INTRODUCTION:

Topical gel formulations are used for skin applications. Fungal infections, which are caused by a variety of pathogenic fungi, are a significant burden on the global healthcare systems and can often lead to chronic illnesses and problems if left untreated. The emergence of drug-resistant fungus strains has made this issue more challenging, necessitating the search for alternative treatments. Gels are essentially semi-solid mixtures with a liquid phase that has been thickened with additional ingredients. ^[1]

Received on 02.05.2025 Revised on 03.06.2025

Accepted on 28.06.2025 Published on 08.07.2025

Available online from July 12, 2025

Asian J. Pharm. Tech. 2025; 15(3):256-260.

DOI: 10.52711/2231-5713.2025.00039

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.

Numerous bodily parts can be invaded by fungi, leading to fungal infections and a range of symptoms and health issues. These diseases can affect the skin, nails, internal organs, and respiratory system. Common types include ringworm, candidiasis, athlete's foot, and fungal nail infections. Fungi can also spread through inhaling fungal spores, contaminated soil, or direct contact with an infected human or animal.²

Fig.1: Fungal Infection.

Herbal remedies have long been recognized to have antibacterial properties; two in particular, tulsi leaf (Ocimum sanctum Linn) and betel leaf (Piper betle), have garnered attention because of their potential to heal fungal infections. Betel and tulsi leaves have long been used in many civilizations for their medicinal properties, which include their capacity to combat fungus. Betel leaf contains bioactive compounds with broad-spectrum antibacterial qualities, including phenols, flavonoids, and tannins.³

Fig.2: Piper betle.

Similarly, tulsi leaves are rich in phenols, flavonoids, and essential oils that have potent antifungal and antibacterial properties.⁴

Fig.3: Ocimum Sanctum.

This study aims to develop and evaluate a herbal gel including extracts from betel and tulsi leaves as a potential treatment for fungal infections. Combining these two herbal ingredients may have synergistic effects that improve the gel's overall antifungal effectiveness while lowering adverse effects.⁵

MATERIALANDMETHODS:

Collection and authentication of plant materials:

In January 2024, the leaves of the plants Piper betle and Ocimum sanctum were gathered from the vicinity of Shahada, Nandurbar, Maharashtra, India. Dr. S.K. Tayade, Head of the Botany Department at P.S.G.V.P. Mandal's Arts, Science, and Commerce College, Shahada, District Nandurbar, respectively, authenticated the plant specimens.

Chemicals: Carbopol 940, Methyl paraben, propyl paraben propylene glycol 400, Triethanolamine, Distilled water, ethanol, methanol, Glycerin.

Equipment’s: Digital balance, PH meter, magnetic stirrer, Digital Water bath, Ultra sonicator, Brookfield viscometer.

The aim of present study is to established the formulation and evaluation of herbal antifungal gel containing extracts of piper betle and ocimum sanctum leaves to treat fungal infection.

Preparation of plant extract:

We start the process of creating plant extract from betel and tulsi leaves. For three to four weeks, the leaves from both were collected, cleaned under running water to remove any dust, and then let to air dry. A 40-mesh screen was used to sift the dried plant components after they had been ground into a coarse powder using a mechanical grinder. Then, to get the proper extracts, the powder was cold macerated with ethanol, methanol, and water. In separate conical flasks, 100 g of powdered dried betel leaves and 100 g of powdered tulsi leaves were macerated with 500 ml of ethanol, methanol, and water for 48 hours. at room temperature, occasionally trembling. after 48 hours.

A simple filtration technique was used to remove the mixture, and the filtrates were gathered in different vessels. A rotary vacuum evaporator operating at 45–50°C was used to extract the solvent from the filtrate under low pressure.⁶

Table1: Ingredients used in formulation.

Sr. No.	Ingredients	Functional category
1	Liquid Extract	Antifungal
2	Carbopol940	Gelling agent
3	PropyleneGlycol400	Solubilizing agent
4	Methyl paraben, propyl paraben	Preservatives
5	Triethanolamine	pH Adjuster
6	Glycerin	Moisturizing agent
7	Rose water	Fragrance
8	Distilled water	Vehicle

Method of Preparation of gel:

Mix 2 grams of carbopol 940 with 80 milliliters of refined water in a measuring glass. Set a container next to the carbopol for 30 seconds. Put 5 milliliters of refined water in a different measuring device, add 0.2 grams of methyl paraben and 0.1 grams of propyl paraben by warming it in a water shower, then cool the mixture. Add 400 milliliters of propylene glycol, mix carbopol, and add refined water and triethanalomine drop by drop to adjust the pH.⁷

Formulation Table:

Table No. 2 contained the formulas and the previously mentioned process. The necessary amounts of piper betle leaf extract and ocimum sanctum leaf extract were added to the antifungal gel to create a control sample gel as well.⁸

Following the testing of four distinct formulation batches, we visually inspected each batch. When the gel was evaluated for homogeneity and the formulation had good gelling properties, the fourth formulation trial was found to have the best visual inspection. After that, the herbal gel was made and put through a number of tests.

