CLIAMS:1. A cosmetic composition for inhibition of melanin transfer from a cell, said composition comprising:
a) an extract of Asparagus racemosus;
b) an extract of Kappaphycus alvarezii; and
c) cosmetically suitable additive(s)
wherein said extract of Asparagus racemosus and Kappaphycus alvarezii is present in a ratio ranging from 1:1 to 50:1.
2. The composition as claimed in claim 1, wherein the extract comprising Asparagus racemosus and Aphanimixis rohituka is present in an amount ranging from 0.1% to 5% (w/w).
3. The composition as claimed in claim 1, wherein said extract of Asparagus racemosus is a root extract.
4. The composition as claimed in claim 1, wherein said cosmetically suitable additive is selected from a group comprising of silicone, vitamin, polyol, vehicle, structurant, emollient, gelling agent, thickener, a structurant, an emulsifying agent, binder, preservative, colorant, perfume, skin-lightening agent, anti-wrinkle agent, antimicrobial, neutralizing agents, free-radical scavengers, antioxidants, anti-ageing active agents, derivative(s) of said compound or combination(s) thereof.
5. The composition as claimed in claim 4, wherein said polyol is selected from a group comprising glycerol, ethylene glycol, propylene glycol, pentaerythritol, diglycerol, polyglycerol, their derivatives or combinations thereof.
6. The composition as claimed in claim 4, wherein said silicone is selected from a group comprising linear, branched, cross linked, silicone oils, volatile and non volatile silicones.
7. The composition as claimed in claim 4, wherein said vitamin is selected from a group comprising vitamin A, vitamin B, vitamin, vitamin D, vitamin E, vitamin K or derivative(s) thereof.
8. The composition as claimed in claim 4, wherein said thickener is selected from a group comprising alkyloamides, carbomer, cetearyl alcohol, cetyl alcohol, gelatin, gums, and magnesium aluminium silicates, ozocarite, paraffin, tragacanth, sodium alginate, Tinovis ADM, Acrylates/C10-30 Alkyl Acrylate Crosspolymer and ammonium Acryloyl- dimethyltaurate / Vinyl pyridine Copolymer/Carbomer or combination(s) thereof.
9. The composition as claimed in claim 4, wherein said structurant is selected from a group comprising fatty acids, fatty alcohols, fatty acid esters, fatty acid amides or combination(s) thereof.
10. The composition as claimed in claim 4, wherein said preservative is selected from a group comprising 2-phenoxyethanol, para-hydroxybenzoic acid esters, formaldehyde-releasing agents, haloalkynyl carbamates, caprylyl glycol, sodium benzoate, N-(3-chloroallyl)-hexaminium chloride, polyhexamethylene biguanide hydrochloride, alkyltrimethylammonium bromides or combination(s) thereof.
11. The composition as claimed in claim 4, wherein said anti-ageing active agent is selected from a group comprising free-radical scavengers, keratolytic agents, vitamins, anti-elastase and anti-collagenase agents, proteins, fatty acid derivatives, steroids, trace elements, bleaching agents, algal and plankton extracts, enzymes and coenzymes, flavonoids, ceramides, tensioning agents, muscle relaxants or combination(s) thereof.
12. The composition as claimed in claim 4, wherein said vehicle/s is selected from a group comprising water, alcohols, oils and combinations thereof.
13. The composition as claimed in claim 1, wherein said composition is in the form of cream, serum, lotion, emulsion.
,TagSPECI:Field of the Invention
The invention relates generally to a synergistic herbal skin care composition. More particularly, the invention provides a herbal skin lightening composition based on the synergistic extract of Asparagus racemosus and Kappaphycus alvarezii.
Background and the prior art
Skin color is the result of melanin, carotene and hemoglobin pigments. Human skin color is determined primarily by the amount and type of melanin present in the skin epidermis. This is accomplished not only by the production of melanin in specialized membrane-bound organelles termed melanosomes but also by transfer of these organelles from melanocytes to the surrounding keratinocytes in upper layers of epidermis.
Due to the complexity and the multi mechanism nature of pigmentation, targeting only melanin biosynthesis is not enough and, therefore, it is increasingly becoming essential in the personal care industry, to target skin lightening through multi-mechanistic approach including inhibition of melanin production as well as transfer.
Conventional skin lightening compositions are based on sunscreens or skin lightening agents. The skin lightening agent(s) are believed to control dispersion of melanosomes or inhibit tyrosinase. However, some of the compounds used as skin lightening agents are also known to have undesirable side effects. Sunscreens alone can not lighten the skin beyond the natural skin colour and their only action is to reduce the ingress of ambient ultraviolet radiation into the skin.
