Herbal Kajal Formulation from Cyanthillium Cinereum and Ocimum Tenuiflorum for the Evaluation of Anti-Microbial Properties

Herbal kajal has been a part of traditional beauty routines for centuries, especially in Indian culture. But it’s more than just a cosmetic when prepared with natural, medicinal ingredient, it becomes medicated herbal kajal, combining the charm of beauty with the power of healing. Unlike synthetic kajal, herbal kajal is made using natural substances like camphor, almond oil, ghee, and soothing herbs. These ingredients not only enhance the appearance of the eyes but also offer various therapeutic benefits cooling the eyes, protecting them from infections, and even improving vision over time.  The eye is a highly sensitive sensory organ responsible for vision and is protected by structures such as the eyelids and conjunctiva. It consists of outer (sclera and cornea), middle (choroid), and inner layers that enable protection, nourishment, and visual function, while the eyelids maintain moisture and protect the ocular surface through blinking. Herbal kajal is considered safer than synthetic kajal due to its reduced chemical content. Hence, the present study focuses on the formulation and evaluation of herbal kajal prepared using medicinal plants Cyanthillium cinereum and Ocimum tenuiflorum, selected for their antimicrobial properties, with soot obtained from plant-extract–soaked cotton wicks serving as the base material.

OBJECTIVES

• Helps protect the eyes from infections and Irritation.
• Naturally cools the eyes, easing strain and tiredness.
•  May support better sleep and help with Insomnia.
• Gives a smooth, long-lasting application without causing irritation.
•  Offers a safe and natural alternative to Chemical-based eye cosmetics. ^[1]

MATERIALS AND METHODS

MATERIALS

1. Cyanthillium Cinereum

Fig.no.1: Cyanthillium Cinereum (L) H. Rob.

Botanical name: Cyanthillium Cinereum (L) H. Rob.

Chemical constituents: flavonoids, phenolics, alkaloids, terpenoids, tannins, saponins, and steroids, which together give it antioxidant, antimicrobial, anti-inflammatory, and wound-healing properties.

Pharmacological properties: Anti-microbial activity, anti-oxidant, analgesics, anti-inflammatory etc.

Uses:

• used in traditional kajal preparations
• applied on skin for rashes, itching and insect bites
• helps in eye cooling and reduces eye irritation
• used for cough cold and bronchitis ^[2]

2. Ocimum Tenuiflorum

Fig.no. 2: Ocimum Tenuiflorum L.

Botanical name: Ocimum Tenuiflorum L.

Chemical constituents: Tulsi composed of approximately 70% of eugenol, other components include ursolic acid, linalool, carvacrol, caryophyllene, Apigenin, Luteolin and Vitamin C, calcium, iron, Zinc.

Pharmacological properties: Anti-microbial, Anti-bacterial, Anti-diabetic activity etc.

Uses:

• Skin infection

• Religious and spiritual

• Respiratory issues
• Good for boosting up the immune system ^[3]

3. Lamp black(soot)

Fig.no. 3: Lamp black (soot)

Source: lamp black is obtained by the controlled burning of cotton cloth impregnated with herbal extract of Cyanthillium cinereum and Ocimum tenuiflorum.

Description: Lamp black is a fine, black carbon powder obtained by the incomplete combustion of oils, ghee, or plant-based materials.

Use: it is used as a natural black pigment in herbal kajal formulation.

4. Ghee

Fig.no.4: Ghee

Description: Ghee is a purified fat. Obtained from butter by removing water and milk solids. it is a clear, stable lipid with a mild nutty aroma and long shelf life.

Use: Emollient and moisturizer in pharmaceutical and cosmetic preparation, improve skin hydration.

5. Camphor

Fig.no. 5: Camphor

Description: It is a crystalline, wax-like, transparent solid that is combustible in nature and possesses a strong, sharp, slightly musty characteristic odor.

Use: Use in various cosmetics and topical formulation due its cooling and soothing effect.

Medicinaly, it is used as an anti-pruritic, analgesics and anti- inflammatory agent for skin application. ^[4]

6. Coconut oil

Fig no.6 : Coconut oil

Description: A white or pale yellow solid or semi solid fatty substance extracted from coconut palm fruit.

Use: Traditionally utilized to nourish the skin and protect it from dryness. Used in cosmetic formulations due to its moisturizing, calming and soothing effect. ^[5]

7. Almond oil

Fig.no. 7: Almond oil

Description: A pale to golden yellow oily liquid with a mild, nutty odor; insoluble in water but soluble in oils and organic solvents.

Use: used as an emollient and carrier oil in creams, lotions and bath oils to improve skin softness and spreadability. ^[6]

8. Castor oil

Fig.no.8: Castor oil

Description: it is a pale yellow to nearly colorless, transparent and viscous liquid with a faint, mild odor and a slightly unpleasant taste.

Use: widely used as a makeup remover and moisturizing agent. It promotes wound healing and exhibits anti-inflammatory, anti-bacterial, and anti- fungal activity. ^[7]

9. Vitamin E

Fig.no. 9: Vitamin E

Description: It is a fats soluble nutrient appearing as a pale yellow, oily liquid with little or no characteristic odor or taste.

