DESC:FORM 2
THE PATENTS ACT, 1970
(39 of 1970)
&
THE PATENTS RULES, 2003

COMPLETE SPECIFICATION
[See section 10, Rule 13]

HAIR CARE COMPOSITION AND A METHOD OF PREPARING THE SAME

BAJAJ CONSUMER CARE LTD., A COMPANY REGISTERED UNDER THE COMPANIES ACT, 1956 AND HAVING ITS REGISTERED OFFICE AT OLD STATION ROAD, SEVASHRAM CHOURAHA, UDAIPUR, RAJASTHAN 313001

THE FOLLOWING SPECIFICATION DESCRIBES THE INVENTION AND THE MANNER IN WHICH IT IS TO BE PERFORMED
FIELD OF INVENTION
The present invention relates to a hair care composition and a method of preparation thereof.
BACKGROUND OF INVENTION
Background description includes information that may be useful in understanding
the present invention. It is not an admission that any of the information provided
herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art. Hair is a complex structure which is made of many components, that provides protection to the scalp and also enhances physical attractiveness of a person. Hair care and styling has become very important for consumers and they use wide ranges of hair cosmetics and products are used for the same. In recent times, one of the major concerns of the consumers is premature and/or early greying of hair. The hair follicles contain specialized cells called melanocytes. Inside these melanocytes are the organelles melanosomes in which the process of melanogenesis takes place. Melanogenesis is the process of producing a pigment known as melanin which renders colour to the hair. There are 2 types of melanin –eumelanin and pheomelanin. Eumelanin is the black/brown pigment while pheomelanin is red-orange pigment. Hair typically undergoes three phases-growth (anagen), transition (catagen), rest (telogen). Melanin production takes place in the anagen phase.

Hair is largely made up of protein known as keratin which in turn is produced by keratinocytes. Each melanocyte is associated with 5 keratinocytes in the hair bulb forming the “hair follicle-melanin unit”. This pigmentary unit is a pear-shaped black structure at the tip of dermal papilla in the pigmented hair. Keratinocytes then fill up melanin from the hair root till the hair tip. The melanin produced in the melanocytes is transferred to these keratinocytes through filipodia. Higher the melanin content darker is the colour of the hair. As a corollary, if the melanin content generated by the hair follicles is lower, the lighter the hair colour will be.

Greying of hair occurs when the melanocytes stop producing melanin which is responsible for giving the hair its colour. In grey hair, the pigmentary unit becomes fuzzy, the melanocytes become few and rounded, and lightly pigmented oligo dendritic melanocytes become visible in the proximal hair bulb. During anagen, there is a marked reduction in the number of melanocytes in the hair follicles. Greying of hair also called canities or achromotricia occurs with age. This age group may differ for different races. However, premature greying of hair refers to untimely greying of hair. Either of the above can have adverse effect on an individual’s self-confidence. With age, oxidative stress leads to increased storage of the inherently produced H2O2 and other oxidants in the melanosome which hinder the process of melanogenesis thereby leading to the loss of melanin pigments. As a result, melanin production reduces thereby reducing the intensity of the colour and eventually loss of hair colour i.e. greying. This is thought to be central in pathogenesis of greying.

