CLIAMS:1. A personal care composition comprising egg shell extract, at least one humectant and cosmetically acceptable excipients.
2. The composition as claimed in claim 1, wherein egg shell extract is used in an amount of 0.0001 wt% to 3 wt%.
3. The composition as claimed in claim 2, wherein egg shell extract is used in an amount of 0.01 wt% to 0.1 wt%.
4. The composition as claimed in claim 1, wherein said humectant is selected from glycerin, propylene glycol, butylene glycol, sorbitol and mixture thereof.
5. The composition as claimed in claim 4, wherein said humectant is glycerin.
6. The composition as claimed in claim 1, wherein humectant is present in 1-5 wt%.
7. The composition as claimed in claim 6, wherein humectant is present at 3wt%.
8. The composition as claimed in any of preceding claims the cosmetically acceptable excipients are selected fromsilicones, vitamin and/or derivatives thereof, polyols, vehicles, structurants, emollient/s, gelling agent/s, a thickening agent/s, a hydrophilic or hydrophobic polymer/s, an emulsifying agent/s, alcohol/s etc.
9. The composition as claimed in claim 1 is selected from gel, serum and emulsions. ,TagSPECI:Field of the invention
This invention relates to topical personal care composition. More particularly the invention relates to a personal care composition comprising chicken egg shell extract and humectants to improve the skin moisturization and appearance of skin.
Background and prior art
One of the commonly encountered skin problems around the world is ‘excessive drying’ and dehydration of skin which occurs due to moisture imbalance. Various ancient plants serve as a natural relief for dry skin and these include aloe vera, jasmine, lavender, sandalwood etc. Humectants from other sources also serve the purpose of hydrating skin. For instance, a popular moisturizing ingredient used in cosmetics, by the name of ‘hyaluronic acid’ is isolated from animal sources, from bacteria and also from vitreous humor of fish.
Due to modern life-style, with increase in pollution and stress, the demand to improve skin care compositions which provide enhanced skin hydration/moisturization has increased. An ideal moisturizing ingredient that could be used in skin care applications should be effective in preventing water loss from the skin, should be inexpensive, biocompatible, and easily degradable, should be non-irritant, should be compatible with cosmetic formulations and preferably impart a pleasing and cooling sensation to the skin.
In literature, reports are there wherein use of chicken egg shell membrane extracts or egg shell powder have been used as hydrating and moisturizing agents.
US3196075A provides the treatment of egg shell membrane in a manner such that it can be converted into forms that are highly useful in clinical management of skin-denuded areas, deep tissue wounds and lesions. To facilitate this, the egg shell membrane is converted into fibrous pulp and then in turn converted to eggshell membrane felted fiber sheets, mats or non-woven fabrics. These sheets may be made into desired shapes. The process for such preparation is like washing the egg-shell membrane and drying. This is achieved by using a high speed homogenizer wherein dried egg-shell membranes are converted into finely divided particles in a fibrous form.
US5415875 provides an external preparation for skin for suppressing the formation of lipid peroxide. According to ‘875 the suppression of formation of lipid peroxide is achieved by using decomposition product of egg shell membrane and tocopherol. The decomposition product of shell membrane is used in an amount of 0.00005 to 0.5 wt%. It has been shown that the said decomposition product is used as catalyst as the desired effect is observed in such a small amount and relatively for long period. Further it has been shown that the decomposition product is not effective beyond the range as mentioned in the specification. ‘875 further provides the process for preparation of decomposition product from egg shell membrane.
US2012114690A1 relates to an external agent which has inhibitory effect on histamine release from mast cells, IL-4, -5 and -13 generation, selective COX-2 along with positive effect on various refractory skin diseases such as atopic dermatitis. According to ‘690 it has been found that combination of extracts of tarajo holly leaves and Japanese knotweed leaves, brown algae extracts and hen egg shell membrane hydrolysate are effective in achieving desired effect as mentioned. Further ‘690 provides a composition comprising tarajo holly leaves and Japanese knotweed leaves, brown algae extracts and hen egg shell membrane derived peptide (ESM-P) having a molecular mass of 3 kDa or higher.