Use: It functions as a potent anti-oxidant and preservative. It helps to sooth and protect the skin, reduce the formation of wrinkles and lightens dark spots. ^[8]

METHEDOLOGY

1. Collection of herbs

The fresh whole plant of Cyanthillium cinereum and Ocimum tenuiflorum were collected from Thrissur district and authenticated.

2. Extraction of juice

The plants were thoroughly rinsed using clean water to remove dirt and impurities. Then leaves were crushed and juice was extracted hygienically. The extract was filtered using a clean cotton cloth to remove solid particles.

3. Soaking

Clean cotton cloth was soaked in the herbal juice and kept in a well closed container for 8 hrs to allow proper absorption of the extract.

4. Drying

The soaked cotton has been removed from the liquid and then placed in natural drying until the product was completely lose the water content in it.

5. Collection of soot

The dried cotton cloth containing extract was used as a wick and was lightened in a mud lamp containing ghee. An inverted copper plate was placed above the flame. The black soot deposited on the copper plate was carefully scraped and collected. ^{[9] [10]}

Fig.no: 10 Lit the lamp and put the inverted copper plate

Fig.no: 11 Black soot is obtained on copper

Fig.no: 12 Collection of soot

Preparation of wax base

Ghee was weighed and taken in a china dish.

6. Preparation of oil phase

Almond oil, castor oil, coconut oil is mixed together in a separate china dish.

7. Formulation of herbal kajal

The wax base and oil phase are mixed together and gently heated using the double-boiling method for a few minutes until the mixture becomes uniform and melts completely. Once the mixture is well-melted, black soot is added slowly and stirred thoroughly to ensure even dispersion of the pigment throughout the base. After proper mixing, adding Vitamin E capsules as a preservative, camphor which provides a cooling and soothing effect on the eyes. The mixture is stirred well until all ingredients are completely blended to form a smooth kajal paste. The prepared kajal is allowed to cool and then transferred into a clean, airtight container.

Fig.no: 13 Formulated herbal kajal

Table no.1: formulation of herbal kajal

RESULT AND DISCUSSION

• Evaluation test for herbal kajal

Physical evaluation test: The formulated product is shiny black color, with a characteristic odour. It was non- gritty and smooth in texture with a semisolid consistency.

Table no. 2: Physical evaluation

• pH determination:1 gram of kajal was dispersed in 25 ml of DMSO and kept for two hours. The pH was measured three times using a pH meter, and the average value was recorded.

Table no. 3: pH

Graph no.1: pH determination

• Skin irritation test: The prepared kajal formulation was applied on the skin. After 24 hours, no irritation or redness was observed. ^[11]

Fig.no. 14: Skin irritation test

Table no. 4: Skin irritation test

• Spreadability test: The kajal sample was applied between the two glass slides and was compressed between the two-glass slide to uniform thickness by placing 100 gm of weight for 5 minutes.

             Fig.no.15: Product spreadability                                   Fig.no. 16: marketed product spreadability

Table no. 5: Spreadability test

• Stability studies: Physical parameters such as colour, odour, texture, and consistency were determined at room temperature &40ºc.

Table no. 6: Stability studies

• Evaluation of base

The evaluation of base that is ghee was evaluated by Acid value and Saponification value.

5. Acid value

The acid value is to neutralize the free acid in 1 g of substance the number of mg of potassium hydroxide is required. Determined by the following method weigh accurately about 10 g of the substance in the 250 ml of conical flask and add 50 ml of alcohol and add 1 ml of phenolphthalein. Warm up on water bath if necessary until substance was dissolved. Titrate with 0.1 N potassium hydroxide. Shake constantly shake until pink colour is obtained. Note the number of ml required and calculate the acid value by using the formula:

Acid value = a x 0.00561 x 1000/W

Where, a = number of ml of 0.1 potassium hydroxide required

W = weight of g of substance taken.

5. Saponification value

The Saponification value is the number of mg of potassium hydroxide required to neutralized fatty acid determined by following method. Add 40 gm of potassium hydroxide in 20 ml water and add sufficient alcohol to make volume 1000ml. Allow it overnight. Weigh 4g of ghee in 250 ml of conical flask add alcoholic solution of potassium hydroxide, attach to the reflux condenser set another reflux condenser as blank with reagents except ghee.1hr boil on water bath. Add 1ml of phenolphthalein. Titrate with 0.5 N HCl. Note the number of ml required and calculate the Saponification value by using the formula:

Saponification value = (b-a) x28.05/W

Where, W = weight in g of substance taken

a = sample solution reading.

b = blank solution reading ^[12]

Table no. 7: Evaluation of base

• Anti-microbial assay / anti-bacterial assay: No microbial growth was observed after incubation at 37 °C for 24 hr. The formulation exhibited significant antimicrobial activity against staphylococcus aureus with a zone of inhibition is more when comparable to a marketed product.

Fig.no. 17: Anti-microbial activity of kajal

Table no. 8: Anti-microbial activity