The present invention relates to pharmaceutical field, be specifically related to a synergistic herbal cosmetics composition for skin care. Generally speaking, the present invention relates to external (topical) skin care compositions and method of preparation.
BACKGROUND
The Cosmetics are used almost regularly and universally in different forms to enhance the beauty. Cosmetics are developed to reduce wrinkles, fight acne, age defying and to control oil secretion. For various types of skin ailments formulations like skin protective, sunscreen, anti-acne, anti-wrinkle and anti-aging are designed using varieties of materials, either natural or synthetic. The development process for cosmetic formulation needs maintenance of quality standard. The quality of a formulation should satisfy the consumer’s need in terms of its performance. The herbs used in cosmetic preparation have varieties of properties like antioxidant, anti-inflammatory, antiseptic and antibacterial etc. These herbal products claim to have no side effects, commonly seen with products containing synthetic agents. Attractiveness of such herbal preparations has socially as well as technologically resulted in flooding of marketplace in India. The literature in Ayurveda, especially CharakSahita, stated numerous medicinal plants in Varnya kashaya. The herbs like chandan, haldi, khas, nagkheshara, manjistha, yastimadhu are used to obtain glowing complexion and arusa, amalabavchi, guduchi, chakmard are mentioned as kustaharan.
These are the cosmetics which are prepared using plant products having cosmetic actions. Recently uses of botanicals in cosmetics have increased mainly due to the mild action and non-toxic nature. In cosmetics, both natural and phyto-ingredients are used. Natural products Include oils, extracts, secretions etc. Phyto-ingredients include pure constituents obtained by various processes.
Classification of Herbal Cosmetics:
• Skin: Powder, lipsticks, creams, lotion, rouges, solution
• Hair: Shampoos, conditioners, creams, bleaches, coloring preparation,
• Nails: Nail-lacquers, lacquer removers
• Teeth: Pastes, powders, gels, dentifrices
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• Eye: Eyeliners, mascaras, eye shadows and eyebrow pencils
CN103237555B discloses a topical skin formulation and method of using the same to increase the integrity of the dermal-epidermal junction in a person's skin by stimulating the production of proteins and enzymes in dermal and epidermal cells that aid in connecting the dermal layer to the epidermal layer. The composition includes an effective amount of a combination of plant extracts comprising argania spinosa kernel extract and cucurbita pepo seed extract; and an effective amount of a combination of amino acids comprising serine and proline.
JP6474084B2 discloses a finish composition applied as a top layer to a lower layer comprising a skin care composition ,10-25% by weight of substantially spherical silicone elastomer particles having a particle size of 2-40 micrometers; A non-volatile oil in which the ratio of non-volatile oil to silicone elastomer particles is 1:10 to 3: 2; 20-85 wt% water, Optionally 1 to 20% by weight volatile oil.
KR20050002581A discloses a cosmetic composition containing herbal medicines. The composition has skin-aging prevention effects such as skin wrinkle prevention and skin moisturization and acne improving effects, such as pore tightening, anti-pigmentation effects, as well as skin-whitening effects. CONSTITUTION: The cosmetic composition for preventing skin aging is characterized by containing 5-15 wt.% of licorice root, 4-15 wt.% of Lonicera japonica Thunb, 5-15 wt.% of Polygonum multiflorum Thunb, 4-15 wt.% of Taraxacum Platycarpum H. dahlst, 3-10 wt.% of aloe ferox, 3-15 wt.% of chestnut soft inner skin, 8-20 wt.% of Angelica dahurica, 2-5 wt.% of Scutellaria baicalensis Geogri, 8-15 wt.% of Poria cocos, 2-18 wt.% of Saururus Chinensis (Lour) Baill, 1-3 wt.% of rice bran, 1-3 wt.% of Plagiorhegma dubium Maxim, and 3-15 wt.% of ostrea gigas.
