Khin Myo Oo 1, Khin Phyu Phyu 3, Mg Mg 4, Nwe Nwe Than 2
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1 Khin Myo Oo 1, Khin Phyu Phyu 3, Mg Mg 4, Nwe Nwe Than 2 1. Pharmacology Department, University of Pharmacy, Mandalay 2. Pharmacology Department, University of Medicine, Mandalay 3. Department of Medical Research 4. Myanmar Pharmaceutical Factory (Pyin-Oo-Lwin)
2 Introduction Background information Objective Materials and method Results Discussion Conclusion Acknowledgement References
3 Introduction
4 Exposure to solar radiation is recognized to have negative effects on human skin The harmful effects of solar radiation are caused predominantly by the ultraviolet (UV) region of electromagnetic spectrum, UVA, from 320 nm to 400 nm, UVB, from 290 nm to 320 nm UVC, from 200 nm to 290 nm. UVC radiation is filtered out by the ozone layer
5 UVA radiation reaches the deeper layer of epidermis and dermis and provokes the premature ageing of the skin but it is weak and can affect only after prolong exposure The UVB radiation is largely responsible for erythema of sunburn, skin aging and skin cancer UVB causes most of the skin problems related to sun exposure: like aging, wrinkles and cancer
6 Photoaging and photocarcinogenesis are adverse biological effects caused by the photochemical reactions which are generated by excitation of electrons because of ultraviolet radiations Photoprotection could be achieved by the use of sunscreens and antioxidants Natural herbal phenolic compounds also act as antioxidants The function of sunscreen is based on its ability to absorb, reflect or scatter the sun's rays
7 The efficacy of sunscreens is characterized by the sun protection factor (SPF) SPF value = MED (PS) MED (US) Higher SPF sunscreens offer greater protection from sunburn.
8 Natural substances extracted from plants have recently been considered as potential sunscreen resources because of their ultraviolet absorption in the UV region and their antioxidant activity
9 Although synthetic sunscreen formulations are available in market, they have potential adverse effects for some sensitive persons Formulation of herbal photoprotective cream and evaluation of its sun protection activity become important in cosmetic production
10 Punica granatum Linn. (Family: Punicaceae) is commonly known as Pomegranate (Tha-le) From the ancient Ayurveda system of medicine, the pomegranate has extensively been used as a source of traditional remedies for thousands of years
11 Numerous studies on the antioxidant, anticarcinogenic, and anti-inflammatory properties of pomegranate constituents have been published, the majority of research has focused on pulp and juice of fruit New scientists from the institute of hygiene and environmental medicine have reported that the peel offers high yields of phenolics, flavonoids and proanthocyanidins than the pulp
12 Ellagic acid, a polyphenol antioxidant found in large quantities in pomegranates Helps in healing sunburns and reverse sun damage The incredible antioxidant activity in pomegranates may reduce the harmful effects of ultraviolet radiations and inhibit the growth of skin tumors
13 To evaluate sunscreen activty of creams containing fruit peel extract of Punica granatum Linn.
14 Materials and Methods
15 The fresh fruits of ripe Punica granatum Linn. were collected from Pyin Oo Lwin township in June, 2015 and 2016 The fruits were identified and confirmed by the authorized botanist at the Department of Botany, University of Mandalay
16
17 The fruit peels were separated manually and air-dried in the shade for a week Dried fruit peels were powdered in a grinder to get 40 mesh-sized powder
18 The fruit peel powder was extracted and macerated with 50% ethanol solution The extract was macerated for two days, filtered and the marc was then macerated for another two days, filtered and concentrated under vaccum at 50 C Finally the liquid extract was dried at 70 C on the water bath The extract was pooled and kept in desiccators for further analysis and formulations
19 All materials used in formulations and evaluation of formulations were analytical grade
20 Sunscreen creams were developed by using two types of beeswax-borax system of cold cream bases (F-I and F-II) with 5% of fruit peel extract F-I.a, F-II.a (5%) 10% of fruit peel extract F-I.b, F-II.b (10%) 20% of fruit peel extract F-I.c, F-II.c (20%)
21 1g of Cream Dissolved in 100 ml of deionized water, sonication for 5 mins and filtered 10,000 ppm solution of cream 5 ml of 10,000 ppm solution was diluted to 50 ml with deionized water 1000 ppm solution of cream 5 ml of 1000 ppm solution was diluted to 25 ml with deionized water 200 ppm solution
22 The aliquots prepared were scanned between nm and the obtained absorbance values were multiplied with the respective EE (λ) values Then, their summation was taken and multiplied with the correction factor (10) Ref: Mansur et al. (1986)
23 SPF spectrophotometric = CF ΣEE(λ ) I (λ ) Abs((λ ) Where CF=Correction factor (10), EE (λ) = Erythmogenic effect of radiation with wavelength λ, Abs (λ) = Spectrophotometric absorbance values at wavelength λ. The values of EE I are constant, they are determined by Sayre et al. (1979).
24 Wave length (λ nm) EE(λ) x I(λ) (normalized) Ref: Sayre et al. (1979).
25
26 Sunscreen activity was evaluated by in vitro method through recording transmission spectrum of formulations in the range of 290 to 400 nm The analysis was carried out in duplicate and the values were recorded as mean value of two readings
27 Sample was applied on polyvinyl chloride (PVC) sheet strip, spread uniformly with the help of capillary to form a thin film The strip was then placed inside UV-Vis cuvette in such that the formulation touches transparent side of cuvette It was allowed to equilibrate for 15 mins to ensure leveling of the formulation between PVC strip and wall of the cuvette
28 The prepared cuvette was placed inside UV spectrophotometer (Shimadzu) and a transmission spectrum was recorded from 290 to 400 nm, using air as reference The data was appropriately processed to calculate UVA and UVB protection factors using the following formulae
29 (1) Average transmittance spectrum of sunscreen in either region is averaged in order to produces one value, which describes UVA or UVB blocking. 100 T (UVA) or T (UVB) gives % blocking or % protection against UVA or UVB.
