ANTI-AGING. Rosemaryeco.

Transcription

1 ANTI-AGING Rosemaryeco

2 Rosemary eco BOTANY Rosmarinus officinalis L. is a woody aromatic shrub belonging to the Labiatae (Lamiaceae) family. It grows approximately one meter tall. The young branches are pubescent and turn woody on maturation. The leaves are simple, opposite, sessile, lineal, coriaceous, perennial and their edges roll up. The upper and lower surfaces of the leaves are covered by secretory hairs that give the plant a pleasant camphoraceous aroma. The flowers are small, bilabiate, pale blue or lilac with violet spots, arranged in dense axillary or terminal clusters. They appear by the end or the spring or beginning of the summer. The fruit is a bright brown tetra-achene. Rosemary grows in the dry warm regions of southern Europe, especially the Mediterranean area. The wild forms grow on every kind of substrate up to meters altitude. The major rosemary producers are Spain, Tunisia, Morocco and, to a lesser degree, Portugal, Turkey, India and former Yugoslavia. Rosemary-Eco extract is produced from the leaves of Rosmarinus officinalis. V 03-04/

3 CHEMISTRY Flavonoids and Phenolic acids Flavonoids: heterosides of luteolol, diosmetol and methoxylated flavones in C-6 and/or C-7; phenolic acids (2-3%), especially caffeic derivatives: caffeic acid, chlorogenic acid and rosmarinic acid. Terpenoids Rosemary also contains tricyclic diterpenes: carnosic acid and carnosol (mainly), rosmanol (in concentrations higher that 4%) and triterpenes: ursolic acid and oleanolic acid (2-4%). Essential oils The leaves of rosemary contain between % essential oil. Such composition may markedly vary according to the chemotype and the development stage at which the plant has been harvested. The main constituents are α-pinene (25%), 1,8-cineole (variable concentration 12-50%), camphor (10-25%), camphene (5-10%), borneol (1-6%), bornyl acetate (1-5%) and α-terpineol (12-24%). Fig.1. Major components of Rosmarinus officinalis (Armengol R. & Betés C., 1994). Fig.2. Rosmarinic acid V 03-04/

4 Active principles Components Concentration(%) ESSENTIAL OILS α-pinene, 1,8-cineole, camphor, camphene, borneol bornyl acetate, α-terpineol % POLYPHENOLS Flavonoids heterosides of luteolol, diosmetol and methoxylated flavones in C-6 and/or C-7 - Phenolic acids Caffeic acid, chlorogenic acid and rosmarinic acid 2-3% TERPENOIDS Diterpenes Carnosic acid and carnosol (mainly), rosmanol, epirosmanol > 4% Triterpenes Ursolic acid and oleanolic acid 2-4% Table 1. Major compounds in the leaf of Rosmarinus officinalis. TRADITIONAL USES Rosemary is one of the aromatic plants, which have been much sought after since ancient times owing to its medicinal, edible and aromatic properties. The popular uses of rosemary include its digestive, choleretic, emmenagogue, sedative, anti-spasmodic, hypertensive, diaphoretic, wound healing, antiseptic and analgesic properties and as capillary growth stimulant (topical applications). In China it is recommended for headaches, insomnia and mental fatigue. In India it is used for its carminative and antimigraine properties. In Ecuador they make rubs with bunches of rosemary flowers (cooked in vegetable oil) as a palliative for rheumatic pains. The leaves are used in the food industry to flavour meats or stews and in the production of liqueurs, such as Benedictine. Its antioxidant properties make it useful in the preparation of sausages. The aromatic properties of rosemary oil make it useful in the production of bath products, detergents and insecticides. V 03-04/

5 COSMETIC PROPERTIES Antioxidant activity Zeng H. et al. (2001) studied the antioxidant acticity of rosemary and its action mechanism. They specifically studied the capacity of the carnosol, rosmanol and epirosmanol phenolic diterpenes of rosemary to inhibit lipid peroxidation of the low density lipoproteins (LDL) in the blood and the cell membranes, as well as the anti-radical action of these diterpenes. The results obtained showed that carnosol, rosmanol and epirosmanol had the capacity to inhibit the Cu 2+ mediated oxidation of LDL and cell membranes. They also determined that this activity was due to the anti-radical activity of these diterpenes. Chen Q. et al., (1992) measured the antioxidant activity of R.officinalis extract, as well as the antioxidant activity of its purified components, using the Rancimat method. It was observed that the antioxidant activity of R.officinalis extract depended mainly on its carnosol and carnosic acid content. These authors also studied the inhibition of the lipoxygenase enzyme activity and demonstrated that the rosemary extract had an inhibitory effect on the metabolism of arachidonic acid. In this study they tested the capacity of the rosemary extract, and its isolated components, to inhibit the lipoxygenase enzyme. IC 50 values of between 1.3 and 2.6 µg were observed for rosemary extract with 15-lipoxygenase enzyme from soya. Carnosic acid was more effective in inhibiting lipoxygenase than carnosol. Ursolic acid also showed a substantial inhibitory effect on this enzyme. Sample Induction Time Lipoxigenase Inhibition (IC 50 ) (h) (µm) (µg) Control Butter 1.65± V 03-04/

6 ± ± ± ± ± ±0.09 Carnosol 25.40± ± ±0.01 Carnosic acid 30.60± ± ±0.03 Ursolic acid 2.47± ± ±0.04 Table 2. Antioxidant and lipoxygenase inhibitory activity of rosemary extract and its three major components (Samples: 1: extract with hexane; 2: extract with acetone; 3: extract with methanol). (Chen Q. et al., 1992). Hydroalcoholic extracts of medicinal plants with high hydroxycinnamic derivatives content (rosmarinic acid accounting for more than 3-6% of the dry weight) showed significant antioxidant activity by scavenging the 1,1-diphenyl-2-picryl-hydrazyl (DPPH) free radical. This antioxidant action was partially attributed to the high rosmarinic acid content of these plants (Al-Sereiti MR et al., 1999). For all these reasons, the extract of rosemary-eco is of great utility when formulating cosmetic products aimed at preserving the integrity of the skin and hair from oxidative processes. Anti-inflammatory activity Rosmarinic acid had anti-inflammatory activity in rat models with carrageenin-induced plantar oedema. It was experimentally shown that this acid acts on the formation of prostaglandins (PGE 2 ), in a way similar to non-steroidal anti-inflammatories. Additionally it was observed to inhibit the complement factor C3, a mediator in the inflammatory process not involving the cyclooxygenase pathway or prostacyclin-synthase activity. Likewise, it was shown to reduce the production of leukotriene B4 in human polymorphonuclear leucocytes. It must be remembered that free radicals also take part in the inflammatory process (Alonso J., 2004). Al-Sereiti MR. et al (1999) also arrived at these conclusions. Manez S. et al. (1997) verified that ursolic acid reduced the chronic inflammation and neutrophil infiltration produced by repeated applications of TPA (12-O-tetradecanoylphorbol-13-acetate). This activity is closely linked to the structure of this compound. In summary, rosemary-eco extract is highly recommended when formulating cosmetic products for sensitive or irritated skin. Antimicrobial activity V 03-04/

7 Methanol extract containing 30% of carnosic acid, 16% of carnosol and 5% of romsarinic acid was the most effective antimicrobial against gram positive bacteria (MIC between 2 and 15 μg/ml), Gram negative bacteria (MIC between 2 and 60 μg/ml) and yeast (MIC of 4 μg/ml). By contrast, water extract containing only 15% of rosmarinic acid showed a narrow activity. MIC value of the methanol and water extracts is in good correlation with the values obtained with pure carnosic acid and rosmarinic acid, respectively (Moreno S, et al., 2006). Different extracts of rosemary have demonstrated inhibitory activity on cultures of Staphylococcus aureus, S. albus, Escherichia coli, Corynebacterium spp., Bacillus subtilis, Micrococcus luteus, Salmonella spp., Listeria monocytogens and Vibrio cholerae. Two of its components, carnosol and ursolic acid, are responsible for this antimicrobial effect, which also includes the HIV-1 virus, fungi of the Aspergillus sp, Penicillium sp, Alternaria sp genera and other food fermenting microorganisms such as Lactobacillus brevis, Pseudomonas fluorescens, Rhodotorularia glutinis and Kluyveromyces bulgaricus (Alonso J., 2004). The essential oils of rosemary showed antibacterial and antifungal activity against a range of organisms tested (E.S.C.O.P., 1992). Therefore, the active ingredients with bactericidal activity from rosemary-eco extract are ideal when formulating purifying and antiseptic cosmetic products. Finally, we should mention that in the reference publication Plant preparations used as ingredients of cosmetics products. 1 st edition. Council of Europe, 1994 there are one monograph dedicated to the glycolic extract of rosemary leaves. The intended cosmetic effects and recommended maximum concentrations in cosmetics products attributed to this extract are the following: tonic, deodorant, purifying, reactivating up to 5% in stimulant and purifying hair lotions, baths, shampoos, showers, tooth pastes products for greasy and toneless skins other possible effects: antiseptic, stimulant, analgesic, rubifacient, antioxidant, flavouring BIBLIOGRAPHIC EFICCACY TEST V 03-04/

8 Rosemary (Rosmarinus officinalis) contains a number of antioxidant compounds, most of them polyphenols. The antioxidant agents in this species are the carnosic and caffeic acids, as well as their derivatives, such as the powerful antioxidant rosmarinic acid. A number of biological actions have been reported for rosmarinic acid, for example, antioxidant, antiviral and antibacterial actions, and others (Moreno S. et al, 2006). 1. Experimental methods Rosmarinus officnalis were investigated by a combination of bioassays and biochemical analysis to identify bioactive compounds. The 2,2-diphenyl-2-picrylhydracyl hydrate (DPPH) radical scavenging method, Folin- Ciocaulteau method and HPLC cromatography were used to study the distribution and levels of antioxidants (Moreno S. et al, 2006). 2. Results Moreno S et al (2006) investigated the polyphenol composition of fresh leaves, flowers and branches, from the plant R.officinalis. To quantify the rosmarinic acid, carnosic acid and carnosol presenting each part of the plant, it was used the HPLC chromatography. The amount of each compound was expressed as grams of the compound per 100 g of dry weight of rosemary extract. Results are shown in table 3. Compound Yield (g/100g extract) in Leaves Flowers Branches RA 7.9 ± ± ± 0.1 COH 8.5 ± ± 0.9 NF CA 29.3 ± ± 1.3 NF Table 3. Quantification of rosmarinic acid (RA), carnosol (CHO) and carnosic acid (CA) in leaves, flowers and branches extracts after HPLC chromatography (Moreno S et al, 2006). NF= Not founf. Leaves and flowers showed the highest amount of the main active polyphenols. A higher yield of rosmarinic acid and carnosic acid was found in leaves in comparison to flowers, whilst branches did not show significant presence of these polyphenol compounds. Table 4 shows the total phenolics estimated by the Folin-Ciocaulteau method. Extracts Yield (g/100 g extract) of RA CA COH Total phenolic content (g GAE/100 g extract) V 03-04/

9 Acetone 4.0 ± ± ± ± 8 Methanol 5.5 ± ± ± ± 5 Water 14.5 ± 1.4 NF NF 3 ± 2 Table 4. Quantification of rosmarinic acid (RA), carnosic acid (CA) and carnosol (COH) and total phenolic content in different extracts of flowering plants (Moreno S. et al, 2006). The antioxidant activity (AOX) measured by the DPPH method was assayed in extracts from leaves, flowers and branches. Figure 3 shows that leaves and flowers exhibited lower EC 50 values indicative of a high AOX activity, comparable to the AOX activity of commercial BHT, whereas branches presented higher EC 50 values indicative of a low activity. EC 50 = concentration of sample necessary to decrease by 50% the initial DPPH absorbance (Moreno S. et al., 2006). BHT 20 Flow ers 33 Leaves 46 Branches EC 50 (μg/ml) Fig.3. AOX activity in leaves, flowers and branches extracts (Moreno S. et al., 2006). When AOX activity was measured in the HPLC chromatography fractions after separation of the methanol extract of leaves, a high activity was associated with fractions containing rosmarinic acid, carnosol and carnosic acid. Figure 4 shows the AOX activity in the fractions corresponding to the elution of rosmarinic acid, carnosol and carnosic acid after separation of the extract leaves by HPLC chromatography. V 03-04/

10 % Inhibition DPPH RA COH CA Fig.4. AOX activity in the fractions corresponding to the elution of rosmarinic acid, carnosol and carnosic acid after separation of the extract leaves by HPLC chromatography (Moreno S. et al. 2006). These results showed that there was a good correlation between the antioxidant activities of rosemary extracts and its content in rosmarinic acid, carnosic acid and carnosol. COSMETIC APPLICATIONS Action Active Cosmetic Applications Antioxidant Anti-inflammatory Anti-microbial Diterpenes Triterpenes Phenolic acids Phenolic acids Triterpenes Essential oils Diterpenes Triterpenes -Anti-ageing -Photo-protection -Hair colour protection -Sensitive irritated skin -Purifying -Antiseptic RECOMMENDED DOSE The recommended dose is between 0.1% and 2.0%. V 03-04/

11 BIBLIOGRAPHY Abelló R. & Foix C. Desodorantes y Antitranspirantes. El Farmacéutico 1994; 139:33-37 (ref.5475). Alonso, Jorge. Tratado de Fitofármacos y Nutracéuticos. Barcelona: Corpus, 2004, p: (633.8 ALO). Al-Sereiti MR. et al. Pharmacology of rosemary (Rosmarinum officinalis Linn.) and its therapeutic potentials. Indian J. Experim. Biol. 1999, 37 p: (ref.949). Armengol R. & Betés C. Antioxidant Action of a natural Extract. Cosmetic Uses. SÖFW-Journal 1995, p:14-18 (ref.210). Balanehru S. et al. Protective Effect of Oleanolic Acid and Ursolic Acid Against Lipid Peroxidation. Biochem. Internat. 1991, 24(5) p: (ref.3724). Bruneton J. Farmacognosia. Zaragoza: Ed. Acribia, 2001; 249,462,533 (651*1 BRU). Calabrese V. et al. Biochemical Studies On A natural Antioxidant Isolated From Rosemary. J Appl Cosmetol, 1998; p: (ref.4083). Council of Europe. Plant preparations used as ingredients of cosmetics products.1 st edition. Belgium: Council of Europe Press, 1994; p: (68*1 PAT). Chen Q. et al. Effects of Rosemary Extracts and Major Constituents on Lipid Oxidation and Soybean Lipoxygenase Activity. JAOCS, 1992; 69 (10): (ref.1995). Del Baño MJ. et al. Phenolic Diterpenes, Flacones,and Rosmarinic Acid Distribution during the Development of Leaves, Flowers, Ítems, and Roots of Rosmarinus officinalis. Antioxidant Activity. J Agric. Food Chem. 2003, 51 p: (ref.6945). ESCOP Monographs (Rosmarini Folium Cum Flore).Fascicule 1. Exeter (UK): ESCOP (ed), 1996 (633.8(031)ENC). Kuklinski C. Farmacognosia. Barcelona: Ed.Omega,S.A, 2000; p:295-6 (615*1 KUK). Manez S. et al. Effect of selected triterpenoids on chronic dermal inflammation. European Journal of Pharmacology 1997, 334 p:103-5 (ref.3725). V 03-04/

12 Moreno S., Scheyer T., Romano C.S., Vojnov A.A. Antioxidant and antimicrobial activities of rosemary extracts linked to their polyphenol composition. Free Radical Research, (2): Najid A. et al. Characteritzation of ursolic acid as a lipoxygenase and cyclooxygenase inhibitor using macrophages, platelets and differentiated HL60 leukemic cells. FEBS 1992, 299(3) p: (ref.3697). Qinyun C. et al. Effects of Rosemary and Major Constituents on Lipid Oxidation and Soybean Lipoxygenase Activity. JAOCS 1992, 69(10) p: (ref.1995). Schwarz K. et al. Antioxidative constituents of Rosmarinus officinalis and Slavia officinalis. Z lebensm Forsch 1992, 195 p: 95-8 (ref.5163). Wichtl M. Herbal Drugs and Phytopharmaceuticals. London: Norman Grainger Bisset (Ed.),1994, p: (633.8 WIC). Zeng H. et al. Antioxidant properties of phenolic diterpens from Rosmarinus officinalis. Acta Pharmacol. Sin. 2001, 22(12) p: (ref.7049). Web sites: [accessed September 2006]. V 03-04/

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