Hair whitening or greying is a clear sign of aging, often perceived as distressing for people who are affected when they are still young. Enhancing melanogenesis could be a preferable approach for preventing or delaying greying of hair. Topical application products like serums and hair oils that enhance melanogenesis is desirable for the ease of application and convenience.
Currently, the solution provided for grey hair is the use of synthetic hair colorants which coat the hair with synthetic hair dyes. Artificial hair colours can have plausible safety and allergen concerns. The existing solutions are mainly based on the use of colouring products allowing artificial hair colouring of hair fibers. Although dyeing is a method that can be easily selected, the chemical components contained in the dye often adversely affect hair and scalp. Most dyes contain ammonia for effective colour development, which has a low molecular weight that easily penetrates the scalp and can cause skin problems. There is a problem of its high volatility, which causes irritation to the eyes. Toxic hazardous chemicals can potentially be fatal to your health, therefore dyeing products using natural ingredients instead of hazardous chemicals have been introduced.
Many studies have resulted in the development of stable and effective natural hair care compositions prepared from ingredients derived from natural sources. However, these compositions solve the problem of hair fall, and the issue of greying of hair is generally neglected because of the use of readily available hair colorants. Most of the hair care products emphasize on the issues related to hair fall such as alopecia, rather than on hair greying. Numerous products are available in the market to treat hair related problems. There are traditionally known home remedies for preparing oils from herbal extracts, however, are cumbersome and has limitations to upscale and may have short shelf life and low stability. There is a challenge to introduce new products especially from herbal origin in the treatment of hair greying.
Seema Rani discloses management of greying of hairs (Sheeb) and use of hair dyes (Khizaab) in Unani Medicine and focuses on the potential of herbals for dyeing purposes and in the treatment of hair greying. Unani medicine has the potential of generating herbal Unani dyes which can eleviate the system. However, the Unani system in present date is in-attractive and lacks consumer compliance, cosmetic product formulations, official quality control for their products.
Sanjay K. Sharma discloses the chemistry and applications of sustainable natural hair colours and dye products containing henna, black walnut, indigo, Bhringraj, Ratanjot, saffron, etc. There are known hair oils, shampoos, conditioners having natural herbal extracts. However, there isn’t much study on how said natural extracts prevent or delay greying of hair. Murali Krishna C et al, provides a review of ayurvedic herbs with kesharanjana property in the management of caniites (Palitya), and suggest that treatments like purification, palliative care, external applications, internal medications & also preventive aspects of canities have been suggested which are effective, natural & without any side effects.
It is known that alterations of the hair biology may delay or prevent the hair greying effect. Synthetic approaches have been identified in the literature with alpha-MSH mimetics, enzymes or its mimetics of catalase and tyrosinase have been tested for similar activities. Other solutions available are Over-the-counter (OTC) nutraceutical medicines or PABA pills (para-amino benzoic acid) which are available as per prescription for vitiligo and other diseases. However, these drugs have to be taken orally for regular intervals for efficacy.
Traditionally known treatments for treating greying of hair include inhalation of herbs; smearing of paste of herbs on hair or applying oil cooked with herbs. The traditional preparations are cumbersome, and the pastes have to be prepared every time before application. Further, conventionally herbal extracts or juices are also known in the art for treatment of grey hair. The extracts are either applied directly which requires preparation of extracts fresh before application or it is added to the oil before application. Such compositions are often immiscible and tend to separate and the results are not satisfactory.
Hair oils with herbs are conventionally prepared by cooking the herbs such as curry leaves, hibiscus, henna, amla (gooseberry), and other herbs. The process can be cumbersome and product may not be aesthetically pleasing. Also, making such preparation would come with inadequate quality and may not produce desirable results. Further, oils such as mustard oil, coconut oil, sesame oil for preparing such herbal hair oils have a typical odour and are sticky in nature and are not preferred by consumers. Few oils like coconut oil freeze at lower temperatures and hence may not be convenient for use. Some oils with high unsaturation may become rancid over a period of time making them aesthetically unpleasant. Nevertheless, consumers are always in a search for new hair care product range that would not only temporarily treat hair greying, but that would also prevent or delay hair greying without the use of artificial means, and that too by acting on the natural process of pigmentation of the hair fibers.
Thus, there is a need to develop a hair care composition having increased melanogenic property, enhanced antioxidant capacity and that provides anti- greying benefits to the consumers.

