To design and characterization of herbal soap from Manjistha and Mimosa pudica extract for melasma /hyperpigmentation

 Corresponding author: Ankit P. Godane Copyright © 2025 Author(s) retain the copyright of this article. This article is published under the terms of the Creative Commons Attribution License 4.0. To design and characterization of herbal soap from Manjistha and Mimosa pudica extract for melasma /hyperpigmentation Ankit P. Godane *, Vidhi P. Nagbhire, Vaishnavi V. Dorlikar, Mayuri S. Rahangdale and Mauli R. Zirne Manoharbhai Patel Institute of Pharmacy (B. Pharmacy) Kudwa, Gandia (MH)-441614. GSC Biological and Pharmaceutical Sciences, 2025, 33(02), 119-131 Publication history: Received on 26 September 2025; revised on 05 November 2025; accepted on 07 November 2025 Article DOI: https://doi.org/10.30574/gscbps.2025.33.2.0429 Abstract Herbal formulations are gaining increasing popularity due to their minimal side effects and skin-friendly nature. Among them, herbal soaps serve as a promising alternative to synthetic soaps that often cause skin irritation and dryness. This project focuses on the formulation and evaluation of an herbal soap using Rubia cordifolia (Manjistha) and Mimosa pudica (Mimosa), two medicinal plants known for their skin-healing, Skin lightening, anti-inflammatory, and antioxidant properties. The powdered form of the dried roots of Manjistha and leaves of Mimosa was extracted using the Soxhlet method with aqueous and hydroalcoholic solvents. The obtained extracts were incorporated into a standard soap base using the cold process method. The prepared herbal soap was evaluated for various parameters including foaming ability, pH, foam retention, total fatty matter, and irritation. Results indicated that the soap possessed suitable consistency, a skin-friendly pH, and good cleansing and lathering properties. This study concludes that the herbal soap formulated using Manjistha and Mimosa is a safe and potentially effective product for maintaining skin health and managing minor skin issues. Further in vivo studies are needed to confirm its therapeutic benefits. Keywords: Herbal Soap; Hyperpigmentation; Melasma; Manjistha; Mimosa 1. Introduction Herbal soaps are skin-purifying and skin-beautifying products. The primary benefit of utilizing herbal cosmetics is their purity, which leaves the body rich in nutrients and other beneficial minerals instead of causing any negative side effects. Individuals' skin and hair beauty is influenced by their health, lifestyle choices, regular occupation, environment, and upkeep. Summertime dehydration from prolonged heat contact to the skin results in wrinkles, freckles, blemishes, pigmentation, and sunburns.[1] Herbal soap preparation is a medicine that contains antibacterial, anti-aging, antioxidant, and anti-septic properties. It mainly uses parts of plants like seeds, rhizomes, nuts, leaves, flowers, and pulps to treatment for an injury or disease or to achieve good health. Herbal soap does not contain any artificial colour agents, flavour agents, fluorides, etc. [1] Herbs are natural products mostly found in the treatment of almost all diseases and skin problems owing to their high medicinal value, cost-effectiveness, availability, and compatibility. Most common diseases are Eczema, Acne, Rashes, Psoriasis, Allergy, dry skin, urticaria, etc. [2] the herbal remedies used for special skin problems are given in Special skin problem and Herbal Remedies. Soap is a common cleansing agent well known to everyone in the world. Many authors defined soap in different ways. Soap can also be said to be any water-soluble salt of fatty acids containing eight or more carbon atoms Soaps are produced for a variety of purposes likes as washing, bathing, medication, etc.