Description:FIELD OF THE INVENTION

[0001] The present invention relates to a medicinal plants-based hair color and care liquid formulation and method for preparation thereof that restores hair's natural shine, improves texture, and enhances appearance, to make hair look healthier, smoother and shinier.

BACKGROUND OF THE INVENTION

[0002] There is a growing demand for natural and medicinal plants-based hair care solutions due to the potential risks associated with chemical-laden products. Many conventional hair colors and care solutions contain synthetic chemicals, like ammonia, and parabens, which damage hair, causing to lose its natural shine, texture, and health over time. This has led to an increasing interest in medicinal plants-based formulations gaining popularity for safer, more nourishing alternative. Plants-based hair color and care liquids have ability to naturally restore hair’s vibrancy without causing harm. These formulations, derived from medicinal plants, not only color the hair but also improve its texture, promote shine, and enhance overall appearance. Additionally, such formulations are enriched with medicinal plants extracts that possess anti-inflammatory, antifungal, and antioxidant properties, further enhancing the health benefits for the scalp and hair, making hair look healthier, shinier, and more vibrant.

[0003] Currently, there are several medicinal plants-based hair color and care products available, such as henna and indigo, which are popular for their natural coloring properties. However, these products often have limitations, including a lack of variety in shades and the time-consuming application process. Additionally, while henna is known for conditioning properties, but sometimes leave hair feeling stiff or dry, particularly if not properly prepared. Other plant-based alternatives, like herbal hair dyes containing extracts from plants such as chamomile or hibiscus, may offer gentler color, but their effects are often less vibrant or long-lasting than synthetic options. These herbal treatments often work more slowly and may require multiple applications to achieve the desired result, making them less convenient than chemical dyes. While natural formulations are less harsh on the hair and scalp, plant-based dyes often do not provide the same intensity or durability as synthetic alternatives, limiting their appeal for those looking for bold, precise, or permanent color changes.

[0004] WO1999049839A1 discloses a topical liquid composition for promoting hair growth, comprising natural ingredients with no side-effects, which is effective in promoting hair growth and preventing hair loss. More particularly, the invention relates to the topical liquid composition for promoting hair growth, comprising the extracts which are formed by immersing the mixture of powders of Pinelliae Rhizoma, Caryophylli Flos, Rubi Fructus, Zanthoxyli Fructus, Viticis Fructus, Salviae Radix, and Thujae Semen in oil and ethanol, respectively, followed by aging for a fixed period of time. Although WO’839 discloses topical liquid composition for promoting hair growth. However, the cited art lacks in exhibiting protective properties against premature greying by providing a protective barrier to shield the hair from ultraviolet (UV) radiation.

[0005] US6139828A discloses hair care formulations for cleansing the scalp and hair follicle openings are disclosed which comprises black cohosh extract, and a surfactant selected from the group consisting of sodium C14-16 olefin sulfonate, sodium lauryl sulfoacetate, disodium laureth sulfosuccinate and combinations thereof. Although US’828 discloses hair care formulations for cleansing the scalp and hair follicle openings. However, the cited art lacks in preserving the natural or treated color of hair follicles by supporting the pigment-producing melanocytes in the hair follicles.

[0006] Conventionally, many hair-care products are available in the market. However, the available products have certain limitations pertaining to effectively reducing greying of hair by stimulating melanocyte activity within the hair follicles, thus encouraging the production of melanin in view of maintaining original color for longer. The aavailable products also exhibits less vibrant color results, with uneven coverage, making them less convenient for users seeking quick and reliable hair color solutions.

[0007] In order to overcome the aforementioned drawbacks, there exists a need in the art to develop a liquid formulation that improves the appearance of hair by restoring its shine and texture, while preventing premature greying by protecting hair follicles from UV radiation and stimulating melanocytes to encourage melanin production, thus maintaining natural color longer and providing optimum care to hair.

OBJECTS OF THE INVENTION

[0008] The principal object of the present invention is to overcome the disadvantages of the prior art.