Table2: Formula for preparation of gel.

Ingredients	Quantity in gm or ml
Ingredients	F1	F2	F3	F4
Betel leafextract	0.5	0.8	1	1
Tulsileafextract	0.5	0.8	1	1
Carbopol 940	1	1.5	1.8	2
Propyleneglycol400	8	9	10	10
Glycerin	1	1.2	1.4	1.5
Triethanalomine	q.s	q.s	q.s	q.s
Rosewater	q.s	q.s	q.s	q.s
Distilledwater	Maketo100	Maketo100	Maketo100	Maketo100

Evaluation Parameter:

1. Physical evaluation

2. Homogeneity

3. Measurement of pH

4. Spreadability

5. Viscocity

6. Gel strength

7. Stability Study

8. Extrudability

9. Clarity

10. Anti-fungal activity

11. Skin irritation test

Physical Evaluation: Visual inspection was done for physical characteristics such color, consistency, and odor.

Color: Visual inspection was used to verify the formulations' color.

Consistency: By applying the formulations to the skin, their consistency was examined.

Scent: The formulations' odours were assessed by combining the gel with water and monitoring the gel's scent.

Homogeneity:

Before the gels were put inside the container, each manufactured gel formulation was examined visually to make sure it was uniform. Any type of aggregate was used to investigate their appearance and existence. The gel formulation was described as homogenous.¹⁰

Measurement of pH:

The gel formulation's pH was measured with a digital pH meter. One gram of gel should be dissolved in 100 millilitres of distilled water, and the liquid should be separated for two hours. Following that, the glass electrode was completely submerged in the gel system three times to measure the pH of the formulation. The average value was then provided.⁹

Spreadability:

The amount of time it takes for two slides to separate from gel placed between them under a particular load is known as spreadability, and it is measured in seconds. Spreadability is improved if it takes less time to separate two slides. The following formula is used to determine spreadability:

S =M ×L÷T

Where

M=weight tied to upper slide

L = length of glass slide

T = time taken to separate the slides¹²

Spreadability of gel formulation was reported.

Viscocity:

Using a Brookfield viscometer with spindle number 1, the prepared gel's viscosity was determined. at 25°C. The gels rotated at three different speeds: 0.3, 0.6, and 1.5 revolutions per minute. For every speed and beats per minute, the corresponding dial reading was noted. The dial readings of the Brookfield viscometer, which were based on factors found in Brookfield viscometer catalogs, were then used to measure the viscosity of the created gels. After that, viscosity was reported.¹¹

Gel strength:

The number of seconds needed for the weight to penetrate the gel determined the gel's strength. A 10 ml graduated cylinder was filled with 3–5 ml of formulation. The gel surface was subjected to a 250-gram weight. The number of seconds needed for the weight to pierce 2 cm into the gel was used to calculate the gel strength, which represents the gel at physiological temperature. Every measurement was carried out three times.¹⁴

Stability Study:

Collapsible tubes containing the antifungal gel we made were kept at room temperature and 40°C with 75% relative humidity. We checked its appearance, pH, homogeneity, viscosity, and spread ability every month for three months throughout the stability test.¹⁶

Extrudability:

The intended gel was filled into well-known, covered, folding metal tubes and secured with as much creasing as was practical. The cylinders were clamped and placed between two glass slides while the weight of the filled tubes was noted. After placing a 250-gram weight over the slides, the cover was removed to allow for expulsion. Once collected, the amount of gel ejected was measured. still in the air by calculating the amount of gel that was evacuated. Excellent extrudability occurs when it exceeds 90%.

It is considered good extrudability when it exceeds 80%.

Clarity:

The clarity of all four batches was determined by visual inspection.

Anti-fungal activity:

The cup-plate approach has been used to test the antifungal activity of both the optimized formulation and the blank formulation. Candida albicans was used in the antifungal activity test. The prepared nutrition was brought, poured onto sterile petri dishes, and then kept apart to cool and dry. The culture of Candida albicans was then dispersed using a micron wire loop. In the past, a sterile stopper drill with a 6mm measurement was used to drill holes 4 mm deep. Next, insert the gel plan into these holes. After that, the plates were brooded for 48 hours at 27°C. The zone of restraint was then estimated at that location.¹⁷

Skin irritation test:

Applied the herbal antifungal gel formulation on the skin and observed for irritation, redness or Rashes.