US2013315650A1 relates to a cosmetic composition for caring or making up keratin materials comprsing at least one pigment in the form of porous or non-porous, spherical or non-spherical , mineral or organic white particle having a refractive index of greater than or equal to 1.6 and a size by volume of preferably less than 30 µm, and (ii) at least one filler in the form of a porous or non-porous, spherical or non-spherical , mineral or organic white particle having a refractive index of greater than or equal to 2 and a size by volume of preferably less than 30 µm which is distinct from said pigments (i), and characterized in that the total content of fillers is less than or equal to 4% by weight, relative to the total weight of the composition. The said white pigment (i) are chosen from titanium oxide, zinc oxide, optionally bismuth oxychloride, calcium carbonate, calcium sulphate, plaster, natural powders of mineral, animal or vegetable origin, chalk powder, egg-shell or shellfish powders, vegetable ivory and mixtures thereof.
WO2010006260A1 relates to an active component of chicken egg shells having stimulating effect on bone building cells in an ex vivo study. The substance is a polypeptide mixture extracted from egg shells.
JP2002265350 provides a skin care preparation compounded from one or more compounds selected from an [alpha]-hydroxy acid, its salt and its derivative, and/or a protein extracted from egg shell membrane, a nucleic acid and/or its salt. The preparation has excellent skin aging-preventive/improving, skin roughness improving, wound healing, and anti-inflammatory effects given by synergistic effects of fibroblast growth stimulating action, skin moisturizing action, and cutaneous cell activating action, even though each component is compounded in a small amount.
WO 2004080428 relates to cosmetic or pharmaceutical product using eggshell membrane preparation including powder, hydrolysates, and isolates that can be used in effective amounts of about 0.01 to 50 wt % of the preparation. Suitable humectants used according to ‘428 are selected from glycerin, propylene glycol, butylene glycol, urea, sorbitol, sodium PCA, gelatin, polyethylene glycols, sodium lactate, and hyaluronic acid, dipropylene glycol, polypropylene glycol, hydroxypropyl sorbitol, hexylene glycol, 1,3-butylene glycol, 1,2,6-hexanetriol, ethoxylatedglycerin, propoxylated glycerin and mixtures thereof. Most preferably the humectant is glycerin. Amounts of humectant may range anywhere from about 1 to about 50%.
CN1074824 relates to eggshell emulsion in white or light yellow colour, made up by mixing powdered eggshell, water and lactic acid or citric acid together and has the functions of eliminating senile pigmented spots, whitening, eliminating wrinkle, sunscreen and preventing desquamation if it is used to prepare skin-protecting cream. It may be used as additive of both food and feed as it contains ordinary and trace elements necessary for human body and easy to be digested by human body or animal.
JPH04187615 relates to obtaining a scrubbing cream useful for cleaning skin and face, containing eggshell made into a creamy state. Eggshell is made into fine powder and the fine powder is added to a solution of lactic acid. The solution is properly mixed with a humectant, an oil, a neutralizing agent, a surfactant, a perfume and an antiseptic to prepare a scrubbing cream. 70-90 %wt of the scrubbing cream is mixed with 30-10 %wt varnishing cream, prepared by common prescription, while stirring to give the objective scrubbing cream. Eggshell consisting essentially of amino acids is dissolved in a solution of lactic acid, but fine particles of stuck collagen film are insoluble and left in the creamy formed product. These particles are useful for activating skin by frictional action of skin. Eggshell is a waste material and this invention takes advantage of characteristics of eggshell to produce the scrubbing cream.
JPS62270511 provides a cosmetic compound with powder of egg shell and giving durable make-up free from oil-floating and powder-floating. The cosmetic composition according to ‘511 contains 0.1-90wt% egg shell powder. Chicken's egg is most preferable as the raw material because of the availability of uniform material. It may be subjected to protein-removal process, purification process or sieving process during or after the pulverization process. Egg shell powder exhibits high moisture and oil-absorption and is effective in preventing the float of oil and powder when compounded to a cosmetic. According to ‘511, the type of the cosmetic is arbitrary, i.e. solid powder, emulsion, water-oil binary phase system, water-oil-powder ternary phase system, etc., and the cosmetic may be used for various applications.