[0009] An object of the present invention is to develop a medicinal plants-based hair color and care liquid formulation that consists of natural ingredients in view of restoring the natural shine of hair, improving its texture and appearance to make hair look healthier, more vibrant, and well-nourished, providing a smoother and shinier finish.

[0010] Another object of the present invention is to develop a medicinal plants-based hair color and care liquid formulation that exhibits protective properties against premature greying by providing a protective barrier to shield the hair from ultraviolet (UV) radiation that damage hair follicles and reduce melanin production, leading to premature greying.

[0011] Another object of the present invention is to develop a medicinal plants-based hair color and care liquid formulation that shows the ability to delay or prevent the onset of grey hair by stimulating melanocyte activity within the hair follicles, thus encouraging the production of melanin in view of maintaining original color for longer.

[0012] Yet another object of the present invention is to develop a medicinal plants-based hair color and care liquid formulation that is capable of preserving the natural or treated color of hair follicles by means of supporting the pigment-producing melanocytes in the hair follicles, to maintain the richness and vibrancy of the hair color over time.

[0013] The foregoing and other objects, features, and advantages of the present invention will become readily apparent upon further review of the following detailed description of the preferred embodiment as illustrated in the accompanying drawings.

SUMMARY OF THE INVENTION

[0014] The present invention relates to medicinal plants-based hair color and care liquid formulation that is capable of providing a natural color to a user’s hair while providing a complete haircare due to all the natural ingredients that support growth of pigment producing melanocytes, making the hair shinier and healthier.

[0015] According to an embodiment of the present invention, medicinal plants-based hair color and care liquid formulation, comprises of i) Lawsonia Inermis (Henna) leaf extract in the range of 18-22% v/v, ii) Cassia Angustifolia (Senna) leaf extract in the range of 7-10% v/v, iii) Withania Somnifera (Ashwagandha) root extract in the range of 5-8% v/v, iv) Aloe Barbadensis (Aloe Vera) leaf extract in the range of 12-18% v/v, v) Rosmarinus Officinalis (Rosemary) leaf extract in the range of 8-12% v/v, vi) Azadirachta Indica (Neem) leaf extract in the range of 6-10% v/v, vii) Tribulus Terrestris (Gokshura) seed extract in the range of 2-3% v/v, viii) Alizarin (Ratanjot) root extract in the range of 4-7% v/v, ix) Hibiscus Rosa-sinensis (Hibiscus) flower extract in the range of 7-10% v/v, x) Moringa Oleifera (Moringa) leaf extract in the range of 2-4% v/v, xi) Rubia Cordifolia (Manjistha) stem in the range of 3-6% v/v, xii) Cassia Auriculata (Senna Avaram) leaf extract in the range of 6-9% v/v, and xiii) Indigofera Tinctoria (Indigo) leaf extract in the range of 1-3% v/v.