Lohani et al (2019) reported the sun protection factor (SPF) and antioxidant potential of geranium essential oil (GEO) and calendula essential oil (CEO) because having a combination of these two properties moves up the oils as an active ingredient of various cosmeceutical formulations for their preventive and protective properties. Essential oils were obtained by hydro distillation of Pelargonium graveolens leaves (GEO) and Calendula
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officinalis flowers (CEO). The composition and identification of chemical constituents of oils were determined by GCMS analysis. Free radical scavenging activity was measured by nitric oxide scavenging activity and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. It was observed that both GEO and CEO have the potential to reduce or prevent oxidative stress and can be used in skin care regimen to slow down skin aging via its antioxidant properties. In vitro SPF was determined by a very simple and rapid spectroscopic method. SPF value of GEO and CEO was found to 6.45 and 8.36, respectively. The SPF of CEO was higher than GEO, and the results of SPF show that these essential oils can be employed in sunscreen formulations to protect the skin from sunburn. From the results, it can be concluded that the combined antioxidant and SPF property of GEO and CEO can provide synergistic photoprotective effect or lift up the additional value of the cosmeceutical formulation
SUMMARY
This summary is provided to introduce a selection of concepts in a simplified format that is further described in the detailed description of the invention. This summary is neither intended to identify key or essential inventive concepts of the invention and nor is it intended for determining the scope of the invention.
The present invention relates to a herbal cosmetics composition comprising: 02 % (W/V) Guava leaf oil, 02 % (W/V) Wheat germ oil, 0-10 % (W/V) Jojoba oil, 0-10 % (W/V) Hemp seed oil, 5 % (W/V) Rose oil, 70 % (W/V) Aloe vera gel, 5 % (W/V) glycerin and 7% (W/V) nonionic surfactants for skin care by inhibiting the changes in stratum corneum of epidermis upon UV/Sunlight exposure, wrinkling, spongosis, collagen bundle disorientation, ultra structural changes in collagen fibers, photo allergic dermatitis, hyperkeratosis, photo aging of the follicular mouths, Liquefaction degeneration of the basal cell layer of the epidermis region
These as well as additional features and advantages of the present invention will become apparent in the following written description.
To further clarify the advantages and features of the present invention, a more particular description of the invention will be rendered by reference to specific embodiments thereof,
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which is illustrated in the appended figures. It is appreciated that these figures depict only typical embodiments of the invention and are therefore not to be considered limiting of its scope. The invention will be described and explained with additional specificity and detail with the accompanying figures.
OBJECTS OF THE INVENTION
The present invention has been developed in response to the present state of the art, and in particular, in response to the problems and needs in the art that have not yet been fully solved by currently available techniques and processes.
Accordingly, the present invention has been developed to provide a herbal cosmetics composition comprising: 02 % (W/V) Guava leaf oil, 02 % (W/V) Wheat germ oil, 0-10 % (W/V) Jojoba oil, 0-10 % (W/V) Hemp seed oil, 5 % (W/V) Rose oil, 70 % (W/V) Aloe vera gel, 5 % (W/V) glycerin and 7% (W/V) nonionic surfactants for skin care by inhibiting the changes in stratum corneum of epidermis upon UV/Sunlight exposure, wrinkling, spongosis, collagen bundle disorientation, ultra structural changes in collagen fibers, photo allergic dermatitis, hyperkeratosis, photo aging of the follicular mouths, Liquefaction degeneration of the basal cell layer of the epidermis region.
Therefore, the current invention successfully overcoming all of the above-discussed shortcomings present in the art.
1. It is an object of the invention to provide a herbal cosmetics composition comprising: 02 % (W/V) Guava leaf oil, 02 % (W/V) Wheat germ oil, 0-10 % (W/V) Jojoba oil, 0-10 % (W/V) Hemp seed oil, 5 % (W/V) Rose oil, 70 % (W/V) Aloe vera gel, 5 % (W/V) glycerin and 7% (W/V) nonionic surfactants for skin care.
