Formulation And Evaluation Of Polyherbal Gel For Skin Diseases

Fig.No.1:-  Skin Diseases

Kumar et al. (2025): Developed a herbal antibacterial gel in which they studied physiochemical characteristics and stability. Manoj A. Suva et al. (2024): Reported that hydroalcoholic extract exhibits antimicrobial, anti-inflammatory, antioxidant and antiacne activities. Salwa M. Raweh et al. (2024) Evaluated the antimicrobial activity of Azadirachta indica extract against Pseudomonas aeruginosa and Escherichia coli. Dalia A. Pondini et al. (2023): Reported that ethanolic extract of Curcuma longa possesses antioxidant, anti-inflammatory and antimicrobial activities beneficial for skin healing. Gurning Kaste et al. (2023): Rhizomes of Curcuma longa were found to contain phytochemicals such as alkaloids and tannins with performed antimicrobial activity. Femy Safri et al. (2020): Reported that Carbopol 940 is a suitable gelling agent and significantly identified gel pH, viscosity and spreadability. Although individual plant extracts have been studied extensively, there is limited research on polyherbal gels that combine multiple extracts into one formulation. The synergistic potential of combining Curcuma longa, Azadirachta indica, Aloe barbadensis and Allium sativum in a Carbapol-based gel has    Carbopol-based gel has not been systematically explored. Most studies focus on single herbs or isolated compounds, leaving a gap in developing and evaluating a comprehensive polyherbal gel for skin diseases. That’s why my aim of this research paper is To Formulate and Evaluate Polyherbal gel for Skin Diseases and objectives are to identify components required for making the polyherbal gel, to develop the polyherbal gel, to determine the Physiochemical properties, pH, Viscosity, Spreadability of polyherbal gel.

Plant Profile:-

1. Garlic^[4]:-

One of the traditional medicines that can be used to treat skin diseases. Garlic (Allium sativum) contains essential chemical compounds which are good for the body.

Organoleptic Properties:-

Colour: Pinkish white

Odour: Odoriferous

Taste: Soft, Sweet buttery flavour

Size: 1.5-2.5 cm

Biological Functions:-

• Antioxidant Activity
• Anti-inflammatory Activity
• Antimicrobial Activity
• Modulating Immunity System

T

able No.1:- Taxonomical classification of Allium sativum

Fig.No.2:-  Garlic

2. Turmeric^[5]:-

Turmeric (Curcuma longa) is also a traditional medicine which is used in the various skin diseases. It uses as an antiseptic for cuts, burns & brises and as an antibacterial agent.

Organoleptic Properties:-

Colour: Yellow

Odour: Earthy, Woody

Taste: Bitter

Size: 1m

Biological Functions:-

• Antioxidant Activity
• Anti-inflammatory Activity
• Antimicrobial Activity
• Modulating Immune System

Table No.2:- Taxonomical classification of Curcuma longa

Fig.No.3:- Turmeric

3. Aloe Vera^[6]:-

Aloe Vera (Aloe barbadensis) is a semi-tropical with a broad range of applications in the foods, drugs and cosmetic industries. The aloe vera contains various health benefits which are important for the human body.

Organoleptic Properties:-

Colour: Green

Odour: None

Taste: Bitter

Size: 60-100 cm

Biological Functions:-

• Anti-Cancer Activity
• Antiulcer Activity
• Antifungal Activity
• Antidiabetic Activity

Table No.3:- Taxonomical classification of Aloe barbadensis

Fig.No.4:-  Aloe Vera

4. Neem^[7]:-

The Neem is one of the most versatile medicinal plant known for its numerous health benefits. The Neem tree is used for skin care and improve the skins beauty.