[0016] According to another embodiment of the present invention, a method for preparation of the medicinal plants-based hair color and care liquid formulation comprises of the following steps: a) taking Cassia Obovata (Colorless Henna) leaves and demineralized water in a ratio of 1:5 in a round bottom flask and attaching the flask to a heating mantle, b) attaching Clevenger apparatus with water tubing, metal rotating clamp to the support stand rod and collection flask, followed by adjusting temperature of distillation to 80-110oC in order to obtain steam distilled Cassia Obovata leaf extract (condensate 1), c) adding dry Lawsonia Inermis (Henna) leaves for maceration process for a time duration of 70-75 hours to obtain a solution, followed by adding the solution to a round bottom flask, d) setting up steam distillation and Clevenger apparatus and adjusting temperature to 80-110oC in order to obtain steam distilled Lawsonia Inermis leaf extract (condensate 2), e) blending the obtained condensate 2 with the Cassia Angustifolia leaf extract (condensate 3) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 4, f) mixing the obtained condensate 4 with the Withania Somnifera root extract (condensate 5) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 6, g) blending the obtained condensate 6 with the Aloe Barbadensis leaf extract (condensate 7) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 8, h) mixing the obtained condensate 8 with the Rosmarinus Officinalis leaf extract (condensate 9) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 10, i) blending the obtained condensate 10 with the Azadirachta Indica leaf extract (condensate 11) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 12, j) mixing the obtained condensate 12 with the Tribulus Terrestris seed extract (condensate 13) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 14, k) blending the obtained condensate 14 with the Alizarin root extract (condensate 15) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 16, l) mixing the obtained condensate 16 with the Hibiscus Rosa-sinensis flower extract (condensate 17) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 18, m) mixing the obtained condensate 18 with the Moringa Oleifera leaf extract (condensate 19) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 20, n) blending the obtained condensate 20 with the Rubia Cordifolia stem extract (condensate 21) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 22, o) mixing the obtained condensate 22 with the Cassia Auriculata stem leaf extract (condensate 23) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 24; and p) blending the obtained condensate 24 with the Indigofera Tinctoria leaf extract (condensate 25) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 26 i.e. medicinal plants-based hair color and care liquid formulation.

[0017] While the invention has been described and shown with particular reference to the preferred embodiment, it will be apparent that variations might be possible that would fall within the scope of the present invention.

BRIEF DESCRIPTION OF THE DRAWINGS

[0018] These and other features, aspects, and advantages of the present invention will become better understood with regard to the following description, appended claims, and accompanying drawings where:
Figure 1 illustrates a flow chart depicting a method for preparation of a medicinal plants-based hair color and care liquid formulation;
Figure 2 illustrates another flow chart of the proposed method in continuation with the figure 1;
Figure 3 illustrates another flow chart of the proposed method in continuation with the figure 2;
Figure 4 illustrates a graphical representation of cell proliferation rate relative to the control group; and
Figure 5 illustrates a graphical representation of RT PCR analysis of cyclin D1 expression in DP Cells after 12 hrs. of incubation with the indicated concentration of medicinal plants-based hair color and care liquid formulation.

DETAILED DESCRIPTION OF THE INVENTION

[0019] The following description includes the preferred best mode of one embodiment of the present invention. It will be clear from this description of the invention that the invention is not limited to these illustrated embodiments but that the invention also includes a variety of modifications and embodiments thereto. Therefore, the present description should be seen as illustrative and not limiting. While the invention is susceptible to various modifications and alternative constructions, it should be understood, that there is no intention to limit the invention to the specific form disclosed, but, on the contrary, the invention is to cover all modifications, alternative constructions, and equivalents falling within the spirit and scope of the invention as defined in the claims.

[0020] In any embodiment described herein, the open-ended terms "comprising," "comprises,” and the like (which are synonymous with "including," "having” and "characterized by") may be replaced by the respective partially closed phrases "consisting essentially of," consists essentially of," and the like or the respective closed phrases "consisting of," "consists of, the like.

[0021] As used herein, the singular forms “a,” “an,” and “the” designate both the singular and the plural, unless expressly stated to designate the singular only.

[0022] The present invention relates to medicinal plants-based hair color and care liquid formulation and method for preparation thereof to create a liquid formulation containing all herbal ingredients with no side-effects. The prepared formulation enhances the texture and appearance of hair, while protecting against UV-induced damage that accelerates greying and stimulates melanocyte activity to maintain natural hair color.

