorice extract. Journal of
Emerging Technologies and Innovative Research, 9(6),
297-310.
Raghvendra, K. Y. (2021). A review on mouth ulcer and its
various treatment. Research and Reviews: Journal of
Pharmacy and Pharmaceutical Sciences, 10(11), 1-6.
Sharma, P. C., et al. (2005). Database on medicinal plants
used in Ayurveda (1st ed., reprint, Vol. 3, p. 451). New
Delhi: Central Council for Research in Ayurvedic
Sciences (CCRAS).
Neha, R., Jumde, R. D., Dongre, V. G., & Abhijeet, A. B.
(2022). A review of Anacyclus pyrethrum. International
Journal of Scientific Development and Research, 7(5),
378–382.
www.ijzab.com 7
Optimization of batches
After analysis of formulations for their extrudability and gelling strength it is observed that the formulation containing
equal amount of Vitex negundo extract and Anacyclus pyrethrum extract showed good result. The batch F (mixed)
optimized with good pH, viscosity, spreadability, extrudability and gelling strength, hence this formulation is further used
for anti-fungal study.
Table 12. Optimization of batches.
Parameters Optimized batch F (Herbal gel) Optimized batch F (Marketed gel)
Color Yellowish green Dark brown
Odor Characteristics Odorless
pH 6.7 6.9
Viscosity 6200 7100
Spredability 12.31 11.17
Extrudability 86.3 89.5
Gelling strength 28.87 27.19
CONCLUSION
From the mentioned results, it was concluded that the
prepared gel formulations are in good appearance with
suitable pH range. Formulated gel have good homogeneity,
proper gel strength & spread ability. The gel is neither too
thick nor too thin. The all-formulated gels are found to be
stable in closed container as compared to open container.
The formulation showed the antibacterial and antifungal
activities against Staphylococcus aureus and Candida
albicans. It is very good attempt to establish herbal gel of
Vitex negundo extract and Anacyclus pyrethrum extract.
The data presented in this study, it was demonstrated that
the developed herbal gel formulation possess significant,
therapeutically efficacious, suitable vehicle for drug
delivery in low cost but definitely with high potential. The
result showed that due to combination dosage form
developed new herbal gel formulation having good activity
so it is safe, stable and good for mouth ulcer treatment.
ACKNOWLEDGMENT
The authors express sincere thanks to the Head of the
Department of Pharmacognosy, P. E. Society’s Modern
College of Pharmacy, Nigdi, Pune Maharashtra for the
facilities provided to carry out this research work.
CONFLICT OF INTERESTS
The authors declare no conflict of interest
ETHICS APPROVAL
Not applicable
FUNDING
This study received no specific funding from public,
commercial, or not-for-profit funding agencies.
AI TOOL DECLARATION
The authors declares that no AI and related tools are used to
write the scientific content of this manuscript.
DATA AVAILABILITY
Data will be available on request
REFERENCES
Singh, M., & Mittal, V. (2014). Formulation and evaluation
of herbal gel containing ethanolic extract of Ipomoea
fistulosa. International Journal of Scientific Research,
3(6), 1866.
Indhumathi, S., & Siva, K. K. (2020). A review on
medicated chewing gum and its role in mouth ulcers.
Research Journal of Pharmacy and Technology, 13(1),
2020-2023.
Mhaske, S. D., Jagdish, S. V., Holkar, P. U., Gulve, A. S.,
& Tarade, A. S. (2022). Formulation and evaluation of
oral antiulcer gel from liquorice extract. Journal of
Emerging Technologies and Innovative Research, 9(6),
297-310.
Raghvendra, K. Y. (2021). A review on mouth ulcer and its
various treatment. Research and Reviews: Journal of
Pharmacy and Pharmaceutical Sciences, 10(11), 1-6.
Sharma, P. C., et al. (2005). Database on medicinal plants
used in Ayurveda (1st ed., reprint, Vol. 3, p. 451). New
Delhi: Central Council for Research in Ayurvedic
Sciences (CCRAS).
Neha, R., Jumde, R. D., Dongre, V. G., & Abhijeet, A. B.
(2022). A review of Anacyclus pyrethrum. International
Journal of Scientific Development and Research, 7(5),
378–382.
Kalyani Kakad et al. Int. J. Zool. Appl. Biosci., 10(5), 1-8, 2025
www.ijzab.com 8
Nappinnai, M., Pakalapati, S., & Arimilli, R. (2006).
Rofecoxib gels – preparation and evaluation. Indian
Drugs, 43(1), 15–18.
Das, S., Haldar, P. K., & Pramanik, G. (2011). Formulation
and evaluation of herbal gel containing Clerodendron
infortunatum leaves extract. International Journal of
PharmaTech Research, 1(1), 140-143.
Mokashi, M. (2015). Formulation and evaluation of herbal
gel containing methanolic extract of Annona squamosa
leaves. International Journal of Scientific Research, 4(6),
1064-1065.
Thombre, K. P., Sharma, D., & Lanjewar, A. M. (2018).