30 (2) Average UVA protection factor (PF) Arithmetic mean of monochromatic protection factor (MPF) calculated between nm Where, is transmittance at λ MPF is monochromatic protection factor = 1/T is measured wavelength interval (5 nm) [7,9]
31 Results
32 SPF value Formulation ppm 1000ppm 200ppm F-I. a (5%) F-I. b (10%) F-I. c (20%) F-II. a (5%) F-II. b (10%) F-II. c (20%)
33 Formulation % protection against UVA % protection against UVB Average UVA protection factor F-I. a (5%) F-I. b (10%) F-I. c (20%) F-II. a (5%) F-II. b (10%) F-II. c (20%)
34 Discussion
35 Formulated sunscreen creams containing different proportions of fruit peel extract of Punica granatum Linn. were evaluated for sunscreen activity by in vitro methods of absorption and transmission spectroscopy In vitro methods present some limits; it gives accurate and precise result and avoid the exposure of human subjects to harmful ultraviolet radiation
36 Excipients and other active ingredients can also produce UV absorption bands, thus interfering with those of UVA and UVB sunscreen This effect is reflected in a finished formulation There was no indication of an influence of excipients on the absorbance of product however very low effect was realized at high concentration
37 The proof of sunscreen products efficacy is of high importance for the protection of public health as the UVB fraction of solar radiation is the main contributor to skin sunburn, immunosuppression and skin cancer the absorption spectroscopic method was used to measure protection against UVB by using Mansur equation
38 For all of the formulations, different dilutions were carried out for ppm, 1000 ppm and 200 ppm The values of SPF were varied according to the concentrations of final solutions If the skin type of consumer is I and II, ppm of F-I.a and F-II.a containing 5% fruit peel extract of Punica granatum Linn was applicable Only 1000 ppm of formulations containing 10% and 20% fruit peel extract of Punica granatum Linn. was enough for this skin type
39 Other types of skin from III to VI needed the ideal SPF values of 5 or less Different concentrations of each formulation can be used for different types of skin
40 Skin Type Details Ideal SPF I Always burns easily, 8 or more never tans (Sensitive) II Always burns easily, tans 6-7 minimally (Sensitive) III Burns moderately, tans 4-5 gradually (Light brown, Normal) IV Burns minimally, always 2-3 tans well (Moderate brown, Normal) V Barely burns, tans 2 profusely (Dark brown, Insensitive) VI Never burns, deeply pigmented (Insensitive) Not indicated
41 To assess % protections against UVA and UVB and average UVA protection factor of six formulated sunscreen creams, transmission spectroscopy method was also used According to the findings of this study, both F-I.b and F-I.c gave over 95% protections against UVA and UVB and the highest average UVA protection factor It was also found that the sunscreen creams formulated by using cream base F-I offered larger % protection against UVA and UVB than those using cream base F-II
42 Conclusion
43 The formulated creams have potency to protect against UVA and UVB rays indicating sunscreen activity as well The formulations produced by incorporating different concentration of extracts can be applicable for different types of Skin respectively as per SPF value
44 The authors are grateful to pharmacists from the Department of Pharmaceutical Chemistry, Food and Drug Administration, Mandalay, for their kindly help for determination of SPF values
45 References
46 1.Wilkinson, JB and Moore, RJ, (eds) 1994, Harry's Cosmeticology, 7 th edn, Longman Scientic & Techanical, pp Chanchal, D and Swarnlata, S 2009, 'Herbal Photoprotective Formulations and their Evaluation', The Open Natural Products Journal, vol. 2, pp Svobova, A, Psotova, J, and Walterova, D 2003, Natural phenolics in the prevention of UV induced skin damage: A review, Biomed Pap, vol. 147, pp Mishra, AK, Mishra, A and Chattopadhyay, P 2011, Herbal Cosmeceuticals for Photoprotection from Utraviolet B Radiation: A Review, Tropical Journal of Pharmaceutical Research, vol. 10, no.3, pp
47 5.Bambal, V and Mishra, M 2014, 'Evaluation of in vitro sunscreen activity of herbal cream containing extract of Curcuma longa and Butea monosperma', World journal of Pharmaceutical Research, vol. 3. No.2. pp, More, BH, Sakharwade, SN, Tembhurne, SV and Sakarkar, DM 2013, Evaluation of Sunscreen activity of Cream containing Leaves Extract of Butea monosperma for Topical application, International Journal of Research in Cosmetic Science, vol.3, no.1, pp Patil, S, Fegade, B, Zamindar, U and Bhaskar, VH 2015, 'Determination of sun protection effect of herbal sunscreen cream', World Journal of Pharmacy and Pharmaceutical Sciences,vol.4, no. 8, pp
48 8.Bambal, V, Wyawahare, N, Turaskar, A and Mishra M 2011, Study of sunscreen activity of herbal cream containing flower extract of Nyctanthes arbortrirtis L and Tagetes Erecta L, International Journal of Pharmaceutical Sciences Review and Research, vol. 11 (1), pp Patil, S, Fegade, B, Zamindar, U and Bhaskar, VH 2015, 'Determination and active phytocompounds from formulated sunscreen cream containing Pongamia pinnata leaves and Punica granatum extract by high performance thin layer chromatography', World Journal of Pharmacy and Pharmaceutical Research, vol.4, no. 7, pp Sayre, RM, Agin, PP, Levee, GJ and Mariowe, E 1979, Comparison of in-vivo and in-vitro testing of sunscreens formulas, Photochem. Photobio, vol. 29, pp
49
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