[0023] According to an embodiment of the present invention, medicinal plants-based hair color and care liquid formulation, comprises of: i) Lawsonia Inermis (Henna) leaf extract in the range of 18-22% v/v, ii) Cassia Angustifolia (Senna) leaf extract in the range of 7-10% v/v, iii) Withania Somnifera (Ashwagandha) root extract in the range of 5-8% v/v, iv) Aloe Barbadensis (Aloe Vera) leaf extract in the range of 12-18% v/v, v) Rosmarinus Officinalis (Rosemary) leaf extract in the range of 8-12% v/v, vi) Azadirachta Indica (Neem) leaf extract in the range of 6-10% v/v, vii) Tribulus Terrestris (Gokshura) seed extract in the range of 2-3% v/v, viii) Alizarin (Ratanjot) root extract in the range of 4-7% v/v, ix) Hibiscus Rosa-sinensis (Hibiscus) flower extract in the range of 7-10% v/v, x) Moringa Oleifera (Moringa) leaf extract in the range of 2-4% v/v, xi) Rubia Cordifolia (Manjistha) stem in the range of 3-6% v/v, xii) Cassia Auriculata (Senna Avaram) leaf extract in the range of 6-9% v/v, and xiii) Indigofera Tinctoria (Indigo) leaf extract in the range of 1-3% v/v.

[0024] The preferable value of i) Lawsonia Inermis (Henna) leaf extract is 19.00% v/v, ii) Cassia Angustifolia (Senna) leaf extract is 8.50% v/v, iii) Withania Somnifera (Ashwagandha) root extract is 6.50% v/v, iv) Aloe Barbadensis (Aloe Vera) leaf extract is 15.00% v/v, v) Rosmarinus Officinalis (Rosemary) leaf extract is 10.00% v/v, vi) Azadirachta Indica (Neem) leaf extract is 8.00% v/v, vii) Tribulus Terrestris (Gokshura) seed extract is 2.50% v/v, viii) Alizarin (Ratanjot) root extract is 5.50% v/v, ix) Hibiscus Rosa-sinensis (Hibiscus) flower extract is 8.50% v/v, x) Moringa Oleifera (Moringa) leaf extract is 3.00% v/v, xi) Rubia Cordifolia (Manjistha) stem is 4.00% v/v, xii) Cassia Auriculata (Senna Avaram) leaf extract is 7.50% v/v, and xiii) Indigofera Tinctoria (Indigo) leaf extract is 2.00% v/v.

[0025] Each of the extracts used herein (except Lawsonia Inermis (Henna) leaf extract) are prepared by using hydro-distillation with the help of Clevenger apparatus.

[0026] Each of the extracts are prepared by using plants part (leaves/stem/flower/root) and demineralized water in a ratio of 1:5.

[0027] The extraction is carried out at a temperature in the range of 80-110oC for 7-8 hours.

[0028] According to another embodiment of the present invention, a method for preparation of the medicinal plants-based hair color and care liquid formulation comprises of the following steps: a) taking Cassia Obovata (Colorless Henna) leaves and demineralized water in a ratio of 1:5 in a round bottom flask and attaching the flask to a heating mantle, b) attaching Clevenger apparatus with water tubing, metal rotating clamp to the support stand rod and collection flask, followed by adjusting temperature of distillation to 80-110oC in order to obtain steam distilled Cassia Obovata leaf extract (condensate 1), c) adding dry Lawsonia Inermis (Henna) leaves for maceration process for a time duration of 70-75 hours to obtain a solution, followed by adding the solution to a round bottom flask, d) setting up steam distillation and Clevenger apparatus and adjusting temperature to 80-110oC in order to obtain steam distilled Lawsonia Inermis leaf extract (condensate 2), e) blending the obtained condensate 2 with the Cassia Angustifolia leaf extract (condensate 3) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 4, f) mixing the obtained condensate 4 with the Withania Somnifera root extract (condensate 5) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 6, g) blending the obtained condensate 6 with the Aloe Barbadensis leaf extract (condensate 7) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 8, h) mixing the obtained condensate 8 with the Rosmarinus Officinalis leaf extract (condensate 9) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 10, i) blending the obtained condensate 10 with the Azadirachta Indica leaf extract (condensate 11) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 12, j) mixing the obtained condensate 12 with the Tribulus Terrestris seed extract (condensate 13) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 14, k) blending the obtained condensate 14 with the Alizarin root extract (condensate 15) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 16, l) mixing the obtained condensate 16 with the Hibiscus Rosa-sinensis flower extract (condensate 17) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 18, m) mixing the obtained condensate 18 with the Moringa Oleifera leaf extract (condensate 19) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 20, n) blending the obtained condensate 20 with the Rubia Cordifolia stem extract (condensate 21) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 22, o) mixing the obtained condensate 22 with the Cassia Auriculata stem leaf extract (condensate 23) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 24, and p) blending the obtained condensate 24 with the Indigofera Tinctoria leaf extract (condensate 25) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 26 i.e. medicinal plants-based hair color and care liquid formulation.

