Formulation Development and Evaluation of Moringa Oleifera Herbal Shampoo

There has been a dramatic move towards the application of natural and herbal products in personal care over the past few years, propelled by increasing concern over the ill effects of synthetic chemicals on human health and the environment. Of these, herbal shampoos have become increasingly popular as they provide a soft, environmentally friendly alternative to traditional hair care products. Moringa oleifera, also referred to as Moringa, is a highly nutritious plant that is well known for its medicinal and beauty benefits. Moringa is rich in vitamins A, C, and E, along with essential minerals and antioxidants that encourage healthy hair development, relieve inflammation in the scalp, and fight dandruff, which creates a natural lather and cleanses the scalp free of dirt and oil without robbing the scalp of its natural moisture. The blend of Moringa in a shampoo base provides a potent synergy, imparting nourishment as well as cleansing properties. This herbal shampoo not only supports hair strength and luster but also minimizes the use of chemical-based products, making it ideal for all hair types, including sensitive scalp. This project seeks to develop and test a natural herbal shampoo out of Moringa determine its efficacy in cleansing power, conditioning action, and general scalp health. By doing this, we seek to help create environmentally friendly and health-oriented hair care products.

MATERIALS AND METHODS

1.Plant Profile of Moringa oleifera leaves

• Organoleptic Characteristic

a. Color: Bright to dark green (fresh leaves), Pale Green or brown (dried leaves)

b. Odor: Mildly earthy and herbaceous, stronger when dried

c. Taste: Bitter, tangy

d. Texture: Soft and Tender (fresh), Brittle and Coarse (dried)

• Morphological Characteristics

1. Leaf Structure: The leaves of Moringa Oleifera are bi- or tripinnately compound, meaning they are divided into smaller leaflets arranged on either side of a common axis.
2. Leaflets: Each leaflet is obovate (egg-shaped with the narrower end at the base) and measures about 12–18 mm long. The leaflets are hairy and have a smooth texture.
3. Color: The leaves are typically a vibrant green, which can vary slightly depending on the age and health of the plant.
4. Petiole: The petiole (the stalk that attaches the leaf to the stem) is yellow or white without red stripes.

Fig no.1 Morphology of Moringa oleifera leaves

5. Growth: Moringa leaves grow rapidly and are known for their high nutritional content, including vitamins, minerals, and antioxidants^{. [1]}
6. Moringa leaves (Moringa oleifera) exhibit a range of distinct morphological characteristics that contribute to their unique appearance and functionality. The leaves are typically pinnate, featuring a feather-like structure with leaflets arranged along a central stem. Each compound leaf usually consists of 3 to 9 oval to elliptical leaflets, which can vary in size, generally measuring between 1 to 3 inches in length and 0.5 to 1.5 inches in width. ^[2]
7. Young Moringa leaves are a bright green color, which deepens as they mature, while their surfaces are smooth and glossy, often covered with a slightly waxy coating that helps reduce water loss. The leaves are arranged alternately along the stem, each connected by a distinct petiole, or leaf stalk. At the base of the leaf stalk, small leaf-like structures known as stipules may be present. ^[3]

• Taxonomical Characteristics

1. Kingdom: Plantae
2. Clade: Angiosperms
3. Clade: Eudicots
4. Order: Brassicales
5. Family: Moringaceae
6. Genus: Moringa
7. Species: M. oleifera

Moringa Oleifera, also known as the drumstick tree or horseradish tree, is widely recognized for its medicinal and nutritional benefits. Its leaves are used in various traditional medicines and as a dietary supplement due to their rich nutrient profile ^[4]

• Chemical Composition

a. Flavonoids: Major flavonoids include quercetin, kaempferol, myricetin, and rutin. These compounds are known for their antioxidant properties.

 b. PhenolicAcids: Key phenolic acids found are gallic acid, chlorogenic acid, caffeic acid, and ferulic acid.

c. Alkaloids: Various alkaloids have been identified in the leaves such as moringinine and pyrrolemarumine.

d. Glucosinolates: The most abundant glucosinolate is glucomoringin (4-O-(α-L-rhamnopyranosyloxy)- benzyl glucosinolate). e. Saponins and Tannins: These compounds are also present and contribute to the pharmacological properties of the leaves ^[5]

• Extract Collection

The extract of Moringa oleifera used in formulation was collected from Shrusti Herbal Industries, located in MIDC Warud.

2.Reetha (Sapindus mukorossi)

Organoleptic Characteristics

Color: The fruit's skin is typically yellowish-brown when mature and dried, while the seed inside is smooth and black.

Odor: Reetha has a mild, earthy aroma when fresh, which becomes more pronounced and slightly pungent upon drying.

Taste: The fruit is known for its bitter and tangy flavor.