2. It is an object of the invention to provide herbal cosmetics composition which inhibits the changes in stratum corneum of epidermis upon UV/Sunlight exposure, wrinkling, spongosis, collagen bundle disorientation, ultra structural changes in collagen fibers, photo allergic dermatitis, hyperkeratosis, photo aging of the follicular mouths, Liquefaction degeneration of the basal cell layer of the epidermis region
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3. It is an object of the invention to provide process of synthesis of herbal cosmetics composition using the hydrophile-lipophile balance (HLB) technique.
How the foregoing objects are achieved will be clear from the following brief description. In this context, it is clarified that the description provided is non-limiting and is only by way of explanation. Other objects and advantages of the invention will become apparent as the foregoing description proceeds, taken together with the accompanying drawings and the appended claims.
To further clarify the advantages and features of the present invention, a more particular description of the invention will be rendered by reference to specific embodiments thereof, which is illustrated in the appended figures. It is appreciated that this figure depicts only typical embodiments of the invention and are therefore not to be considered limiting of its scope. The invention will be described and explained with additional specificity and detail with the accompanying figure.
BRIEF DESCRIPTION OF FIGURES
These and other features, aspects, and advantages of the present invention will become better understood when the following detailed description is read with reference to the accompanying figures in which like characters represent like parts throughout the figures, wherein:
Figure 1, illustrates a view of Changes in epidermis upon UV exposure for the present invention.
Figure 2, illustrates a view of Changes in epidermis upon Sunlight exposure for the present invention.
Figure 3, illustrates a view of epidermis after giving Formulation F1for the present invention.
Figure 4, illustrates a view of epidermis after giving Formulation F2 for the present invention.
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Further, skilled artisans will appreciate that elements in the figures are illustrated for simplicity and may not have been necessarily been drawn to scale. For example, the flowcharts illustrate the method in terms of the most prominent steps involved to help to improve understanding of aspects of the present invention. Furthermore, in terms of the construction of the device, one or more components of the device may have been represented in the figures by conventional symbols, and the figures may show only those specific details that are pertinent to understanding the embodiments of the present invention so as not to obscure the figures with details that will be readily apparent to those of ordinary skill in the art having benefit of the description herein.
DETAILED DESCRIPTION:
For the purpose of promoting an understanding of the principles of the invention, reference will now be made to the embodiment illustrated in the figures and specific language will be used to describe the same. It will nevertheless be understood that no limitation of the scope of the invention is thereby intended, such alterations and further modifications in the illustrated system, and such further applications of the principles of the invention as illustrated therein being contemplated as would normally occur to one skilled in the art to which the invention relates.
It will be understood by those skilled in the art that the foregoing general description and the following detailed description are exemplary and explanatory of the invention and are not intended to be restrictive thereof.
Reference throughout this specification to “an aspect”, “another aspect” or similar language means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, appearances of the phrase “in an embodiment”, “in another embodiment” and similar language throughout this specification may, but do not necessarily, all refer to the same embodiment.
The terms "comprises", "comprising", or any other variations thereof, are intended to cover a non-exclusive inclusion, such that a process or method that comprises a list of steps does not include only those steps but may include other steps not expressly listed
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or inherent to such process or method. Similarly, one or more devices or systems or elements or structures or components proceeded by "comprises... a" does not, without more constraints, preclude the existence of other devices or other systems or other elements or other structures or other components or additional devices or additional systems or additional elements or additional structures or additional components.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The system, methods, and examples provided herein are illustrative only and not intended to be limiting.
The terms “a” and “an” herein do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced items.
The terms “having”, “comprising”, “including”, and variations thereof signify the presence of a component.
“Administering” or “administration” or “administer” means giving a material to a subject in a manner such that there is a pharmacological result in the subject. This may be direct or indirect administration, such as by inducing or directing another subject, including another clinician or the subject itself, to perform the administration.