Several chemical agent(s) have been used for lightening of skin color like zinc peroxide has been utilized in anhydrous ointments as a bleaching agent. Monobenzyl ether of hydroquinone has also been marketed for its skin lightening effect. But the use of zinc peroxide and hydroquinone has not been found safe.
Ascorbic acid preparations, either pure or made from some natural material, such as lemon juice, were suggested as useful. While seemingly entirely safe, they do not seem to be very effective.
US 4,096,240 refers to niacin as effective in skin lightening. This material is postulated to operate by retarding melanin dispersion or distribution into the epidermis. Since unpleasant skin flushing occurs with niacin, the patent suggests use of niacinamide as a substitute.
EP 0396422 discloses a skin lightening composition, comprising niacinamide, Parsol MCX and Parsol 1789, UV-B and UV-A sunscreens, as well as silicone oil in the skin lightening composition. The composition gives enhanced skin lightening.
However, the composition based upon niacinamide disclosed in the above cited patent(s) is effective, but only to a limited extent.
US 6,123,959 discloses arbutin (a compound present in the leaves of the bearberry plant (Arctostaphylos uva ursi), licorice extract (from licorice root), ascorbic acid, and kojic acid as skin lightening agents that inhibit the tyrosinase mediated production of melanin.
US 5,980,904 disclose that hydroquinone, glutathione, cysteine, and mulberry extract (which also include arbutin) provide such inhibition.
US 6,348,204 discloses a composition for skin whitening, which is made of at least one extract of mulberry, at least one extract of Scutellaria baicalensis, and at least one salicylic acid derivative. However, the composition(s) of US ‘904 and US ‘204 are expected to have adverse side effects, such as cytotoxicity or skin irritation since hydroquinone and arbutin (a derivative of hydroquinone) are cytotoxic and kojic acid have been reported to cause skin irritation. It is also noticed that Kojic acid is unstable and may lose efficacy upon exposure to air and sunlight.
The above referred patent documents deals with the use of chemical agents for achieving the skin lightening benefit(s), and hence suffer with the inherent disadvantage of using the chemical agents on skin.
There are few safe actives known in the prior art which provide the benefit of inhibiting melanin transfer. Actives of herbal origin usually are the formulator’s choice due to expected safety, easy availability and biocompatibility aspects. However, their use is limited by either low efficacy levels or toxicity at higher doses. Therefore, it has been found that however the chemical agent(s) are effective in skin whitening but they tend to have toxic effects, while the herbal composition are safe to use but are not found to be efficient well for causing the skin whitening.
Therefore, there has been felt a strong need for effective herbal skin care composition.
Search in Traditional Knowledge Digital Library reveals that Asparagus racemosus and composition comprising same, has been used for the treatment of Duodenal/gastric ulcer, Hernia, Gout, Diarrhoea, Hyper tension, obesity, headache, cough/bronchitis, dyspnoea, pyrexia.
Prior art search reveals a non-patent literature titled “Screening of some Medicinal Plants for their Antityrosinase and Antioxidant activities” (Nithya Narayanaswamy, Arun Duraisamy and K .P. Balakrishnan), discloses use of aqueous extract of Asparagus racemosa for inhibiting tyrosinase and protect the skin from free radical toxicity.
Another non-patent published literature having Title “Development and assessment of tyrosinase
inhibitory activity of liposomes of Asparagus racemosus extracts” by Narin Therdphapiyanak, Montree Jaturanpinyo, Neti Waranuch, Lalana Kongkaneramit, and Narong Sarisuta, discloses that Asparagus racemosus extract might be a candidate for therapeutic applications in preventing pigmentation disorders and other melanin-related health problems as well as for cosmetic applications for skin whitening effect.
Another non-patent literature having Title “A review on skin whitening property of plant extracts” by Jennifer, C. , Stephie, C.M., Abhishri, S.B. AND Shalini B. U discloses tyrosinase inhibiting potential of aqueous extract of Asparagus racemosus (Liliaceae). Substances having a whitening effect, that is, substances suppressing the production of melanin, are mainly used for the purpose of the prevention and alleviation of such pigment abnormalities. The method of orally administering a large amount of vitamin C, the method of injecting glutathione and the like, the method of topical application of Kojic acid, vitamin C or its derivative, cysteine and the like in the form of an ointment, cream, lotion are known.