[2] The affinity of the hydrocarbon chain to oil and grease, while the carboxylic group to water is the main reason soap is being used mostly with water for cleaning purposes A natural soap may be generally divided based on the production method into: a melt pour soap, a hot process soap and a cold process soap. The hot process soap is called a transparent or translucent soap. The soap has good detergency or cleansing power, good moisturizing effects, long-lasting fragrance, and less of irritant. Herbal soaps are prepared by adding various dried herbs, flowers and stems into soap base. Herbs are the natural products could be found in the treatment of almost all diseases and skin GSC Biological and Pharmaceutical Sciences, 2025, 33(02), 119-131 120 problems owing to their high medicinal value, cost effectiveness, availability and compatibility; hence it can be used in soap base. The attribute of a soap includes gentleness on the skin, rich lather, protection against various skin disorders (including rashes, eczema, scabies) treatment of skin infection (such as ringworm), protection of even skin toning and smoothness of the skin. Herbal soap does not contain the artificial colours, flavours, fluorides etc., when compared to the content of commercial soap.[1] Natural soaps help maintain skin pH levels easily without disturbing the natural balance. Herbs are the natural products mostly found in the treatment of almost all diseases and skin problems owing their high medicinal value, cost effectiveness, availability and compatibility.[4] Hyperpigmentation is a common dermatologic condition that is found in all skin types but is most prominent in skin of colour. Any inflammation or injury to the skin can almost immediately be accompanied by alterations in pigmentation, either hyperpigmentation or hypopigmentation. Post inflammatory hyperpigmentation appears in many skin conditions, including acne, eczema, and contact dermatitis, and treatment can be challenging. The goal is to reduce the hyperpigmentation without causing undesirable hypopigmentation or irritation in the surrounding area. A variety of topical agents are available to treat hyperpigmentation. [3] Figure 1 Melasma 1.1. Advantage Of Herbal Soap • Mild on the skin: Herbal soaps are often kinder and milder on the skin than synthetic soap, which makes them appropriate for skin types that are more sensitive. • Natural ingredient: They frequently include skin nourishing and hydrating natural ingredients including coconut oil, etc. • Chemical-Free: The absence of harsh chemicals, artificial perfumes, and synthetic colours in herbal soaps lowers the possibility of allergic responses and skin irritation. • Environmentally Friendly: Compared to synthetic soaps, the herbal ones are typically biodegradable and environmentally friendly as they are made with natural ingredients. • Aromatherapy advantages: By combining essential oils with herbal soaps, aromatherapy advantages including stress alleviation, mood enhancement, and relaxation can be experienced 2. Plant Profile 2.1. Mimosa pudica • Botanical name-Mimosa pudica. • Kingdom-Plantae • Order-Fables • Family-Fabaceae • Genus-Mimosa • SpeciesM. pudica • Popular Name(s)- Laari, Lajwanti, churmuri, touch me not plant • Part use -leaves • HabitatIt is native to south America and Central America. • Chemicals constitute: - The ethanol extract consisted of alkaloids (9.05%), Flavonoids (8.32%), Steroids (2.49%), Saponins (8.15%), Phenols (1.02%), Tannins (0.083%), Cyanogenic glycosides (0.122%) and anthocyanins (1.913%). GSC Biological and Pharmaceutical Sciences, 2025, 33(02), 119-131 121 2.1.1. Uses • To treat hyperpigmentation and melasma • Removal of dark spot • Used for healthy and glowing skin Figure 2 Mimosa pudica 2.2. Rubia cordifolia • Botanical name-Rubia cordifolia. • Kingdom-Plantae • Order-Gentian ales • Family-Rubiaceous • Genus-Rubia • SpeciesR. cordifolia • Popular Name(s)- Indian madder, common madder • Part use -Roots. • Habitat-It is native to Himalayan region and India. • Chemical Constituent: The ethanol extract consisted of alkaloids (9.05%), Flavonoids (8.32%), Steroids (2.49%), Saponins (8.15%), Phenols (1.02%), Tannins (0.083%), Cyanogenic glycosides (0.122%) and anthocyanins (1.913%). 2.2.1. Uses • It is used in Blood Purification. • Removal of dark spot. • Used for healthy and glowing skin. • Used as Anti-inflammatory properties Figure 3 Rubia cordifolia GSC Biological and Pharmaceutical Sciences, 2025, 33(02), 119-131 122 3. Material and method Table 1 List of ingredients and their sources Sr.no. Chemicals sources 1 Manjistha extract powder Lab prepared 2 Mimosa extract powder Lab prepared 3 Coconut oil Marico Ltd Mumbai 4 Glycerine Soap base Buhari North Delhi Table 2 List of equipment’s Sr.no. Name of equipment’s 1 Soxhlet Apparatus 2 Reflux condenser 3 Hot air oven 4 Hot plate 5 Muffle furnace 6 Beaker, test tube 7 Measuring cylinder 8 Stirrer, porcelain dish 3.1. Methodology of isolation Procedure: -25gm of Manjistha powder was placed into the thimble and placed in the Soxhlet chamber. 