Organoleptic Properties:-

Colour: Yellowish Green

Odour: Indistinct

Taste: Bitter

Size: 15-20 m

Biological Functions:-

• Anti-Cancer Activity
• Antiulcer Activity
• Antifungal Activity
• Antidiabetic Activity

Table No.4:- Taxonomical classification of Azadirachta indica

Fig.No.5:- Neem

MATERIALS AND METHODOLOGY

Materials:-

Ethanol, Nutrient agar, Carbopol-940, Propylene glycol., Methyl paraben, Propyl Paraben, Triethanolamine, Plant extracts (Allium sativum, Curcuma longa, Aloe barbadensis and Azadirachta indica), Rose water, Distilled Water.

Instruments:-

Digital pH meter (alpha-01d), Digital Brookfield Viscometer (DV-E)

Methodology:-

Collection

Leaves of Aloe barbadensis, Azadirachta indica was collected from college premises and dried rhizomes of Curcuma longa and fresh buds of Allium sativum was brought from market of Warthi, Bhandara.

Authentication

Authentication of leaves of Aloe barbadensis, Azadirachta indica, buds of Allium sativum and dried rhizomes of Curcuma longa was done by Dr. Mrs Sayeda Qureshi, Associate Professor and Head of Botany Department, J. M, Patel College, Bhandara.

Extraction:-

By using the process of maceration, the extract of Aloevera, Garlic, Neem and Turmeric was prepared.

1. Aloevera

Aloevera leaves were washed and homogenized using a blender. The homogenate was dissolved in a ratio of 1:1 of distilled water and methanol respectively for 48 hours. The solutions were filtered and the filtrate

was evaporated at 45℃ in an obtain aqueous and methanol extract of Aloevera[8].

2. Turmeric

The Turmeric rhizome was cleaned and cut into small pieces. Samples of black rice bran and turmeric dried in an oven at 35-40℃ to a constant weight, then ground and sieved through a 40- mesh sieve. 500 g of rice bran and 60g of turmeric powder soaked in a 96% ethanol solvent (1:10 weight/volume) for 2 days samples (24h) with occasional stirring. The filtrate with Whatman paper No. Yield (%) of evaporated dried extracts was calculated as Distilled Water extracts/Distilled Water×100% Where Distilled Water sample is the dry weight of the sample and Distilled Water is the weight of the extracts after drying[9].

3. Neem

Neem extraction of mature fresh neem leaves collected from farm in Warthi. The fresh neem leaves were washed under tap water to remove the dust and foreign particles then dried in shade for 4 hours and then in oven at 50℃ for 1 hr. The dried leaves were used to make neem powder. 30 g of neem powder was extracted with 95% ethanol, methanol or distilled water. The solutions were incubated for 24 h in shaker incubator. Subsequently, the mixture was boiled in hot water bath for 30 min and further incubated vin shaker incubator for 24 h. After incubation the solution was filtered under vacuum through Whatman No. 1 filter paper. The organic solvents were removed by rotary evaporator under reduced pressure at 40℃ to give crude extract. The aqueous extract was evaporated by freeze dry machine to remove the solvent[10]

4. Garlic

The garlic was first dried then grind into a fine mash. This mash was soaked in 96% ethanol at a ratio of 1:10 and left to macerate for 24 hours. The process was repeated three times to ensure through extraction. Afterward, the liquid extract was concentrated using a rotary vacuum evaporator, leaving behind a thick garlic extract. To prepare different strengths the concentrated extract was diluted with distilled water to create solutions of 20%, 40%, 60% and 80%. These preparations were then examined for their organoleptic properties such as colour, smell and subjected to phytochemical screening to identify the presence of secondary metabolites in a qualitative manner[11].

Preparation of Polyherbal Gel

Polyherbal gel formulations are made by combining Carbopol-940, Propylene glycol, Methyl paraben, Propyl paraben and Triethanolamine. After that the dried crude extract of all Four plant ingredients was added and the mixture was stirred continuously to ensure the extracts blended evenly throughout the gel resulting in consistent formulation[12].

Table No.5:- Composition of Polyherbal Gel

Phytochemical Screening for Extracts:-

1. Test for Garlic (Flavonoids)

Ferric chloride Test: Add extract and 2ml of ferric chloride which develops black colour and it shows positive test.