SUMMARY OF THE INVENTION
The present invention provides a hair care composition for delaying or preventing greying of hair. The present invention provides a hair care composition with an improved antioxidant property and that increases the melanin content in primary human melanocytes. Both, antioxidant property and increase in melanin content can attribute to anti-greying effect.
In an aspect, the present invention provides a hair care composition comprising 5 wt % to 90 wt % mineral oil, 1 wt % to 45 wt % vegetable oil, and 0.01 wt % to 10 wt % of herbal extracts consisting of henna (Lawsonia inermis), shikakai (Acacia concinna), hibiscus (Hibiscus rosa-sinensis), kalonji (Nigella sativa), and onion (Allium cepa) in a ratio of 1:1:1:1:1.
In an embodiment, the hair care composition comprises 5 wt% of herbal extracts consisting of henna extract (Lawsonia inermis), kalonji extract (Nigella sativa), shikakai extract (Acacia concinna), hibiscus extract (Hibiscus rosa-sinensis) and onion extract (Allium cepa) in a ratio of 1:1:1:1:1 in oil.
In an embodiment, the mineral oil is light liquid paraffin.
In an embodiment, the vegetable oil is selected from sunflower oil, mustard oil, olive oil, canola oil, rapeseed oil, safflower oil, groundnut oil, palm oil, rice bran oil, wheatgerm oil, sesame oil, castor oil, almond oil, soyabean oil, corn oil, coconut oil, or walnut oil.
In another aspect, the hair care composition can comprise 60 wt % to 80 wt % light liquid paraffin, 15 wt % to 40 wt % vegetable oil, and 5 wt% of herbal extracts consisting of henna (Lawsonia inermis), shikakai (Acacia concinna), hibiscus (Hibiscus rosa-sinensis), kalonji (Nigella sativa), and onion (Allium cepa) in a ratio of 1:1:1:1:1.
In yet another aspect, the hair care composition can comprise 70 wt % to 75wt% light liquid paraffin, 20 wt % to 25 wt % vegetable oil, and 5 wt% of herbal extracts consisting of henna (Lawsonia inermis), shikakai (Acacia concinna), hibiscus (Hibiscus rosa-sinensis), kalonji (Nigella sativa), and onion (Allium cepa) in a ratio of 1:1:1:1:1.
In an embodiment, the light liquid paraffin and vegetable oil can be in a ratio of 1:1 to 5:1.
In an embodiment, the vegetable oil can be mustard oil or sesame oil.
In an embodiment, the hair care composition of the present invention can further comprise 0.00001 wt% to 2 wt% antioxidants selected from Vitamin E, sweet almond oil, butylated hydroxytoluene, tertiary butyl hydroquinone.
In an embodiment, the hair care composition of the present invention can further comprise 0.00001wt% to 3 wt% permitted colour and perfumes selected from natural or synthetic colour or natural essential oils or synthetic perfumes.
BRIEF DESCTIPTION OF THE DRAWINGS
Figure 1 is a graph depicting melanin estimation study using individual extracts and the composition for the combination of extracts of the present invention.
Figure 2A is a graph depicting the 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) assay for assessing the antioxidant property of the hair oil composition of the present invention.
Figure 2B is a graph depicting the 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) assay for assessing the antioxidant property of ascorbic acid diluted.
DETAILED DESCRIPTION OF THE INVENTION
The foregoing objects of the present invention are accomplished, and the problems and shortcomings associated with the prior art, techniques and approaches are overcome by the present invention as described below in the preferred embodiments. The present is directed to a hair care composition comprising of mineral oil, vegetable oil, herbal extracts and excipients. The present invention provides a non-sticky, stable hair oil composition that is in liquid form even at lower temperature i.e. anti-freeze. The hair care composition increases the melanin concentration and has significant antioxidant property which relates to anti-greying benefits.
In an embodiment, the present invention provides a hair care composition. The composition comprises 5 wt % to 90 wt % mineral oil, 1 wt % to 45 wt % vegetable oil, and 0.01 wt % to 10 wt % of herbal extracts consisting of henna (Lawsonia inermis), shikakai (Acacia concinna), hibiscus (Hibiscus rosa-sinensis), kalonji (Nigella sativa), and onion (Allium cepa) in a ratio of 1:1:1:1:1.
The herbal extracts consisting of henna extract (Lawsonia inermis), kalonji extract (Nigella sativa), shikakai extract (Acacia concinna), hibiscus extract (Hibiscus rosa-sinensis) and onion extract (Allium cepa) in oil can be in a range of 0.01% to 20%., 0.01 wt % to 10 wt %, preferably 5 wt%.
In an embodiment, the hair care composition comprises 5 wt% of herbal extracts consisting of henna extract (Lawsonia inermis), kalonji extract (Nigella sativa), shikakai extract (Acacia concinna), hibiscus extract (Hibiscus rosa-sinensis) and onion extract (Allium cepa) in a ratio of 1:1:1:1:1 in oil. The melanin estimation studies showed that the hair care composition of the present invention comprising the combination of all 5 extracts (Hibiscus + Onion+ Kalonji + Shikakai+ Henna) at 5% in ratio 1:1:1:1:1 in base oil chassis showed synergistic anti-greying properties (i.e melanin increase) compared to individual extracts at a 5% or 1% in base oil chassis. The present invention provides significantly improved anti-greying properties (i.e melanin increase) at a lower concentration of the herbal extracts than the individual extracts at a higher concentration.
The herbal extracts of the present invention can preferably be oil extracts. The oil extracts solubilize in the oil thereby providing an even hair care composition.
In addition the herbal extracts such as henna (Lawsonia inermis), kalonji (Nigella sativa), shikakai (Acacia concinna), hibiscus (Hibiscus rosa-sinensis), and onion (Allium cepa), the oil composition can further include one or more extracts of ritha (Sapindus mukorossi), amla (Phyllanthus emblica), Bhringraj, methi, brahmi, mulethi, triphala, harad, manjishth, jatamansi, nagkesar, naringenin, gingo biloba, stevia, grapes (Vitis vinifera), blueberry, mulberry, pinewood, peasprout, peas, green tea, larch tree leaves, walnut, capsicum, carrot, turmeric, rose, jasmine, aloe vera, sunflower. Preferably, the herbal extracts can be henna (Lawsonia inermis), kalonji (Nigella sativa), shikakai (Acacia concinna), hibiscus (Hibiscus rosa-sinensis) and onion (Allium cepa). The amount of additional herbal extracts to be added can be optimized based on the amount of the extracts of the present invention and would be apparent to a person skilled in the art.
In an embodiment, the mineral oil can be selected from light liquid paraffin or any hydrocarbon cosmetic grade solvents while are non-volatile, miscible with other vegetable oils and can provide lubrication, or any partially hydrogenated oils, any pharmaceutical grade white oils which is acid treated, de-waxed and contain extremely low levels of aromatics or low polarity or non-polar materials. The mineral oil can preferably be light liquid paraffin. In an embodiment, the liquid paraffin can be in a range of 5 wt % to 90 wt %, preferably 60 wt % to 80 wt %, more preferably 70 to 75%.
In an embodiment, the vegetable oil is selected from sunflower oil, mustard oil, olive oil, canola oil, rapeseed oil, safflower oil, groundnut oil, palm oil, rice bran oil, wheatgerm oil, sesame oil, castor oil, almond oil, soyabean oil, corn oil, coconut oil, or walnut oil. In an embodiment, the vegetable oil preferably can be mustard oil. In an embodiment, the mustard oil can be in a range of 1 wt % to 45 wt %, preferably 15 wt % to 40 wt %, more preferably 20 wt % to 25 wt %.