[0029] After preparation of each of the condensates, 0.5% preservative is added to increase shelf life.

[0030] The preservative used herein is preferably sodium benzoate.

[0031] After each step of extraction, contents of extraction tank are cooled to ambient temperature by circulating cool water through the jacket of the extraction tank.

EXAMPLE

[0032] Referring to Figure 1, a flow chart depicting a method for preparation of a medicinal plants-based hair color and care liquid formulation is illustrated. Referring to Figure 2, an another flow chart of the proposed method in continuation with the figure 1 is illustrated. Referring to Figure 3, an another flow chart of the proposed method in continuation with the figure 2 is illustrated.

[0033] The first step includes collection of the parts of each of the ingredient used for preparation of the medicinal plants-based hair color and care liquid formulation. The ingredients used in the present invention comprises of, Cassia Obovata (Colorless Henna Leaves), Lawsonia Inermis (Henna Leaves), Cassia Angustifolia (Senna Leaves), Withania Somnifera (Ashwagandha Roots), Aloe Barbadensis (Aloe Vera Leaves), Rosmarinus Officinalis (Rosemary Leaves), Azadirachta Indica (Neem Leaves), Tribulus Terrestris (Gokshura Seeds), Alizarin (Ratanjot Roots), Hibiscus Rosa-Sinensis (Hibiscus Flower), Moringa Oleifera (Moringa Leaves), Rubia Cordifolia (Manjistha Stems), Cassia Auriculata (Senna Avaram Leaves), and Indigofera Tinctoria (Indigo Leaves).

[0034] After collection, the ingredients are pre-treated by cleaning, drying, and increasing the surface of area of the ingredients, followed by drying all the ingredients through air drying and heating. After the ingredients are ready for extraction, the extraction was performed of each ingredient by Hydro-distillation with the help of Clevenger apparatus, where the dry weight ratio of each ingredient and water for distillation was taken about 1:5 (w/w in the g/g) for the liquid herbal extraction, keeping the temperature of the extraction about 80° C-105° C for about 7-8 hours.

[0035] Hydro-distillation is a laboratory technique to heat the plant material with water to extract volatile compounds, primarily essential oils, by vaporizing them and then collecting the condensed vapor in a separate chamber, allowing the oil to separate from the water due to its different density to isolate essential oils from plants. The Clevenger apparatus typically consists of a distillation flask, a condenser, a collection flask, and a heating source.

[0036] As mentioned in the previous step, 100gms of Dry of Cassia Obovata (Colorless Henna) leaves and 500gm of demineralized (DM) water (in a ratio of 1:5) was taken in a round bottom flask which was attached to a appropriate sized heating mantle, followed by attaching Clevenger apparatus with water tubing, metal rotating clamp to the support stand rod and collection flask. The temperature of distillation was adjusted to 80oC in order to obtain steam distilled Cassia Obovata leaf extract that was labelled as Condensate No. 1.

[0037] Then, extraction of Lawsonia Inermis (Henna) leaves was done through steam distillation and Clevenger apparatus at 80oC in order to obtain steam distilled Lawsonia Inermis leaf extract labeled as Condensate No. 2. The Senna Leaves liquid extract was prepared using distillation unit with Clevenger for extraction labelled as Condensate No. 3. Further, the obtained Condensate No. 2 was blended with condensate 3 by stirring using a stirrer for 10 minutes in order to obtain a Condensate No. 4.