JPH0956363 relates to obtaininga composition containing egg shell powder and a cyclodextrin. This egg shell calcium composition is obtained by compounding (A) egg shell powder and (B) a cyclodextrin (preferably, [alpha]-cyclodextrin, [beta]- cyclodextrin or [alpha]- or [beta]-cyclodextrin together with [gamma]-cyclodextrin, etc.) preferably in an amount of 0.5-15 pts.wt. of the component B based on 100 wt% of the component A. The composition is prepared by washing shells of poultry eggs sufficiently with water, crushing them after drying to obtain the component A, mixing the component A with a pasty material obtained by adding water to the component B in an amount of several or several ten times the component B. Alternatively the said amount of water is added to a mixture of the components A and B. Successively, the mixture is dried and then pulverized.
Although it has been shown in prior art that egg shell powder or membrane extracts are useful as hydration agents, the present inventors have demonstrated that, egg shell extract alone does not exhibit moisturizing activity, on the contrary, in certain cases, the effect of egg shell extract alone in a composition has negative effects on the hydration efficacy of the composition (Figure 1a). Further, egg shell extract when used with high concentrations of humectants does not also provide the desired effect (Figure 1b). And, none of the prior arts suggest use of egg shell extract with humectants in the right proportions that synergistically provides an effective hydration to the skin.
Thus there is a need for an effective topical or personal care composition which imparts high levels of moisturization using low levels of humectants, as increasing humectant levels alter the aesthetics of the formula or make the composition impart an undesired oily feel on skin.
Objects of the invention
It is an objective of the present invention to overcome the drawbacks of the prior art.
It is an objective of the present invention to provide a personal care composition comprising chicken egg shell extract.
It is another objective of the present invention to provide a personal care composition comprising chicken egg shell extract and at least one humectant that exhibits potent hydration activity on skin in a synergistic manner.
It is further an objective of the present invention to provide a personal care composition wherein common cosmetic humectants having an enhanced moisturizing efficacy with very low levels of egg shell extract to obtain synergistic effect of hydrating skin.
It is yet another objective of the present invention to provide a process for preparation of the composition in various forms like gels, serums, emulsions etc.
Summary of the Invention
The present invention relates to a personal care composition comprising egg shell extract, at least one humectant and cosmetically acceptable excipients.
Description of the Accompanying Figures
Figure 1a illustrates the hydration efficacy of egg shell extract dosed in a cream formulation at 1% in absence of humectant.
Figure 1b illustrates effect of egg shell extract in presence of high concentration of humectant.
Figure 2 illustrates the SDS-PAGE profile of the crude extract and purified fractions of the extracts of the egg shell according to the present invention.
Figure 3 illustrates determination of hydration activity of neat egg-shell extract using corneometer.
Figure 4 illustrates determination of MMP inhibition activity of neat egg-shell extract when tested in vitro using a commercial MMP assay kit
Figure 5 illustrates determination of hydration efficacy of chicken egg shell extract dosed in a hydrogel using corneometer. A: Untreated, B: Formulation+ 3% glycerin , C: Formulation + 3% glycerin + 0.01% egg shell extract, D: Formulation + 3% glycerin + 0.1% egg shell extract, E: Formulation + 3% glycerin + 1% egg shell extract, F: Formulation+ 3% glycerin + 5% egg shell extract.
Figure 6 illustrates the effect of various concentration of egg shell extract on the viscosity of the composition.
Figure 7 illustrates Comparison of hydration activity of various concentrations of glycerin (1-5%) with/without 0.01% egg shell extract dosed in hydrogel formulation.
Figure 8 illustrates comparison of hydration activity of hydrogel formulation dosed with/without 0.01% egg shell extract and 3% glycerin.
Detailed Description of the Invention
The present invention relates to a personal care composition comprising chicken egg shell extract and at least one humectant.
According to the present invention, egg shell extract may be present in the composition in an amount of about 0.00001% to about 3% by weight of the composition. Preferably, the amount of the extract is about 0.001% to about 1%, more preferably about 0.01% to about 0.1%, to achieve optimum hydration activity.