Texture: Fresh Reetha fruits are soft and tender, whereas dried fruits become brittle and coarse. ^[6-8]

Morphological Characteristics

Tree Size: Sapindus mukorossi is a deciduous tree that can grow up to 20 meters tall, with a trunk diameter ranging from 60 to 80 cm.

Leaves: The leaves are compound, measuring 15–35 cm in length, with 4–8 pairs of lanceolate to oblong leaflets.

Leaflets: Each leaflet is 6–15 cm long and 3–5 cm wide, glabrous, and lanceolate to oblong in shape.

Petiole: The petiole is glabrous and narrowly bordered.

Flowers: Small, greenish-yellow flowers are borne in terminal, pubescent panicles.

Fruit: The fruit is a globose, fleshy drupe, 1.5–2.5 cm in diameter, yellowish-brown when mature, containing a single smooth black seed

Fig no.2 Sapindus mukorossi

Reetha is widely used in preparations like shampoo. The dried fruit powder is used as a foaming agent in shampoos. It cleans the oily secretions in the skin and can be used as a cleanser for hair and a hair tonic as it forms a natural lather. It is also used for removing lice from hair. ^[9-11]

Traditional and Modern Uses

Reetha has been used for centuries in Ayurvedic medicine and as a natural detergent. Modern applications include: 

- Natural surfactant in cosmetics and shampoos 

- Antimicrobial agent in traditional medicine 

- Potential insecticidal properties 

- Source of bioactive compounds for pharmaceutical research ^[12-13] 

3. Sodium Lauryl Sulfate (SLS)

Sodium lauryl sulfate is made by reacting lauryl alcohol, which comes from either petroleum or natural sources like coconut or palm oil, with sulfur trioxide, oleum, or chlorosulfuric acid. This creates hydrogen lauryl sulfate, which is then neutralized with sodium carbonate or sodium hydroxide to form SLS. The process can be adjusted to produce high-purity SLS and may include steps such as concentration, crystallization, and drying. In shampoos, the two most common sulfates are sodium lauryl sulfate (SLS) and sodium laureth sulfate (SLES). Both are surfactants, which help to create foam and clean the hair, but SLES is usually considered milder than SLS. These ingredients are mainly used for their lathering ability, which helps emulsify and wash away oil and dirt. A rich lather in shampoo is often a sign that it contains sulfates. ^[14-16]

4.  Methyl Cellulose (HPMC and CMC)

Hydroxypropyl methylcellulose (HPMC) and carboxymethyl cellulose (CMC) are plant-based polymers made from cellulose. They are added to shampoos to make them thicker, stabilize the mixture, and improve the quality of the foam. These ingredients help prevent separation of the formula and create a more stable, longer-lasting lather by forming thin films around bubbles. ^[17-19] HPMC also helps retain moisture and makes hair easier to comb by reducing friction. It works well with salts and alcohols. CMC enhances foam stability, especially when used with surfactants like SLS. Both HPMC and CMC are biodegradable, gentle on the skin, and improve the texture and feel of shampoo, giving it a smooth, silky consistency. ^[20-22]

5. Rose Water

Rose water adds a natural, soothing floral scent to herbal shampoos, enhancing the sensory experience while promoting relaxation. Unlike synthetic fragrances, it's gentle and ideal for sensitive scalps. Its calming aroma also offers mild mood-lifting benefits, making hair care feel more like a spa ritual. Rose water is valued in hair care for its moisturizing and pH-balancing qualities. It can be used to dilute shampoo or as a final rinse after washing hair. It helps cleanse the scalp, adds shine, and makes hair softer and easier to manage. ^[23-25]

2. Formulation of herbal shampoo:

Table no. 1 Formulation of herbal shampoo

Procedure:

1. Preparation of extract

Weigh the required quantities of Moringa oleifera (drumstick) leaves extract and Sapindus mukorossi (Reetha) extract, prepare extract by decoction method then use in the formulation.

2. Addition of Surfactant

After the combination of all of the filtrates obtained, sodium lauryl sulfate (SLS) was added in a gradual way while we stirred constantly. The mixture was stirred for a time until all of the SLS was dissolved fully. This even distribution of the surfactant within the formulation was thereby ensured.

3. Thickening Process

With constant stirring, methyl cellulose was slowly incorporated into the surfactant-herbal mixture. This was done in order to adjust the viscosity of the formulation. As this process continued, a uniform, semi-solid consistency was achieved.

4. Ph Adjustment

Citric acid was added bit by bit while constantly stirring to change pH to a skin-safe level. In this process, the pH was checked to stay in a good range for hair application to the scalp.

5. Final Touch & Mixing

The mixture had rose water added for a natural fragrance enhancer. The entire formulation was then thoroughly mixed to ensure homogeneity, also this resulted in a smooth and stable semi-solid herbal shampoo

6. Storage

Transfer the prepared shampoo into a sterile container. Store in a cool, dry place away from direct sunlight.  

