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WO2008119852A1 - Extract of the leaves of couroupita guianensis, method for obtaining same and cosmetic use thereof as an antioxidant/antirradical/uv filter - Google Patents

Extract of the leaves of couroupita guianensis, method for obtaining same and cosmetic use thereof as an antioxidant/antirradical/uv filter Download PDF

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Publication number
WO2008119852A1
WO2008119852A1 PCT/ES2008/000178 ES2008000178W WO2008119852A1 WO 2008119852 A1 WO2008119852 A1 WO 2008119852A1 ES 2008000178 W ES2008000178 W ES 2008000178W WO 2008119852 A1 WO2008119852 A1 WO 2008119852A1
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Prior art keywords
guianensis
extracts
couropita
extraction
abs
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PCT/ES2008/000178
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Spanish (es)
French (fr)
Inventor
Ana María MARTÍNEZ MORÁN
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Caroi'line Cosmética, S.L.
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Publication of WO2008119852A1 publication Critical patent/WO2008119852A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/16—Emollients or protectives, e.g. against radiation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/08—Anti-ageing preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q5/00—Preparations for care of the hair
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
    • A61K2800/52—Stabilizers
    • A61K2800/522—Antioxidants; Radical scavengers

Definitions

  • Oxidative stress is defined as the situation of cellular damage that results when the generation and / or exogenous incidence of RLO exceeds the capacity of the different physiological mechanisms of the organism to prevent or intercept its accumulation.
  • Natural antioxidants in addition to protecting the body from damage caused by free radicals, responsible for degenerative diseases such as cancer, arteriosclerosis, arthritis and neurodegenerative and aging processes; they can present antibactericidal, antiviral, antimutagenic, antiulcer or anticarlogenic activity.
  • UV radiation can be classified in UV-C (wavelength less than 280 nm); UV-B (wavelength between 280 nm and 320 nm) and UV-A (wavelength between 320 nm and 400 nm). Of these ultraviolet radiation, the most lethal is UV-C radiation, although most of it is absorbed by ozone. Therefore, those that may have the greatest influence on the skin are UV-A and UV-B radiation.
  • Couroupita guianensis is a plant used in India for the treatment of skin diseases [a) A. S. R. Anjaneyulu et al, Indian Journal of Chemistry, Section B; 1998, 37B (4), 382-386. b) The useful plants of India; Publications & Information
  • indole alkaloids of the indole type [2, 1-6] quinazolo-6,12-diones, isolated from the fruits of Couroupita guianensis, and in particular of tryphantrin and its derivatives.
  • These alkaloids have antibacterial, antituberculous activity [a) L. A. Mitscher et al., Puré & Applied Chemistry; 1998, 70 (2), 365-371. b) WO951380], antimalarial [US2006160827 Al] and anti-inflammatory activity, for acting as inhibitors of prostaglandins and leukotrienes [H. Danz et al., Planta Medica; 2001, 67, 411-416].
  • Citrus, malic and isocitric acids have been isolated from the fruits of Couroupita guianensis [a) E. K. Nelson et al., Journal of the American Chemical Society; 1937, 59, 2499-2500. b) T. R. Soderstrom, American Journal of Botany; 1962, 49 (8), 850-855], chlorogenic acids [P. V. Pontes et al, Journal of the Science of Food and Agriculture; 2002, 82 (10), 1177-1181], stigmaesterol and camfesterol sterols; and the tryptanthrine, couropitin A, couropitin B or indirubin, indigo and isatin [alkaline] alkaloids [a) J. Bergman et al.
  • the present invention describes extracts of Couroupita guianenis leaves, their method of obtaining and their cosmetic use as antioxidants / anti-radicals / UV filters (ultraviolet).