The humectants in the composition according to the present invention are selected from glycerin, propylene glycol, butylene glycol and sorbitol.The humectant is present in the composition at a concentration of 2-5wt%, more preferably at 3% by wt.
The whole eggshell is composed of 95% of calcium carbonate in the calcite form. It also contains a 3.5 % of organic matter in the eggshell membranes and cuticle together with some being present in the calcified part of the shell...” – [Ref: Daengprok W, Issigonis K, Mine Y, Pornsinpatip P, Garnjanagoonchorn W, Naivikul O: Chicken eggshell matrix proteins enhance calcium transport in the human intestinal epithelial cells, Caco-Journal of agricultural and food chemistry 2003, 51:6056-6061]. Eggshell membrane proteins when extracted comprise of elastin, desmosine, lysozyme, ovotransferrin, acetylglucosaminidase, collagens such as Type I collagen, Type V collagen, Type X collagen, etc. They also contain polysaccharides such as hyaluronic acid, chondrotin and glucosamine.
The egg shell is a highly specialized and mineralized structure comprising of minerals and low quantities of proteins. The egg shell powder as cited in the prior art comprises of >99% minerals and very little organic matrix. In cosmetic composition the use of egg shell powder has been explored as exfoliant/scrubbing agents (friction related properties).
On the other hand the egg shell extract according to the present invention is prepared from only the eggshell (only the calcified layer) wherein the proteinaceous membrane component has been removed.
The egg shell extract of the present invention consists of organic composition; especially the extract is a proteinaceous matrix (figure 2) with minimal components (lysozyme-14 kDa and OC17-17 kDa). The organic matrix from the eggshell amounts to only ~ 0.15% by weight. This amount of organic matrix is even lower as compared to the extract from the eggshell membrane and calcified matrix which amounts to ~3.5%. Thus the composition of egg shell extract is very different from egg shell powder and as well as from egg shell membrane extract. The present invention makes use of eggshell extract containing proteinaceous components (without the mineral portion or the matrix proteins from eggshell membrane) to provide a cosmetic formulation that imparts high levels of moisturization to skin.
Glycerin in itself is a good moisturizing agent; however, adding higher concentration of it into the formulation might make the formula too sticky, affects the rheological properties of the product and result in poor absorption onto the skin. Hence, the inventors have found that adding very little of eggshell extract to optimal levels of humectantsin the formulation can result in significantly enhanced hydration efficacy and help maintain the product sensorials. In fact the inventors of the present invention have further found that the egg shell extract when used at concentrations above 3% tends to pull down the viscosity of the product. Thus combination of egg shell extract and humectants at right concentration is important to achieve the desired hydration efficacy.
The personal care composition further comprises cosmetically acceptable excipients.
The cosmetically acceptable excipients can be selected from silicones, vitamin and/or derivatives thereof, polyols, vehicles, structurants, emollient/s, gelling agent/s, a thickening agent/s, a hydrophilic or hydrophobic polymer/s, 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 personal care composition according to the present invention can be in the form of gels, serums and emulsions.

The present invention contemplates the process for preparing the egg shell extract comprises the steps of washing and peeling off the membrane. This is followed by grounding the egg shells to a fine powder. The powdered egg shell is decalcified using 20% acetic acid (10 ml of acetic acid/gram egg shell). The turbid solution is then centrifuged and the supernatant is dialyzed in a dialysis membrane after which the crude extract is lyophilized. The process for preparation of the egg shell extract generally follows the protocol as described in FEBS Letter, 582 (2008),699-704 and Poultry Science (2000) 79:580–588.
Thepresent invention provides a process of preparation of the composition in the form of serum such that a weighed amount of chelating agent and humectant are mixed with water to form a premix, and a thickener is added to this premix to solubilize for 15-20 minutes. This is then followed by addition of active (egg shell extract) and fragrance.
The composition in the form of hydrogel is prepared by solubilizing the chelating agent in water followed by addition of required quantity of structurants. The humectant is added to said mixture and then the neutralizer is added. Lastly the active is added and stirred.