US20030096023 discloses tyrosinase inhibitors obtained from several dicotyledonous plant species thus leading to skin lightening. The plants are those belonging to the family Polygonaceae, Rosaceae, and Onagraceae.

However, compounds suppressing the activity of tyrosinase, other than hydroquinone, are extremely mild in the manifestation of the effect, so the effect of the improvement of skin pigmentation is insufficient. On the other hand, hydroquinone has a recognizable effect, but has sensitivity, so the general use thereof is restricted.

Efforts are still going on for obtaining an effective herbal skin care composition, particularly a skin whitening composition.
The present Invention intends to provide an effective herbal composition for skin lightening.
Objects of the Invention
An object of the present invention is to overcome the drawback / disadvantages of the present invention.
Yet another object of the present invention is to provide a synergistic herbal personal care composition with enhanced skin lightening benefits.
Yet another object of the present invention is to provide a herbal composition comprising synergistic extract of Asparagus racemosus with an alga, usefulfor the inhibition of melanin transfer from melanocyte(s).
Yet another object of the present invention is to provide a herbal composition comprising synergistic herbal combination of an aqueous extract of Asparagus racemosus and Kappaphycus alvarezii.
Yet another object of the present invention is to provide a herbal skin care composition that gives consumer acceptable sensorial properties.
Yet another object of the invention is to provide a herbal skin care composition which does not have any side effect(s) on the skin.
Summary of the Invention
The following presents a simplified summary of the invention in order to provide a basic understanding of some aspects of the invention. This summary is not an extensive overview of the present invention. It is not intended to identify the key/critical elements of the invention or to delineate the scope of the invention. Its sole purpose is to present some concept of the invention in a simplified form as a prelude to a more detailed description of the invention presented later.
In an aspect of the present Invention, there is provided a topical herbal skin care composition for inhibition of melanin transfer, said composition essentially comprising:
a. an extract of Asparagus racemosus;
b. an extract of Kappaphycus alvarezii; and
c. cosmetically suitable additive(s)
such that said extract(s) of Asparagus racemosus and Kappaphycus alvarezii is present in a ratio ranging from 1:1 to 50:1.
Brief Description of the Accompanying Drawing
Figure 1: shows percent melanin transfer Inhibition of A. racemosus, Kappaphycus alvarezii, combined extract of A. racemosus and Kappaphycus alvarezii.
Detailed Description of the Invention
Accordingly, the present invention provides a high performing, potent, safe and cost effective herbal skin care composition. The herbal composition is useful in lightening of skin color.
It has been surprisingly found that the composition based on extract made of Asparagus racemosus and Kappaphycus alvarezii, provides synergistic effect(s) in terms of inhibition of melanin transfer from melanocytes. The root extract of Asparagus racemosus has been used for the present Invention. Kappaphycus alvarezii is a species of red alga, which is one of the most important commercial sources of carrageenans. The extract of Asparagus racemosus and Kappaphycus alvarezii is combined in a ratio ranging from 1:1 to 50:1 where the concentration of Asparagus racemosus extract ranges from 5ppm to 50ppm while Kappaphycus alvarezii is present in the concentration ranging from 1ppm to 10ppm.
The composition comprises herbal extract and cosmetically acceptable additive(s). The composition may be a single phase or a multiphase system.
The additive(s) comprises components like preservative, a rheology modifier, a neutralizing agent, fragrance, water or any combination of mentioned component(s) may also be used.
The cosmetically acceptable additives as may be added to the composition of the present invention include but are not limited to silicones, vitamin and/or derivatives thereof, polyols, vehicles, structurants, emollient/s, gelling agent/s, a thickening agent/s, a hydrophilic or hydrophobic polymer, an emulsifying agent/s, alcohol/s etc. Examples of these ingredients include but are not limited to such substances as binders, emollients, preservatives (such as methyl paraben), colorants, perfumes, skin-lightening agents, anti-wrinkle agents, antimicrobials and the like.

The polyols used are selected from glycerol, ethylene glycol, propylene glycol, pentaerythritol, diglycerol, polyglycerol, their derivatives and combinations thereof. In a preferred embodiment the invention relates to compositions wherein the concentrations of polyol compounds varies from about 0.1 to 10.0 percent by wt, preferably between 0.5 and 5.0 percent by wt, most preferably between 1.0 and 4.0 percent by wt.

The silicone used in the invention is selected from linear, branched, cross linked, silicone oils, volatile and non volatile silicones such as dimethicone copolyol, dimethylpolysiloxane, diethylpolysiloxane, high molecular weight dimethicone, mixed C1-C30 alkyl polysiloxane, phenyl dimethicone, dimethiconol, cyclopentasiloxane, dimethicone, dimethiconol, mixed C1-C30 alkyl polysiloxane, and mixtures thereof. The concentration of silicones is about 0.01 to 5.0 percent; preferably about 0.1 to 3.0 percent, more preferably about 0.5 to 2.0 percent by wt.

The vitamin used in the composition is selected from a group comprising vitamin A, vitamin B (1-12), vitamin C, vitamin D (2-4), vitamin E, vitamin K, their derivatives, such as acetates, propionates, palmitates, phosphates, alone on in combinations thereof. The concentrations of the vitamin and/or its derivative(s) may be about 0.01 to 5.0 percent, preferably about 0.05 and 3.0 percent, and more preferably between 0.5 and 2.0 percent by wt.

