Cleansing composition and method of preparing the same thereof

Abstract

The present invention discloses a cleansing composition comprising a combination of Alpha Hydroxy Acids (AHA) organic; a salt of a hydroxy acid; a natural humectant; rheology modifier; a natural preservative, and a solvent. The present invention also provides a method of preparing the cleansing composition. The disclosed invention performs the function of skin cleansing effectively with a simpler formulation without the use of complex surfactant blends and with natural or nature identical ingredients that readily biodegrade in nature after usage, free from harsh chemicals and suitable for all skin types.

Description

CLEANSING COMPOSITION AND METHOD OF PREPARINGTHE SAME THEREOFFIELD OF THE INVENTION

[0001] The present invention relates to a cleansing composition. More particularly, the present invention relates to water-based liquid skin cleansing compositions comprising a combination of Alpha-Hydroxy Acids (AHAs) as active ingredients, a salt of hydroxy acid and an organic rheology modifier.BACKGROUND OF THE INVENTION

[0002] 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.

[0003] Cleansing compositions are products designed to clean and purify the skin. These compositions typically come in various forms, such as liquid cleansers, foaming cleansers, creams, gels, or solid bars, and they are used to remove dirt, excess oil, makeup, and impurities from the skin. The specific ingredients and formulations of these compositions can vary widely depending on the type of skin they are designed for and the desired effects.

[0004] Common ingredients found in skin cleansing compositions include water, surfactants, emollients and humectants along with some preservatives in few cases to extend the shelf life of the product and prevent the growth of harmful microorganisms. Nevertheless, several skin cleansing compositions include Alpha Hydroxy Acids (AHAs) as an active ingredient along with various types of surfactants. Few non-foaming rinse-off formulations contain non-ionic surfactants. There are other non-foaming formulations known in the art which are not waterbased and rely on alcohols as active ingredients and as a solvent base.

[0005] Most of the current rinse-off cleansing composition for topical application contain active ingredients in the form of surfactant. For instance, US patent US5030374A discloses a non-foaming rinseable cleanser formulation for facial skin application comprising of mild nonionic surfactant, wound cleanser, moisturizer / humectant, moisturizer / emollient, gelling agent, viscosity stabilizer, pH adjuster, substantive emollient, preservative or bacterial inhibitor, solubilizer, and masking agent.

[0006] Another patent document, EP3324926B1, discloses a composition for cleansing a keratin material comprising amino acid surfactant and calcium alginate.

[0007] It is understood that surfactants are one of the important ingredients for most of the cleansing compositions. However, potential side effects have been reported for the cleansing compositions comprising surfactants. For instance, surfactants can be harsh on the skin, especially if used in high concentrations. They can strip away the skin's natural oils, leading to dryness, tightness, and irritation, which can be especially problematic for people with already dry or sensitive skin. Nevertheless, surfactants, especially more potent ones, can cause skin irritation, redness, and inflammation, particularly for individuals with sensitive or reactive skin. Some people may be sensitive or allergic to specific surfactants, leading to allergic reactions like itching, redness, or hives.

[0008] Therefore, there is a dire need to develop a cleansing composition without the use of any surfactant with providing additional benefits beyond the basic cleansing effects brought about by the traditional liquid soap product.OBJECTIVE OF THE INVENTION

[0009] An objective of the present invention is to provide a cleansing composition comprising Alpha Hydroxy Acids (AHAs) along with a salt of a hydroxy acid which provide the cleansing efficacy similar to surfactant-based skin cleansers.

[0010] Another objective of the present invention is to provide a cleansing composition with a pH matching the skin's natural pH to avoid irritation or disruption of the skin's protective barrier.

[0011] Another objective of the present invention is to provide a method of preparing a cleansing composition without the use of complex surfactant blends.

[0012] Another objective of the present invention is to provide a method of preparing a cleansing composition with natural or nature identical ingredients that readily biodegrade in nature after usage.

[0013] Another objective of the present invention is to provide a cleansing composition free from harsh chemicals and suitable for all skin typesSUMMARY OF THE INVENTION

[0014] The disclosed invention performs the function of skin cleansing effectively with a simpler formulation without the use of complex surfactant blends and with natural or nature identical ingredients that readily biodegrade in nature after usage.

[0015] In an embodiment, the present invention discloses a cleansing composition comprising;i. a combination of Alpha Hydroxy Acids (AHA) organic; ii. a salt of a hydroxy acid; iii. a natural humectant iv. rheology modifier; and v. a natural preservative, and vi. a solvent.