The preservatives according to the present invention can be chosen from a group comprising, but are not limited to dimethyloldimethylhydantoin, parabens, BHT, EDTA, phenoxy ethanol phenoxy ethanol, sorbic acid, phenoxy ethanol and combination thereof. The amount of preservative ranges from 0.01 to 0.5 wt%., preferably from 0.02 - 0.2 wt%.
The fragrances suitable for the present invention can be chosen from a group comprising, but are not limited to essential oils, natural fragrance, synthetic fragrances or combination thereof. The amount of fragrance required ranges from 0.1 to 1.5 wt%, preferably from 0.5 - 1.3 wt%.
The present invention may further require 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.
The term “neat extract” referred herein as the use of the crude extract (without chromatographic purification) as such without a matrix (i.e., not dosed in any formulation but used as such on skin).
The invention is now further illustrated by non limiting examples.
Example 1
I. Preparation of chicken egg shell extracts
1. The egg shell was washed with distilled water and dried. The membrane of the egg shell was peeled off manually. The egg shell without membrane was ground to a fine powder using pestle and mortar.

2. The powdered egg shell was decalcified using 20% acetic acid (10 ml of acetic acid/gram eggshell) at 4°C on a magnetic stirrer for 48 hrs.
3. The turbid solution was centrifuged at 5,000 rpm for 5 minutes. The supernatant was dialyzed in a dialysis membrane at 4° C for 72 hrs after which the solution was lyophilized to obtain a powdered form of the crude extract.

II. Characterization of eggshell extract
The crude extract obtained in step I was further subjected to chromatography using an Akta Purifier (RP-HPLC, Jupitor C18 analytical column, with a gradient of 30% to 70% of buffer B for 30 min, where Buffer A is 0.1% TFA in water and Buffer B is 0.1% TFA in 80% acetonitrile). The SDS-PAGE profile (Figure 2) shows that the purified extract according to the present invention contains two major protein bands corresponding to OC-17 (~17 kDa) and Lysozyme (14 kDa).
Example 2
1. Effect of neat chicken egg shell extract on the skin hydration activity.
Chicken egg shell extract was applied on the fore-arm of the human volunteers and hydration activity of neat extract was compared with untreated skin and skin treated with 10% Glycerin(Figure 3) using a corneometer. The result shows that the neat extract provided negative effect as skin hydrating agent when used alone.
2. Effect of neat chicken egg shell extract on the skin MMP activity.
Chicken egg shell extract at different concentrations weretested for MMP inhibition using the MMP-1 Colorimetric Drug Discovery assay kit from Enzo-lifesciences following the supplier’s assay protocol (Figure 4). The result shows that the neat extract at all concentrations provided negative effect.
Principle and protocol of the assay: MMP (matrix metalloproteinase) inhibition assay aims to measure the protease activity of MMP-1 using thiopeptide as a chromogenic substrate. The MMP cleavage site peptide bond is replaced by a thioester bond in thiopeptide. Hydrolysis of this bond by an MMP produces a sulfhydryl group, which reacts with Ellman’s reagent to form 2-nitro-5-thiobenzoic acid, which can be detected by its absorbance at 412nm. 20µL of chicken egg shell extract of different concentrations was incubated with 20µL of MMP-1 enzyme, 10µL of substrate and 50µL of assay buffer. The reaction kinetics was read at 412A against a suitable blank for 30 minutes. NNGH [N-Isobutyl-N-(4-methoxyphenylsulfonyl) glycylhydroxamic acid] was used as a standard inhibitor.It was found that egg shell extract did notexhibit MMP inhibition activity.
3. Determination of moisturizing efficacy/hydration activity of hydrogel formulation (containing chicken egg shell extract and glycerin) on human volunteers using corneometer.
The egg shell extract was used at different concentration in combination with a humectant such as glycerin, wherein glycerin concentration was ranging from 1-5%.
Figure 5shows the effect of egg shell extract at various concentration (0.01-5%) when used in combination with 3% glycerine.
Table I below demonstrates different concentrations of egg shell extract were tested for its hydration activity. The results are presented in Table I.
