ANTI-AGING. Edelweisseco.

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1 ANTI-AGING Edelweisseco

2 Edelweiss-eco BOTANY Leontopodium alpinum belongs to the Asteraceae family and its common name is edelweiss, snow flower or silver star (due to its star-shape). It is a species that grows in the alpine meadows and high rocky ridges of European mountain ranges. It grows up to 30 cm tall and it has thick and fleshy bracts, covered with a fine fluff. It is white in color, with greenish or yellowish tones. It hides under the appearance of a single flower (between cm in size), but in reality it is a set of tiny flowers that have evolved and grow as a group to survive. The central yellow elements are capitula, where both male and female flowers are grouped; and what appears to be white petals are actually bracts, structures that protect it during its development. A vegetable fiber protects it from frost and ultraviolet radiations. It blossoms between July and September and its leaves can be white, grayish or slightly yellowish. Its apparent fragility hides an incredibly resilient flower, which is able to survive at heights of more than 3,000 meters and withstand the extreme temperatures of alpine mountains. In fact, those are the appropriate conditions for its development, as it only grows naturally at heights over 1,500 meters, on limestone walls and slopes or on rocks, in those grooves that receive a small amount of sunlight. V 02-04/

3 The edelweiss flower originates from the area of the Austrian and Swiss Alps and it is native to the European mountain regions. Its habitat extends from the Carpathians to the Pyrenees, but it can also be found in some very high mountain ranges in Asia, such as the Himalayas. This flower is a symbol of high altitudes and, therefore, it has been avidly collected, having disappeared from many areas. In consequence, the edelweiss is now a protected species. Since the year 2000 it is grown organically at lower elevations, in alpine plantations in Switzerland. Selected and adapted seeds are used to achieve the same quality of the wild flower, and its harvest takes about eight days. Edelweiss Eco extract is obtained from the leaves and flowers of Leontopodium alpinum, from organic farming. CHEMISTRY The edelweiss aerial parts contain leontopodic acid, typical of edelweiss and one of its most abundant components (Schwaiger, S. et al, 2005), and a large amount of chlorogenic acid. In addition, it has flavonoids (luteolin and its derivatives, traces of apigenin and its glucoside derivatives), tannins, phenolic acid, linoleic acid, linolenic acid, terpenes, bisabolene derivatives, phenylpropanoids and sitosterol (Dweck, A.C., 2004; Schwaiger, S. et al, 2006). The following table shows the concentrations found in 2006 by Schwaiger, S. et al in the aerial parts of edelweiss (percentage of dry weight). V 02-04/

4 Table 1. Quantification of different aerial parts of Edelweiss. Concentrations below 0.05% were recorded as traces and compounds that could not be detected were recorded as n.d. (Schwaiger, S. et al, in 2006). TRADITIONAL USES The binomial Leontopodium alpinum comes from Greek and it means "lion foot of the Alps". In German, the term Edelweiss means "noble white" or "pure white" and it also means "White Nobility". Edelweiss is considered as the symbol of nature protection, purity and immortality. It represents honor, the world of dreams and eternal love, which will never dry out. Its image is seen as the perfect reflection of a strange and silent beauty. In the language of flowers, edelweiss means "write to me". It is also an icon of courage and bravery. Its beauty and strength have fueled a living legend that says that men who needed to show their love had to venture into the high peaks to find an edelweiss flower and take it to their beloved. It is also said that it is able to escape from the men who chase it, rising increasingly higher on the mountains, and that it took its color from the moon. Nowadays it is the symbol of Switzerland and also Austria, and it is said it was the favorite flower of the famous Empress Sissi. Apart from these two countries, today edelweiss is cultivated in many more countries V 02-04/