[0038] The prepared condensate No. 4 was mixed with Withania Somnifera root extract (Condensate No. 5) by stirring using stirrer for 10 minutes in order to obtain a Condensate no. 6. Then, the obtained condensate no. 6 was blended with Aloe Barbadensis leaf extract (Condensate no. 7) by stirring using stirrer for 10 minutes in order to obtain a Condensate no. 8. The obtained condensate 8 was then mixed with the Rosmarinus Officinalis leaf extract (Condensate no. 9) by stirring using stirrer for 10 minutes in order to obtain a Condensate no. 10. The condensate no. 10 was blended with the Azadirachta Indica leaf extract (Condensate no. 11) by stirring using stirrer for 10 minutes in order to obtain a Condensate no. 12.

[0039] After that, the obtained condensate no. 12 was mixed with the Tribulus Terrestris seed extract (Condensate no. 13) by stirring to obtain a Condensate no. 14. The condensate no. 14 was mixed with the Alizarin root extract (Condensate no. 15) by stirring using stirrer for 10 minutes in order to obtain a Condensate no. 16. The condensate no. 16 was added to the Hibiscus Rosa-sinensis flower extract (Condensate no. 17) by stirring using stirrer for 10 minutes in order to obtain a Condensate no. 18. The condensate no. 18 was mixed with the Moringa Oleifera leaf extract (Condensate no. 19) by stirring using stirrer for 10 minutes in order to obtain a Condensate no. 20.

[0040] Furthermore, the obtained condensate no. 20 was added with the Rubia Cordifolia stem extract (Condensate no. 21) by stirring using stirrer for 10 minutes in order to obtain a Condensate no. 22. The obtained condensate no. 22 was mixed with the Cassia Auriculata stem leaf extract (condensate no. 23) by stirring using stirrer for 10 minutes in order to obtain a condensate no. 24. The obtained condensate no. 24 was then blended with the Indigofera Tinctoria leaf extract (Condensate no. 25) by stirring using stirrer for 10 minutes in order to obtain a Condensate no. 26 i.e. medicinal plants-based hair color and care liquid formulation.

[0041] The Withania Somnifera root extract (Condensate No. 5), Aloe Barbadensis leaf extract (Condensate no. 7), Rosmarinus Officinalis leaf extract (Condensate no. 9), Azadirachta Indica leaf extract (Condensate no. 11), Tribulus Terrestris seed extract (Condensate no. 13), Alizarin root extract (Condensate no. 15), Hibiscus Rosa-sinensis flower extract (Condensate no. 17), Moringa Oleifera leaf extract (Condensate no. 19), Rubia Cordifolia stem extract (Condensate no. 21), Cassia Auriculata stem leaf extract (condensate no. 23), Indigofera Tinctoria leaf extract (Condensate no. 25) was prepared using the Hydro-distillation method with the help of Clevenger apparatus as mentioned earlier.

CHARACTERIATION

[0042] The efficacy of the prepared medicinal plants-based hair color and care liquid formulation was tested through Primary Irritation Patch Test, Melanin content or Anti-greying effect, and Cell Proliferation Assay Using Cyclin D1 Expression Marker in Dermal Papilla.

Primary Irritation Patch Test

[0043] For conducting the patch test, 24 healthy adult humans were selected as subjects. The evaluation of the product to be tested was carried out versus a positive control: 1% (w/w) SLS, and versus a negative control: 0.9% isotonic solution. Out of 24 subjects screened, none were found screen failures, none were dropouts. 24 subjects underwent the Trial. The mean age of the subjects was 36.4 years. The mean height of the subjects was 161cm. The mean weights of the subjects were 67 kg. Data from all 24 subjects who completed the study were analyzed. There were no subjects lost to follow up. There was no subject who withdrawn the consent. Result: The medicinal plants-based hair color and care liquid formulation was assessed by qualified dermatologists and was found to be non-irritating and zero-allergic during the 24-hour Primary Irritation Patch Test. Starting on day 1st, followed by 2nd day. A follow up visit was done after 1 week of patch removal, and the formulation was still found non-irritating and showed zero-allergy.