Emulsifying agents which may be optionally added to the compositions of the present invention include but are not restricted to oxyalkylenated fatty acid esters of polyols, for example polyethylene glycol stearates, for instance PEG-100 stearate, PEG-50 stearate and PEG-40 stearate; and mixtures thereof, mixture of glyceryl monostearate and of polyethylene glycol stearate (100 EO) (Simulsol 165), oxyalkylenated fatty acid esters of sorbitan comprising, for example, from 20 to 100 EO such as Tween 20 or Tween 60, oxyalkylenated (oxyethylenated and/or oxypropylenated) fatty alcohol ethers; alkoxylated or non-alkoxylated sugar esters, such as sucrose stearate and PEG-20 methylglucose sesquistearate; sorbitan esters such as the sorbitan palmitate (Span 40), esters of diacid and of fatty alcohol, such as dimyristyl tartrate; mixtures of these emulsifiers, for instance the mixture of glyceryl stearate and of PEG-100 stearate (Arlacel 165), and mixtures comprising these emulsifiers, such as the mixture of dimyristyl tartrate, cetearyl alcohol, Pareth-7 and PEG-25 laureth-25, (Cosmacol PSE) , steareth – 2, steareth 21, PPG-15 stearyl ether. The concentrations of the emulsifying agents may be about 0.1% to about 8.0%; preferably about 0.4% to about 4.0%.
Thickeners which may be used in the instant invention include but are not restricted to alkyloamides, carbomer 934,940,941,960,961, cetearyl alcohol, cetyl alcohol, gelatin, gums, and magnesium aluminium silicates, ozocarite, paraffin, tragacanth, sodium alginate, Tinovis ADM, Acrylates/C10-30 Alkyl Acrylate Crosspolymer and ammonium Acryloyl - dimethyltaurate / Vinyl pyridine Copolymer/Carbomer, Hydroxyethyl Acrylate / Sodium Acryloyldimethyl Taurate Copolymer (and) Isohexadecane (and) Polysorbate 60 and the like. The concentrations of the thickeners may be about 0.1% by wt to about 2% by wt; preferably about 0.5% by wt to about 1% by wt.

Structurants which may be used in compositions of the present invention include those materials which are well known in the art and include fatty acids, fatty alcohols, fatty acid esters, and fatty acid amides, having fatty chains of from 8 to 30 carbons atoms. Preferably the structurant used is stearic acid. The concentrations of the structurants may be about 0.1% by wt to about 8.0% by wt; preferably about 0.4% by wt to about 4.0% by wt.
Various emollients as are known to a person skilled in the art and as are available in the market can be employed as an optional constituent in the instant invention. These include but are not restricted to the group consisting of lanolin, octyl dodecanol, hexyl decanol, oleyl alcohol, decyl oleate, isopropyl stearate, isopropyl palmitate, isopropyl myristate, hexyl laureate, dioctyl cyclohexane, PPG-15 stearyl ether, isohexadecane, stearic acid, cetyl alcohol, mineral oil etc. The concentrations of the emollients may be about 0.1 to 10 percent by wt; preferably about 1 and 5 percent by wt.

Various chelating agents may be used as optional constituents of the instant invention. These may be selected from a group consisting of but not limited to Dimercaptosuccinic acid (DMSA), Dimercapto-propane sulfonate (DMPS), Alpha lipoic acid (ALA), Calcium disodium versante (CaNa2-EDTA), Disodium EDTA, Dimercaprol (BAL). The concentrations of the various chelating agents may be about 0.1 to 1 percent by wt; preferably about 0.2 to about 0.5 percent by wt.

Various cosmetically and dermatologically suitable preservatives may be added to the instant composition. These may be selected from the group consisting of but not restricted to 2-phenoxyethanol, para-hydroxybenzoic acid esters, also known as parabens, for instance methyl para-hydroxybenzoate (methyl paraben), ethyl para-hydroxybenzoate (ethyl paraben) and propyl para-hydroxybenzoate (propyl paraben) and mixtures thereof; formaldehyde-releasing agents, for instance imidazolidinylurea or diazolidinylurea; haloalkynyl carbamates, for instance 3-iodo-2-propynyl butyl carbamate (IPBC); caprylyl glycol, also known as 1,2-octanediol; sodium benzoate; N-(3-chloroallyl)-hexaminium chloride (or Quaternium-15); polyhexamethylene biguanide hydrochloride (CTFA name: polyaminopropyl biguanide); alkyltrimethylammonium bromides, for instance dodecyltrimethylammonium bromide, myristyltrimethylammonium bromide and hexadecyltrimethylammonium bromide, and mixtures thereof. The concentrations of the various cosmetically and dermatologically suitable preservatives may be about 0.05 to 1.5 percent by wt; preferably about 0.1 and 1 percent by wt.