5 and its risk of extinction has decreased in comparison to previous decades (1960), in which the species became very popular and people were after it with greater interest due to, among other reasons, the film The sound of music, in particular due to the song entitled Edelweiss. Because of its great beauty, the edelweiss flower is used as an image for different decorative motifs. It is the image of many badges and medals awarded to outstanding climbers, and curiously it also appears on the Austrian euro-cent coins. In popular medicine, this species is recommended in cases of stomach disorders, dysentery, poor circulation, pain and as a relaxant. Edelweiss is primarily used to produce cosmetics, because it has multiple benefits for the skin. It contributes to slow down skin aging, it has antioxidant, anti-inflammatory, relaxing and purifying properties, it helps protect skin from solar radiation (it absorbs UV rays), it stimulates cell regeneration, it reduces aging signs and it provides moisturization, elasticity and freshness. COSMETIC PROPERTIES Antioxidant activity The antioxidant activity of edelweiss and its components has been confirmed in several studies. Leontopodic acid contained in the aerial parts of edelweiss has in vitro protective effects against induced oxidative reactions. The results from the study by Costa, S. et al (2009) showed that this compound protected cells against the consequences of exposure to toxic agents, with an antioxidant effect. Previously, Schwaiger, S. et al (2005) had isolated this acid from the aerial parts, being one of their main components. It was found that it had important antioxidant effects in different in vitro methods (BR and TEAC). It was also demonstrated in vitro (3D model) that it protected cellular DNA against oxidative damage. Other studies also confirmed the antioxidant effectiveness of chlorogenic acid, a compound present in the flowers of edelweiss. Bouayed, J. et al (2007) verified that, in vitro, this acid protected granulocytes from V 02-04/

6 induced oxidative stress. In 2009, Verma, A.R. et al also observed the antioxidant capacity of a phenolic fraction containing chlorogenic acid, protecting DNA from in vivo induced oxidative damage. Therefore, Edelweiss Eco is highly recommended for cosmetic products with antioxidant and anti-aging activity and to protect against oxidative damage. In traditional medicine, edelweiss has been used as an anti-inflammatory for years and, for this reason, in 2004 Dobner, M.J. et al wanted to verify its anti-inflammatory activity. In this study, anti-edema activity of the aerial parts of the plant when applied topically on an induced dermatitis was analyzed in vivo. Results showed an important reduction of edema, which confirms the anti-inflammatory activity of edelweiss and its components. In a later study (Speroni, E. et al, 2006), the anti-inflammatory effects caused by oral administration of the aerial parts of edelweiss were analyzed on animal models. Animals pretreated with the extract showed a significant reduction of edema and inflammatory response. In consequence, Edelweiss Eco is very convenient for cosmetic products with anti-inflammatory and soothing activity. Antibacterial activity The antibacterial and purifying activity of edelweiss and its components was analyzed by Dobner, M.J. et al (2003). Minimum inhibitory concentrations against some microorganisms were determined and the results showed that edelweiss inhibited varieties of Staphylococcus aureus, Enterococcus faecium, Escherichia coli, Pseudomonas aeruginosa, Streptococcus pneumoniae and Streptococcus pyogenes. Therefore, Edelweiss Eco is very useful to formulate cosmetic products with a purifying and antibacterial activity. BIBLIOGRAPHIC EFFICACY STUDY V 02-04/

7 Edelweiss is known for its antioxidant effect and this activity is partly due to the activity of leontopodic acid, a compound typical of this species. 1. Experimental method In 2009, Costa, S. et al conducted an in vitro study to determine the protective action of leontopodic acid against cell damage induced by two mycotoxins: aflatoxin (AFB1) and deoxynivalenol (DON). It is believed that the protective mechanism is related to the activation of antioxidant enzymes such as glutathione peroxidase (GPx). For this reason, the effect of pretreatment with leontopodic acid was determined on the viability of two cell lines (HepG2 for AFB1 and U937 for DON) exposed to toxins (compared to control) and also on the formation of reactive oxygen species (ROS) and the activity of the antioxidant enzyme. The two cell lines were cultured, exposed to the corresponding toxin and, in some cases, pre-treated with leontopodic acid, at three different concentrations. Measurement of ROS production was performed by fluorescence and increased GPx activity was also calculated by absorbance (400 nm), measuring the chromophoric product of a complex chemical reaction in which this enzyme is involved. 2. Results Viability results clearly showed that the U937 cells pretreated with leontopodic acid had a higher survival rate, increasing this parameter by 30-41% depending on the concentration of LA applied. Regarding GPx, results indicated that leontopodic acid (LA) increases the activity of this enzyme, thereby decreasing the production of ROS caused by toxins and contributing to cell survival (picture attached). Finally, toxins increased production of ROS in both studied cell lines, but when these cells were pretreated with leontopodic acid, it was noted that its production decreased significantly and at all concentrations tested. Figure 1 shows the results. V 02-04/