[0016] In another embodiment, said organic acid combination is selected from, but not limited to, Lactic acid, Glycolic acid, Malic acid, Citric acid, Tartaric acid, Mandelic acid, Ascorbic acid in a range of 0. 1% to 5%; and

[0017] In yet another embodiment, said salt of a hydroxy acid is selected from, but not limited to, Monosodium citrate, Disodium citrate, Trisodium citrate in a range of 0.1% to 5%.

[0018] In still another embodiment, said natural humectant is selected from, but not limited to, Glycerin, Sorbitol, Hyaluronic acid, Sodium lactate, Honey or a combination thereof.

[0019] In yet another embodiment, said rheology modifier is selected from, but not limited to, Tara Gum, Xanthan Gum, Alginates, Microcrystalline Cellulose, Hydroxypropyl Methylcellulose, Glycerin (and) Cellulose (and) Cellulose Gum or a combination thereof.

[0020] In still another embodiment, said natural preservative is selected from, but not limited to, Triethyl Citrate, Glyceryl Caprylate, Benzoic Acid Benzyl Alcohol, Benzoic Acid, Dehydroacetic Acid, Tocopherol, Potassium Sorbate, Lactobacillus Ferment, Caprylhydroxamic Acid, Glyceryl Caprylate, Glycerin, Gluconolactone, Sodium Benzoate, Sodium benzoate, Glyceryl Caprylate, Glyceryl Undecylenate or a combination thereof; and

[0021] In yet another embodiment, said carrier solvent is water in the percentage of >90%.

[0022] In another embodiment, the present invention discloses a method of preparing the cleansing composition as claimed in claim 1, comprising the steps of: i. dispersing and neutralizing xanthan gum in an amount of 0.72% in 90.57% ml deionized water ii. adding citric acid and lactic acid in a range of 0. 1% to 5%; with constant stirring to obtain mixture 1 ; iii. providing trisodium citrate in the range of 6-8% and glycerin in a range of 1.5- 2.5% to mixture 1; iv. adding preservatives in an amount of 0. 1-0.3%. v. adding additional optional ingredients comprising viscosity adjusting agent, a skin conditioner, an antibacterial agent, an antioxidant, a fragrance, a colorant, achelating agent, a humectant, an emollient, an insoluble material, a pearlescent material, a diluent, and a combination thereof; and vi. obtaining the cleansing composition.

[0023] In another embodiment, the present invention discloses a method of using the cleansing composition as claimed in claim 1, comprising the steps of: i. applying the composition to the target area; ii. massaging the composition for a specific duration; and iii. rinsing the composition from the skin or hair to reveal improved skin condition.

[0024] Various objects, features, aspects, and advantages of the inventive subject matter will become more apparent from the following detailed description of preferred embodiments.BRIEF DESCRIPTION OF FIGURES

[0025] FIG. 1 displays the graph for showing the IC50 values for cytotoxic assessment

[0026] FIG. 2 shows the amplification curve, melt curve and Melt peak of GAPDH gene

[0027] FIG. 3 shows the amplification curve, melt curve and Melt peak of TNF-q gene

[0028] FIG. 4 shows the Amplification curve, melt curve and Melt peak of IL-6 gene

[0029] FIG. 5 shows the relative normalized expression of TNF-q gene

[0030] The accompanying drawings are included to provide a clear understanding of the present invention and a detailed description, and they constitute a part of this complete specification.DETAILED DESCRIPTION OF THE INVENTION

[0031] The following is a full description of the disclosure's embodiments. The embodiments are described in such a way that the disclosure is clearly communicated. The level of detail provided, on the other hand, is not meant to limit the expected variations of embodiments; rather, it is designed to include all modifications, equivalents, and alternatives that come within the spirit and scope of the current disclosure as defined by the attached claims. Unless the context indicates otherwise, the term "comprises" and variants such as "comprises" and "comprising" throughout the specification are to be read in an open, inclusive meaning, that is, as "including, but not limited to."

[0032] When "one embodiment" or "an embodiment" is used in this specification, it signifies that a particular feature, structure, or characteristic described in conjunction with theembodiment is present in at least one embodiment. As a result, the expressions "in one embodiment" and "in an embodiment" that appear throughout this specification do not necessarily refer to the same embodiment. Furthermore, in one or more embodiments, the specific features, structures, or qualities may be combined in any way that is appropriate.