Formulation and evaluation of pharmaceutical aqueous
gel of powdered Cordia dichotoma leaves with guava
leaves. American Journal of PharmTech Research, 8(3),
269-274.
Yogi, D. (2015). Formulation and evaluation of gel
containing amlexanox for mouth ulcer. International
Journal of Applied Pharmaceutical and Biological
Research, 4(3), 356–364.
Kaur, L. P., Garg, R., & Gupta, G. D. (2010). Development
and evaluation of topical gel of minoxidil from different
polymer bases in application of alopecia. International
Journal of Pharmacy and Pharmaceutical Sciences, 2(3),
43-47.
Mendhekar, S. Y., Thorat, P. B., Bodke, N. N., & Jadhav,
S. L. (2017). Formulation and evaluation of gel
containing neem, turmeric, aloe vera, green tea and lemon
extract with activated charcoal and honey. European
Journal of Pharmaceutical and Medical Research, 4(3),
439-444.
Pandey, N. (2011). Formulation and evaluation of in vitro
antimicrobial activity of gel containing essential oils and
effect of polymer on their antimicrobial activity.
International Journal of Pharmacy and Pharmaceutical
Sciences, 3(2), 234-238.
Koland, M. (2011). In vitro and in vivo evaluation of
chitosan buccal films of ondansetron hydrochloride.
International Journal of Pharmaceutical Investigation,
1(3), 164-171.
Satish Kumar, A., Ravindra, A., Gopi Krishna, C. H.,
Vijaya Prakash, C. H., & Jansi Rani, M. (2016).
Formulation and evaluation of an herbal mouth gel
containing methanolic extract of Psidium guajava tender
twigs for treating oral mucositis. Journal of Global
Trends in Pharmaceutical Sciences, 7(3), 3009-3012.
Sharma, R., et al. (2013). Phytochemical investigation and
in-vitro antioxidant activity of Vitex negundo L. leaves.
Journal of Chemical and Pharmaceutical Research, 5(7),
56-62.
Sharma, P. K., et al. (2012). Phytochemical investigation
and pharmacognostic standardization of Anacyclus
pyrethrum DC. Asian Pacific Journal of Tropical
Biomedicine, 2(1), S329-S332.
www.ijzab.com 8
Nappinnai, M., Pakalapati, S., & Arimilli, R. (2006).
Rofecoxib gels – preparation and evaluation. Indian
Drugs, 43(1), 15–18.
Das, S., Haldar, P. K., & Pramanik, G. (2011). Formulation
and evaluation of herbal gel containing Clerodendron
infortunatum leaves extract. International Journal of
PharmaTech Research, 1(1), 140-143.
Mokashi, M. (2015). Formulation and evaluation of herbal
gel containing methanolic extract of Annona squamosa
leaves. International Journal of Scientific Research, 4(6),
1064-1065.
Thombre, K. P., Sharma, D., & Lanjewar, A. M. (2018).
Formulation and evaluation of pharmaceutical aqueous
gel of powdered Cordia dichotoma leaves with guava
leaves. American Journal of PharmTech Research, 8(3),
269-274.
Yogi, D. (2015). Formulation and evaluation of gel
containing amlexanox for mouth ulcer. International
Journal of Applied Pharmaceutical and Biological
Research, 4(3), 356–364.
Kaur, L. P., Garg, R., & Gupta, G. D. (2010). Development
and evaluation of topical gel of minoxidil from different
polymer bases in application of alopecia. International
Journal of Pharmacy and Pharmaceutical Sciences, 2(3),
43-47.
Mendhekar, S. Y., Thorat, P. B., Bodke, N. N., & Jadhav,
S. L. (2017). Formulation and evaluation of gel
containing neem, turmeric, aloe vera, green tea and lemon
extract with activated charcoal and honey. European
Journal of Pharmaceutical and Medical Research, 4(3),
439-444.
Pandey, N. (2011). Formulation and evaluation of in vitro
antimicrobial activity of gel containing essential oils and
effect of polymer on their antimicrobial activity.
International Journal of Pharmacy and Pharmaceutical
Sciences, 3(2), 234-238.
Koland, M. (2011). In vitro and in vivo evaluation of
chitosan buccal films of ondansetron hydrochloride.
International Journal of Pharmaceutical Investigation,
1(3), 164-171.
Satish Kumar, A., Ravindra, A., Gopi Krishna, C. H.,
Vijaya Prakash, C. H., & Jansi Rani, M. (2016).
Formulation and evaluation of an herbal mouth gel
containing methanolic extract of Psidium guajava tender
twigs for treating oral mucositis. Journal of Global
Trends in Pharmaceutical Sciences, 7(3), 3009-3012.
Sharma, R., et al. (2013). Phytochemical investigation and
in-vitro antioxidant activity of Vitex negundo L. leaves.
Journal of Chemical and Pharmaceutical Research, 5(7),
56-62.
Sharma, P. K., et al. (2012). Phytochemical investigation
and pharmacognostic standardization of Anacyclus
pyrethrum DC. Asian Pacific Journal of Tropical
Biomedicine, 2(1), S329-S332.