“Amount effective” in the context of a composition or dose for administration to a subject refers to an amount of the composition or dose that produces one or more desired responses in the subject, e.g., preventing or treating a disease or disorder of the liver as is described herein, preventing or treating liver toxicity. Therefore, in some embodiments, an amount effective is any amount of a composition or dose provided herein that produces one or more of the desired therapeutic effects and/or preventative responses as provided herein. This amount can be for in vitro or in vivo purposes. For in vivo purposes, the amount can be one that a clinician would believe may have a clinical benefit for a subject in need thereof. Any one of the compositions or doses, including label doses, as provided herein can be in an amount effective.
Now the present invention will be described below in detail with reference to the following
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embodiment.
Materials
All or few of the following ingredients may be used: Guava leaf oil, Rose oil, Virgin coconut oil, Calendula oil, Wheat germ oil, Hemp seed oil, Jojoba oil, Avocado oil, Olive oil, Neem oil, Xanthan gum, Aloe Vera gel, Non-ionic surfactants (Tween 80 and Span 80) etc. All other chemicals used will be of analytical grade (AR). Ethics declaration The present investigation was conducted according to the ethical principles and protocol will be approved by the Institutional Animal Ethical Committee, School of Pharmaceutical Sciences, I.F.T.M. University Moradabad, India.
Methods GCMS analysis of essential oils The identification of the components of the essential oil samples was carried out by gas chromatography equipment (Shimadzu QP-2010 Plus with Thermal Desorption System TD 20, Shimadzu, Kyoto, Japan); AB-INN Wax column (60 m length × 0.25 mm id × 0.25µm thickness) was used under the conditions: column oven temperature was 50.0°C, injection temperature was 260.0°C, pressure was 69.0 k Pa, total flow was 125.2mL/min, and column flow was 1.21mL/min. Injected oil sample volume was 0.1µL, split ratio was 100.0, ion source temperature was 230.00°C, interface temperature was 270.0°C, and the mass spectrometer was scanned over the 40-650 m/z. Identification of individual oil component was done by comparing the mass data of individual oil component peaks with the standard library database (Wiley 7 NIST 05 mass spectral database). Preparation of emulsions Various emulsion formulations will be prepared using the hydrophile-lipophile balance (HLB) technique. Briefly, the oil phase comprising of essential oil(s), fixed oil, hydrophobic
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non ionic surfactants and other excipients will be mixed with the aqueous phase made up of emulsion stabilizers containing a hydrophilic non ionic surfactant using a mechanical stirrer until the mixture becomes homogenous (Table 5). The volume will be made up using deionized water, and the emulsions will be kept in wide mouth air tight containers53.
Table 1: Composition of emulsion formulation Name of ingredients Concentration (%W/V) Formulation F1
Formulation F2
Guava leaf oil
02
02
Wheat germ oil
02
02
Jojoba oil
10
----
Hemp seed oil
----
10
Rose oil
5
5
Glycerin
5
5
Nonionic surfactants
7
7
Aloe vera gel
70
70
In vitro sun protection factor determination of cosmetic emulsions The in vitro SPF measurement techniques represent an admissible and fast tool for shortening in vivo experiment numbers and risks. A quantity of 1.0 g of emulsion formulation was transferred to a 100-ml volumetric flask and diluted to the required volume with ethanol. Furthermore, it was ultrasonicated for 5 min and filtered through a cotton filter, discarding the first 10 ml. A 5.0-ml aliquot was transferred to a 25-ml volumetric flask