[0033] Unless the content clearly demands otherwise, the singular terms "a," "an," and "the" include plural referents in this specification and the appended claims. Unless the content explicitly mandates differently, the term "or" is normally used in its broad definition, which includes "and / or."

[0034] All processes described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided with respect to certain embodiments herein is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention.

[0035] The headings and abstract of the invention provided herein are for convenience only and do not interpret the scope or meaning of the embodiments.

[0036] All publications herein are incorporated by reference to the same extent as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. Where a definition or use of a term in an incorporated reference is inconsistent or contrary to the definition of that term provided herein, the definition of that term provided herein applies and the definition of that term in the reference does not apply.

[0037] Groupings of alternative elements or embodiments of the invention disclosed herein are not to be construed as limitations. Each group member can be referred to and claimed individually or in any combination with other members of the group or other elements found herein. One or more members of a group can be included in, or deleted from, a group for reasons of convenience and / or patentability. When any such inclusion or deletion occurs, the specification is herein deemed to contain the group as modified thus fulfilling the written description that follows, and the embodiments described herein, is provided by way of illustration of an example, or examples, of particular embodiments of the principles and aspects of the present disclosure. These examples are provided for the purposes of explanation, and not of limitation, of those principles and of the disclosure.

[0038] It should also be appreciated that the present invention can be implemented in numerous ways, including as a system, a method or a device. In this specification, theseimplementations, or any other form that the invention may take, may be referred to as processes. In general, the order of the steps of the disclosed processes may be altered within the scope of the invention.

[0039] Various terms as used herein are shown below. To the extent a term used in a claim is not defined below, it should be given the broadest definition persons in the pertinent art have given that term as reflected in printed publications and issued patents at the time of filing.

[0040] In a general embodiment, the disclosed invention performs the function of skin cleansing effectively with a simpler formulation without the use of complex surfactant blends and with natural or nature identical ingredients that readily biodegrade in nature after usage. A combination of Alpha Hydroxy Acids (AHAs) along with a salt of a hydroxy acid delivered in a gel provide the cleansing efficacy similar to surfactant-based skin cleansers. In accordance with a general embodiment of the present invention, the liquid cleanser comprises a combination of Alpha Hydroxy Acids (AHAs), a salt of a hydroxy acid, a humectant, an organic rheology modifier a combination of preservatives, a colorant, fragrance and deionized water.

[0041] In an embodiment, the present invention discloses a cleansing composition comprising; i. a combination of Alpha Hydroxy Acids (AHA) organic; ii. a salt of a hydroxy acid; iii. a natural humectant iv. rheology modifier; and v. a natural preservative, and vi. a solvent.

[0042] In another embodiment, said organic acid combination is selected from, but not limited to, Lactic acid, Glycolic acid, Malic acid, Citric acid, Tartaric acid, Mandelic acid, Ascorbic acid in a range of 0. 1% to 5%; and

[0043] In yet another embodiment, said salt of a hydroxy acid is selected from, but not limited to, Monosodium citrate, Disodium citrate, Trisodium citrate in a range of 0.1% to 5%.

[0044] In still another embodiment, said natural humectant is selected from, but not limited to, Glycerin, Sorbitol, Hyaluronic acid, Sodium lactate, Honey or a combination thereof.

[0045] In yet another embodiment, said rheology modifier is selected from, but not limited to, Tara Gum, Xanthan Gum, Alginates, Microcrystalline Cellulose, Hydroxypropyl Methylcellulose, Glycerin (and) Cellulose (and) Cellulose Gum or a combination thereof.

[0046] In still another embodiment, said natural preservative is selected from, but not limited to, Triethyl Citrate, Glyceryl Caprylate, Benzoic Acid Benzyl Alcohol, Benzoic Acid, Dehydroacetic Acid, Tocopherol, Potassium Sorbate, Lactobacillus Ferment, Caprylhydroxamic Acid, Glyceryl Caprylate, Glycerin, Gluconolactone, Sodium Benzoate, Sodium benzoate, Glyceryl Caprylate, Glyceryl Undecylenate or a combination thereof; and

[0047] In yet another embodiment, said carrier solvent is water in the percentage of >90%.

[0048] In still another embodiment, said deionized water is used in the ratio percentage of 90.5%, Xanthan Gum is used in the ratio percentage of 0.72%, Citric Acid is used in the ratio percentage of 2.7%, Lactic Acid is used in the ratio percentage of 0.81%, Trisodium Citrate is used in the ratio percentage of 2.7%, Glycerin is used in the ratio percentage of 2.1% and Preservative is used in the ratio percentage of 0.2%.

