Formulation and Evaluation of Herbal Handwash using Trigonella foenum graecum

 

Amruta Anil Mali, Dipali.N. Hagir

1Student, Navsahyadri Institute of Pharmacy, Pune.

2Guide, Navsahyadri Institute of Pharmacy, Pune.

*Corresponding Author E-mail: maliamruts@gmail.com, dipalihagir5@gmail.com

 

ABSTRACT:

A herbal handwash made using the aqueous extract of Trigonella foenum-graecum (fenugreek) seeds—a plant well known for its therapeutic qualities, such as antibacterial, anti-inflammatory, antioxidant, and antifungal activities—is the goal of the current study. The main objective was to create a natural handwash that was safe, efficient, and kind to skin. The seeds of fenugreek were extracted with distilled water and mixed with glycerin (a moisturizer), sodium lauryl sulfate (a foaming agent), lemon juice (an antibacterial), rose oil (a fragrance), and propyl paraben (a preservative). Alkaloids, flavonoids, glycosides, saponins, tannins, proteins, amino acids, and phenolic compounds were among the bioactive components found in the extract according to preliminary phytochemical screening, all of which support the formulation's potential for therapeutic use. During the skin irritancy test, participants did not exhibit any irritation, indicating that it was stable and compatible with the skin. Using the agar well diffusion (cup plate) method, antimicrobial efficacy was evaluated. A zone of inhibition of 12 mm against Bacillus subtilis indicated good antibacterial activity. Furthermore, the dirt dispersion test verified a moderate level of particle matter removal efficacy. The herbal handwash made from fenugreek provides a powerful, natural substitute for store-bought synthetic solutions, according to the study's findings. In addition to promoting the growing demand for herbal, eco-friendly personal care products, it guarantees cleanliness and skin safety. Additional investigation may improve its effectiveness and economic feasibility.

 

KEYWORDS: Fenugreek, Handwash, Antimicrobial, Phytoconstituents, Cleanser, Lemon, Herbal.

 

 

INTRODUCTION:

The annual leguminous plant Trigonella foenum-graecum L. is primarily grown as a spice in many parts of the world. T. foenum-graecum is also recognized as one of the first therapeutic plants to be recorded in history. T. foenum-graecum's leaves and seeds are frequently used to make extracts and powders for medicinal purposes.1

 

As an expectorant, anti-inflammatory, and antispasmodic, this plant is used in traditional medicine to treat oral and dental infections; it is used as a mouthwash and gargle; in West Africa, fenugreek gum is boiled while the face is held over a heated kettle to relieve eye discomfort; it is thought to have mucilaginous demulcent, laxative, nutritional, expectorant, orexigenic, and vulnerary qualities. Today, fenugreek's antifungal and antibacterial properties are used to preserve food in addition to flavoring it.2,3

 

The practice of cleaning one's hands with soap or hand wash and water is known as hand washing or hand hygiene. This procedure is essential for getting rid of bacteria, viruses, microbes, grime, oil, and other things that can stick to the skin.4

 

 

Herbal medicine, also known as phytomedicine or botanical therapy, is the use of different parts of plants, such as seeds, roots, leaves, bark, flowers, and aerial parts, for medicinal purposes. Protecting the skin from pathogenic risks is crucial because it is the organ most exposed to the elements. Traditional herbal therapy has been used for many years to treat and manage a variety of illnesses.5,47

 

Regular hand washing is an essential step in preventing the spread of germs and lowering the risk of disease. In addition to reducing the need for antibiotics, this decrease in illness also lowers the chance of antibiotic resistance. According to research, good hand hygiene can prevent about 20% of respiratory diseases, including the common cold, and about 30% of diarrheal illnesses.6

 

Advantages:7

1.     Promotes maintaining personal hygiene in a variety of settings.

2.     Uses natural ingredients to lessen the environmental impact.

3.     Addresses skin issues caused by artificial hand soap.

4.     Makes use of natural extracts to offer additional skin benefits.

5.     Its safety and effectiveness have been thoroughly investigated.

6.     It has few negative effects when being tested.

 

Disadvantages:5,8

1.     Chronic harm to the skin.

2.     Itchy contact dermatitis and eczema.

3.     There has been little research done on the effectiveness of polyherbal handwash compared to other types of handwash or hand sanitizer.

4.     The effectiveness of polyherbal handwash is determined on the caliber and care of the herbal extract utilized.

5.     Hard to modify.

6.     The herbs in polyherbal handwash may cause skin irritation or other adverse reactions in certain people who are allergic to them.

 

Herbal hand wash benefits:9

1.     No unanticipated consequences.

2.     There might be less contamination on our hands.

3.     It also helps get rid of fungus and antiseptic skin problems.

4.     It also makes it easier to remove oil and debris from the skin.

5.     Easy to handle compared to soap and water.

6.     The simplest method is to eradicate microbes.

7.     Cleaning our hands rids our bodies of germs.

 

Physical Properties:10

·       Ras-Katu suggests a strong flavor, which adds to its medicinal qualities.

·       The heating potency shown by Virya-Usna may improve metabolic processes.

·       Guna: Snigdha (unctuous) and Laghu (light), indicating a well-balanced impact on the body.

·       Vipaka-katu describes the post-digestive effect as being strong and impacting the digestive process as a whole.

·       Doshas: Kaphaghna and Vataghna.

·       Used in part: seeds and the full plant.

·       Dosage: 1–3grams of powdered seed or leaf.

·       CaturbijaCurna, Methimodaka, and PancaJirakapaka are formulations.

 

Morphology:11,12

Appearance: The seeds are rhomboidal, firm, and 3 to 5 mm long with a thickness of 2mm.

Color: golden brown to light brown.

Odor: distinctly fiery.

Taste: Mucilaginous and harsh in taste.

 

The annual leguminous plant fenugreek is distinguished by the lack of aneuploidy and its diploid chromosome number (2n = 16). In terms of morphology, it resembles giant clover species and is an upright, fragrant annual. The tall, cylindrical stem is reddish in color and ranges in length from 30 to 60cm. It has large, finger-like structures that make up its root system. Pinnate and trifoliate, fenugreek leaves have long stalks, toothed, lanceolate stipules, and triangular, obovate to oblanceolate leaflets. The plant produces axillary, sessile, hermaphrodite blooms that are white to yellowish-white in color and dependent on insect pollination.

 

Stem:

Among the several steroidal sapogenins found in fenugreek, diosgenin is only found in the oily embryo. In contrast, the stem includes nicotinic acid, trimethylcoumarin, trigocoumarin, and the alkaloid trigonelline. One important part of the seeds is mucilage. According to later research, the stem is composed of roughly 28% mucilage, 5% of a more bitter and strong fixed oil, 22% proteins, a volatile oil, two alkaloids, and a substance that gives it its yellow hue.13

 
Seed:

About 76% of the seeds, according to the statistics, have lengths between 3.44 and 4.75mm, while about 80% have widths between 2.02 and 2.73mm. Additionally, nearly 74% of the seeds have a seed mass ranging from 0.0122g to 0.0197g, and 77% of the seeds have a thickness between 1.27mm and 1.73mm, measured at a dry basis with a moisture content of 8.9%.14

Alkaloids

trimethylamine, neurin, trigonelline, choline, gentianine, carpaine betain.

Amino acids

isoleucine, 4-hydroxyisoleucine, histidine, leucine, lysine, L-tryptophan, argenine.

Saponins

graecunins, fenugrin B, fenugreekine, trigofoenosides A-G.

Steroidal

yamogenin, diosgenin, smilagenin, sarsasapogenin, tigogenin.

Flavonoids

quercetin, rutin, vitexin, isovitexin.

Fibres

gum, neutral detergent fibre.

Lipids

triacylglycerols, diacylglycerols, monoacylglycerols, phosphatidylcholine, phosphatidylethanoamine, free fatty acids.

Other

coumarin, lipids, vitamins, minerals. 28% mucilage; 22% proteins; 5% of a stronger-swelling, bitter fixed oil.

 

Small and golden-yellow in color, fenugreek seeds have a rigid, four-faced structure that resembles stone. These seeds' wide range of bioactive chemical components, which serve as building blocks for the creation of numerous hormonal and medicinal drugs, are primarily responsible for their varied biological and pharmacological characteristics.15

 
Leaves:

Glaekunins are seven saponins found in the leaves. Diosgenin glycosides are what these substances are. About 86.1% of the leaves are made up of water, 4.4% protein, 0.9% fat, 1.5% minerals, 1.1% fiber, and 6% carbs. The leaves contain calcium, zinc, iron, phosphorus, riboflavin, carotene, thiamine, niacin, and vitamin C, among other minerals and vitamins.16

 

Phytochemical constituents of Fenugreek Seed:17,18

Literature review:

Sharma et al. (2017) investigated the antibacterial properties of petroleum ether and methanolic extracts made from callus tissues and fenugreek seeds. According to the findings, the petroleum ether extracts showed significant action against a number of bacteria, such as Candida albicans, Pseudomonas aeruginosa, Bacillus subtilis, Staphylococcus aureus, Escherichia coli, and Aspergillus niger. This implies that fenugreek has potent antibacterial properties.