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and the volume was adjusted with ethanol. The absorbance spectra of the sample in the solution form were obtained in the range of 290–320 nm, at every 5 nm interval, and three determinations were made at each point, followed by the application of Mansur equation 54-55. Where EE(I) is the erythemal effect spectrum; I(I) is the solar intensity spectrum; Abs is the absorbance of the sunscreen product; and CF is the correction factor (10). The value of EE × I is constant. The present study was designed with an objective to in vitro determine the SPF, using an ultraviolet (UV)–visible spectrophotometer (Shimazdu-1800, Japan), of the investigational age-defying formulations. Determination of Antioxidant activities of Cosmetic emulsions
1, 1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity
To evaluate the antioxidant capacity of formulations, the oil samples were allowed to react with methanolic solution of DPPH. DPPH is a stable nitrogen centered free radical which changes the color from dark violet to yellow in the presence of compounds that are capable of either donating hydrogen or transferring an electron. During the reaction, the reduction in DPPH can be monitored by the decrease in its absorbance at a characteristic wavelength (?max: 515-517 nm). The free radical scavenging activities of formulations were estimated by DPPH using the method described by Elmastas et al 56-57. Different dilutions of oil samples and standard (ascorbic acid) in the concentration range of 10-250µg/mL were prepared in methanol. 0.1 mmol/L solution of DPPH was prepared in methanol, and 3mL of this solution was added to the equal volume of diluted sample and standard solutions, mixed well, and the tubes were incubated for 30 minute in dark at 30°C. The absorbance of each
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dilution was measured at 517 nm using UV-Visible Spectrophotometer. DPPH radical scavenging capability was calculated using the following equation:
Where AbC is absorbance of control and AbS is absorbance of sample/standard. % Inhibition of each sample dilution was calculated and a graph was plotted to obtain a linear regression equation, by taking concentration (µg/mL) on x-axis and % Inhibition on y-axis. From the linear regression equation, IC50 value was calculated. IC50 value is the concentration of the sample required to scavenge 50% DPPH-free radical. The experiment was done in triplicate.
Nitric oxide scavenging activity
Sodium nitroprusside decomposes in aqueous physiological solution (pH-7.4) and generates nitric oxide. Nitric oxide reacts with oxygen under aerobic conditions and generated nitrite ions that can be estimated by using Griess reagent (1 part of 0.1% napthylethylenediaminedihydrochloride in distilled water and 1 part of 1% sulfanilamide in 5% phosphoric acid)58. Nitric oxide scavengers compete with oxygen leading to reduced production of nitrite ions. In this method, 10mmol/L sodium nitroprusside was prepared in phosphate buffer (pH 7.4) and 2mL of this solution was added to oil sample and standard (ascorbic acid) dilutions (10-250µg/mL) in methanol. The tubes were incubated at 25°C for 2 hours. After incubation, 0.5mL Griess reagent was added to the incubated tubes and absorbance was measured at 546 nm using UV-Visible Spectrophotometer. The amount of nitric oxide radical was calculated by following the equation:
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Where AbC is absorbance of control and AbS is absorbance of sample/standard. % Inhibition of each sample dilution was calculated and a graph was plotted to obtain a linear regression equation, by taking concentration (µg/mL) on x-axis and % Inhibition on y-axis. From the linear regression equation, IC50 value was calculated. IC50 value is the concentration of the sample required to scavenge 50% Nitric oxide-free radical. The experiment was done in triplicate.
Animal studies
The study was conducted after obtaining institutional animal ethical committee (IAEC) clearance. Albino rats (Wistar strain) were employed in the present study. The animals were maintained at controlled temperature under alternating light and dark conditions, relative humidity (60±5%) and housed in polypropylene cages. Standard food pellets ad libitum and drinking water were provided.
Experimental Groups
To evaluate the effect of cosmetic emulsion formulation on animal skin of albino rats against UV-B radiation, the four groups each of six albino rats each were prepared and assigned Gr1, Gr2, Gr3 and Gr4 (Table 6).