[0049] In yet another embodiment, the pH of said composition ranges from 4 to 5.

[0050] In still another embodiment, the kinematic viscosity of said composition ranges from 500 to 2000 centistokes (cSt).

[0051] In yet another embodiment, the shelf of life of said composition is 2-3 years the room temperature.

[0052] In still another embodiment, said composition is in the form of solid, liquid or gel.

[0053] In still another embodiment, said cleansing composition further comprises, a viscosity adjusting agent, a skin conditioner, an antibacterial agent, an antioxidant, a fragrance, a colorant, a chelating agent, an insoluble material, a pearlescent material, a diluent, and combinations thereof. The Trisodium citrate helps in adjusting the pH and no other component was used to adjust the pH of the composition.

[0054] In yet another embodiment, said cleansing composition is dermatologist-tested and clinically proven.

[0055] In another embodiment, the present invention discloses a method of preparing the cleansing composition as claimed in claim 1, comprising the steps of: i. dispersing and neutralizing xanthan gum in an amount of 0.72% in 90.57% ml deionized water ii. adding citric acid and lactic acid in a range of 0. 1% to 5%; with constant stirring to obtain mixture 1 ; iii. providing trisodium citrate in the range of 6-8% and glycerin in a range of 1.5- 2.5% to mixture 1; iv. adding preservatives in an amount of 0. 1-0.3%.v. adding additional optional ingredients comprising viscosity adjusting agent, a skin conditioner, an antibacterial agent, an antioxidant, a fragrance, a colorant, a chelating agent, a humectant, an emollient, an insoluble material, a pearlescent material, a diluent, and a combination thereof; and vi. obtaining the cleansing composition.

[0056] In yet another embodiment, all the steps are performed at room temperature.

[0057] In still another embodiment, the optional ingredients are used in an amount ranging from 0. 1-0.2 %.

[0058] In another embodiment, the present invention discloses a method of using the cleansing composition as claimed in claim 1, comprising the steps of: i. applying the composition to the target area; ii. massaging the composition for a specific duration; and iii. rinsing the composition from the skin or hair to reveal improved skin condition.

[0059] While the foregoing describes various embodiments of the disclosure, other and further embodiments of the disclosure may be devised without departing from the basic scope thereof. The scope of the invention is determined by the claims that follow. The invention is not limited to the described embodiments, versions, or examples, which are included to enable a person having ordinary skill in the art to make and use the invention when combined with information and knowledge available to the person having ordinary skill in the art. EXAMPLES

[0060] The present invention is further explained in the form of the following examples. However, it is to be understood that the following examples are merely illustrative and are not to be taken as limitations upon the scope of the invention.MaterialsAlpha Hydroxy Acids (AHAs)

[0061] The AHAs are the active ingredients of the composition and are known to have antimicrobial and cleansing properties. Care should be taken that the concentration of the AHAs should not exceed beyond the optimal point to make them non-irritant to the skin. The present composition also takes into consideration that AHAs can lead to skin sensitization under sunlight at elevated concentrations. Hence, care has been taken to standardize the concentration of the selected AHAs as to be as safe and effective as possible.

[0062] A combination of two Alpha Hydroxy Acids (AHAs) are selected from the following:Lactic acidGlycolic acidMalic acidCitric acidTartaric acidMandelic acidAscorbic acidSalt of a Hydroxy Acid

[0063] Salts of Hydroxy Acids are widely used in cleaning composition as pH adjuster and cleansing enhancers. The salts are used to carefully adjust he pH of the composition to suit the nominal skin pH of 4.5 to 5.5. the salts combine with the AHAs to provide improved cleansing performance against stains.

[0064] A salt of a Hydroxy Acid is chosen from the following list:Monosodium citrateDisodium citrateTrisodium citrateHumectant

[0065] Humectants can be used in topical dosage forms to increase the solubility of a chemical compound's active ingredients, increasing the active ingredients' ability to penetrate skin, or its activity time. This hydrating property can also be needed to counteract a dehydrating active ingredient.

[0066] A natural humectant for the present composition is selected from the below list:GlycerinSorbitolHyaluronic acidSodium lactateHoneyRheology Modifier

[0067] The viscosity of the composition needs to be sufficiently high as to make the product not to run-off of the skin when applied and help spread the product in the absent of a surfactant. The current composition achieves the desired viscosity without compromising on clarity.