Melanin content or Anti-greying effect

[0044] The study aimed to evaluate the effect of the medicinal plants-based hair color and care liquid formulation on melanin production in melanocytes (cells responsible for pigmentation). Melanin is the natural pigment responsible for the color of hair, skin, and eyes. As people age, the melanocytes in hair follicles produce less melanin, leading to grey or white hair. By increasing melanin production, the formulation could potentially reduce or reverse greying.

[0045] The study used the WM 266-4 melanocyte cell line, a human cell line often used in research related to pigmentation and melanin production. This model allows for controlled testing of the formulation's effects on melanocytes. The formulations contain plant-based ingredients that can support or stimulate melanin production or pigmentation. The study compared melanocyte cells treated with the herbal formulation to a control group that did not receive the treatment. This comparison helps determine the impact of the herbal product.

[0046] Results: The results showed that treatment with the herbal formulation led to a significant increase in melanin content in the WM 266-4 melanocyte cells. Specifically, the melanin content in the treated cells increased by 49.65% compared to the control cells. This means that the herbal formulation nearly doubled the amount of melanin produced by the melanocytes, suggesting a substantial effect on the pigmentation process. This increase suggests that the formulation could enhance or restore melanin production in hair follicles.

Cell Proliferation Assay Using Cyclin D1 Expression Marker in Dermal Papilla

[0047] The study was conducted to observe effects of medicinal plants-based hair color and care liquid formulation on the proliferation of Dermal Papilla Cells (DPCs), which are crucial for regulating hair growth. Cyclin D1 was used as a marker to assess cell proliferation. Cyclins are proteins that regulate the progression of cells through the cell cycle. Increased expression of Cyclin D1 typically indicates higher cell proliferation, suggesting that the cells are dividing and proliferating at an accelerated rate. The study was tested at two concentrations of the medicinal plants-based hair color and care liquid formulation, one at 5% and the other at 10%, and outcomes were compared with a control group, which did not receive the extract.

[0048] Result: Referring to Figure 4, a graphical representation of cell proliferation rate relative to the control group is illustrated. Referring to Figure 5, a graphical representation of RT PCR analysis of cyclin D1 expression in DP Cells after 12 hrs. of incubation with the indicated concentration of the medicinal plants-based hair color and care liquid formulation is illustrated. At the 5% concentration, the extract resulted in a 41.62% increase in cell proliferation relative to the control group. This indicates a moderate enhancement in cell division and growth in the presence of the extract.

[0049] However, the 10% concentration showed significant increase, with cell proliferation rising by 96.8% when compared to the control group. This suggests that at higher concentrations, the extract has a pronounced stimulatory effect on cell growth and division. Both concentrations of the henna extract promote cell proliferation, but the 10% concentration has a substantially stronger effect, nearly doubling the rate of cell proliferation compared to the control.

[0050] Although the field of the invention has been described herein with limited reference to specific embodiments, this description is not meant to be construed in a limiting sense. Various modifications of the disclosed embodiments, as well as alternate embodiments of the invention, will become apparent to persons skilled in the art upon reference to the description of the invention. , Claims:1) A medicinal plants-based hair color and care liquid formulation, comprising:

i) Lawsonia Inermis (Henna) leaf extract in the range of 18-22% v/v;
ii) Cassia Angustifolia (Senna) leaf extract in the range of 7-10% v/v;
iii) Withania Somnifera (Ashwagandha) root extract in the range of 5-8% v/v;
iv) Aloe Barbadensis (Aloe Vera) leaf extract in the range of 12-18% v/v;
v) Rosmarinus Officinalis (Rosemary) leaf extract in the range of 8-12% v/v;
vi) Azadirachta Indica (Neem) leaf extract in the range of 6-10% v/v;
vii) Tribulus Terrestris (Gokshura) seed extract in the range of 2-3% v/v;
viii) Alizarin (Ratanjot) root extract in the range of 4-7% v/v;
ix) Hibiscus Rosa-sinensis (Hibiscus) flower extract in the range of 7-10% v/v;
x) Moringa Oleifera (Moringa) leaf extract in the range of 2-4% v/v;
xi) Rubia Cordifolia (Manjistha) stem in the range of 3-6% v/v;
xii) Cassia Auriculata (Senna Avaram) leaf extract in the range of 6-9% v/v; and
xiii) Indigofera Tinctoria (Indigo) leaf extract in the range of 1-3% v/v.

2) The formulation as claimed in claim 1, wherein each of said extracts (except Lawsonia Inermis (Henna) leaf extract) are prepared by using hydro-distillation with the help of Clevenger apparatus.

3) The formulation as claimed in claim 1, wherein each of said extracts are prepared by using plants part (leaves/stem/flower/root) and demineralized water in a ratio of 1:5.

4) The formulation as claimed in claim 1, wherein extraction is carried out at a temperature in the range of 80-110oC for 7-8 hours.

5) A method for preparation of said medicinal plants-based hair color and care liquid formulation as claimed in claim 1, comprising the steps:
a) taking Cassia Obovata (Colorless Henna) leaves and demineralized water in a ratio of 1:5 in a round bottom flask and attaching said flask to a heating mantle;
b) attaching Clevenger apparatus with water tubing, metal rotating clamp to the support stand rod and collection flask, followed by adjusting temperature of distillation to 80-110oC in order to obtain steam distilled Cassia Obovata leaf extract (condensate 1);
c) adding dry Lawsonia Inermis (Henna) leaves for maceration process for a time duration of 70-75 hours to obtain a solution, followed by adding said solution to a round bottom flask;
d) setting up steam distillation and Clevenger apparatus and adjusting temperature to 80-110oC in order to obtain steam distilled Lawsonia Inermis leaf extract (condensate 2);
e) blending said obtained condensate 2 with said Cassia Angustifolia leaf extract (condensate 3) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 4;
f) mixing said obtained condensate 4 with said Withania Somnifera root extract (condensate 5) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 6;
g) blending said obtained condensate 6 with said Aloe Barbadensis leaf extract (condensate 7) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 8;
h) mixing said obtained condensate 8 with said Rosmarinus Officinalis leaf extract (condensate 9) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 10;
i) blending said obtained condensate 10 with said Azadirachta Indica leaf extract (condensate 11) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 12;
j) mixing said obtained condensate 12 with said Tribulus Terrestris seed extract (condensate 13) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 14;
k) blending said obtained condensate 14 with said Alizarin root extract (condensate 15) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 16;
l) mixing said obtained condensate 16 with said Hibiscus Rosa-sinensis flower extract (condensate 17) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 18;
m) mixing said obtained condensate 18 with said Moringa Oleifera leaf extract (condensate 19) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 20;
n) blending said obtained condensate 20 with said Rubia Cordifolia stem extract (condensate 21) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 22;
o) mixing said obtained condensate 22 with said Cassia Auriculata stem leaf extract (condensate 23) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 24; and
p) blending said obtained condensate 24 with said Indigofera Tinctoria leaf extract (condensate 25) by stirring using stirrer for 5-15 minutes in order to obtain a condensate 26 i.e. said medicinal plants-based hair color and care liquid formulation.

6) The method as claimed in claim 5, wherein after preparation of each of said condensates, 0.5% preservative is added to increase shelf life.

7) The method as claimed in claim 5 and 6, wherein said preservative is preferably sodium benzoate.

8) The method as claimed in claim 5, wherein after each step of extraction, contents of extraction tank are cooled to ambient temperature by circulating cool water through the jacket of the extraction tank.