Table 2: Animal groupings to evaluate the effect of cosmetic formulation
Groups (n=4)
Treatment Group I Control Group II UV B irradiated group Group III UV B irradiated group + Formulation F1 Group IV UV B irradiated group + Formulation F2
Skin irritation testing
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This study was conducted using Wistar strain, albino rats weighing (150–200 g), which were kept for an acclimation period of 7 days in order to gain surety for their usage in experiments. The animals were taken and their backs (dorsally) were shaved. An experimental study was done after 2 days of hair removal. Approximately 0.1 g of each test formulation was applied over the shaved skin (25 × 25 mm area) of the animals from each group Approximately 0.1 g of each test product was applied over the shaved skin (25 × 25 mm area) of the animals from each group. The formulation applied was left over for 30 min and then washed off using tap water. Experimented animals were observed for erythema and edema after 24 and 72 h, respectively. On the basis of the observation, the formulations were scored for primary irritation, so as to confirm the extent of irritation and the comparison of effectiveness 59-60. Exposure to UV B irradiation
All rats in various groups were shaved four days before the initiation of the experimentation. Two UV lamps (Toshiba Company) were used to induce photo damage on the skin surface of rats. The emission peak was observed at 298 nm, and the spectral output was in the range of 290–310 nm. The dose for radiation exposure (mJ/cm-2) was calculated from irradiance (mW cm-2) × time of exposure (min). The time of exposure was calculated as per Organization for Economic Co-operation and Development (OECD) guidelines. Thirty minutes prior to irradiation, 0.1g/cm-2 of cream formulations were applied to the back skin of animals61. In the present study, animals were irradiated with a radiation exposure dose of 500 mJ/cm-2 with an exposure time of 12 min per day (daily) for 1 month. The distance from the lamps and the backs of mice was 20 cm -1.
Histopathological studies
After 1 month of study, the animals were sacrificed. The shaved dorsal skin samples were carefully dissected. Tissue specimens from the back of the skin for all the groups were
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thoroughly rinsed using physiological saline, diluted 10 times with distilled water, stored at -70°C in 40% formalin solutions61-62. The tissue specimens were then studied under microscope for following histopathological observations:
Table 3: Histopathological observations to be made in animal tissues
Groups (n=4)
Histopathological observations
Group I
Normal histopathological characteristics of epidermis Group II
Changes in stratum corneum of epidermis upon UV/Sunlight exposure: wrinkles, spongosis, collagen bundle disorientation, Ultra structural changes in collagen fibers, photo allergic dermatitis and hyperkeratosis and photo aging of the follicular mouths. Liquefaction degeneration of the basal cell layer of the epidermis region. Group III
Improvement/changes in above symptoms after treatment with formulations Group IV
Improvement/changes in above symptoms after treatment with formulations
Selection of formulation ingredients
Guava (Psidium guajava L.) leaf oil ( GLO)
Chemical constituents of GLO
Key constituents of Guava leaf oil
Chemical constituents Percentages-
limonene 53–58%
1,8-cineole 30–34%
?- caryophyllene 2.3–3.4
a-pinene 1.4–1.9%
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a-terpineol 1.4–1.9%
Aloe Vera Gel (AVL)
Carrier oils
Jojoba oil (JO)
Hemp (Cannabis sativa L.) seed oil (HCO) Characterization of essential oils
Guava (Psidium guajava L.) leaf oil( GLO)
SPF determination of cosmetic emulsions
UV radiation from the sun is transmitted at three wavelengths – UVA, UVB, and UVC. UVC does not penetrate the earth’s atmosphere. UV irradiation in the form of UVA is associated with skin aging. UVA affects the elastin in the skin and leads to wrinkles and sun-induced skin aging (e.g., coarse wrinkles, leathery skin, and brown pigmentation), as well as skin cancer. UVA can penetrate window glass and penetrates the skin more deeply than UVB.
UVB is the form of UV irradiation most responsible for sunburn and has strong links to malignant melanoma and basal cell carcinoma risk (types of skin cancer) 71-73. SPF stands for “Sun Protection Factor” which theoretically refers to the amount of time you can stay in the sun without getting sunburned compared to going without sun block. It is the ratio of UV energy required to produce a minimal erythemal dose (MED) in protected skin to unprotected skin. When the SPF is 15, it provides > 93% protection against UV-B, and a SPF of +30 provides 97% protection from UV-B. Here, the SPF values of formulations (F1 & F2) were found to be 5.6 & 12.4 respectively. The SPF of formulation F2 was significantly better than formulation F1. This could be attributed to the presence of Pumpkin seed oil in the formulation F2
Table 4: Determination of SPF in cosmetic emulsions
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Cosmetic emulsion formulations
SPF
F1
5.6
F2
12.4
Skin irritation testing
Skin irritation study was designed to observe the extent or the degree of skin irritation caused by the formulation. Skin irritation of a newly made formulation is very essential in order to check any non-immunogenic inflammatory reaction that arises locally that usually diminishes or finishes soon after some time of application. Some of the symptoms, such as the presence of pain, itching, erythema, or oedema, signify the presence of skin irritation56. These symptoms could be taken as proof for any cellular, neural, biochemical, or vascular response to the start of the irritation over the skin (Table 9). The formulation F1& F2 showed a 0.613 and 0.489 primary irritation index (PII), which indicates mild (F1) to negligible irritancy (F2).