[0068] A rheology modifier is chosen from the below list:Tara GumXanthan GumAlginatesMicrocrystalline CelluloseHydroxypropyl MethylcelluloseGlycerin (and) Cellulose (and) Cellulose GumPreservatives

[0069] Natural preservatives are used in the composition to protect it from development of microbes and to prolong the shelf life.

[0070] A combination of preservatives selected from the following:Triethyl Citrate, Glyceryl Caprylate, Benzoic AcidBenzyl Alcohol, Benzoic Acid, Dehydroacetic Acid, TocopherolPotassium SorbateLactobacillus FermentCaprylhydroxamic Acid, Glyceryl Caprylate, GlycerinGluconolactone, Sodium BenzoateSodium benzoateGlyceryl Caprylate, Glyceryl Undecylenate

[0071] In addition to the foregoing ingredients, the composition can include other optional adjuncts conventionally used in liquid cleansers. These include, for example, one or more preservatives, one or more viscosity adjusting agents, one or more skin conditioners, antibacterial agents, one or more antioxidants, one or more fragrances, one or more colorants, one or more chelating (sequestering) agents and one or more insoluble materials. These optional materials are described in more detail below:Example 1 : Method of preparing the cleansing composition

[0072] Prerequisites:1. The process is conducted at room temperature throughout.2. Sufficient quality measures are in place to minimize contamination during the process.Stepl: neutralize 7.2 g xanthan gum in 905.7 ml of D.I water at room temperature with constant stirring.Step2: after all the xanthan gum has neutralized add citric acid (27 g) and lactic acid (8.1 g) with constant stirring.Step3: Add Trisodium citrate (27 g) to the mixture with constant stirring.Step4: Add Glycerin (21 g) with constant stirring.Step5: Add colorant and fragrance (Approx. 2g) with constant stirring.Step6: Add preservative mixture of potassium sorbate and sodium benzoate of (2 g) and stir until completely dissolved.Final Specifications:Appearance: transparent liquidOdor: characteristic of the fragrance usedKinematic Viscosity: 500 to 2000 cSt pH: 4 to 5Final water content: >90%

[0073] Xanthan gum is dispersed in Deionized water until completely neutralized. Citric acid and Lactic acid are added with constant stirring. Trisodium Citrate is added followed by Glycerin. Suitable colorant and Fragrance are added as required. Preservatives are added to the composition at the end.1000 ml sampleRaw Material Percentage weightDeionized Water 90.5% 905.7gXanthan Gum 0.72% 7.2gCitric Acid 2.7% 27gLactic Acid 0.81% 8.1gTrisodium Citrate 2.7% 27gGlycerin 2.1% 21gFragrance 0.2% 2gColorant 0.002 ,02gPreservative 0.2% 2gKey Specifications:- pH: 4 to 5- Water content: >90%Surfactants: 0%- Appearance: ClearKinematic Viscosity: 500 to 2000 cstIt is interesting to note that the composition is valid only if the above key specifications are met.Example 2: Assessment of anti-inflammatory activity of test product upon LPS induction in THp-1 celts Gell line THP-1 (ATCC No: TIB-202rM- Human monocytes)Reagents and Materials required1" RPMI (Gibco)2. Fetal Bovine Serum (Gibco)3. Trypsin 0.25%4. PMA (Sigma Aldrich)5. B-mercaptoethanol (SigmaAldrich)6. L-Glutamine (Sigma Atdrich)7. T-25 and r-75cm2 cell culture flaks (Farcon and NUNC respectively)8. 96 well plate, 12-well plate (Coming)9. LPS10. MTT reagent11. RNase free environment, after cleaning pipettes and table with DEpC water. 12. DEPC treated water13. TRlzol reagent (Life technologies, Cat. No. 15596026)14. Chloroform15. Chilled isopropanol16. Chilled 70% ethanol17. Ice cold Phopshate buffered saline (pBS, pH 7. a)18. DEPC treated, autoclaved, oven dried 1.5 And 2 ml micro-centrifuge tubes19. cDNA synthesis kit or Prime script RT Reagent kit (TAKARA, Cat. No. RR037A)20. SyBR Green I (PCR Biosystems, Cat. No. pB20.1S)21. Dimethyl Sulphoxide (DMSO)22. Target specific forward and reverse primers etc.

[0074] Cell culture:Cell culture THP-1, a promonocytic cell line was obtained from the ATCC. Cells were cultured in RpMl -1640 supplemented with 10% inactivated fetal bovine serum, 50 pM 2- mercaptoethanol, 2 mM LGlutamine penicillin (100 lU / ml) in a humidified atmosphere of 5o / o COz at 37oC until confluent.