Anti-ageing active agents which may optionally added to the compositions of the instant invention may be chosen from free-radical scavengers, keratolytic agents, vitamins, anti-elastase and anti-collagenase agents, proteins, fatty acid derivatives, steroids, trace elements, bleaching agents, sea weed and plankton extracts, enzymes and coenzymes, flavonoids, ceramides, tensioning agents and muscle relaxants, and mixtures thereof. The concentrations of the Anti-ageing active agents may be about 0.001 to about 10 wt %, preferably from about 0.01 to about 5 wt %.

Free-radical scavengers and antioxidants which may be optionally added to the compositions of the instant invention include but are not restricted to phosphonic acid derivatives such as ethylenediaminetetra (methylenephosphonic acid), methylene phosphonic acid, methylenephosphonic acid and salts thereof, in particular the sodium salts thereof; ethylenediaminetetraacetic acid and its salts, such as the sodium salt; guanosine; superoxide dismutase; tocopherol (vitamin E) and its derivatives (acetate); ethoxyquine; lactoferrin; lactoperoxidase, and nitroxide derivatives; superoxide dismutases; glutathione peroxidase; plant extracts with free-radical-scavenging activity, such as the aqueous extract of wheatgerm (Detoxiline), green tea, and mixtures thereof. The concentrations of the Free-radical scavengers and antioxidants may be about 0.001 to about 2 wt %, preferably from about 0.01 to about 1 wt %.

The composition of the present invention may further include one or more neutralizers, such as, for example, strong and weak bases. Any suitable neutralizer can be selected, as will be appreciated by one of ordinary skill in the art. Exemplary neutralizers suitable for use in the compositions of the present invention included sodium hydroxide, potassium hydroxide, ammonium hydroxide, diethanolamine, triethanolamine, 2-dimethylamino-2-methyl-1-propanol (DAMP), 2-aminomethyl-1propanol (aminomethyl propanol) (AMP), and the like, or combinations thereof. The neutralizer, if present, may be provided in any amount, e.g., an amount sufficient to achieve a desired pH for the composition. In this respect, the composition preferably has a pH of from about 4-9, more preferably, from about 5-8, and still more preferably from about 5.5-7. Typically, the neutralizer may be present in an amount of from about 0.01% -10% by weight of the composition.

Essential oils and extracts that may be used in the present invention include but are not limited to mentha, jasmine, camphor, white cedar, bitter orange peel, ryu, turpentine, cinnamon, bergamot, citrus unshiu, calamus, pine, lavender, bay, clove, hiba, eucalyptus, lemon, starflower, thyme, peppermint, rose, sage, sesame, ginger, basil, juniper, lemon grass, rosemary, rosewood, avocado, grape, grapeseed, myrrh, cucumber, watercress, calendula, elder flower, geranium, linden blossom, amaranth, seaweed, ginko, ginseng, carrot, guarana, tea tree, jojoba, comfrey, oatmeal, cocoa, neroli, vanilla, green tea, penny royal, aloe vera, menthol, cineole, eugenol, citral, citronelle, borneol, linalool, geraniol, evening primrose, camphor, thymol, spirantol, penene, limonene, rose, lemon grass, peach, honey, almond oil, olive oil, shea butter, olive butter, bearberry, black currant, rosemary, blue lotus, white lily, and terpenoid oils.

The composition of the invention may utilize a fragrance composition comprising a blend of essential oils and synthetic aroma compounds. The blend is often diluted with a carrier like propylene glycol, vegetable oil, or mineral oil. Some examples of synthetic aroma compound that are suitable for soap bar compositions of the present invention include, but are not limited to benzaldehyde, citral, vanillin, ethyl acetate, fructone, octyl acetate, pentyl butanoate, pentyl pentanoate, methyl salicylate, isoamyl acetate, limonene, citronellol, and mixtures thereof. Preferably, the fragrance containing the essential oil is present in the composition of the invention in an amount between approximately 0.1% to approximately 2% by weight.
Various cosmetically acceptable vehicles may be used for the preparation of compositions as per the instant invention. These may be selected from the group consisting of but not restricted to water, alcohols, oils and combinations thereof.

Accordingly, when the surface contemplated is skin, the composition of this invention may contain ingredients, which are added to known creams, lotions, ointments, gels or medicaments, which are physiologically acceptable to skin and which do not contain ingredients that reverse or retard the action of skin-lightening agent(s).