200 ml of selected solvent (distilled water) were placed in a round bottom flask and assembled for Soxhlet extractor then the distillation process was begun. After completed the extraction process, the solvent and extractor were placed on water bath to evaporate the solvent.[6] Table 3 Extraction of Manjistha Soxhlet extraction Extraction of Manjistha Extract powder Actual weight = 1.969 gm Theoretical yield=2.5 gm Percentage yield =7.88% Procedure: -25gm of mimosa powder was placed into the round bottom flask 250 ml of selected solvent (water) were placed in that, then the distillation process was begun in reflux condenser. After completed the extraction process, the solvent and extractor were filtered then placed on water bath to evaporate the solvent.[5] GSC Biological and Pharmaceutical Sciences, 2025, 33(02), 119-131 123 Table 4 Extraction of Mimosa Reflux condenser Extraction of mimosa Extract powder Actual weight=1.479 Theoretical yield=2.5 Percentage yield=5.916% 3.2. Preparation of hyperpigmentation herbal soap Table 5 Procedure STEP -1 Solidified basic glycerine soap was weighed and broken in to smaller pieces. STEP -2 It was transferred in to vessel and heated in water bath to melt the soap base and stirred STEP -3 The powder compositions were added to the base after soap base was liquefied. The mixture was stirred continuously. STEP -4 Homogenous mixture was removed from the water bath, oil was added. STEP-5 The homogenous mixture was added the soap mould and dried overnight. Table 6 Formulation of soap Ingredients F1 F2 F3 Manjistha 0.50gm 1.00gm 1.50gm Mimosa 0.50gm 1.00gm 1.50gm Coconut oil 2.00ml 2.00ml 2.00ml Essential oil 0.75ml 0.75ml 0.75ml Glycerine soap base q, s qasr qasr Total 20.00 gm 20.00 gm 20.00 gm Figure 4 Soap formulation GSC Biological and Pharmaceutical Sciences, 2025, 33(02), 119-131 124 3.3. Reformulation studies 3.3.1. Foreign Matter Test Table 7 Foreign matter test Sr.no. Name of Ingredients Observation inference 1 Manjistha No foreign matters are found 2 Mimosa No foreign matters are found Table 8 Determination of Loss on Drying (Lod) Manjistha Initial weight=36.406m Mimosa Initial weight=36.988 gm Weigh of sample after 1ˢᵗ drying = 36.316gm Weigh of sample after 1ˢᵗ drying =36.941gm Weight of sample after 2ⁿᵈ drying =36.282 gm Weight of sample after 2ⁿᵈ drying =36.847 Difference between both drying=0.034gm Difference between both drying=0.09 gm Percentage yield = 7.61% Percentage yield =0.338% Table 9 Determination of Total Ash Value Manjistha Weight of empty dish (x) = 30.854 gm 1.Mimosa: - Weight of empty dish (x) = 29.868 gm Weight of dry powder taken (y) = 2.004 gm Weight of dry powder taken (y) =2 gm Weight of dish + ash (z) = 31.011 gm Weight of dish + ash (z) =31.857 gm Weight of ash = 0.157 gm Weight of ash =0.247 gm Total cash value of Manjistha = 7.83% Total cash value of Mimosa =12.35% Table 10 Determination of Acid Insoluble Ash Value Manjistha Sample weight wₛ = 2 gm mimosa Sample weight wₛ =2gm Crucible weight w = 30.845 gm Crucible weight w = 30.781 gm Crucible + ash weight wᶠ = 30.847 gm Crucible weight + ash weight wᶠ = 30.880 gm Acid insoluble ash == 0.1% Water soluble ash =4.9 % GSC Biological and Pharmaceutical Sciences, 2025, 33(02), 119-131 125 4. Determination of extractive value/ extractive matter 4.1. Determination Of Alcohol Soluble Extract Table 11 Percentage of extractive value of alcohol soluble extract Sr.no. Name of powder Extractive value 1 Manjistha 11.4% 2 Mimosa 11.2% 4.2. Determination Of WaterSoluble Extract Table 12 Percentage of extractive value of water-soluble extract Sr.no. Name of powder Extractive value 1 Manjistha 10.9% 2 Mimosa 10.6% Mimosa Manjistha Cold maceration Figure 5 Cold maceration and extractive value 5. Preliminary Phytochemical Screening [8] 5.1. Tests For Carbohydrates 5.1.1. General test • Molisch’s test (General test): To 2–3 ml aqueous extract, add a few drops of α-naphthol solution in alcohol, shake, and add conc. H₂SO₄ from the sides of the test tube. • Violet ring is formed at the junction of two liquids. 