8 3. Conclusions Figure 1. ROS production results in both cell lines with two concentrations of AFB1 (5 and 10 µm) and DON (80 and 160 µm) toxins, depending on the applied concentration of leontopodic acid (LA). The analyzed parameters clearly show that leontopodic acid directly reduces the production of reactive oxygen species, increases cell viability and intensifies the action of glutathione peroxidase, an enzyme that helps reduce the concentration of ROS. Therefore, it is confirmed that this acid has a global action slowing cellular oxidation and damage caused by oxidation in the cells. In consequence, it is confirmed that Edelweiss Eco, due to its significant leontopodic acid content, is highly recommended and useful for cosmetic products with antioxidant and anti-aging activity, both for the face and body. It is also very suitable for photo-protective products. V 02-04/

9 COSMETIC APPLICATIONS Action Active Ingredient Cosmetic Application Antioxidant Anti-inflammatory Antibacterial Extract Leontopodic acid Chlorogenic acid Extract Extract -Anti-aging -Antioxidant -Photo-protection -Protection of hair coloration -Anti-edema -Soothing -Sensitive skin -Purifying -Antibacterial RECOMMENDED DOSE Recommended dosage ranges between 0.5% and 5.0%. BIBLIOGRAPHY Bouayed, J., Rammal, H., Dicko, A., Younos, C., Soulimani, R. Chlorogenic acid, a polyphenol from Prunus domestica (Mirabelle), with coupled anxiolytic and antioxidant effects. Journal of the Neurological Sciences, Nov 2007; 262 (1-2): Costa, S., Schwaiger, S., Cervellati, R., Stuppner, H., Speroni, E., Guerra, M.C. In vitro evaluation of the chemoprotective action mechanisms of leontopodic acid against aflatoxin B1 and deoxynivalenol-induced cell damage. J Appl Toxicol., 2009 Jan; 29(1): Dobner, M.J., Sosa, S., Schwaiger, S., Altinier, G., Della Loggia, R., Kaneider, N.C., Stuppner, H., Antiinflammatory activity of Leontopodium alpinum and its constituents. Planta Medica, 2004 Jun; 70 (6): Dweck, A.C. A Review of Edelweiss. SÖFW-Journal, 2004 (9); 130: V 02-04/

10 Schwaiger, S., Cervellati, R., Seger, C., Ellmerer, E.P., About, N., Renimel, I., Godenir, C., André, P., Gafner, F., Stuppner, H. Leontopodic acid - a novel highly substituted glucaric acid derivative from Edelweiss (Leontopodium alpinum Cass.) and its antioxidative and DNA protecting properties. Tetrahedron, May 2005; vol 61 (19): Schwaiger, S., Seger, C., Wiesbauer, B., Schneider, P., Ellmerer, E.P., Sturm, S., Stuppner, H. Development of an HPLC-PAD-MS Assay for the Identification and Quantification of Major Phenolic Edelweiss (Leontopodium alpinum Cass.) Constituents. Phytochem. Anal., 2006; 17: Speroni, E., Schwaiger, S., Egger, P., Berger, A.T., Cervellati, R., Govoni, P., Guerra, M.C., Stuppner, H. In vivo efficacy of different extracts of Edelweiss (Leontopodium alpinum Cass.) in animal models. J Ethnopharmacol., 2006 May 24;105(3): Verma, A.R., Vijayakumar, M., Mathela, C.S., Rao, C.V. In vitro and in vivo antioxidant properties of different fractions of Moringa oleifera leaves. Food and Chemical Toxicology, Sep 2009; 47 (9): Websites: anel.pubmed_rvdocsum&ordinalpos=1 anel.pubmed_rvdocsum&ordinalpos=8 anel.pubmed_rvdocsum&ordinalpos=2 anel.pubmed_rvdocsum&ordinalpos=9 1&_user=10&_coverDate=11%2F15%2F2007&_alid= &_rdoc=1&_fmt=high&_orig=search&_cdi V 02-04/

11 =4854&_sort=r&_docanchor=&view=c&_ct=2291&_acct=C &_version=1&_urlVersion=0&_userid =10&md5=832c7b9da6b54af f3cbf6123 V 02-04/

12 PROVITAL. S.A. Pol. Ind. Can Salvatella Gorgs Lladó, Barberà del Vallès Barcelona (España) Tel. (+34) V06-02/ ,12310,12340,12350,12710,

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