The term “water phase” is defined as and includes those cosmetically suitable additives as are hydrophilic and/or soluble in water. These additives may be selected from a group comprising Di sodium EDTA, magnesium sulphate, acrylates/C10-30 Alkyl Acrylate Crosspolymer and ammonium Acryloyl - dimethyltaurate / VP Copolymer/Carbomer or any combination of mentioned component(s) may also be used.
The term “oil phase” is defined as and includes those cosmetically suitable additives as are soluble in oil. The oil phase of the skin care composition is selected from a group comprising of C12-15 Alkyl Benzoate, Potassium Cetyl Phosphate (and) Hydrogenated Palm Glycerides, Cetostearyl alcohol and Glyceryl Stearate and PEG-100 Stearate or any combination of mentioned component(s) may also be used.
The skin care composition has the topical application and may be formulated in the form of a, emulsion, cream, serum/gel, lotion etc.
While, the neutralizing agent or fragrances used is Triethanolamine, which acts as neutralizer in the composition.
The invention will now be explained with the help of following examples. However, the scope of the invention should not be limited to these examples as the person skilled in the art can easily vary the proportion of the ingredients and combinations.
Experimental Details:
The invention will now be explained with the help of following examples. However, the scope of the invention should not be limited to these examples as the person skilled in the art can easily vary the proportion of the ingredients and combinations.
• For Determining synergism of the skin care composition:
The extract of Asparagus racemosus root and sea weed i.e. Kappaphycus alvarezii is prepared by the following procedure:

1. 10 gm of extract (of Asparagus racemosus root and sea weed powder) was boiled in water at 60°C for 30 minutes for obtaining 10% aqueous extract.
2. The boiled extract was brought to room temperature (RT) and centrifuged at 5000 rpm for 5 minutes.
3. The supernatant was vacuum filtered.
4. The filtrate was subjected to rotor evaporation.
5. The dry extract was scrapped using a spatula and weighed.

The extract of Asparagus racemosus root Kappaphycus alvarezii was mixed in a ratio ranging from 1:1 to 50:1 to obtain the final composition. The extract has been used at 0.1% to 5% of the final formulation which may be in the form of cream/lotion/gel.

The below Table 1 shows percent (%) inhibition of melanin transfer by Asparagus racemosus and sea weed (Kappaphycus alvarezii) individually.
Extract % Inhibition of melanin transfer
Vehicle Control 0
A. racemosus 50 ppm 9
Kappaphycus alvarezii 10 ppm 20
A. racemosus 5 ppm 0
Kappaphycus alvarezii 1 ppm 0
A.racemosus 1 ppm 0
Kappaphycus alvarezii 20 ppm 0
A.racemosus 60 ppm 0
Kappaphycus alvarezii 0.5 ppm 0
Kappaphycus alvarezii 5 ppm 6
A. racemosus 10 ppm 1

Table 1
While, Table 2 shows percentage (%) inhibition of melanin transfer by the herbal extracts in combination (of Asparagus racemosus and Kappaphycus alvarezii) at predetermined concentration ranges. Asparagus racemosus is present in the concentration ranging from 5 ppm to 50 ppm while Kappaphycus alvarezii is present in the concentration ranging from 1 ppm to 10 ppm.

Extract Expected Additive % inhibition of melanin transfer Obtained synergistic % Inhibition of melanin transfer Effect
Vehicle Control 0 0 N.A.
A. racemosus (50 ppm) + Kappaphycus alvarezii (1 ppm) 9+0=9 22.30 Synergy
Kappaphycus alvarezii (10 ppm) + A.racemosus (5 ppm) 20+0=20 39.15 Synergy
Table 2