RESULTS AND DISCUSSIONS:

1. Physical evaluation

2. Homogeneity

3. Measurement of pH

4. Spreadability

5. Viscocity

6. Gel strength

7. Stability Study

8. Extrudability

9. Clarity

10. Anti-fungal activity

11. Skin irritation test

1. Physical evaluation:

When the herbal antifungal gel formulation was applied, it was discovered to have a yellowish green color, a pleasant odor, and a smooth, nice consistency.

2. Homogeneity:

Every gel that was created was meticulously inspected visually for homogeneity and to look for any lumps, flocculates, or aggregates. All formulations were found to be sufficiently consistent. The herbal gel formulation has good uniformity.

3. Measurement of pH:

These antifungal gel formulations have a pH of 5.43, however the pH range of antifungal herbal gel formulations is 4–7. which is suitable for the skin, ensuring that the herbal gel composition is compatible with the skin. Figure No. 4 displays the pH of the antifungal gel.

4. Spreadability:

Spreadability is crucial for patient compliance and promotes uniform gel application. A high-quality gel will have outstanding spreadability and require less time to spread. The rbal antifungal gel formulation has a spreadability of 9.80g Cm/sec.

5. Viscosity:

Viscosity of gel was determined using Brookfield viscometer. Viscosity is an essential requirement for distinguishing the gels as it influences the spreadability, extrudability, and release of the drug. The viscosity of herbal antifungal gel is 4600cps.

6. Gel strength:

One crucial factor is the gel strength. The herbal antifungal gel has a gelling strength of 25±0.11.

7. Stability Study:

For three months, stability tests were performed on each formulation. Physical appearance, pH, odor, homogeneity, and spreadability at room temperature were among the established limits that showed no discernible alterations.

8. Extrudability:

When extruded from the metallic collapsible tube, the gel formulation exhibits good extrudability. The herbal gel formulation has an extrudability of 89.2.

9. Clarity:

Visual inspection was used to determine the gel formulation's clarity. The formulation of the herbal antifungal gel was quite clear.

10. Anti-fungal Activity:

Candida albicans was used in the antifungal activity test. After that, the diameter of the zone of inhibition—which is 20 mm—was measured.

11. Skin irritation test:

The prepared herbal antifungal gel formulation was applied on skin of hand and exposed to sunlight for 4-5 min. It was discovered to be non-irritating and skin compatible.

Fig.4: Zone of inhibition

RESULT AND DISCUSSION:

Following batch examination of formulations for evaluation characteristics such as pH, viscosity, spreadability, extrudability, and gelling strength, it was found that the formulation with an equal amount of extract from betel and tulsi leaves performed well. These formulations are further utilized for antifungal research because they are tailored with good pH, viscosity, spreadability, extrudability, and gelling strength (figure 3).

CONCLUSION:

In conclusion, there is encouraging potential for treating fungal infections with the creation of a herbal gel that contains extracts of powdered betel leaf (Piper betle) and tulsi leaves (Ocimum sanctum Linn). In vitro, the combination of these two medicinal plants, which are well-known for their antifungal qualities, showed notable effectiveness in preventing fungal growth. During the assessment process, the gel formulation also demonstrated good skin compatibility and stability. Given the rising incidence of drug-resistant strains and the adverse effects of traditional antifungal drugs, the study emphasizes the significance of investigating alternative treatments for fungal infections. The herbal gel formulation offers a more comprehensive approach to healthcare by providing a safer and maybe more sustainable option for treating fungal infections.

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Received on 05.05.2025 Revised on 06.06.2025

Accepted on 26.06.2025 Published on 08.07.2025

Available online from July 12, 2025

Asian J. Pharm. Tech. 2025; 15(3):247-255.

DOI: 10.52711/2231-5713.2025.00038

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.

Sr. No.	Formulation code	Color	Odour	pH	Homogeneity	Viscosity
1	F1	Yellowish green	Characteristics	5.43	Sufficient consistency	4600
2	F2	Yellowish green	Characteristics	5.43	Sufficient consistency	4600
3	F3	Yellowish green	Characteristics	5.43	Sufficient consistency	4600
4	F4	Yellowish green	Characteristics	5.43	Sufficient consistency	4600

Sr. No.	Formulation code	Spreadability	Extrudability	Gelling strength	Clarity	Skin irritation
1	F1	9.8	89.2	25±11	Quite Clear	No irritation
2	F2	9.2	85.2	25±12	Quite Clear	No irritation
3	F3	9.4	88.2	25±13	Quite Clear	No irritation
4	F4	9.9	87.2	25±14	Quite Clear	No irritation