The mineral oil, vegetable oil and the excipients of the composition of the present invention together form a base oil chassis. The base oil chassis can be a mix of 5 wt % to 90 wt % mineral oil, 1 wt % to 45 wt % vegetable oil, and excipients selected from 0.00001 wt% to 2 wt% antioxidants namely, vitamin E, sweet almond oil, butylated hydroxytoluene, tertiary butyl hydroquinone and 0.00001wt% to 3 wt% permitted colour and perfumes selected from natural or synthetic colour or natural essential oils or synthetic perfumes. In an aspect, the present invention provides a base oil chassis comprising the mineral oil and the vegetable oil in a specific ratio of 1:1 to 5:1. The specific weight ratio of the mineral oil and vegetable oil provides the anti-freeze characteristics of the composition making it light and non-sticky. The hair care composition of the present invention remains stable and in liquid form at a temperature of 5°C to 45°C. Preferably, the present invention provides a hair care composition having light liquid paraffin and mustard oil can be in a ratio of 1:1 to 5:1 In another aspect, the present invention provides a hair care composition having light liquid paraffin and sesame oil in the ratio of 1:1 to 5:1. The base oil chassis can further include excipients selected from vitamin E, sweet almond oil, butylated hydroxytoluene, tertiary butyl hydroquinone natural or synthetic permitted colours, natural essential oils or synthetic perfumes.
In an embodiment, the hair care composition can comprise 60 wt % to 80 wt % light liquid paraffin, 15 wt % to 40 wt % vegetable oil, and 5 wt% of herbal extracts consisting of henna (Lawsonia inermis), shikakai (Acacia concinna), hibiscus (Hibiscus rosa-sinensis), kalonji (Nigella sativa), and onion (Allium cepa) in a ratio of 1:1:1:1:1.
In yet another embodiment, the hair care composition can comprise 70 wt % to 75wt% light liquid paraffin, 20 wt % to 25 wt % vegetable oil, and 5 wt% of herbal extracts consisting of henna (Lawsonia inermis), shikakai (Acacia concinna), hibiscus (Hibiscus rosa-sinensis), kalonji (Nigella sativa), and onion (Allium cepa) in a ratio of 1:1:1:1:1.
In yet another embodiment, the hair care composition can comprise 70 wt % to 75wt% light liquid paraffin, 20 wt % to 25 wt % mustard oil, and 5 wt% of herbal extracts consisting of henna (Lawsonia inermis), shikakai (Acacia concinna), hibiscus (Hibiscus rosa-sinensis), kalonji (Nigella sativa), and onion (Allium cepa) in a ratio of 1:1:1:1:1.
In a further embodiment, the hair care composition can comprise 70 wt % to 75wt% light liquid paraffin, 20 wt % to 25 wt % sesame oil, and 5 wt% of herbal extracts consisting of henna (Lawsonia inermis), shikakai (Acacia concinna), hibiscus (Hibiscus rosa-sinensis), kalonji (Nigella sativa), and onion (Allium cepa) in a ratio of 1:1:1:1:1.
The hair care composition of the present invention can further comprise 0.00001 wt% to 2 wt% antioxidants selected from Vitamin E, sweet almond oil, butylated hydroxytoluene, tertiary butyl hydroquinone. The composition of the present invention not only provides increased melanogenesis but also enhanced antioxidant capacity of the composition probably attributing to the anti-greying benefit of the composition.
The hair care composition can include permitted colours and perfumes. The permitted colours can be natural or synthetic colours. The perfumes can be natural essential oils or synthetic perfumes. In an embodiment, the hair care composition of the present invention can further comprise 0.00001wt% to 3 wt% permitted colour and perfumes selected from natural or synthetic colour or natural essential oils or synthetic perfumes.
In an aspect, the present invention also provides a method for preparing a hair care composition. The method comprising the steps of preparing a blend of antioxidants, permitted colours and perfume; adding herbal extracts to the said blend to form a homogenous mixture; adding antioxidants to the said mixture; preparing a blend of mineral oil and vegetable oil; adding the blend of said homogenous mixture to said blend of mineral oil and vegetable oil to form a homogenous and transparent liquid composition.
The process of the present invention provides a homogenous, clear transparent liquid composition.
The herbal extracts can be selected from henna (Lawsonia inermis), kalonji (Nigella sativa), shikakai (Acacia concinna), hibiscus (Hibiscus rosa-sinensis) , onion (Allium cepa) , ritha (Sapindus mukorossi), amla (Phyllanthus emblica, Bhringraj, methi, brahmi, mulethi, triphala, harad, manjishth, jatamansi, nagkesar, naringenin, gingo biloba, stevia, grapes (Vitis vinifera), blueberry, mulberry, pinewood, peasprout, peas, green tea, larch tree leaves, walnut, capsicum, carrot, turmeric, rose, jasmine, aloe vera, sunflower. The herbal extracts can be oil extracts. The oil extracts of henna extract (Lawsonia inermis), kalonji extract (Nigella sativa), shikakai extract (Acacia concinna), hibiscus extract (Hibiscus rosa-sinensis) and onion extract (Allium cepa) can be in the ratio of 1:1:1:1:1 and added one by one to the said blend.
The addition of herbal extract in the method of the present invention can also be by preparing a herbal extract mix by adding oil extracts of henna extract (Lawsonia inermis), kalonji extract (Nigella sativa), shikakai extract (Acacia concinna), hibiscus extract (Hibiscus rosa-sinensis) and onion extract (Allium cepa) in ratio of 1:1:1:1:1, followed by adding the herbal extract mix to the mineral oil and vegetable oil blend.
The hair care composition of the present invention provides a method of delaying or preventing greying of hair by applying the hair care composition to the hair. The said invention increases the melanin content in primary human melanocytes and has significant antioxidant activity. Both these can attribute to anti-greying effect.
EXAMPLES
While the invention is broadly as defined above, it will be appreciated by those persons skilled in the art that it is not limited thereto and that it also includes embodiments of which the following description gives examples. The following examples are included to demonstrate examples of certain preferred embodiments of the invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples that follow represent approaches the inventors have found function well in the practice of the invention, and thus can be considered to constitute examples of preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments that are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention.
Example 1: Process of preparation of hair care composition
Butylated Hydroxytoluene, Tertiary butylhydroquinone and colours –CI No. 47000 and CI No. 26100 were stirred with the perfume until completely dissolved. Henna extract, onion extract, shikakai extract, kalonji extract and hibiscus extract were weighed and added to the perfume blend one by one and stirred to achieve a homogenous mixture. Vitamin-E and sweet almond oil were added while stirring continuously to blend completely. In a separate beaker/vessel, light liquid paraffin and refined bleached mustard oil were combined. While stirring at an ambient RPM using a stirrer to create a vortex formation, perfume blend was added in constant flow without any loss and stirred until the homogenous, clear, transparent liquid was achieved.