Table I: Hydration effects of various concentration of egg shell extract in presence of 3% glycerin
Hydrogel dosed with varying concentrations of Egg Shell extract % improvement in hydration against placebo
0.0001% 14.93
0.001% 28.64
0.01% 89.99
0.10% 63.51
1.0% 35.76
5.0% 9.14

4. Effect of higher concentration of egg shell extract on the viscosity of the composition.
50 µL of hydrogel formulation containing varying levels of eggshell extract was spotted on a glass slideand network structure of the gel is observed. The results are shown in table II and depcted in figure 6.

Table II: Visosity of the composition depends on the concentration of egg shell extract
Hydrogel formulated with 3% glycerin containing different concentrations of eggshell (ES) extract Viscosity(cps)
Placebo (without ES extract) 32,500
0.01% ES extract 32,800
0.1% ES extract 31,400
1% ES extract 18,600
3% ES extract 13,300
5% ES extract 2,100
6% ES extract 1,300

It can be seen from the table shown above that at concentrations greater than 3%, the viscosity of the product drops down immensely (<10000 cps). Figure 6further substantiates the finding that the egg-shell extract beyond 3% concentration is not suitable according to the present invention.
5. The composition was prepared using varied concentration of glycerin as humectant and egg shll extract as hydrating agent.
Figure 7 demonstrates the effect of varied concentration of glycrine(1-5%) on the hydration activity when used with 0.01% egg shell extract compared to the palcebo containing no such extract. The results show that 3% glycerin is providing best result compared to any other concentartion of such humectant.
Further figure 8 demonstrates that a combination of 0.01% egg shell extract and 3% glycerine provides the best synergisticresult compared to the placebo formulation made without egg shell extract.

Example 3
Table III: Personal care composition (hydrogel) comprising egg shell extract
Sl.No INCI Name % w/v Function
1 Water 95.28
2 Disodium EDTA 0.05 Chelating agent
3 CarbopolUltrez 10 0.4 Structurant
4 Pemulen TR2 0.1 Structurant
5 Glycerine 3 Humectant
6 Phenoxetol 0.2 Preservative
7 Triethanolamine (99%) 0.75 Neutralizer
8 Egg shell extract 0.01 Active
9 Perfume (any) 0.02 Fragrance

Table IV: Personal care composition in an emulsion form
Sl.No INCI Name Weight (%) Function
1 Distilled water 92.29
2 Disodium EDTA 0.1 Chelating agent
3 Pemulen TR2 0.05 Structurant
4 CarbopolUltrez 0.1 Structurant
5 Glycerine 3.0 Humectant
6 Steareth-1 1.2 Emulsifier
7 Steareth-2 0.6 Emulsifier
8 Triethanolamine 0.15 Neutralizer
9 Egg shell extract 0.01 Active
10 Phenoxyethanol 1 Preservative
11 Perfume 1.5 Fragrance

Table V: Personal care composition in a serum form
Sl No Description Weight % Function
1 Pasteurized water 95.69
2 Disodium EDTA 0.1 Chelating agent
3 Glycerine 3 Humectant
4 Xanthan gum 0.5 Thickener
5 Egg shell extract 0.01 Active
6 Phenoxyethanol 0.7 Preservative

Example 4
I. Process of preparation of serum:
i. Weighed quantity of chelating agent and humectantswere mixed with water to form a premix.
ii. Thickener was added to solubilize the premix of step 1 and stirred for about 15 – 20 minutes.
iii. To this, preservative, followed by active and fragrance were added to obtain the final serum formulation of the present invention.
II. Process of preparation of hydrogel:
i. The chelating agent was solubilized in water and required quantities of surfactants were added to mixture and mixed for 15-20 minutes.
ii. To the mixture of step (i), required quantity of humectantwas added and stirred for another 15 – 20 minutes, followed by addition of a neutralizer.
iii. After that required quantity of active was added to the formula and stirred for 5 to 10 min.
iv. Finally, preservative and fragrance were added at optimum levels to obtain the hydrogel formula of the present invention.