2. Test for Turmeric (Alkaloids)

Mayer Test: Add extract and 2 ml of Mayer reagent which develops  cream colour precipitate and it shows positive test.

Hager Test: Add extract and 2 ml of Hager Reagent which gives white precipitate and it shows positive test.

3. Test for Aloevera (Glycoside)

Modified Borntrager Test: Add extract and 2 ml of ferric chloride,2 ml of dilute Hydrochloric acid and heat it for 5 minutes for boiling then filter, cool and add benzene which separates organic layer and due to ammonia, it develops pink colour it shows positive result.

4. Test for Neem

Biuret Test: Add extract and water, 2ml of 1% potassium hydroxide, few drops of copper sulphate which develops violet colour and it gives positive test[13].

Anti-microbial Activity of Extract:-

Test Micro-organisms

In this work, two prevalent pathogenic micro-organisms the gram-negative bacteria Pseudomonas aeruginosa and Escherichia coli were employed. Antibacterial activity was assessed using both bacterial strains that were acquired from an Anurag College of Pharmacy in Warthi, Bhandara.

Anti-microbial Activity Screening

The anti-microbial activity of the four extracts was determined according to the agar well diffusion method. In this method pure isolates of micro-organisms were subcultured on the recommended specific medium at 37℃ for 24 hours. One hundred microliters of inoculum of each test organism was spread onto the specific media plates (containing Mueller Hinton Agar) and 6 mm diameter wells were bored with a sterile borer in the inoculated agar plates 100 microliters of each extract were injected directly into wells (in triplicates) of inoculated specific media agar plates for each test organism. The plates were left to stand for 10 minutes to allow diffusion of the extract and incubated at 37℃ for 24 hours. Sterile Distilled water and 20% DMSO were used as negative controls and 0.2% Chlorhexidine as a positive control. The antimicrobial activity was indicated by the presence of an inhibition zone around the well containing the extract. The zone of inhibition was measured in millimeters by means of vernier caliper[14].

Evaluation of Polyherbal Gel:-

1. Visual Appearance:-

The prepared polyherbal gels were inspected for their colour, greasiness, consistency and odour[15].

2. pH Determination:-

The pH of all formulated herbal gels was measured by using digital pH meter[16].

3. Spreadability of Gel Formulations:-

The spreadability of the formulated gels were assessed using the glass slide technique, a widely adopted method for evaluating topical preparations. In this procedures, approximately 0.5 g of gel was carefully placed between two clean glass slides. A weight of125 g was applied for five minutes to allow the gel to spread evenly, forming a uniform thin film. Following this, any excess gel was removed and a smaller weight of 20 g was attached to upper slide. The time required for the two slides to separate under this load was recorded as an indicator of spreadability. The spreadability value was then calculated using the standard equation providing a quantitative measure of the ease with which the gel can be applied over a surface.

S= M / T

Where,

S= Spreadability (g/s)

M= Applied weight (g)

T = Time taken for slide separation (s)[17].

4. Viscosity:-

Gels viscosity was studied using Brookfield Viscometer[18].

5. Homogeneity:-

The homogeneity of the gel formulations was examined visually after the gel set in their containers. The evaluation focused on their overall appearance of any visible aggregates with a smooth and uniform textures indicating good homogeneity[19].

6. Washability:-

The formulations were applied on the skin and then ease and extent of washing with water were determined manually[1].

7. Skin Irritancy Test:-

The formulation was applied to a marked area on the dorsal surface of the skin and observed for erythema, oedema or irritation over 2 hours[20].