Table 5: determination of Primary Irritation Index (PII)
Cosmetic emulsion formulation
Primary Irritation Index (PII)
F1
0.613
F2
0.489
Determination of Antioxidant activities of Cosmetic emulsions
1, 1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity
The antioxidant activities were measured using the free stable radical, 1, 1-diphenyl-2- picrylhydrazyl is characterized as a stable free radical by virtue of the delocalization of the spare electron over the molecule as a whole, so that the molecules do not dimerise , as would be the case with most other free radicals. The delocalization also gives rise to the
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deep violet colour, characterized by an absorption band in ethanol solution centered at about 520 nm.When a solution of DPPH is mixed with that of a substance that can donate a hydrogen atom, then this gives rise to the reduced form (2) with the loss of this violet colour (although there would be expected to be a residual pale yellow colour from the picryl group still present).
Table 6: Determination of DPPH radical scavenging activity
S.No
Concentration
(v/v)
F1
Percent inhibition
F2
Percent inhibition
1.
10
18.14 ± 1.51
19.16 ± 1.11
2.
20
21.01 ± 1.39
22.34 ± 1.21
3.
40
26.87 ± 2.15
28.43 ± 1.98
4.
60
30.56 ± 3.12
32.26 ± 3.15
5.
80
36.97 ± 3.33
38.19 ± 3.89
6.
80 (ASC)
68.8 ± 1.51
The result of DPPH scavenging activity of cosmetic formulations showed that the prepared formulations have the power to diminish the dark violet color of DPPH radical to the yellow diphenylpicrylhydrazine radical. The highest DPPH radical scavenging activity obtained by standard Ascorbic acid (ASC) was 68.8 ± 1.51% in a concentration of 80 µg/ml, while in the same concentration; the DPPH scavenging activity of formulations was 36.97 ± 3.33% (F1), and 38.19 ± 3.89 % (F2) when recorded after 30 min of incubation. The percent inhibition was found in concentration dependent manner almost same in both assay methods.
Nitric oxide scavenging activity
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Nitric oxide (NO) is generated from amino acid L-arginine by vascular endothelial cells, phagocytes, and certain cells of the brain. Nitric oxide is classified as a free radical because of its unpaired electron and displays important reactivity with certain types of proteins and other free radicals. The toxicity of NO becomes adverse when it reacts with superoxide radical, forming a highly reactive peroxynitrite anion (ONOO-)75. In this experiment Nitric oxide was generated from sodium nitroprusside. Sodium nitropruside in aqueous solution at physiologic pH (7.4) spontaneously generates nitric oxide, which interact with oxygen to produce nitrite ions, which can be determined by use of Griess reagent. Ascorbic acid was used as a standard. Ascorbic acid inhibits induction of nitric oxide synthase and is a naturally occurring direct scavenger of nitric oxide. It reduces the amount of nitrite formed between oxygen and nitric oxide generated from sodium nitroprusside. The absorbance was measured at 596 nm and the percentage antioxidant activity calculated. Table 7: Determination of Nitric oxide scavenging activity
S.No
Concentration
(v/v)
F1 (2%)
Percent inhibition
F2 (4%)
Percent inhibition
1.
10
18.89 ± 1.24
17.86 ± 1.09
2.
20
23.17 ± 1.79
21.29 ± 1.25
3.
40
25.11 ± 2.11
29.88 ± 2.31
4.
60
31.23 ± 2.45
31.24 ± 2.16
5.
80
38.17 ± 2.87
37.16 ± 2.63
6.
80v/v (ASC)
62.2 ± 0.25
The maximum nitric oxide scavenging activity shown by ASC was 62.2 ± 0.25% at 80 µg/ml, while at the same concentration; the emulsion formulations exhibited 38.17± 2.87 % (F1) and 37.16 ± 2.63 % (F2) nitric oxide scavenging activity. From the results, it can be
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observed that both formulations scavenge DPPH and nitric oxide with efficiency near to that of 60% of the standard antioxidant.
Histopathological evaluation
UV rays affect the epidermis of mammals, contributing to the aging of skin. All these negative effects justify the studies on the relation between the UV rays and epidermis. The purpose of this study i.e., the histopathology was to investigate the effects of UV radiation on the stratum corneum of Wistar rat’s epidermis. Wistar rats were divided into four groups (n = 4). After treatment of 30 days with topical preparations, the photomicrographs were observed. The histopathological observations are as follows:
H & E stained tissue section show lining stratified squamous epithelium. Dermis show well arranged collagen fibers with fat, muscle tissue and hair follicles in various stages. No abnormal changes seen.
H & E stained tissue section show mild increased thickness of lining stratified squamous epithelium with increase in melanin production areas in epidermis. Dermis shows disorientation of collagen fibres with mild infiltration by mononuclear cells. Muscle and fat tissue with scattered hair follicles also seen.
H & E stained tissue section show mild increased thickness of lining stratified squamous epithelium. Dermis show mild disorientation of collagen fibers with no infiltration by mononuclear cells. Muscle and fat tissue with scattered hair follicles also seen.
H & E stained tissue section show mild increased thickness of lining stratified squamous epithelium. Dermis shows mild disorientation of collagen fibers. Muscle, fat tissue with scattered hair follicles also seen.
H & E stained tissue section show normal thickness of lining stratified squamous epithelium.
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Dermis show well arranged collagen fibers. Muscle, fat tissue with scattered hair follicles also seen.

WE CLAIM:

1. A herbal cosmetics composition comprising: 02 % (W/V) Guava leaf oil, 02 % (W/V) Wheat germ oil, 0-10 % (W/V) Jojoba oil, 0-10 % (W/V) Hemp seed oil, 5 % (W/V) Rose oil, 70 % (W/V) Aloe vera gel, 5 % (W/V) Glycerin and 7% (W/V) Nonionic surfactants for skin care.
2. The herbal cosmetics composition as claimed in claim 1, wherein said Jojoba oil and Hemp seed oil are optional ingredients.
3. The herbal cosmetics composition as claimed in claim 1, wherein said composition inhibits the changes in stratum corneum of epidermis upon UV/Sunlight exposure, wrinkling, spongosis, collagen bundle disorientation, ultra structural changes in collagen fibers, photo allergic dermatitis, hyperkeratosis, photo aging of the follicular mouths, Liquefaction degeneration of the basal cell layer of the epidermis region.
4. The herbal cosmetics composition as claimed in claim 1, wherein said composition has SPF of 5.6 to 12.4.
5. The herbal cosmetics composition as claimed in claim 1, wherein said composition showed a 0.613 and 0.489 primary irritation index (PII).
6. The process of synthesis of herbal cosmetics composition as claimed in claim 1 comprising the step of:
(i) 02 % (W/V) Guava leaf oil, 02 % (W/V) Wheat germ oil, 0-10 % (W/V) Jojoba oil, 0-10 % (W/V) Hemp seed oil, 5 % (W/V) Rose oil, 70 % (W/V) Aloe vera gel, 5 % (W/V) Glycerin and 7% (W/V) are mixed with the
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aqueous phase made up of emulsion stabilizers containing a hydrophilic non ionic surfactant using a mechanical stirrer until the mixture becomes homogenous,
(ii) Making the volume as per desire using de-ionized water,
(iii) stored in wide mouth air tight containers.