 

Sindhusha and Rajasekar et al. (2023) tested the antibacterial efficacy of a fenugreek-derived gel against a range of oral pathogens, such as Lactobacillus, Enterococcus faecalis, Streptococcus mutans, and Candida albicans. Fenugreek's promise in antimicrobial applications was highlighted by the results, which showed that the gel produced a more significant zone of inhibition than traditional antibiotics.

 

Al-Habori et al. (2023) evaluated fenugreek seed extracts' antibacterial capabilities against a range of harmful bacterial pathogens. Their findings supported the potential use of fenugreek extracts in the manufacture of antimicrobial products by showing that they have strong antibacterial activity, especially against Gram-positive bacteria.

 

Sajad and Pradyuman et al. (2018) have discovered important bioactive substances in fenugreek. According to the review, fenugreek has several uses, such as gum, emulsifier, glue, and food stabilizer. It has been used to produce a variety of extruded goods and baked goods. Given the many health benefits outlined in the study and backed by earlier scientific studies, fenugreek is advised to be a part of the daily diet because it is safe to use in large quantities and has a number of health benefits. According to the studies cited, fenugreek's medicinal, nutritional, and functional qualities merit more research in order to produce healthier food items.

 

Krishnapura Srinivasan et al. (2006) According to reports, the average adult Indian consumes between 0.3 and 0.6grams of fenugreek seeds every day. Doses 50–100 times higher than this typical dietary level have been used in numerous research showing the health benefits of fenugreek in both human and animal models. Such high dietary intakes can be attained by some cooking techniques that use large amounts of fenugreek seeds, which are especially common in southern India. Because of its high fiber content and several bioactive substances, fenugreek has been used extensively in these diets and has been proven to be safe. It may be easily incorporated to take advantage of its health-promoting qualities.

 

Nasim, Mojtaba, Masoumeh, Abolfazl, and Amir e al. (2015) The therapeutic and functional properties of fenugreek seeds have been the subject of several studies. Protein and fiber abound in fenugreek, and its important bioactive components point to possible medicinal uses. Fenugreek has been shown to have a number of important medical qualities, such as antidiabetic, antioxidant, anticarcinogenic, hypoglycemic, and hypocholesterolemic benefits. It is recommended that we incorporate fenugreek into our regular diet and incorporate it into food products to create functional foods because of its many health advantages.

 

Patel Priyanka and Rukhsar Ansari et al. (2022) According to research, fenugreek powder includes phytochemicals that may have antioxidant and antibacterial effects. The results imply that some human infections are vulnerable to fenugreek extracts' antibacterial properties. Therefore, it is crucial to do additional research in order to determine the active chemicals that are in charge of this plant's biological functions. It has been discovered that fenugreek powder's methanolic extract contains phenolic chemicals, alkaloids, saponins, glycosides, tannins, and carbohydrates. According to certain research, fenugreek powder has the potential to be a powerful natural antibacterial and antioxidant. More extraction, purification, and analysis of this plant's bioactive components are necessary to investigate potential therapeutic uses in medicine as well as additional public health advantages.

 

MATERIAL AND METHOD:

Fenugreek:19

 

Kingdom: Plantae

Clade: Tracheophytes

Clade: Angiosperms

Clade: Eudicots

Clade: Rosids

Order: Fabales

Family: Fabaceae

Subfamily: Faboideae

Genus: Trigonella

Species: T. foenum-graecum

 

Chemical constituents:

Various vitamins and minerals, alkaloids such as trigonelline and choline, amino acids such as 4-hydroxyisoleucine and lysine, saponins, and flavonoids.

 

Uses: For over 6,000 years, fenugreek has been utilized as a medicinal herb. It has been used as a laxative or tonic in many cultures, as well as to promote milk supply during breastfeeding to treat kidney stones, menopause symptoms, and menstrual cramps.