Table 1 shows that A. racemosus (50 ppm) when tested alone, inhibits about 9% of melanin transfer while Kappaphycus alvarezii (10 ppm) when tested alone, inhibits about 20% of the melanin transfer. Similarly, Kappaphycus alvarezii (1 ppm) when tested alone, does not show any melanin transfer inhibition. The expected effect(s) for the inhibition of the melanin transfer when A. racemosus (50 ppm) and K. alvarezii (1 ppm) is combined should be 9% (9% + 0%). While, Table 2 shows the actual obtained % inhibition of melanin transfer achieved by the herbal extract of A. racemosus and K. alvarezii is 22.30% (when the concentration of A. racemosus is 50 ppm and of K. alvarezii is 1 ppm). Against the expected additive effect(s) of 9%, the Percent (%) Inhibition of melanin transfer obtained is 22.30%, which shows the synergistic behavior of the present herbal skin care composition in inhibiting the melanin transfer.
Again, Table 1 shows that A. racemosus (5 ppm) when tested alone, does not show inhibition of melanin transfer while Kappaphycus alvarezii (10 ppm) when tested alone, inhibits about 20% of the melanin transfer. Therefore, the expected effect(s) for the inhibition of the melanin transfer when A. racemosus (5 ppm) and K. alvarezii (10 ppm) is combined should be 20% (0% + 20%=20%). While, Table 2 shows the actual obtained % inhibition of melanin transfer achieved by the herbal extract of A. racemosus and K. alvarezii is 39.15% (when the concentration of A. racemosus is 5 ppm and of K. alvarezii is 10 ppm), which shows the synergistic behavior of the present herbal skin care composition in inhibiting the melanin transfer. The result(s) have been illustrated graphically in Figure 1.
Another experiment was conducted for determining the synergistic effect achieved by the combination of A. racemosus with K. alvarezii in a particular ratio. Below mentioned Table 3 shows that there has been achieved the synergistic % inhibition of melanin transfer of 27% against the expected % inhibition of melanin transfer of 15%. Similarly, there has been achieved the synergistic % inhibition of melanin transfer of 30.32% against the expected % inhibition of melanin transfer of 21%.
Extract Expected Additive % inhibition of melanin transfer Obtained synergistic % Inhibition of melanin transfer Effect
Vehicle Control 0 0 N.A.
A. racemosus (50 ppm) + Kappaphycusalvarezii (5 ppm) 9+6=15 27 Synergy
A. racemosus (10 ppm) + Kappaphycus alvarezii (10 ppm) 1+20=21 30.32 Synergy
Table 3
• For Determining the effective amount of the herbal active(s) present in the skin care composition:

Further, experiments have been conducted to study and evaluate the percentage (%) inhibition of melanin transfer by using combination of extracts of Asparagus racemosus and Kappaphycus alvarezii over and below the predetermined ranges.

Combination of Extract Expected Additive % inhibition of melanin transfer Obtained % Inhibition of melanin transfer Effect
A.racemosus (1 ppm) + Kappaphycus alvarezii (10 ppm) 0+20=20 16.5 No Synergy
A.racemosus (50 ppm) + Kappaphycus alvarezii (0.5 ppm) 9+0=9 2 No Synergy
A.racemosus (5 ppm) + Kappaphycus alvarezii (20 ppm) 0+0=0 0 No Synergy
A.racemosus (60 ppm) + Kappaphycus alvarezii (1 ppm) 0+2=2 0 No Synergy
Table 4
Table 4 shows that no synergistic feature is obtained in terms of skin whitening, when the amount of A. racemosus is used less than 5ppm and above 50ppm or when the amount of K. alvarezii is used below 1 ppm and over 10ppm.
Therefore, the best result(s) is achieved only when A. racemosus (having concentration from 5ppm to 50ppm) is combined with K. alvarezii (having concentration from 1 ppm and 10ppm). Even, if one herbal extract is not present in the determined range, synergistic feature towards lightening the skin color will not be obtained.
Further, the referred 'ppm' levels are relevant only for the cell culture experiments, wherein the upper limit is determined by cytotoxicity levels for cells and the lower limits are defined as those required for minimum biological efficacy.
In the topical formulation, the extract comprising Asparagus racemosus and Kappaphycus alvarezii, is present in an amount ranging from 0.1 to 5% w/w, more preferably 0.1 to 3% w/w, most preferably 0.1 to 1% w/w, that shows significant melanin transfer inhibitory activity.
The final formulation may be in the form of a cream/lotion/gel.