[0075] Example 3: Evaluation of cytotoxic effects of given test compounds against THp-1 and 816F10 cells:

[0076] Procedure for THP 1 cells

[0077] The cell suspension was collected from T25 flask, cells were counted and seeded 50,000 cells per well in 96 well plate using respective media containing 10% FBS. Aler 24 hrs, plate was centrifuged at 1000 rpm for 5 mins, medium was aspirated gently and cells were treated with 100 pl of different test concentrations of test drugs in respective wells of micro titer plates. Further, the plate was incubated for 24 hrs at standard condition of 5o / o COz atmosphere at 37"C. After incubation, 1 0 pl of Ml_f (4 mg / 1 ml of MTT dissolved in PBS) was added to each well. The plate was incubated for 4 hrs at 37oC in 5o / o COz atmosphere. After 4 hrs of incubation, plate was centrifuged at 1000 rpm for 5 mins, medium aspirated and 100 pl of DMSO added onto the formazon crystals.

[0078] The plate was measured using a micro-plate reader at a wavelength of 590 nm. The percentage inhibition was calculated using the following formula and generated the concentration of test drug needed to inhibit cell groMh by 50% (1C50) value from the doseresponse curves using Graphpad prism S.O (FIG. 1).

[0079] Procedure for Bl 6F 10 cells:

[0080] To each well of the pre-labelled 96-well microtiter plate, 100 pl of the prepared cell suspension (50,000 cells / well) was added and incubated at 37oC with 5o / o COz. After 24 hrs of incubation, the supernatant was removed, and the monolayer was rinsed with respective media. To each predesignated well, 100 pl of test drugs at various concentrations were added and incubatedfor24 hrs. After incubation, the test solutions in the wells were discarded and 100 pl of MTT reagent (S mg / 10 ml of MTT in PBS) was added to each well. The plates were incubated for 4 hrs at 37 o C in 5o / o COz. The supernatant was removed and 100 pl of DMSO was added and the plates were genily shaken to solubilize the formazan crystals. The absorbance was measured using a microplate reader at 590 nm wavelength using a multimode plate reader, Spectramax i3X, Molecular devices.

[0081] Calculation for percentage Inhibition% Inhibition = ((oD of control - oD of sample) / oD of controt) x 100Example 4: Preparation of test solutions

[0082] Standard: Doxorubicin 3.7 mM stock of Doxorubicin was taken. Further Serial twofold dilutions were prepared from 100 pM to 1.56 pM using DMEM plain media for treatment.Sample 2: Liquid HandwashResults

[0083] The results suggested that Test Sample Liquid hand wash did not show marked cytotoxicity in THP-1 cells and B16F10 cells respectively. While on other hand, reference standard Doxorubicin have shown the 1C50 value of 6.23 and 4.303 pM against THP-1 cells and 816F10 cells. Based on cytotoxicity experiment study, we selected 0.3% and 1.1% concentration of liquid hand wash sample which are not toxic to cells for further gene expression study.Example 5 : Differentiation of THP-I cells to Macrophage using PMA as differentiating agent and induction of inflammation using LPS

[0084] The cells were aspirated from the 80% confluence culture flask and centrifuged at 1500 rpm for 5 mins. The cell pellet was then resuspended in 1 ml of RPMI complete growth medium and 2 X 105 cells / well was seeded to each well of the 12-well microtiter plate. After 24 hrs of incubation, pMA (25 ng / ml) was added to'12-well plate to differentiate THP-1 cells and incubated for 24 hrs. After incubation, differentiated cells were pretreated for 4 hrs with various concentrations of test compounds followed by LPS (10 ng / ml) stimulation for 12 hrs. Dexamethasone (2 pg / ml) was used as a positive control. Post incubation, cells were harvested for gene expression study further.

[0085] Evaluation of gene regulation of cytokines in differentiated cells Cellular Markers: (FIG. 3 and FIG. 5)1. I TNF-q2. l tL-6

[0086] Sample Preparation and RNA Isolation:

[0087] Treated cells were dissociated and rinsed with sterile IX PBS and centrifuged. The supernatant was decanted and 0.1 ml of TRlzol was added and gently mixed by inversion for 1 min. Samples were allowed to stand for 10 minutes at room temperature. To this 0.75 ml chloroform was added per 0.1 ml of TRlzol used. The contents were vortexed for 15 seconds. The tube was allowed to stand at room temperature for 5 mins. The resulting mixture was centrifuged at 12,000 rpm for 15 mins at 4"C. Upper aqueous phase was collected to a newsterile micro-centrifuge tube to which added and gently mixed by inverting the contents for 30 seconds and incubated at -20"C for 20 minutes. The contents were centrifuged at 12,000 rpm for 10 minutes at 4'C. Supernatant was discarded and the RNA pellet was washed by adding 0.25 ml olTOo / o ethanol. The RNA mixture was centrifuged at 12,000 rpm at 4"C. Supernatant was carefully discarded and the pellet was air dried. The RNA pellet was then resuspended in 20 pl of DEPC treated water. Total RNA yield was quantified using Spectra drop (Spectramax i3x, Molecular devices, USA).Table 3; Tefal RNA yieldExample 6: qPCR analysis and cDNA synthesis

[0088] The cDNA was synthesized from 500 ng of RNA using the oDNA synthesis kit from Prime script RT reagent kit (TAKARA) with oligo dT primer according to the manufacturer's instructions. The reaction volume was set to 20 pl and cDNA synthesis was performed at 50 oC for 30 min, followed by RT inactivation at 85oC for 5 min using applied biosystems, Veritii. The cDNA was further used for realtime PCR analysis.Example 7 : Primers and qPCR analysis

[0089] The PCR mixture (final volume of 20 pl) contained 1 .4 pl of cDNA, 10 pL of SyBr green Master mix and 1 pM of respective complementary forward and reverse primers specific for respective target genes. The reaction was carried out with enzyme activation at 95oC for 2 minutes followed by 2 step reaction with initial denaturation and annealing cum extension step at 95"C for 5 seconds, annealing for 30 seconds at appropriate respective temperature amplified for 39 cycles followed by secondary denaturation at 95 "C for 5 seconds, 1 cycle with melt curve capture step ranging from 65'C to 95'C for 5 secs each. The obtained results were analyzed and fold expression or regulation was calculated.Table 4: Primer DetailsResults: Table 5: Cq data of GAPDH, TNF-o, IL-6 gene (FIG. 2)Table 6: Relative normalized expression of TNF-o geneTable 7: Relative normalized expression of IL-6 gene (FIG. 4)

[0090] The RT PCR analysis of target genes TNF-o and IL-6 were carried out and fold regulation was determined. The results suggested the increase in TNF-o and IL-6 with 11.16- and 14.72-fold expression respectively when exposed to LPS as compared to control differentiated cells (FIG. 1 and 5). On other hand, there is decrease in expression of TNF-o with 7.96 and 6.55 when cells were treated with Sample at 0.3% and 1.1 % respectively. Likewise, the gene expression of IL-6 also got downregulated by 0.58 and 0.57-fold when cells were treated with lower and higher concentration of Liquid hand wash sample (FIG. la and 15). While reference positive control Dexamethasone at 2 pglml concentration showed significant down regulation of both TNF-o and 1L6 by 2.98 and 0.21fold respectively as compared to disease control (FIG. 3 and FIG. 4).

[0091] Evaluation of Antimicrobial Activity by AATCC 100- 2012:1. Name of the test: Evaluation of AntimicrobialActivity by AATCC 100- 2012.2. Appearance: White colour liquid.3. Temperature: 25"C t2"C.4. Agar Used: NutrientAgar.5. Test Organisms:5.1. Staphylococcus aureusATCC 6538 (1.23 X 108cfu / mL)5.2. Escherichia coliNICC 8739 (1.20X 108cfu / ml)5.3. Pseudomonas aeruginosa ATCC 15442 (1.15 X 108cfu / ml)5.4. Candida albicansATCC 10231(1.13 X 108cfu / mL)5.5. Aspergillus nrgerATCC 6275 (1.24X 108cfu / mL)5.6. Staphylococcus epidermisATCc 12228 (1.37 X 108cfu / ml)6. Results: Hand wash Liquid 290gms in contact with individual test cultures for 48 hrs. at 37"C,25"C tor 3 days to 5 days showed the following results:7. Remarks: 7.1. cfii- colony forming unit = Number of microorganisms. B-A 7.2. Percentage Reduction of Microorganism (R) = 100 ,8. Interpretation: Hand wash Liquid 290gms has shown >99.999, >99.999, >99.999, >99.999, >99.999 & >99.999, antimicrobial activity, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Aspergillus niger & Staphylococcus epidermr.s respectively when analyzed as per AATCC 100-2012 test method.