 

Rose oil:20

 

Kingdom: Plantae

Division: Magnoliophyta

Class: Magnoliopsida

Order: Rosales

Family: Rosaceae

Genus: Rosa

Species: Centifoli

Uses of rose-

Rose oil may be used as perfuming agent.

 

Lemon:21

Kingdom: Plantae

Clade: Tracheophytes

Clade: Angiosperms

Clade: Eudicots

Clade: Rosids

Order: Sapindales

Family Rutaceae

Genus: Citrus

Species: C. limon

Chemical Constitute:  citric acid, ascorbic acid, flavonoids, minerals, and essential oils.

 

Uses: Antioxidant properties of citrus flavonoids, particularly lemon-hesperidin, improve cellular defenses against oxidative stress. According to research, gram-negative Shigella soneni and gram-positive bacteria including Enterococcus faecalis and Bacillus species are inhibited by lemon fruit. Pharmaceuticals and cosmetics use lemon oil as a natural preservative and flavoring due to its antibacterial and fungistatic qualities.

 

Chemicals: Sodium lauryl Sulphate, Glycerin, Propyl Paraben, Rose Oil.

Chemical

Use

Sodium lauryl sulphate

Foaming agent

Glycerine

Moisturizing agent

Rose oil

Perfuming agent

Propyl paraben

Preservative

 

Plant Collection:

Seeds of Trigonella foneum are collected from the Nasrapur market.

 

 

Plant Extract:

Five grams of commercially purchased fenugreek seeds were pulverized and mixed with one hundred milliliters of distilled water to create the fenugreek extract. For 30 minutes, the resultant solution was heated to 70°C.22

 

Preparation of Fenugreek based handwash:23

·       The formulation was made by mixing 20 milliliters of lemon juice with 20milliliters of the filtrate of the aqueous extract made from T. foenum-graecum seeds.

·       After that, 40milliliters of glycerin and 6 grams of sodium lauryl sulfate (SLS) were added to the filtrate.

·       In addition, 5ml of rose oil and 0.3g of propyl paraben were added to the mixture.

·       Using filtered water, the preparation's overall volume was brought down to 100 milliliters.

 

Ingredients

Quantity

Action

Extract of T. foenum-graecum

20ml

Antimicrobial

Lemon Water

20ml

Antiseptic agent

SLS

6ml

Foaming agent

Glycerine

40ml

Moisturizing agent

Rose oil

5ml

Perfuming agent

Propyl paraben

0.3gm

Preservatives

Distilled water

Upto 100 ml

Solvent

 

 

Evaluation of Handwash39,44

Physio -Chemical Evaluation24,46

Color: A visual evaluation was conducted49.

Odor: A manual evaluation was conducted.

Appearance: A visual evaluation was conducted.

Homogeneity: A visual evaluation was conducted.

Fragrance test: Its acceptability was assessed based on individual observations.

Five people were questioned about whether they found the scent to be acceptable, and their answers were noted.

 

The following criteria were also used to evaluate aroma.

A.   The fragrance was agreeable and comparable to that of the reference hand wash.

B.    The scent was comparable to the reference hand wash, although not being particularly potent.

pH Determination:

The pH was measured with a digital pH meter. One gram of the produced hand wash was dissolved in 100milliliters of purified water. The hand wash's pH was changed with a 40% NaoH solution.25,45

 

Fig: PH determination

 

Determination of Viscosity:

Herbal handwash's viscosity was measured using a digital Viscolead viscometer.26,27.

 

Foam Height:

Fifty milliliters of distilled water were combined with a one-gram sample of hand wash gel. A measuring cylinder with a capacity of 500ml was filled with the mixture. To reach the 100ml capacity, water was added. A total of twenty-five strokes were given and removed. The foam's height above the aqueous volume was observed.28,48

 

Fig: Foam height.

 

Foam Retention:

50ml of herbal hand wash was put into a 250ml graduated cylinder and shaken ten times. Every minute, the amount of foam was measured. Retention should be consistent for at least five minutes.29,30

 

Fig: Foam retention.

 

Skin Irritancy Test:

Five individuals in good health were selected. After washing the palms of five healthy subjects, the time was noted. We looked at the parameters of itching, dryness, redness, and irritation.31,50

 

Dirt dispersion test:

Ten milliliters of distilled water and one milliliter of Polyherbal Hand Wash Gel were used in a test tube for the dirt dispersion experiment. After adding a drop of Indian ink, the test tube was sealed and shaken. It was determined that the ink content of the foam was either light, moderate, or significant.32,43

 

Antimicrobial test:40,41,42

Cup plate method:

The cup plate method, also known as the agar well diffusion method, is a widely used technique for assessing a formulation's antibacterial activity. This is a general overview of how to evaluate a formulation's antibacterial activity using the cup plate method33,34.