Table 1: Composition:
Ingredients Ex-1A
(wt%) Ex-1B
(wt%) Ex-1C
(wt%) Ex-1D
(wt%) Ex-1E
(wt%) Ex-1F
Base oil chassis
Light Liquid Paraffin 72.66 72.66 56.52 5 72.66 72.66
Mustard oil 21 21 37.15 88.21 0 21
Sesame Oil 0 0 0 0 21 0
Excipients* 1.7866 1.7866 1.7866 1.7866 1.7866 1.7866
Herbal Extracts
Henna Extract 0.1 1 1 1 1 0
Shikakai Extract 0.1 1 1 1 1 0
Hibiscus Extract 1.6 1 1 1 1 0
Kalonji Extract 1.6 1 1 1 1 0
Onion Extract 1.6 1 1 1 1 0
*(Vitamin E, Butylated hydroxytoluene, permitted colours, fragrance, sweet almond oil)
Example 2: Melanin estimation study
Example 2A: Melanin estimation study with respect to mustard oil in base chassis
Hair care compositions with an active delivery system of the herbal extract mix in mineral oil and vegetable oil blend or a combination thereof at an optimum maximum added concentration of 5 wt% were selected to be tested for melanin estimation study on human primary melanocytes.
The human primary melanocytes were seeded in 6-well cell culture plates at a density of 50,000 cells/ well in complete basal medium.70 to 80% confluency of the cells after 24 hours were maintained. The cells were treated with appropriate samples with respective concentrations in triplicates. 100mM alpha- MSH was added as positive control. The medium was removed from the plate after 72 hours of incubation with the samples. Each well was washed twice with PBS. 0.5ml of 2% TritonX- 100 was added and the cells were scrapped from the wells. The lysate was transferred to 2ml tube and incubated for 1 hour on ice. Pellets were collected by centrifuging the tubes at 10,000 rpm at 4°C for 10 minutes. Pellets were washed with 0.5ml ethanol: ether at 1:1 ratio and centrifuged the tubes at 10,000 rpm at 4°C for 10 minutes. The supernatant was removed. 0.5 ml NaOH / DMSO solution (0.2N NaOH and 2% DMSO) was added to the pellets and incubated at 80°C for 5 minutes to dissolve the insoluble pellets containing melanin. Unknown melanin contents synthesized from different samples was estimated. The samples were screened for cytotoxic effect on the cells at 6 different concentrations and only the proposed non-toxic concentrations were taken for melanin estimation study at 2 concentrations – 100 times dilution and 500 times dilution. The base oil chassis in the formulation included light liquid paraffin, mustard oil, Butylated hydroxytoluene (BHT), tertiary butyl hydroquinone (TBHQ), Colours, Vitamin E, sweet almond oil, except the extracts.
The sample oil compositions included:
a) Oil compositions containing individual extracts of henna (Lawsonia inermis), shikakai (Acacia concinna), hibiscus (Hibiscus rosa-sinensis), kalonji (Nigella sativa), and onion (Allium cepa) at 5 % each in base oil chassis;
b) Oil composition containing individual extract of hibiscus (Hibiscus rosa-sinensis) at 1% in base oil chassis;
c) Oil compositions containing combination of extracts of hibiscus (Hibiscus rosa-sinensis), and onion (Allium cepa) at 1% each in base oil chassis;
d) Oil compositions containing combination of extracts of henna (Lawsonia inermis), hibiscus (Hibiscus rosa-sinensis), kalonji (Nigella sativa), and onion (Allium cepa) at 1% each in base oil chassis;
e) Oil compositions containing combination of extracts of shikakai (Acacia concinna), hibiscus (Hibiscus rosa-sinensis), kalonji (Nigella sativa), and onion (Allium cepa) at 1% each;
f) the hair oil composition containing a combination of all above 5 extracts in the ratio 1:1:1:1:1 cumulatively at 5% in the composition was performed simultaneously.
The sample details along with results are indicated in Table 2A below:
Sample Details Melanin estimation assay results
0.01x* 0.002x
(100 times dilution) (500 times dilution)
Henna extract @ 5% in base oil chassis 39.09 21.41
Shikakai extract @ 5% in base oil chassis 10.69 10.79
Hibiscus extract @ 5% in base oil chassis 13.19 9.76
Kalonji extract @ 5% in base oil chassis 20.03 11.8
Onion extract @ 5% in base oil chassis 14.37 10.03
Hibiscus extract @ 1% in base oil chassis 19.3 18.06
Hibiscus + Onion extract @ 1% each in base oil chassis 21.4 18.7
Hibiscus +Onion + Kalonji +Henna extract @1% each in base oil chassis 16.53 18.56
Hibiscus + Onion+ Kalonji + Shikakai extract @1% each in base oil chassis 21.7 21.4
Present invention - combination of all 5 extracts (Hibiscus + Onion+ Kalonji + Shikakai+ Henna) @ 5% in ratio 1:1:1:1:1 in base oil chassis – (Example 1B) 45.74 33.36
Base Oil Chassis- (Example 1F) 34.9 21.9