Evaluation

Organoleptic Evaluation

Table no. 2 Organoleptic Evaluation

2. Physicochemical Evaluation

Each of the five herbal shampoo formulations (F₁–F₅) was subjected to a series of physicochemical tests to assess parameters relevant to product quality, stability, and performance. The evaluation procedures and their significance are outlined below. The respective results are presented in Table no. 3.

 pH 

The pH of each formulation was measured using a calibrated digital pH meter at room temperature, which is crucial for determining scalp condition and product performance. Acid-balanced shampoos, with pH values of 5-6, maintain the scalp's natural acidity, reduce eye and skin irritation, and encourage hair cuticle closure, resulting in smoother hair and improved scalp homeostasis.

Fig.no.3 Digital ph meter

Viscosity

The viscosity of a shampoo was measured using a Brookfield viscometer to determine its flow properties and structural homogeneity. A suitable viscosity ensures efficient dispensement from packaging, adequate hair coverage, product stability upon storage, and consumer usability.

Foaming Ability

The foaming capacity and stability of preparations were assessed through lathering ability and foam retention over time, which are crucial for user experience and cleaning performance. Uniform foam with consistent bubble size indicates effective surfactant action and acceptable formulation performance, while foam persistence over time indicates naturally occurring surfactants sustaining lather quality.

Table no. 3 Physicochemical Properties

Fig no.4 Foaming Ability

3. Antimicrobial Activity

Experimental Method

The Kirby-Bauer disc diffusion technique is a widely used method for determining the antibiotic susceptibility of bacterial isolates. In this method, a standardized bacterial suspension, typically adjusted to match 0.5 McFarland turbidity standards, is uniformly spread over the surface of a Mueller-Hinton agar plate using a sterile cotton swab. After inoculation, antibiotic-impregnated paper discs are placed on the agar surface at appropriate distances. The plates are then incubated at 35–37°C for 16–18 hours to allow bacterial growth and diffusion of the antibiotics into the surrounding medium. Following incubation, zones of inhibition—clear areas where bacterial growth has been prevented—are measured in millimeters. These measurements are compared with standard interpretation charts provided by organizations such as CLSI (Clinical and Laboratory Standards Institute) or EUCAST (European Committee on Antimicrobial Susceptibility Testing) to determine whether the organism is susceptible, intermediate, or resistant to each antibiotic tested. The Kirby-Bauer method is valued for its simplicity, reliability, and cost-effectiveness, making it a standard procedure in both clinical diagnostics and microbiological research ^[26-27]

1.Name of Assay: Kirby-Bauer disc diffusion technique.

Test organisms: 1. Escherichia coli (ATCC 25922), 2. Staphylococcus aureus (ATCC 6538P)

Positive control: Enoxacin (10 µg/disc).

Negative control: Moringa oleifera leaf extract (1 mg/mL in water).

Test compounds: 20 µL/disc (Samples 1, 2, 3, and 4 – as provided) ^[28]

2. Preparation of Agar Medium

• Mueller-Hinton Agar (MHA) was prepared from the dehydrated medium according to the manufacturer’s instructions using sterile distilled water.
• The medium was completely dissolved by boiling with continuous stirring and sterilized by autoclaving at 121°C for 15 minutes.
• The agar medium was cooled to 40–50°C and poured into sterile plastic Petri dishes on a flat surface aseptically to a uniform depth of 4 mm. The plates were allowed to solidify. ^[29]

3. Inoculum Preparation

• An actively growing fresh bacterial culture incubated at 37°C with a turbidity of 0.5 McFarland standard was used.

4. Inoculation of Plates and Incubation

• 100 µL of inoculum was spread uniformly on MHA plates.
• An antibiotic disc (standard), solvent disc (control), and sample discs were placed on the surface of the culture-inoculated plate using sterile forceps.
• The plates were incubated in an inverted position at 37°C until visible growth was observed (approximately 7–8 hours).
• The visible zone of inhibition (ZOI) was measured using a zone measurement scale. ^[3

Table no. 4 Antimicrobial Activity

Observation & Interpretation (as shown in the figure below):

1. Both the test organisms were found susceptible to the standard broad-spectrum antibiotic, Enoxacin.
2. The negative control sample did not show any zone of inhibition (ZOI) against either of the test organisms.
3. All four test samples exhibited antimicrobial activity against the test organisms.
4. The results can be interpreted based on the standard ZOI values for Enoxacin: a) Resistant: ZOI < 14 mm, b) Intermediate Sensitive: ZOI = 15–17 mm c) Sensitive: ZOI > 18 mm

Fig no.5 Antimicrobial Activity

Table no.5 Samples