Asparagus racemosus (Satavar, Shatavari, or Shatamull) is a species of asparagus common throughout Sri Lanka, India and the Himalayas. Kappaphycus alvarezii is a species of red alga. This seaweed is an introduced species and now is commonly found in India growing quickly without natural controls. The A. racemosus was obtained from external supplier – K. Patel Phyto Extractions Ltd. while K. alvarezii was obtained from the commercial sources in Tamilnadu, India.
Process for the preparation of herbal formulation:
The process comprises steps of:
a) weighing all water soluble ingredients along with thickeners and making its neutralized premix in water;
b) oil soluble ingredients are then weighed and melted to obtain a molten oil phase;
c) the neutralized premix and the molten oil phase are homogenized together to form the emulsion.
d) natural extract(s) of A. racemosus and Kappaphycus alvarezii along with emotives, colorants and fragrance are added to the emulsion to obtain the personal care formulation of the present invention.
Example 1:
Preparation of the extract:
The method adopted for preparation of the herbal extract is following:
1) The herbal powder (comprising powdered A.racemosus root and K.alverezii seaweed whole powdered) was boiled in the water. Particularly, about 10 gm of herbal powder was boiled in water at 60°C for 30 minutes for preparing 10% aqueous herbal extract.
2) The boiled extract resulting from above step was brought to room temperature and further centrifuged at 5000 rpm for 5 minutes.
3) The supernatant resulting from above step was vacuum filtered.
4) The filtrate was subjected to rotor evaporation.
5) The dry extract was scrapped using a spatula and weighed.
Example 2:
In this experiment, the personal care formulation has been prepared in the cream form. The formulation comprises the extract of Asparagus racemosus and Kappaphycus alvarezii as the herbal actives.
S.No. Ingredient Wt/wt% Function
1 Pasteurized water 84.8 Base
2 Carbomer 0.2 Thickener
3 Disodium EDTA 0.1 Chelating agent
4 Magnesium sulfate 0.7 Stabilizer/Absorbent
5 C12-15 Alkyl Benzoate 3 Structurant
6 Potassium Cetyl Phosphate (and) Hydrogenated Palm Glycerides 2 Thickener
7 Cetostearyl alcohol 1.5 Thickener
8 Glyceryl Stearate (and) PEG-100 Stearate 2 Thickener
9 Triethanolamine 0.2 Neutralizer
10 Hydroxyethyl Acrylate / Sodium AcryloyldimethylTaurate Copolymer (and) Isohexadecane (and) Polysorbate 60 3 Thickener
11 Asparagus racemosus and Kappaphycusalvarezii(1:1) 1 Skin Lightening Agent
12 Phenoxyethanol 0.7 Preservative
13 Fragrance 0.8 Perfume
Table 5
Example 3:
In this experiment, the personal care formulation has been prepared in the serum/gel form having the extract of Asparagus racemosus and Kappaphycus alvarezii as the active component.
S.No. Ingredient Function Wt/wt%
1 Pasteurized water Base 87.8
2 Disodium EDTA 98% (powder) Chelating agent 0.1
3 Amaze XT (28-029 A) Structurant 1
4 Glycerine CP Humectant 2
5 Allantoin Anti-inflammatory agent 0.1
6 HydramolTMTGL ester Solubilizer 3
7 Asparagus racemosus and Kappaphycusalvarezii (10:1) Skin lightening agent 1
8 Fragrance Fragrance 0.2
9 Dow Corning 2501 cosmetic wax Feel enhancer 4
10 Phenoxetol Preservative 0.8
Table 6
Example 4:
In this experiment, the personal care formulation has been prepared in the lotion form having the extract of Asparagus racemosus and Kappaphycus alvarezii as the active component.
S.No. Ingredietnt Wt/wt% Function
1 Distilled water 87.8 Base
2 Disodium ethylenediaminetetraacetic acid 0.05 Chelating Agent
3 Glycerine CP 2 Humectant
4 Carbomer 0.2 Thickener
5 Steareth-21 1.8 Emulsifier
6 Steareth-2 0.9 Emulsifier
7 Stearic acid 1 Structurant
8 Cetyl Alcohol 0.5 Emollient
9 Methyl Paraben 0.16 Preservative
10 Propyl Paraben 0.04 Preservative
11 Triethanolamine 0.2 Neutralizer
12 Cyclopentasiloxane, dimethiconol,dimethiconecrosspolymer,phenyltrimethicone blend 3.5 Feel enhancer
13 2 -Phenoxyethanol 0.2 Preservative
14 Asparagus racemosus and Kappaphycus alvarezii (10:1) 1 Skin Lightening Agent
15 Acrylates/ Acrylamide copolymer (and) mineral oil (and) polysorbate 85 (and) aqua (Novamer) 0.3 Thickener
16 Fragrance 0.35 Fragrance
Table 7

Example 5:
The synergistic combination of natural extracts have been further used for testing the evaluation of melanin transfer. The method of testing the efficacy of herbal extract for inhibiting the melanin transfer is following:

1. HaCaT cells were cultured and seeded into carboxylate modified 96-well plates and incubated overnight at 37º C.
2. The cells were treated with Asparagus racemosus and Kappaphycus alvarezii (single/combination) at >80% cell-confluency levels in the wells for 24 hrs.
3. The cells were incubated with carboxylate-modified fluorescent microspheres (polystyrene beads) of 0.2 micron, suspended in a medium free from calcium and Magnesium ions for 6h.
4. The cells were washed twice with phosphate buffered saline free of calcium and Magnesium ions.
5. The fluorescence signal was quantified by using microplate reader.
6. Total uptake /transfer inhibition was calculated by:

The tendency of Kappaphycus alvarezii (10ppm) for inhibiting melanin transfer between cells as calculated in accordance with the above formula is mentioned below:
Table 8

S.No. Fluorescence Signal Control Treated
1 Replicate 1 0.3356 0.2737
2 Replicate 2 0.3509 0.2467
3 Replicate 3 0.3388 0.2726
Average 0.34325 0.264333

The experiment was repeated thrice for calculating the % melanin transfer inhibition of Kappaphycus alvarezii. The average of the three experiments for % melanin transfer inhibition was found to be 19.64183351.