Table 2A: Melanin Estimation Study: *The term ‘x’ is defined as number of times the neat sample has been diluted.
It was observed that as the concentration was increased i.e. the dilution decreased (500X ? 100X); the oil formulation with the combination of extracts in the ratio 1:1:1:1:1 cumulatively at 5% in the composition showed a synergistic effect with significant increase in the melanin estimation as compared to all the other oil formulations containing individual extracts. Such a combination of all 5 extracts was able to produce significantly more melanin pigments in the primary human melanocytes at 45.74 units than the untreated cells at 34.90 units.
A synergistic effect was observed when all the five herbal extracts (Hibiscus + Onion+ Kalonji + Shikakai+ Henna) were included in a formulation in the ratio of 1:1:1:1:1 at total concentration of 5% v/s individual extracts each of which was tested at same concentration level i.e. 5% as well as combination of extracts at 1%. Figure 1 showed melanin estimation study using individual extracts and the composition for the combination of extracts of the present invention. The graph (Figure 1) showed a sharp increase in the melanin produced for the formulation of combination of extracts.
Example 2B: Melanin estimation study with respect to sesame oil in base chassis
Hair care composition with an active delivery system of the herbal extract mix in mineral oil and vegetable oil blend or a combination thereof at an optimum maximum added concentration of 5 wt% were selected to be tested for melanin estimation study on human primary melanocytes.
The human primary melanocytes were seeded in 6-well cell culture plates at a density of 50,000 cells/ well in complete basal medium.70 to 80% confluency of the cells after 24 hours were maintained. The cells were treated with appropriate samples with respective concentrations in triplicates. 100mM alpha- MSH was added as positive control. The medium was removed from the plate after 72 hours of incubation with the samples. Each well was washed twice with PBS. 0.5ml of 2% TritonX- 100 was added and the cells were scrapped from the wells. The lysate was transferred to 2ml tube and incubated for 1 hour on ice. Pellets were collected by centrifuging the tubes at 10,000 rpm at 4°C for 10 minutes. Pellets were washed with 0.5ml ethanol: ether at 1:1 ratio and centrifuged the tubes at 10,000 rpm at 4°C for 10 minutes. The supernatant was removed. 0.5 ml NaOH / DMSO solution (0.2N NaOH and 2% DMSO) was added to the pellets and incubated at 80°C for 5 minutes to dissolve the insoluble pellets containing melanin. Unknown melanin contents synthesized from different samples was estimated. The samples were screened for cytotoxic effect on the cells at 6 different concentrations and only the proposed non-toxic concentrations were taken for melanin estimation study at 2 concentrations – 100 times dilution and 500 times dilution. The base oil chassis (Example 1F) in the formulation included light liquid paraffin, sesame oil, Butylated hydroxytoluene (BHT), tertiary butyl hydroquinone (TBHQ), Colours, Vitamin E, sweet almond oil and excipients, except the extracts.
The sample oil composition included the hair oil composition containing a combination of all above 5 extracts in the ratio 1:1:1:1:1 cumulatively at 5% in the composition was performed simultaneously.
The sample details along with results are indicated in Table 2B below:
Sample Details Melanin estimation assay results
0.01x* 0.002x
(100 times dilution) (500 times dilution)
Example 1E - combination of all 5 extracts (Hibiscus + Onion+ Kalonji + Shikakai+ Henna) @ 5% in ratio 1:1:1:1:1 in base oil chassis 31.63 21.10
Table 2B: Melanin Estimation Study: *The term ‘x’ is defined as number of times the neat sample has been diluted.
Example 3: Anti-oxidant study using 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) assay.
1g test samples were mixed with the 100% methanol (100mg/ml). Homogenous solution of the test samples were prepared by vigorously shaking for 5 minutes. The above cycle was repeated for 5 to 6 times at room temperature. The samples were then centrifuged at 5000 rpm for 10 minutes. The intermittent layer was collected in a separate tube. 200 µl of the clear intermittent solution was diluted serially at 6 different concentrations. 80 µl and 500 µM DPPH in ethanol was added in 96 well clear bottom plate. The plate was incubated for 30 minutes in dark at room temperature. Post incubation the plate was read at 517 nm and DPPH inhibition activity was determined. Ascorbic acid, a known DPPH inhibitor was used as a reference control.
Referring to Figure. 2A and 2B, Figure. 2A included hair care oil composition (Example 1B of Table 1) samples diluted from 1000, 100, 10, 1,0.1, and 0.01 mg/ml concentrations.

Compound ID Date of experiment Compound Type IC50,
mg/mL % Max response Max inhibition concentration, mg/mL No of points used to derive the DRC
Combination of 5 extracts at 5% at ratio 1:1:1:1:1 (Example 1B) Aug 21 2019 TEST 41.5 84 100 6

Figure. 2B included ascorbic acid diluted from 1000, 100, 10, 1,0.1, and 0.01 mg/ml concentrations.
Compound ID Date of experiment Compound Type IC50,
mg/mL % Max response Max inhibition concentration, mg/mL No of points used to derive the DRC
Ascorbic acid Aug 21 2019 REF 0.25 96 10 6

The free radical scavenging activity of 2, 2-Diphenyl- 1-picrylhydrazyl (DPPH) was investigated using the protocol as described above. The inactivation of free radicals represents the total antioxidant capacity of tested oil samples. Different concentration of test samples (the present invention) ranging from 100mg/ml to 0.3mg/ml (1000 mg/ml to 0.01mg/ml) were evaluated to determine the concentration in mg/ml required to inhibit the radical oxidation 50% (IC50). It was observed that the composition of the present invention showed 84% inhibition of DPPH at a concentration of 100mg/ml. Based on the concentration dependent inhibition curve, the IC50 determined was 41.5 mg/ml. It was observed that the composition of the present invention showed good antioxidant activity.
Example 4: Comparative study on the sensorials of the hair care composition.
A comparative study on the sensorials of the hair care composition of the present invention was observed.
Prototypes for testing the sensorials on subjects were:
• A- Example 1D of Table 1
• B- Example 1B of Table 1
Study type was Qualitative paired comparison. Number of respondents/ subjects considered were 10. Feedback was collected from the subjects on both the prototypes A (Ex. 1D) and B (Ex. 1B). The feedback regarding non-stickiness, spreadabilty, sensorials and overall preference feedback were collected from the subjects.
It was observed that 100% of respondents found prototype B to be more non-sticky than prototype A. 90% of respondents found prototype B easier to spread than prototype A. 70% of respondents found prototype B lighter than prototype A. 100% of respondents preferred prototype B over prototype A.

Example 5: Effect of storage conditions and temperature on the stability of the hair care composition.
The objective was to conduct a stability study of the hair care oil composition of the present invention (Example 1B of Table 1). The parameters studied at different temperatures were organoleptic property -color, odor, Viscosity (cP) @ LVDV1 - 100 rpm (in 500ml beaker), specific gravity, acid value, peroxide value, and microbiological examination. The above parameters were studied at temperature 5°C, 25°C, 37°C and 45°C & relative humidity (RH) 70%. Below is the table depicting the parameters and respective test values.
Sr. No. Parameters Example 1B Example 1E
1. Organoleptic property - Color Clear orange coloured oily liquid Clear orange coloured oily liquid
2. Organoleptic property - Odour To match with the standard To match with the standard
3. Viscosity (cP) @ LVDV1 - 100 rpm (in 500ml beaker) 15-30 cP confirm

15-30 cP confirm

4. Specific gravity 0.845-0.855 0.845-0.865
5. Acid Value NMT 1 mg/gm NMT 1 mg/gm
6. Peroxide value NMT 7.5 m.eq./kg NMT 20 m.eq./kg
7. Microbiological Examination <1 cfu/g <1 cfu/g

It was observed that the composition of the present invention passed all the stability parameters at different temperatures.
With the above-mentioned scientific tests, the anti-greying benefit– increase in melanin content and reduction of oxidative stress, of the present invention can be implied. The novel composition of all the five herbal extracts, preferably at 1:1:1:1:1 ratio of the five aforementioned herbal extracts show the best efficiency in increasing melanin at the said concentration mix. The uniqueness of the invention not only lies in the observation of the increased melanogenesis of the aforesaid composition but also the enhanced antioxidant capacity of the composition probably attributing to the anti-greying benefit of the composition.
The foregoing description of the invention has been set merely to illustrate the invention and is not intended to be limiting. Since modifications of the disclosed embodiments incorporating the spirit and substance of the invention may occur to person skilled in the art, the invention should be construed to include everything within the scope of the disclosure.
The embodiments were chosen and described in order to best explain the principles of the present invention and its practical application, to thereby enable others, skilled in the art to best utilize the present invention and various embodiments with various modifications as are suited to the particular use contemplated.
It is understood that various omission and substitutions of equivalents are contemplated as circumstance may suggest or render expedient, but such are intended to cover the application or implementation without departing from the spirit or scope of the present invention.
,CLAIMS:WE CLAIM:
1. A hair care composition comprising 5 wt % to 90 wt % mineral oil, 1 wt % to 45 wt % vegetable oil, and 0.01 wt % to 10 wt % of herbal extracts consisting of henna (Lawsonia inermis), shikakai (Acacia concinna), hibiscus (Hibiscus rosa-sinensis), kalonji (Nigella sativa), and onion (Allium cepa) in a ratio of 1:1:1:1:1.
2. The hair care composition as claimed in claim 1, wherein comprises 5 wt% of herbal extracts consisting of henna extract (Lawsonia inermis), kalonji extract (Nigella sativa), shikakai extract (Acacia concinna), hibiscus extract (Hibiscus rosa-sinensis) and onion extract (Allium cepa) in a ratio of 1:1:1:1:1 in oil.
3. The hair care composition as claimed in claim 1, wherein the mineral oil is light liquid paraffin.
4. The hair care composition as claimed in claim 1, wherein the vegetable oil is selected from sunflower oil, mustard oil, olive oil, canola oil, rapeseed oil, safflower oil, groundnut oil, palm oil, rice bran oil, wheatgerm oil, sesame oil, castor oil, almond oil, soyabean oil, corn oil, coconut oil, or walnut oil.
5. The hair care composition as claimed in claims 1 to 4, wherein comprises 60 wt % to 80 wt % light liquid paraffin, 15 wt % to 40 wt % vegetable oil, and 5 wt% of herbal extracts consisting of henna (Lawsonia inermis), shikakai (Acacia concinna), hibiscus (Hibiscus rosa-sinensis), kalonji (Nigella sativa), and onion (Allium cepa) in a ratio of 1:1:1:1:1.
6. The hair care composition as claimed in claim 5, wherein comprises 70 wt % to 75wt% light liquid paraffin, 20 wt % to 25 wt % vegetable oil, and 5wt% of herbal extracts consisting of henna (Lawsonia inermis), shikakai (Acacia concinna), hibiscus (Hibiscus rosa-sinensis), kalonji (Nigella sativa), and onion (Allium cepa) in a ratio of 1:1:1:1:1.
7. The hair care composition as claimed in claim 1 to 6, wherein comprises light liquid paraffin and vegetable oil in a ratio of 1:1 to 5:1.
8. The hair care composition as claimed in claims 1 to 7, wherein the vegetable oil is mustard oil or sesame oil.
9. The hair care composition as claimed in claims 1 to 8, further comprises 0.00001 wt% to 2 wt% antioxidants selected from Vitamin E, sweet almond oil, butylated hydroxytoluene, tertiary butyl hydroquinone.
10. The hair care composition as claimed in claims 1 to 9, further comprises 0.00001wt% to 3 wt% permitted colour and perfumes selected from natural or synthetic colour or natural essential oils or synthetic perfumes.

Dated this 31st day of January 2020

For Bajaj Consumer Care Ltd.
By their Agent

ANSHUL SUNILKUMAR SAURASTRI) (IN/PA 3086)
KRISHNA & SAURASTRI ASSOCIATES LLP