 

Preparation of Agar plate:35

Component

Quantity in grams per 100ml

Beef extract

0.3 g

Peptone

0.5g

Sodium chloride

0.5g

Agar

1.5g

Distilled water

100ml

 

Inoculation:

Make a suspension of a microbiological culture. Using a sterile cotton swab, apply the test organism over the whole surface of the hardened agar plate36.

 

Prepare Wells (Cups):

Usually 4–5 wells per plate, punch wells in the agar using a sterile cork borer (6–8mm in diameter). Carefully remove the agar plugs33.

 

Add Test Solutions:

Fill each well with a specific volume (usually 50–100 µL) of the test sample37.

 

Incubation:

Incubate the plates at room temperature for a few minutes to enable the formulation to permeate into the agar. The plates should next be incubated upside down at the appropriate temperature for a specified period of time, usually overnight35,38.

 

Observation:

Measure the zone of inhibition (clear area around the wells) using a ruler or caliper34,37.

 

Fig: Antimicrobial testing

 

Fig: Phytochemical screening test

 

Fig: Prepared Fenugreek based Handwash

 

Phytochemical test

Result

 

Mayer

Positive

Alkaloids

Wagner

Positive

Hager

Positive

Dragendorff

Positive

 

Flavonoids

Lead acetate

Positive

Ferric chloride

Positive

Shinoda test

Positive

Carbohydrate

Molish

Positive

Benedict

Positive

Glycosides

Legal

Positive

Keller killiani

Positive

Protein and Amino acid

Ninhydrin

Positive

Biuret

Positive

Millon

Positive

 

 

Phenolic Compound and Tannins

Ferric chloride

Positive

Lead acetate

Positive

Gelatin solution

Positive

Acetic acid solution

Positive

Dil.HNO3

Positive

Dil. Iodine solution

Positive

Saponins

Foam test

Positive

Mercuric chloride

Positive

Steroid

Salkowski reaction

Positive

Liebermann Burchard reaction

Positive

 

 

 

 

RESULT AND DISCUSSION:

Phytochemical screening Evaluation of fenugreek handwash:

Sr. No

Parameters

Observation

1

Color

Pale Yellow

2

Odor

Rosy fragrance

3

Appearance

Opaque

4

Homogeneity

Good and homogenous

5

PH

6.8

6

Viscosity

1947 CP at 100RPM R7 spindle

7

Foam height

6 cm

8

Foam retention

65 ml

9

Fragrance test

Not very strong, compare to reference hand wash

10

Stability

Stable

11

Dirt dispersion test

Medium

12

Skin irritation test

No irritation

 

Cup plate Method:

Species

Zone of inhibition

Bacillus subtilis

12mm

 

CONCLUSION:

The herbal handwash with fenugreek extract has antibacterial qualities and is of excellent quality. It contains glycerin to retain moisture, sodium lauryl sulfate as a surfactant, and rose oil for scent. pH, color, foaming efficiency, viscosity, odor, and after-feel were among the factors used to evaluate the handwash. A Viscolead viscometer was used to assess its middle viscosity, and it was also evaluated for stability, non-irritancy, spreadability, and washability. The outcomes show that the product works well and is safe for human consumption. As a result, it can be said that this herbal handwash formulation is a good and popular substitute for conventional handwashes, offering both natural antibacterial properties and a delightful and revitalizing sensory experience.

 

FUTURE PROSPECTIVE:

By mixing it with other therapeutic herbs, more research can increase its effectiveness and create synergistic antimicrobial effects. These products can be successfully marketed for a range of uses, such as sensitive skin formulations, healthcare settings, and general usage, provided that they are properly standardized, clinically validated, and dermatologically safe. Its market potential may also be increased by regulatory support for sustainable sourcing methods and traditional herbal remedies, opening the door for creative and ecologically friendly product development.

 

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Received on 13.06.2025      Revised on 17.07.2025

Accepted on 16.08.2025      Published on 08.10.2025

Available online from October 17, 2025

Asian J. Pharm. Tech. 2025; 15(4):331-338.

DOI: 10.52711/2231-5713.2025.00049

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