8. Antimicrobial Study:-

1. Isolation of Bacterial Strains

Pseudomonas aeruginosa and Escherichia coli strains were produced from the microbiology laboratory and cultured on nutrient agar medium (Merck Millipore). Thon nutrient agar medium (Merck Millipore). The plates were incubated under anaerobic conditions for 48 hours to promote bacterial growth. For the human swab sample, a volunteer clinically diagnosed with acne vulgaris was selected. The volunteer’s face was rinsed with distilled water and dried using a sterile cotton wipe. A visible comedone was carefully punctured with a sterile acne needle,  A visible   acne needle, and the exudate was collected using a sterile cotton swab. This swab was then immersed in 5 ml of distilled water for five minutes, After which the suspension was evenly spread onto freshly prepared agar plates. The plates were incubated at 37℃ for 48 hours, allowing distinct bacterial colonies to develop.

2. Evaluation of Antibacterial Activity

The antibacterial efficacy of the gel formulation was tested using the disc diffusion method against the two bacterial stains. Bacterial stain cultures were standardized in Brain Heart Infusion broth to achieve a density of approximately 10⁸  cells /ml. From this suspension , 10 ml was uniformly spread onto agar plates. Sterile filter paper discs (1 cm diameter) were soaked for five minutes in either the prepared polyherbal gel or a Standard Clindamycin gel. The treated discs were then placed at the centre of the inoculated plates and incubated for 72 hours. After incubation the zones of inhibition were measured in millimeters, starting from the edge of the disc to the boundary where bacterial  growth was halted[21].

9. Thin Layer Chromatography (TLC):-

1. TLC of Garlic Extract:-

The polar mobile phase was formulated using equal volumes of glacial acetic acid, propanol, water and ethanol (20:20:20:20, v/v/v/v). For the non-polar mobile  phase, a mixture of hexane and acetone in the ratio of 70:30 was prepared and adjusted to a final volume of 100 ml. Silica gel60 Tlc plates served as the stationary phase. After development, the plates were examined under UV light at 254 nm, and Rf values were determined. To visualize amino acid derivatives, ninhydrin and anisaldehyde were applied as spraying reagents[22].

2. TLC of Turmeric Extract and Neem Extract:-

A standard solution was prepared by dissolving 10 mg of the reference compounds in 10 ml of methanol. The mixture was filtered to eliminate any insoluble particles and the clear filtrate was subsequently used for spotting onto silica gel plates. The test solution was prepared by dissolving 0.5 g of the extract in 100 ml of methanol. The mixture was then filtered to remove any insoluble material and the resulting filtrate was applied for spotting onto silica gel plates After development, the plates were examined under UV light at 256 nm, and Rf values were determined.[23].

3. TLC of Aloevera Extract:-

TLC plates were prepared using silica gel as absorbent. To make the coating slurry 15 g of silica gel-G was blended with 30 ml of distilled water and promptly spread onto the plates. These were left to air-dry for 1 hour followed by heat treatment at 100℃ for 90 minutes to fix the layer. Using a micropipette approximately 10 microliters of extract were carefully applied to the plates and allowed to dry. Development was carried out with two solvent systems: system 1 composed of chloroform and methanol (12:2) while system 2 consisted of ethyl acetate, toluene and formic acid in the ratio of 2:2:1:1:1:1 After development, the plates were examined under UV light at 254 nm, and Rf values were determined.[24].

RESULT AND DISCUSSION:-

Evaluation Test for Extract

1. Determination of Phytochemical screening for extract:-

Table No.6:- Phytochemical screening for extracts

Note: (+++) Highly present, (++) Moderately present, (+) Weakly Present, (-) Absent.

2. Determination of  TLC of Extracts:-

Table No.7:- Rf value of Extracts

Fig.No.6:- Rf Value of Extracts

Evaluation Test for Polyherbal Gel

1. Determination of Physiochemical Properties:-

Table No.8:-  Physiochemical Properties of Polyherbal Gel

2. Determination of Spreadability:-

Table No.9:- Spreadability of Polyherbal Gel

3. Determination of Viscosity:-

Table No.10:- Viscosity of Polyherbal Gel

4. Determination of Antimicrobial Activity

Table No.11:-  Antimicrobial Activity of Polyherbal gel

Fig.No.07:- Zone of Inhibition of Extracts

Fig.No.08:- Antimicrobial Activity of Polyherbal gel