[0092] The foregoing examples are merely illustrative and are not to be taken as limitations upon the scope of the invention. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art. Such changes and modifications may be made without departing from the scope of the invention.ADVANTAGES OF THE PRESENT INVENTION

[0093] The disclosed invention performs the function of skin cleansing effectively with a simpler formulation without the use of complex surfactant blends and with natural or nature identical ingredients that readily biodegrade in nature after usage.

Claims

aim:

1. A cleansing composition comprising; i. a combination of Alpha Hydroxy Acids (AHA) organic; ii. a salt of a hydroxy acid; iii. a natural humectant iv. rheology modifier; and v. a natural preservative, and vi. a solvent wherein said organic acid combination is selected from, but not limited to, Lactic acid, Glycolic acid, Malic acid, Citric acid, Tartaric acid, Mandelic acid, Ascorbic acid in a range of 0.1% to 5%; and wherein said salt of a hydroxy acid is selected from, but not limited to, Monosodium citrate, Disodium citrate, Trisodium citrate in a range of 0. 1% to 5%. wherein said natural humectant is selected from, but not limited to, Glycerin, Sorbitol, Hyaluronic acid, Sodium lactate, Honey or a combination thereof. wherein said rheology modifier is selected from, but not limited to, Tara Gum, Xanthan Gum, Alginates, Microcrystalline Cellulose, Hydroxypropyl Methylcellulose, Glycerin (and) Cellulose (and) Cellulose Gum or a combination thereof. wherein said natural preservative is selected from, but not limited to, Triethyl Citrate, Glyceryl Caprylate, Benzoic Acid Benzyl Alcohol, Benzoic Acid, Dehydroacetic Acid, Tocopherol, Potassium Sorbate, Lactobacillus Ferment, Caprylhydroxamic Acid, Glyceryl Caprylate, Glycerin, Gluconolactone, Sodium Benzoate, Sodium benzoate, Glyceryl Caprylate, Glyceryl Undecylenate or a combination thereof; and wherein the carrier solvent comprises >90% of water.

2. The cleansing composition as claimed in claim 1, wherein the deionized water is used in the ratio percentage of 90.5%, Xanthan Gum is used in the ratio percentage of 0.72%, Citric Acid is used in the ratio percentage of 2.7%, Lactic Acid is used in the ratio percentage of 0.81%, Trisodium Citrate is used in the ratio percentage of 2.7%, Glycerin is used in the ratio percentage of 2.1% and Preservative is used in the ratio percentage of 0.2%.

3. The cleansing composition as claimed in claim 1, wherein the pH of said composition ranges from 4 to 5.

4. The cleansing composition as claimed in claim 1, wherein the kinematic viscosity of said composition ranges from 500 to 2000 centistokes (cSt).

5. The cleansing composition as claimed in claim 1, wherein the shelf of life of said composition is 2-3 years at the room temperature.

6. The cleansing composition as claimed in claim 1, further comprising a viscosity adjusting agent, a skin conditioner, an antibacterial agent, an antioxidant, a fragrance, a colorant, a chelating agent, an insoluble material, a pearle scent material, a diluent, and combinations thereof in an amount of 1-2% by wt.

7. The cleansing composition as claimed in claim 1, wherein said cleansing composition is dermatologist-tested and clinically proven.

8. A method of preparing the cleansing composition as claimed in claim 1, comprising the steps of: i. dispersing and neutralizing xanthan gum in an amount of 0.72% in 90.57% ml deionized water ii. adding citric acid and lactic acid in a range of 0. 1% to 5%; with constant stirring to obtain mixture 1 ; iii. providing trisodium citrate in the range of 6-8% and glycerin in a range of 1.5- 2.5% to mixture 1; iv. adding preservatives in an amount of 0. 1-0.3%. v. adding additional optional ingredients comprising viscosity adjusting agent, a skin conditioner, an antibacterial agent, an antioxidant, a fragrance, a colorant, a chelating agent, a humectant, an emollient, an insoluble material, a pearlescent material, a diluent, and a combination thereof; and vi. obtaining the cleansing composition.

9. The method of preparing the cleansing composition as claimed in claim 9, wherein all the steps are performed at room temperature.

10. The method of preparing the cleansing composition as claimed in claim 9, wherein the optional ingredients are used in an amount of 1-2% by w / v.

11. A method of using the cleansing composition as claimed in claim 1, comprising the steps of: i. Applying the composition to the target area. ii. Massaging the composition for a specific duration. iii. Rinsing the composition from the skin or hair to reveal improved skin or hair condition.

Images