5.1.2. Tests for reducing sugars: • Fehling’s test: Mix 1 ml Fehling’s A and 1 ml Fehling’s B solutions, boil for one minute. Add an equal volume of test solution. Heat in boiling water bath for 5–10 min. Yellow to brick-red ppt. is observed. • Benedict’s test: Mix equal volumes of Benedict’s reagent and test solution in a test tube. • Heat in boiling water bath for 5 mi. Solution appears green, yellow, or red depending on the amount of reducing sugar. 5.1.3. Test for non-reducing polysaccharides (starch): • Iodine test: Mix 3 ml test solution and a few drops of dilute iodine solution. Blue colour appears; disappears on boiling and reappears on cooling. GSC Biological and Pharmaceutical Sciences, 2025, 33(02), 119-131 126 • Tannic acid test for starch: With 20% tannic acid, the test solution gives ppt. 5.1.4. Tests for proteins • Biuret test (General test): To 3 ml test solution add 4% NaOH and a few drops of 1% CuSO₄ solution. Violet or pink color appears. • Millon’s test (for proteins): Mix 3 ml test solution with 5 ml Millon’s reagent. Warm. White ppt. turns brick red, or ppt. dissolves giving red-colored solution. 5.2. Tests for glycosides 5.2.1. Tests for Cardiac Glycosides • Baljeet’s test • A section shows yellow to orange colors with sodium picrate. 5.2.2. Liebermann’s test (for guaianolides) Add 2 ml plant extract to 2 ml acetic anhydride, cool in ice, add H₂SO₄ carefully along test tube sides. Color change from violet to blue-green indicates steroid nucleus. 5.2.3. Tests for Anthraquinone Glycosides • Borntrager’s test: To 3 ml extract, add dilute H₂SO₄, boil, and filter. • Add equal volume benzene or chloroform, shake, separate organic layer, add ammonia. • Ammoniacal layer turns pink or red. 5.2.4. Tests for Saponin Glycosides: • Foam test: Shake drug extract or dry powder vigorously with water. Persistent stable foam observed. • Tests for Cyanogenetic Glycosides: Add 3% aqueous mercuric nitrate solution to dry drug powder or extract. Metallic mercury forms. 5.3. Tests For Tannins and Phenolic Compounds • To 2–3 ml of aqueous or alcoholic extract, add few drops of the following: • 5% Fecal₃ solution: Deep blue-black color. • Lead acetate solution: White ppt. 5.4. Tests for alkaloids • Evaporate extracts (aqueous, alcoholic, chloroform) separately. Add dilute HCl, shake, filter, and perform • Drage Dorff’s test: Add a few drops of Drage Dorff’s reagent to filtrate. Orange-brown ppt. formed. • Mayer’s test: Add a few drops of Mayer’s reagent to filtrate. Cream or white ppt. formed. 5.5. Tests For Amino Acids • Ninhydrin test (General test): Heat 3 ml test solution with 3 drops 5% Ninhydrin solution in boiling water bath for 10 min. purple or bluish color appears. • Test for Tyrosine: Heat 3 ml test solution with 3 drops Millon’s reagent. Dark red color appears. 5.6. Test For Flavonoids • Shinoda test: To dry powder of extract, add 5ml 95% ethanol, few drops conc. HCL and 0.5g magnesium turnings. Orange, pink, red to purple color appears. • Sulphury acid test: On addition of sulphury acid Flavones and flavone dissolve into it. deep yellow color appears. 5.7. Test for steroids • Salkowski reaction: To 2ml of extract, add 2ml chloroform and add 2ml con.H2SO4 Shake well. Chloroform layer appears red and acid layer shows greenish yellow fluorescence. • Liebermann-Burchard reaction: Mix 2ml extract with chloroform. Add 1-2 ml acetic anhydride and 2 drop cons. H2SO4 from the side of test tube. First red then blue and finally green colour appears. GSC Biological and Pharmaceutical Sciences, 2025, 33(02), 119-131 127 Table 13 Phytochemical Screening of Ethanolic Extract Sr.no. Test Inference Mimosa Manjistha Tests for carbohydrates 1 Molisch’s test (general test) +ve +ve 2 Fehling’s test +ve +ve 3 Benedict’s test +ve +ve 4 Iodine Test +ve +ve Test for proteins 1 Biuret Test (General test) +ve -ve 2 Millon's Test +ve -ve Tests for cardiac glycosides 1 Baljeet’s Test: +ve +ve 2 Liebermann's Test (for bufadienolides) +ve +ve Tests for anthraquinone glycosides 1 Borntrager's Test -ve +ve Tests for saponin glycosides 1 Foam Test +ve +ve Test for cyanogenetic glycoside 1 Sodium Picrate Test -ve +ve Tests for tannins and phenolic compound 1 Lead Acetate Solution +ve +ve Tests for alkaloids 1 Dragendorff’s test +ve +ve 2 Mayer’s test +ve +ve Tests for amino acids 1 Ninhydrin test (General test +ve -ve 2 Test for tyrosine +ve -ve Test for steroids 1 Salkowski reaction +ve ---- 2 Liebermann-Burchard reaction: +ve ---- Test for flavonoids: