Title of invention:
BROAD SPECTRUM SUN PROTECTIVE TOPICAL FORMULATION
Field of Invention
The present invention relates to a sunscreen formulation having a broad spectrum photoprotective and antiaging action on skin.
Background
UV radiations are a part of the electromagnetic (light) spectrum that reaches the earth from the sun. These wavelengths are classified as UVA (320-400 nm), UVB (290-320nm) and UVC (200-290nm). UVC is absorbed by the ozone layer and does not reach the earth. However, UVA and UVB penetrate the atmosphere and play an important role in conditions such as premature skin aging (photoaging), eye damage (including cataracts) and skin cancers (Moyal 2012). They also suppress the immune system of the body (Schwarz 2005).
Sunscreens are topical products capable of protecting the skin from these damages. However, most of the sunscreens in the market contain chemical filters. These products generally include a combination of two to six of the following active ingredients: oxybenzone, avobenzone, octisalate, octocrylene, homosalate and octinoxate.
A large number of in vitro and in vivo studies have reported endocrine disrupting effects of sunprotectants which are chemical in nature (Krause et al. 2012). According to the Centre for Disease Control and Prevention (CDC), benzophenone-3 (BZ3) or oxybenzone is very hazardous. It has been found in the list of Registry of Toxic Effects of Chemical Substances (RTECS) because the chemical is absorbable through the skin and causes endocrine disruption. It is also reported that some chemical UV filters cause skin reactions like contact urticaria, contact dermatitis and other photo allergic reactions (Pustišek, Lipozencic, and Ljubojevic 2005).
Thus, it raises important questions about unintended human health consequences arising from frequent sunscreen application. Therefore, there is a high demand to explore and develop safer and natural alternatives for skin photoprotection.
The response of the dermal cells and organelles to the detrimental effects of sunrays varies from one type of ethnic skin to another. As per Fitzpatrick scale, the skin of the Indians is classified under type IV and V, based on the presence of melanin, genetic predisposition and its sensitivity to UV rays. Since the percentage of Indian females going outdoors to study or work is increasing day by day, skin care has become a necessity to protect them from photoaging. But ironically, the sunscreen products present in the Indian market claim irrelevant SPF values and they are loaded with chemical sunscreen ingredients.
SPF (Sun Protection Factor) is an indicative value of protection from only UVB rays. That means, the product is not meant to protect the consumer from the effect of UVA rays. Additionally, if the product is blocking UVB rays completely, it will indirectly block the production of Vitamin D in the body which is dependent on the absorption of UVB rays (Holick 2014).
Besides this, sunscreen which gives protection from UVA alongwith UVB is more desirable because the person is protected from probability of having skin aberrations like photoaging, photoimmunosupression and photocarcinogenesis. Ideally speaking, a broad spectrum sunscreen product with moderate SPF is optimal for Indian skin. It would be highly advantageous if the same sunscreen also has antioxidant and anti-inflammatory properties which would extrapolate the role of a sunscreen agent to an antiaging product. Due to frequent exposure to sunlight, matrix metalloproteinases enzymes in dermal layer of the skin gets activated and cause degradation of collagen, elastin and hyaluronic acid which leads to sagging and wrinkling of skin (Imokawa, Nakajima, and Ishida 2015). Therefore, it is desirable to have a natural ingredient in a delivery vehicle that could penetrate the skin up to the dermal layer and inhibit the said enzymes. A suitable delivery system is also required to overcome the other disadvantages such as photo instability, toxicity, skin penetration, high dose and poor efficiency.
For the above reasons, it would be highly desirable to formulate a topical sunscreen whose ingredients are from natural source, non-irritating, non-allergenic, safe, photostable, non-toxic, suitably textured, evenly-blending with the skin tone and able to provide broad spectrum UV protection along with antiaging and moisturizing effects on skin.
Object of the Invention:
The aim of the invention is to design and develop well textured, moisture and pH balanced sunprotective topical formulation containing natural photoprotective minerals and antiaging bioorganic molecule(s) to meet the unique requirement of the Indian skin type.
Another object of the invention is to formulate a topical sunscreen whose ingredients are natural, non-irritating, non-allergenic, safe, non-toxic and photostable. The formulation must blend evenly with the skin colour and capable of providing broad spectrum UV protection along with antiaging and moisturizing effect on skin.
Another object of this invention is to design and develop well textured, moisture and pH balanced sunprotective topical formulation containing natural photoprotective minerals and antiaging bioorganic molecule(s) particularly suitable for the Indian skin type.
Another object of this invention is to formulate a sunscreen product with all-natural ingredients for Type IV and V skin type with good antioxidant and photoprotective properties.
Another object of this invention is to develop a suitable delivery system to overcome the disadvantages such as photo instability, toxicity, skin penetration, high dose, less efficacy and immediate release.
Another object of this invention is to develop a novel broad spectrum sunscreen formulation containing natural bioorganic molecule(s) having antioxidant, anti inflammatory, anti elastase, anti collagenase and anti hylauronidase activity which would expand the role of a sunscreen agent to an antiaging product also.
Another object of this invention is to develop a non-whitening broad spectrum sunscreen with skin coloured ingredients.
Another object of this invention is to develop a novel broad spectrum sunscreen formulation having spherical solid lipid nanoparticles so that permeation can be enhanced into the dermal layers of the skin providing antiaging benefits.
Another object of the invention is to develop antisolar product containing dyed zinc oxide particle for better skin aesthetics.
Another object of this invention is to provide synergistic combination of nanolipidic carriers of safranal with other sunscreen agents.
Another object is to provide broad spectrum UV protection (UV-A and UV-B both).
Another object of this invention is inclusion of all natural agents into the topical formulation which contributes to antiaging and/or antioxidant and/or skin nourishing effects.
Statement of invention:
The present invention discloses a novel broad spectrum sunscreen composition having a photoprotective and antiaging action on skin comprising safranal entrapped spherical solid lipid nanoparticles, micron-sized coloured zinc oxide, colored pearl powder (using Pterocarpus santalinus extract used as natural colorant), gum acacia, aloe vera gel, shea butter, egg oil, other emollient, antioxidant, emulsifier, chelating agent, preservative, skin nourishing and skin protective agents. Said sunscreen formulation possesses a mild SPF, particularly suited for Indian skin type and does not give the white clown like effect, as noticed in many sunscreen products.
Summary of Invention
In a first aspect, the present invention is directed to a composition including but not limited to a broad spectrum topical sunscreen formulation containing spherical solid lipid nanoparticles of Safranal, a physical sunscreen agent, a natural inorganic reflectant, a natural colorant, a natural dispersing agent, a natural anti-inflammatory compound, a natural emollient, skin nourishing agent, a skin protective oil, skin protective ingredient, emulsifier, chelating agent, preservatives.
In another aspect of the invention the composition of the present invention comprises Safranal entrapped spherical solid lipid nanoparticles, coloured zinc oxide, coloured pearl powder, dye santalin, gum acacia, aloe vera gel, shea butter, egg oil, other emollient, antioxidant, emulsifier, chelating agent, preservative, skin nourishing and skin protective agents.
In another aspect of the invention the composition comprises a well textured, moisture and pH balanced sunprotective topical formulation containing natural photoprotective minerals and antiaging bioorganic molecules typically suited for the Indian skin type.
In another aspect of the invention the compositions comprises a broad spectrum sunscreen product with moderate SPF which is optimal for Indian skin.
In another aspect of the invention the broad spectrum sunscreen compositions provides photoprotection along with antiaging and skin nourishing effects.
In another aspect of the invention the compositions of the present is directed towards broad spectrum sunscreen formulation comprising safranal show antioxidant, anti inflammatory, anti elastase, anti collagenase and anti hylauronidase activity.
In yet another aspect of the invention the composition is directed towards broad spectrum sunscreen formulation where spherical solid lipid nanoparticles were designed so that they can permeate into the dermal layers to provide antiaging benefits.
In yet another aspect of the invention the composition includes pearl powder as an inorganic antioxidant natural compound which provides skin nourishment.
In yet another aspect of the invention the composition uses extract from Pterocarpus santalinus to colour zinc oxide and pearl powder for augmentation of aesthetic value.
In yet another aspect of the invention the composition uses redwood extract to provide colour to the skin similar to natural tanning.
In yet another aspect of the invention the sunscreen formulation comprises a micron-sized inorganic zinc oxide particle which removes its skin whitening effects.
In yet another aspect of the invention the sunscreen formulation eliminates skin whitening effect of physical sunscreening agent; zinc oxide by incorporating coloured zinc oxide particles.
In yet another aspect of the invention the sunscreen formulation contains pearl powder and zinc oxide colored with extract from Pterocarpus santalinus which adds to the antioxidant activity of safranal synergistically.
In yet another aspect of the invention the sunscreen formulation contains established source of phospholipids, triglycerides, cholesterol, lutein and zeaxanthin with antisolar properties.
Detailed description of the invention
The rationale behind this research work was to develop a sunscreen cream which provided sunprotection from both UV-A and UV-B rays, avoiding the use of harmful chemical sunscreen moieties as found in the commercial products. Before starting the project, a survey was carried out on a sample population mainly comprising of working men and women in the NCR region of India, in order to have an idea about the awareness regarding sunscreen products. After analysis of the survey data, it was very evident that the Indian users lacked awareness about sunscreen related terms and find themselves in a quandary before buying a sunscreen product. Since sunscreen market is a very rapidly growing segment nowadays, a scientific and ethical approach for selection of appropriate product is the need of the hour. Considering the mind-set of the present Indian consumers and need of sunscreen as per their skin type, the formulation was optimized and all natural ingredients were added to it. An approach was to utilize indigenous sources of the country. Each and every ingredient was assayed, optimized and added with a specific purpose as given in table 1.
Example 1: SUNCREEN COMPOSITION
This composition includes:
A. Safranal entrapped nanopearls.
B. One physical sunscreen agent.
C. Natural inorganic reflectant, with antiaging properties.
D. Naturally existing colourant with antityrosinase effect.
E. Natural dispersing agent with antiaging properties.
F. Natural anti-inflammatory compound.
G. Natural emollient and skin nourishing agent.
H. Effective amount(s) of one or more skin protective ingredients such as antioxidants, vitamins and fixed oils.
I. Additional components: emulsifier, emollient, chelating agent, preservative, or any combinations thereof.
Solid lipid Nanoparticles of Safranal
Safranal proved to possess great potential against photo damage. But as this herbal compound is volatile and light sensitive in nature, it became essential to entrap it into a suitable delivery system. After extensive search of literature and based on preformulation studies, solid lipid nanoparticles were chosen as the most suitable delivery system for Safranal, because of their innate UV blocking potential.
SLN consists of biodegradable and physiological substances similar to skin constituents and hence they are free from adverse effects. They play an important role in cosmetics due to their submicron-size, physical stability and pearl-like nature (Dingler et al. 1999). It is a lipidic carrier system composed of biodegradable and physiological lipids designed to release the drug at controlled release manner.
• Solid lipid Nanoparticles (SLN) of Safranal; nanopearls.
• Composed of physiological and biodegradable lipids.
• The crystalline structure of the lipid has UV scattering properties which makes SLN itself a UV protectant.
• Able to provide photostability to the sunscreen molecule.
The present research work aims at designing and optimization of SLN delivery system, by QbD approach to achieve maximum entrapment efficiency and least particle size. This was done using Central Composite design (CCD). Screening of various excipients was done prior to experimental model. Amount of lipid(X1), Surfactant concentration (X2) and Stirring time (X3) were chosen as independent variables. Entrapment efficiency (Y1), Particle size (Y2) were the responses observed. Design-Expert®7.0 was used to generate the combinations of these factors at different levels using Central Composite Design (on face). A total number of 20 experiments were performed as per the design software. The best model among the linear, two-factor interaction model and quadratic model due to the analysis of variance (ANOVA) F-value was chosen.
Egg oil was extracted and SPF of egg oil was determined and was found to be 5.1
EVALUATION OF THE FORMULATION
The Sunscreen cream is a topical skin preparation which should meet some defined specifications like pH, occlusiveness, total fatty acid substance and other properties including spreadibility and texture for consumer acceptance. These official tests were carried out in accordance with B.I.S. (Bureau of Indian Standard). Other tests were carried out as per defined procedures and results of all the tests were found to be well in the limits.
Physical Evaluation
Physical parameters of the SLN containing sunscreen cream like colour, odor and pH was observed. The parameters were of satisfactory as per consumer acceptance. The cream had skin tone colour despite of zinc oxide added to it because zinc oxide and pearl powder had been previously colored using natural dye; Pterocarpus santalinus (Lal Chandan)
• pH -5.6 Limit (4-9)
• Occlusion Factor-45.6+1.45 S.D Petroleum Jelly (32.13) Limit (0-100)
• Total Fatty acid substance-4.84+0.9% (Limit upto 5%)
Total fatty acid substance comes out to be 4.84+0.9% which is lower than the limit of B.I.S (5%) (Committee 2004).
• Determination of Residue 39.2%. (Minimum 10%)
• Rheology studies -Shear thinning behaviour
Sample was found to be sensitive to the given deformations and found to show shear thinning behavior across the given experimental conditions due to the orientation of the microstructure in the direction of given deformation.
• Texture Analysis
Texture profile analysis was done using Texture Pro CT V1.3 Build 15 to analyze the textural properties of creams. Uniformity of curve shows smooth texture and absence of grittiness. The value of spreadibility was 230 g.sec and that of hardness was 9754± 143 g. Smooth texture is an important attribute of a topically applied formulation. This sunscreen cream exhibited smooth texture and acceptable value of spreadibility so that it can easily spread on the epidermis when applied.
SPF and Broad spectrum determination
The sun protection factor was determined through instrument UV-2000S Ultraviolet Transmittance Analyzer, Lab sphere. The instrument measured SPF based on the sample transmittance which are in good correlation with current research hypothesis. This all natural sunscreen cream is made particularly for Indian skin.
Indian skin generally falls into category of skin type IV and V (Pai, Vinod, and Krishna 2002). As value of SPF is indirectly related to MED, Indian skin which has a higher value of MED requires lesser SPF (Mehta et al. 2004). However, there are no guidelines in the BIS for sunscreen standards. Therefore, sunscreen cream in the current research work has been made according to the requirements of USFDA. Over-the-Counter (OTC) Panel of the USFDA recommended consumers sunscreen products suitable as per their skin type (J. B. W. and R. J. M. Moore 1982). As per these guidelines, SPF requirement of Skin type IV and V is 2-3. However, these guidelines have been revised from time to time. As per the current guidelines by USFDA, the value of SPF of the sunscreen cream in this research study is mild. The value of PA++ indicated moderate protection level as per Japanese cosmetic Industry (Latha et al. 2013). A value of *** indicated high UVA protection (USFDA 2007). The product has superior (UVA/UVB of 0.756) absorption at wavelengths throughout the UVA/UVB radiation spectrum (290-400nm).
Stability Studies of Sunscreen formulation
Accelerated stability studies have been carried out as per ICH Guidelines for 6 months (Zone III and Zone IV) at temperature of 40+/- 2oc and Humidity level of 75+/- 5%. No change was observed in the following parameters at the end of 3 months and 6 months. No microbial growth was observed. Consistency of the product changes insignificantly. Product became little more viscous. Stability stuides show that product is stable throughout 6 months.The formulation containing Safranal loaded SLNs is an aestheically acceptable sunscreen cream with broad spectrum UV protection.Sunscreen cream when applied to skin was devoid of any skin whitening effect. Pterocarpus santalinus extract could conceal the skin whitening effect of micron sized zinc oxide powder and pearl powder.
Example-2 Preparation of formulations complete experimental details
Formulation of SLN containing sunscreen
Prior to formulation, various excipients were optimized for their concentration to be used.
Step I- Making of Aqueous Phase (A)
Taken 20 ml of distilled water in a clean and disinfected beaker and heated the water to 40-450c. Sprinkled 3-7g of gum acacia into it and stirred with high speed on a magnetic stirrer. Then added 0.2g of EDTA, 3-12 g of aloe vera gel, and 2-5 g glycerin to it. Stirred all the things well.
Step-II Making of Oily phase (B)
Taken another dry beaker and placed it on hot plate at round 40-500c. Added 2-4 g of cetyl alcohol and melted it. Added 3-6g of anhydrous lanolin to this. Added 6 gm of shea butter and stirred it well. At this stage, added 0-5g almond oil, 1-4g of hen egg oil and given quantity of Span-80 and Tween-80. Then added 5-15 coloured zinc oxide and 1g coloured pearl powder and stirred again with overhead stirrer. 1g vitamin E was added and stirred till uniform dispersion was obtained.
Step III mixing of phases
Both the aqueous and oily phases were mixed thoroughly at same temperature and then added 0.5g of ascorbic acid. When temperature cooed to around 350c, added preservatives namely 0.1g propyl paraben and 0.1g methyl paraben and 30µl of safranal as perfume.
Example 3
Formulation of drug delivery system
Screening of various excipients was done prior to experimental model. Amount of lipid (A), Surfactant concentration (B) and Stirring time (C) were chosen as independent variables. Drug loading (Y1), Particle size (Y2) were the responses observed. Design- Expert ® 7.0 was used to generate the combinations of these factors at different levels using Central Composite Design (on face). The effect of various independent variables on the responses was calculated by ANOVA. The results were further analyzed by ANOVA and the significance of (p value) different variables for Y1 and Y2 was calculated which proved that the model is highly statistically significant (P < 0.05).The Model F-value also implies that the model is significant.
Particle Size analysis of SLN:
Dynamic light scattering (ZetaSizer Nano-ZS90; Malvern Instruments, UK) method was used to assess the mean particle size, zeta potential and polydispersity index of SLN formulations. All measurements were performed in triplicate at a temperature of 25 ± 2 0C.
DSC of optimized batch of SLN :
DSC studies were done using TA InstrumentsQ10 (Waters LLC). An empty aluminium pan was used as reference. Plain drug, lipid, physical mixture of the two and SLN were scanned from 10o C to 200oC at a rate of 50 C/min under nitrogen atmosphere (20 ml/min). A sharp endothermic peak of the pure safranal was observed indicating crystalline nature of the drug. Physical mixture of drug and lipid showed distinct peaks for drug and lipid at their respective melting points. DSC thermogram of drug loaded SLN does not show peak of drug. This is attributed to the molecular inclusion of safranal in the lipid matrix of stearic acid. This also suggested that safranal exists in amorphous state in SLNs.
Entrapment Efficiencyof SLN:
The entrapment efficiency of the SLN was determined indirectly by calculating the unentrapped drug.10-15ml of dispersion was centrifuged at rpm at 15oC for 1hr. The amount of the drug in supernatant was determined. The supernatant was decanted and filtered through 0.45µ nylon filter. The amount of the safranal was determined at 310nm using UV Spectrophotometer.
Entrapment Efficiency
FTIR Studies of optimized batch of SLN
The IR analysis of drug, lipid and SLN was done using Bruker FTIR spectrometer. About 1–2 mg of sample was mixed with dry potassium bromide and the samples were examined at transmission mode over wave number range of 4,000 to 400 cm-1. FTIR peaks of Safranal, stearic acid (lipid) and optimized batch of SLN have been compared. The characteristic peaks confirmed the presence of both the compounds, safranal and lipid suggesting that there is no significant interaction between the drug and the lipid.
TEM Analysis of optimized batch of SLN
The morphology of optimized safranal loaded SLN was investigated using TECNAI G 2 HR-TEM 200KV, G 20 (FEI, HOLAND). The samples were diluted in ultrapure Milli-Q® water, and approximately 5 µL of each sample was deposited on carbon-coated copper grids (200 mesh). After drying at room temperature, the grids were observed under TEM. All of the SLN formulations exhibited nanometric size and spherical shape when observed under TEM.
Sphericity Index of SLN:
The sphericity Index of the Solid lipid nanoparticles was done using “Image J” software. The image was selected and analysed using “set scale” command of the software. The analysed image dimension was measured.
Dyeing zinc oxide and pearl powder:
Taken 8-12ml of Pterocarpus solution and added to zinc oxide powder in a petridish. Allowed solvent to evaporate at room temperature. The coloured zinc oxide was obtained and passed through sieve before use. Similarly, pearl powder was coloured.
Taken hot distilled water in a clean and disinfected beaker. Sprinkled gum acacia into it and stirred with high speed. Added EDTA, aloe vera gel, glycerin. In another beaker, added shea butter, cetyl alcohol, anhydrous lanolin, Span-80, Tween -80, almond oil, hen egg oil and Vitamin E. Added sieved and coloured zinc oxide and pearl powder to the melted mixture and stirred with high speed until the solids dispersed. Removed the two phases from heat and added phase A to phase B. Mix thoroughly and cooled to 400c. Added SLN dispersion at this stage. Finally added Phase C and stirred again.
Example: 4 Evaluation of Sunscreen cream
Determination of spreadability/ Texture Analysis.
Texture profile analysis is a popular double compression test for determining the textural properties of creams. Texture analysis was performed using Texture Pro CT V1.3 Build 15. Uniformity of curve shows smooth texture and absence of grittiness. The value of spreadibility was 230g.sec and that of hardness was 9754± 143 g. Research has shown that the firmness and energy required to deform a sample to a defined depth grades samples is in order of spreadibility. Lower peak load (firmness) value coupled with a lower hardness work done value in this sunscreen sample indicates a good spreadibility. Smooth texture is an important attribute of a topically applied formulation. This sunscreen cream exhibited smooth texture and acceptable value of spreadibility so that it can easily spread on the epidermis when applied.
2. Determination of Occlusive factor
The occlusion factor of sunscreen cream determined was 45.6+1.45 S.D. (n=3) as compared to petroleum jelly which is used as a standard and had a value of 32.13.The B.I.S limit of occlusion factor is 0-100 (Committee 2004). High occlusion factor provided an impermeable layer on the skin which prevented loss of moisture from the epidermal layer.
3. Rheology
Sample was found to be sensitive to the given deformations and found to show shear thinning behaviour across the given experimental conditions due to the orientation of the microstructure in the direction of given deformation. In the given experimental conditions, all the sample was found to be dominated by their solid character i.e. G’ (loss modulus) > G’’ (elastic modulus) and known as viscoelastic solids due to the presence of prominent 3D network. The LVER in the sample was observed till 0.14 % strain. Immediately after the LVER sample yields and give a crossover where G’=G’’ followed by a region where viscous character starts overpowering i.e. G’’>G’.
SPF and Broad spectrum determination
The sun protection factor was determined through instrument UV-2000S Ultraviolet Transmittance Analyzer, Lab sphere. The instrument measures SPF which is based on the sample transmittance. Transpore tape in the size of (4*4 cm2) was placed over the quartz input optics of the transmittance analyzer and the intensity of radiation transmitted through the tape was de- termined in 5-mn intervals from 290 to 400 nm (Diffey and Robson 1989). The tape sample was then stuck onto a piece of stiff photographic film over a circular open aperture 2 cm in diameter and analysis was done.
The value of PA++ indicates moderate protection level as per Japanese cosmetic Industry. The value of SPF is 9.22 which mean this that the developed sunscreen provides Sun protection from UV rays around 90%. (Between 87.5-93.3% (Schalka and Reis 2011) sufficient for mild sunprotection and typically meant for Indian skin. As value of SPF is indirectly related to MED, Indian skin which has a higher value of MED requires lesser SPF.
5. Accelerated stability studies
The optimized formulation was subjected to accelerated stability studies as per ICH guidelines (WHO 2018). The sample was subjected to 40 + 2°C and 75 + 5% relative humidity conditions for 6 months. The formulations were evaluated at initial, 30, 60 and 90 days for the parameters such as colour, texture, appearance, pH, spreadibility and microbial growth. No change was observed in the following parameters at the end of 3 months and 6 months. No microbial growth was observed. Consistency of the product changes insignificantly. Product became little more viscous.
Stability stuides show that product is stable throughout 6 months.The formulation containing safranal loaded SLNs is an aestheically acceptable sunscreen cream with broad spectrum UV protection.
SUPPORTING PROOF FOR ITS ANTI-INFLAMMATORY AND SKIN NOURISHING EFFECT.
Anti-inflammatory effect
UV radiation when falls on the skin produces reactive oxygen species (ROS) which causes inflammation. Phytochemicals protect against UV radiation-inflicted damage by their ability to act as direct and indirect antioxidants, as well as anti-inflammatory action (Dinkova-Kostova 2008). Safranal also possesses anti-inflammatory activity. Anti-inflammatory activity is determined by various in-vitro methods.
Anti-inflammatory activity of safranal
The reaction mixture consisted of 0.2ml of egg albumin, 2.8ml of phosphate buffer saline (pH 6.4) and 2 ml of varying concentration of sample or control in the range 0-100 µg/ml. These were incubated at 37+20c for 15 minutes and heated upto 700c for 5 minutes. After cooling, their absorbances were measured at 660nm. Aspirin was taken as standard.
% denaturation was calculated as: (Abs Control –Abs Sample) X 100/ Abs control.
The drug had significant value of anti-inflammatory activity in comparison to standard.
Skin nourishing effect
The almond oil used in this sunscreen contains fatty acids along with the unsaponifiable fraction, which usually is mainly composed of squalane, tocopherol and phytosterols which provides nutrition to the skin. To confirm the presence of various components of almond oil, GCMS of almond oil was done.
GCMS of Almond Oil
Prior to GCMS analysis, transesterification of oil was done. The GCMS spectrum of the oil shows the fatty acid profile of the almond oil.
More than 90% of the fatty acids in almond oil are unsaturated (oleic and linoleic).The oleic acid is present in ester form up to 49% followed by linoleic acid in the concentration (24%). The high oleic acid content makes almond oil more resistant to oxidation than other oils with higher amounts of polyunsaturated fatty acids. Octadecanoic acid also exists in 4.07% followed by stearic acid, palmitoleic acid and linolenic acid. Linoleic acid plays an important role in prostaglandin synthesis. It has also been proved that the ceramides present in epidermis contain several fatty acids derivatives, 41% of them being linoleic acid. Additionally, oleic acid has been found to improve transportation of PUFAs into skin as it promotes skin penetration through a mechanism including softening of the horny layer. Polyunsaturated fatty acids or physiological lipids, locally applied, may reach lower skin layers thus enhancing skin properties.
Antioxidant activity of almond oil
Almond oil also has antioxidant effects. Antioxidant activity of the oil was performed using DPPH scavenging method. Tocopherol was taken as standard and results were comparable as shown below.
The antioxidant property of almond oil is useful in preventing photoaging. Topical application of almond oil has been reported in preventing the structural damage caused by UV irradiation.
Hen egg oil is a good source of phospholipids, triglycerides, cholesterol. It is also a good carrier for sunprotective agents where carotenoids such as lutein and zeaxanthin are stabilized with emollients. Additionally, the presence of phosvitin, and free aromatic amino acids make it a very good antioxidant. Being protein free, it is a safer product for the patients allergic to egg products.
Supporting experiment/reference to prove its skin moisturising properties.
Skin when exposed to UV radiations become dry and loses moisture. Dry skin has been demonstrated to contain reduced linoleic acid amount, which suggests that this fatty acid is required for a correct barrier function. Almond oil contains 24% oleic acid as seen in its GCMS spectra.
Skin tends to lose its moisture when exposed to Sun for longer periods. The moisture content of the skin can be retained by the substances which prevent it from losing by forming an impermeable layer on the skin. This can be done by checking occlusiveness of a formulation. The sunscreen cream has good skin occlusive properties as proved by its occlusion factor.
Shea butter
Due to its semi-solid characteristics and buttery consistency, Shea butter is great emollient and moisturizer for the skin. In a study by Poelman and co workers a cream containing 5% Shea Butter versus a placebo cream were applied to the forearms of 10 volunteers. Short-term moisturization was observed; it peaked after 1 hour and persisted for 8 hours. For all subjects, a daily application maintained a very good moisturization of the superficial layers of the skin. Shea butter has also been shown to be superior to mineral oil at preventing transepidermal water loss (TEWL). In a test where participants’ arms were washed in ethanol, it was found that Shea butter was able to help the skin totally recover from TEWL within two hours. One study showed that it worked as an emollient for eczema. Using a scale from zero to five — zero denoting clear and five denoting very severe disease — Shea butter took a three down to a one, while Vaseline only took a three down to a two.
Vitamin E and C
1. Together vitamin E and vitamin C both have photoprotective activity.
2. Both have antioxidant properties. (Proved and used as standards in various DPPH assay procedures)
Synergistic effect due to combination of ZnO with Lal chandan
As a result of combination of lal chandan and zinc oxide , two synergistic effects were observed
1. The addition of lal chandan extract to ZnO changes the colour of the sunscreen formulation from white to skin which is aesthetically desirable.
2. The SPF value of zinc oxide cream increased from 15.9 to 17.1 by this combination. This was proved in SPF assay of zinc cream and coloured zinc cream where the content of zinc oxide was 32%.
Zinc oxide powder when coloured using 1mg/ml extract of Pterocarpus santalinus gave a pink tinge to the powder similar to skin tone. The coloured zinc oxide was further used to create a new formulation which matches the colour of the skin. This improves the aesthetic value of the cream. Besides, Synergism was observed SPF of zinc cream and coloured zinc cream determined by transmittance method was compared.
Formulation of zinc cream and its SPF determination
Zinc cream, and coloured zinc cream were formulated as per the following formula. Both the creams were analyzed for their SPF values.
SPF determination of creams
The SPF value of zinc oxide cream determined by absorbance method comes to be 15.9 whereas that of coloured zinc oxide cream comes to be 17.1. The synergy was shown in efficacy of zinc cream and in improvement of its aesthetic properties. Besides, the presence of lal chandan extract in the formulation added to the antioxidant power of the whole cream.
Synergistic effect due to combination of safranal with colored ZnO
Safranal is a biomolecule with radical scavenging and antiphotoaging. Zinc oxide is a physical sunscreen agent which reflects UV-A rays. In order to provide complete protection, a sunscreen should have a broad spectrum of protection. This could be achieved by combination of a UV reflector and UV absorber. Colored zinc oxide is extract of lal chandan adsorbed on zinc oxide. This zinc oxide does not have skin whitening effect.
Example: 5
SAFRANAL IN-SILICO STUDIES
This biomolecule docked with antiaging enzymes namely elastase, collagenase and hyaluronidase using Schrodinger v11.5 exhibited its antiphotoaging activity by interacting with amino acid residues of receptor through intermolecular hydrogen bond interactions. Hydrogen bond interactions of safranal with collagenase (Ala182 and Leu181) and hyaluronidase (Asn290 and Asn580) were observed. ADME parameters of Safranal were determined using online software SwissADME in which it displayed drug likeliness as per Lipinski’s rule of five and showed considerable values of tissue permeability and bioavailability.The ligand interaction diagram represented that aldehyde group of safranal formed H-bond with the Ala182 and Leu181 amino acid residues of the collagenase receptor.
Example: 4
Augmentation of Aesthetic and Antisolar attributes of a sunscreen cream by methanolic extract of Raktchandan (Pterocarpus santalinus)
Red sandalwood or Raktchndan or Pterocarpus santalinus is an indigenous drug mentioned in “Ayurveda”. Heartwood of the tree is used in Ayurveda, Siddha, Unani,Tibetian and folk medicines.(Forests n.d.) This heartwood is in high demand in domestic and international market and is predisposed to illegal harvest, trade and habitat destruction. It is used as astringent. A paste of the powder is used as a cooling application in hedache. In pharmacy, chiefly used as a colouring agent. Phytochemical investigation revealed the presence of various bioactives such as alkaloids, phenols, saponins, sterols, tannins, glycosides, flavonoids and triterpenoides. The Red Sandalwood contains natural dye called santalin, which is used as a colouring agent in pharmaceutical preparations and in food industry. The unique red colour of the wood has been added to dermatological preparation to give natural colour of tanning. This invention has been patented in cosmetic composition comprising at least one Santalin or Santarubin.(Patrick Darmenton, Delphine Allard 2001) .
Recent phytochemistry and pharmacological experiments have indicated that santalin, which is the major active component of Pterocarpus santalinus has anti-tyrosinase activity. Santalin inhibited melanin formation process by down regulation of a critical transcription factor; microphthalmia-associated transcription factor (MITF), tyrosinase, tyrosinase related protein 1 (TRP-1) and tyrosinase related protein 2(TRP-2). This study indicated the use of Pterocarpus santalinus as depigmentation agent. In the year 2007, a patent has been granted to a topical composition containing extract of Pterocarpus santalinus and showing skin whitening effect. This action is attributed to inhibitory effect on the tyrosinase activity and melanin biosynthesis through inhibiting the enzymes namely MMP-1 (collagenase) and MMP-2 (gelatinase).
In a recent study, ethanolic extract of Pterocarpus santalinus exhibited potential effect against UV-B induced photoaging. The wood extract inhibited the upregulation of matrix metalloproteinases (MMP’s) and interleukin(IL)-6 which exposure is caused by UVB irradiation, and suppresses UVB-induced phosphorylation of extracellular signal-regulated kinase.
Identification,authentication and extraction of lal Chandan (Pterocarpus santalinus)
The heartwood of the drug was obtained from local supplier and authenticated by NISCAIR, New Delhi (NISCAIR/RHMD/Consult/2018/3174-23) The heartwood of the Pterocarpus santalinus was dried at room temperature was crushed into powder by electric blender .The wood powder was extracted by three methods.
Soxhelation For the extraction of phytochemicals, wood powder was subjected to extraction in soxhlet apparatus at 55-85°C for 12-24 h using 1:1 acetone and and absolute ethanol. The extracts obtained were then dried using freeze dryer, and further freeze dried to get the solid extracts which were preserved in refrigerator for further analysis.
Maceration 5gm of powder of Pterocarpus santalinus wood was macerated with 100 ml of methanol for 24 hours, shaken frequently and allowed to stand for 24 hours. Thereafter, filtered, evaporated the filtrate and residue was freeze dried and yield was calculated.
Microwave Extraction Natural dye from the plant was extracted using microwave assisted extraction. (Hemanthraj, Desai, and Bisht 2014) 1g of powder of stem wood of the Pterocarpus santalinus was sieved and was mixed with 250 ml volume of methanol and water (80:20) solvent to give a solid to liquid ratio ranging from 1:250 (g/ml) and allowed for gentle mixing in container and then placed in micro treatment chamber (oven) with extraction time for 20 minutes at ranging temperature of 45°C. The microwave power of 800W was used for the extraction. The extract so obtained was filtered and filtrate was evaporated in rotary evaporator at 60°C of water bath temperature. The filtrate was further freeze dried.
Antioxidant activity determination
Antioxidant activity of the extracts was performed using DPPH scavenging method (Molyneux 2004). 1 ml of 0.1 mM solution of DPPH in methanol was added to 3 ml of the solution of all the extracts in methanol at different concentrations (5, 20, 40, 60, 80, and 100 µg/ml). The mixtures were shaken and were allowed to stand in dark room for half an hour. Then, the absorbance was measured at 517 nm. Finally, scavenging capability of DPPH radical was determined by the formula:
Scavenging effect = [(A0-A1)/A0]×100
Where, A0 is the absorbance of the control,
A1 is the absorbance in the presence of all of the extract samples,
A graph was plotted between scavenging effect and concentration, and a regression equation was obtained to calculate inhibitory concentration 50 (IC50).
FTIR Assay of zinc oxide and coloured zinc oxide
The FTIR of zinc oxide and coloured zinc oxide was done using Bruker FTIR research spectrometer 12060280. About 1–2 mg of sample was mixed with dry potassium bromide and the samples were examined at transmission mode over wavenumber range of 4,000 to 400 cm-1.The data handling was done using OPUS 7.2.139.1294.spectroscopic software.
Transmission analysis of zinc oxide and coloured zinc oxide
Transmission analysis of zinc oxide, coloured zinc oxide and calamine was carried out using UV-3600 Plus. The data obtained was plotted and compared.
SPF determination of Lal Chandan
SPF of Pterocarpus santalinus was determined by absorbance method given by Mansur et al (Kaur and Saraf 2010). 0.1% hydroalcohlic solution of Pterocarpus santalinus was made and its absorbance was noted from 290-320nm. The value of absorbance was higher than 1 and hence solution was diluted ten times. Absorbance of 0.01% hydroalcohlic solution was carried out in triplicate from 290-320nm and the obtained absorbance values were multiplied with the respective EE (?) values. Then, their summation was taken and multiplied with the correction factor.
Coating of zinc oxide with methanolic extract of Lal Chandan (MELC)
Taken 100mg of Pterocarpus santalinus extract in a 100 ml volumetric flask. Added methanol and dissolved. Taken 9ml of this 1mg/ml solution and added to 5gms of zinc oxide powder in a petridish. Allowed solvent to evaporate at room temperature. The coloured zinc oxide was obtained and passed through sieve before use.
Formulation of cream and its SPF
Zinc cream, coloured zinc cream and calamine cream was formulated as per the following formula. Both the creams were analyzed for their SPF values. In the case of latter, coloured zinc oxide was used instead of white zinc oxide.
The SPF value of all the three creams was determined by transmission method. Transpore tape was used as a substrate and each sample was applied in concentration of 2mg/cm2. Transmission data was noted in the range from 290-400nm using UV 3600 Plus. In –vitro SPF was calculated as per formula given Diffey and Robson.(Diffey and Robson 1989)
SPF= 290 -400? E*e/ 290 -400? E*e/MPF
E is the spectral irradiance of terrestrial sunlight under defined conditions and e is the relative effectiveness of UVR at wavelength nm in producing delayed erythema in human skin equivalent to "erythema action spectrum". MPF is monochromatic protection factor. It is calculated as ratio of transmission of blank to the transmission of sample.
Calculation of yield
The extraction yield of Pterocarpus santalinus varied with the technique used. The yield of extract made by soxhelation, maceration and microwave extraction was 19.68%, 16.56% and 0.077% respectively. The highest yield was given by soxhelation process.
Antioxidant value determination
The soxhlet extract exhibited the maximum antioxidant activity.
FTIR Results
The FTIR peaks of zinc oxide and coloured zinc oxide reveals that there is no difference in the structure of powder.
Transmission spectra
The graph shows transmittance spectra of zinc oxide, calamine and coloured zinc oxide.
SPF Determinationof Pterocarpus extract
The SPF value of 0.01% hydroalcohlic solution comes out to be 3.3.
Comparison of aesthetic attributes between dyed and non-dyed zinc oxide
Zinc oxide powder when coloured using 1mg/ml extract of Pterocarpus santalinus gave a pink tinge to the powder as that of calamine. The coloured zinc oxide was further used to formulate zinc cream which is devoid of forming a white layer on the skin.
SPF determination of zinc creams
The SPF value of zinc oxide cream comes to be 15.9 whereas that of coloured zinc oxide cream comes to be 17.1 and that of calamine cream come out to be 8.3. This higher SPF is due to UV protectant nature of lal chandan.
Modern personal care products often contain nanosized particles. Sunscreens are also composed of nano sized particles of inorganic sunscreens such as zinc oxide and titanium dioxide. The purpose of nanosizing was to remove the cloud like skin whitening effect of these dispersive powders. Nanoparticles in sunscreen offer advantages like improved properties in context of their application, aesthetics, performance and use. However, these non-biodegradable mineral particles are suspected of entering into systemic circulation. Although a large number of studies states that zinc oxide nanoparticles do not penetrate into the human skin but still some published studies data suggested cell uptake, oxidative cell damage or genotoxicity due to nanoparticles. Sunscreens are often applied topically when going out in the sun. The colour of sunscreen should match the skin. This can be achieved by either incorporating suitable colorant or calamine into it. Calamine is zinc oxide alongwith with a small proportion of ferric oxide. As per Indian Pharmacopeia, it contains not less than 98.0 per cent and not more than 100.5 per cent of ZnO, calculated on the ignited basis. However, the use of natural calamine has been outdated in modern cosmetics. This is due to inconsistency in its colour and has been replaced by synthetically prepared material coloured with a form of ferric oxide.(Allam and Kumar 2011) Besides this, it is also associated with incompatibility issues related to iron oxide.
Zinc oxide is a broad spectrum safe inorganic sunscreen which prevents the skin from deleterious effects of ultraviolet rays by transmitting or scattering UV light. However, it has skin whitening effect when applied onto the topical layer. In this research study, methanolic extract of Pterocarpus santalinus has been used to colour zinc oxide particles to give natural colour similar to calamine. The transmission data of coloured zinc oxide powder was comparable to calamine. In the UV-B range (290-320), this drug offered more transmission and hence higher prevention. It also showed higher transmission values in UV-A range (320-400nm) as well. However, the values at wavelength 390 and 400nm were comparably lesser than zinc oxide or calamine. This is due to the lighter colour which is neither white nor pink but matches the skin tone. The zinc oxide cream formulated with the same offers synergistic combination of a physical sunscreen and a natural UV absorber; Pterocarpus santalinus. It exhibited a higher value of SPF of 17.1 in comparison to 15.9 of zinc oxide cream or 8.3 of calamine cream. This indigenous drug is also a natural potential depigmentation agent. The drug has been patented in dermatopharmaceutical composition due to its skin coloring effect. But no studies have reported its UV-absorbing properties. In this work, this indigenous drug has been explored to provide synergistic combination of physical and chemical sunscreen.
India is bestowed with rich inheritance of medicinal plants of extensive diversity, which have been used by local population since ages for the cure of multiple diseases. Ayurveda; traditional medicine system of India includes one such plant; Raktchandan or Chandan lal or Pterocarpus santalinus, which is supplemented with extensive medicinal characteristics. Pterocarpus santalinus Linn. or Red sanders is a deciduous tree which belongs to the leguminosae family. The therapeutic properties and various potential applications of heartwood of Pterocarpus santalinus have been well established. The bioactive component of plant santalin is known for its natural colouring properties since ages. Various pharmacological studies of Pterocarpus santalinus have indicated its use in wound healing, hepatoprotective, anti-inflammatory, anti-diabetic, anti-bacterial therapies. Its antiaging property has also been reported. However, no study has stated its antisolar property. The current research study established its photoprotective and antioxidant potential. In this work, an attempt has been made to mask the skin whitening effect of zinc oxide by colouring it with natural dye. The extract of Pteroocarpus santalinus has been used for the same. The heartwood of Pterocarpus santalinus has been extracted using various methods. The extract exhibited photoprotective potential with a significant SPF value. The zinc oxide thus coloured resembled calamine and was free from any chelating effect of ferrous oxide present in calamine. This was formulated into zinc cream which showed higher SPF in comparison to zinc cream. It can be concluded that Pterocarpus santalinus can be recommended as a natural photoprotective agent. This ethnopharmacological agent has a great potential to be used in modern day natural sunscreen products. Its skin colouring, depigmentation properties along with its SPF can be of great significance for dermatological purposes.
Abstract
Composition of a broad spectrum topical sunscreen formulation comprising of safranal entrapped spherical solid lipid nanoparticles, coloured zinc oxide, coloured pearl powder, pterocarpus santalinus extract, gum acacia, aloe vera gel, shea butter, egg oil, other emollient, antioxidant, emulsifier, chelating agent, preservative, skin nourishing and skin protective agents is disclosed. Said composition comprises all natural compounds and is non-irritating, non-allergenic, safe, photostable, non-toxic, suitably textured and devoid of whiteness produced due to the presence of zinc oxide. Disclosed formulation evenly blends with the skin tone and is able to provide broad spectrum UV protection along with antiaging and moisturizing effects on skin. The novel sunprotective formulation exhibits good texture, excellent rheological properties, optimum pH and stability. The product shows broad spectrum sunscreen protection with UV-A ++ and *** Boot Star Rating and SPF value of 9.22, which is appropriate for Indian skin type.
References
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Hemanthraj, K P M, Sudhanva M Desai, and Surendra Singh Bisht. 2014. “Optimization of Extraction Parameters for Natural Dye from Pterocarpus Santalinus by Using Response Surface Methodology.” Int. Journal of Engineering Research and Applications 4(9(3)): 100–108.
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CLAIMS
Claims:
1 A broad spectrum sunscreen composition, comprising safranal entrapped spherical solid lipid nano-particles, a physical sunscreen agent, an inorganic reflectant, natural colourant, natural dispersing agent, natural anti-inflammatory compound, natural emollient and skin nourishing agent, one or more skin protective antioxidants, vitamins and fixed oils, emulsifier, emollient, chelating agent, preservative, or any combinations thereof.

2. The broad spectrum sunscreen composition of claim 1, further comprising antioxidant, anti inflammatory, sunprotection , anti-elastase, anti-collagenase and anti-hylauronidase activity.

3. The broad spectrum sunscreen composition of claim 1, wherein inorganic reflectant with antiaging properties is colored pearl powder.

4. The broad spectrum sunscreen composition of claim 1, wherein the physical sunscreen agent is coloured zinc oxide.

5. The broad spectrum sunscreen composition of claim 4, wherein micron-sized coloured zinc oxide is used to conceal its skin whitening effect.

6. The broad spectrum sunscreen composition of claim 3, 4 and 5 wherein the pearl powder and micron-sized zinc oxide is colored using Pterocarpus santalinus extract.

7. The broad spectrum sunscreen composition of claim 1, having an SPF of 9.22 suited for skin type IV and V on the Fitzpatrick scale.

8. The broad spectrum sunscreen composition of claim 1 comprises at least one UVA sunscreen agent and at least one UVB sunscreen agent.

9. The broad spectrum sunscreen composition of claim 8, wherein zinc oxide is an inorganic sunscreen acting by scattering of UV (A&B) rays and Safranal is a natural organic sunscreen which acts by quenching UV B rays.

10. An ultraviolet radiation protective composition, comprising a mixture of oily phase and aqueous phase components, the oily phase comprising shea butter; 1.5-4.5%, almond oil; 3-6%, hen egg oil; 0.5-2.5%, vitamin E; up to 1%, anhydrous lanolin; 2-6%, cetyl alcohol; 1-5%, zinc oxide; 8-15%, pearl powder; 0.5-2.5%, span-80; 1-3%, tween-80; 2-6%, the aqueous phase comprising water; 25-35%, safranal entrapped SLN dispersion;10-20%,gum acacia; 4-8 %, EDTA;0.001%, aloe vera gel; 5-8%, glycerin; 1.5-4%, lal chandan extract; 0-1%, Vitamin C; 0-1%, Preservatives and Perfume; q.s.