  • Extracts of Couroupita guianensis obtained by the described procedure, that is, the extracts of Couroupita guianensis, characterized by presenting, among others, high phenolic content and relevant antioxidant, anti-radical and UV protection properties, which makes them possible to be used for skin and / or hair care, for example as antioxidants or radical inhibitors, as agents to combat aging or sunscreens. Extracts frequently have higher antioxidant activity than synthetic antioxidants such as BHT (butyrohydroxytoluene), BHA (butyrohydroxyanisole) or Trolox (soluble equivalent of ⁇ -tocopherol or vitamin E).
  • BHT butyrohydroxytoluene
  • BHA butyrohydroxyanisole
  • Trolox soluble equivalent of ⁇ -tocopherol or vitamin E
  • the dried leaves of ground and sifted Couroupita guianensis are used. They are subjected to a grind, preferably in a blade mill, and screened, preferably at a particle size of 0.6 mm. approximately.
  • Liquid solid ratios are used high, preferably 30 g / g.
  • Water and / or hydroalcoholic and / or alcoholic solvents are used as solvent to be extracted.
  • Ci to C 4 alcohols and, preferably, ethanol or methanol are advantageously chosen.
  • a 1: 1 ethanol / water mixture is advantageously chosen.
  • the extraction temperature is the reflux of the solvent or mixture of extraction solvents and is in the range of 60-120 0 C and preferably between 85-95 0 C.
  • the extraction time is in the range of 3 hours to 21 days
  • the extractive process is protected from light to avoid possible alterations.
  • the yield of the extractive process ranges from 15% to 40% (g / g).
  • High liquid: solid ratios are used, preferably 99 g / g.
  • water and / or hydro-glycol and / or glycol solutions are used.
  • Cz to Ce glycols are advantageously chosen.
  • these glycols it is preferred to use propylene glycol and butylene glycol.
  • a 1: 1 butylene glycol / water mixture is advantageously chosen.
  • the extraction temperature is room temperature, approximately 20 0 C.
  • Extraction time is prolonged, from 3 to 21 days.
  • UV (ultraviolet) radiation is classified as UV-C (wavelength less than 280 nm); UV-B (wavelength between 280 nm and 320 nm) and UV-A (wavelength between 320 nm and 400 nm), therefore, if a product absorbs radiation between 250 to 400 nm it can be used as a sunscreen.
  • the absorbency (Abs) of the filtered liquid extracts of Couroupita guianensis is measured at the concentration of 100 ppm in ethanol between the wavelengths of 250 to 400 nm against an ethanol blank. All have maximum absorption in the UV (ultraviolet) region. Thus at the wavelength of 250 nm they present Abs>1,000; at 280 nm they present Abs>0.800; at 320 nm they present Abs> 0.300 and at 400 nm they present Abs> 0.100.
  • ii) Determination of the phenolic content of the filtered liquid extracts of Couroupita guianensis The phenolic compounds have a high antioxidant activity [YS Velioglu, op.
  • the phenolic content of the liquid extracts of Couroupita guianensis is determined according to the Folin Ciocalteu's method [A. Escarpa et al., Analytica Chinaca Acta; 2001, 427 (1), 119-127].
  • the absorbance at 765 nm of a solution of 0.5 mL of the antioxidant extract, 3.75 mL of distilled water, 0.25 mL of the reagent is measured Ciocalteau's Folin (1: 1 with distilled water) and 0.5 mL of 10% Na 2 CO 3 ; after 1 hour at room temperature, against a white without extract.
  • the phenolic content of the extract is determined by comparing the absorbency with a straight standard of gallic acid (0-100 ppm).
  • the filtered liquid extracts of Couroupita guianensis obtained according to the extraction procedure, have, according to the described procedure, a high phenolic content, between 25-35%.
  • the IC 50 or concentration that inhibits 50% of the DPPH * radical is measured.
  • the filtered liquid extracts of Couroupita guianensis obtained according to the extraction procedure, have an IC 50 between 200 and 400 ppm.
  • the IC 50 of the BHA, measured according to the same procedure, is 240 ppm and that of the BHT of 2790 ppm.
  • ⁇ -carotene pro-vitamin A
  • linoleic acid a solution of ⁇ -carotene (2 mg in 10 mL of chloroform) is prepared. 1 mL of this solution is added in a tube with 20 mg of linoleic acid and 200 mg of the Tween 40 emulsifier. It is homogenized. Chloroform is removed in vacuo. 50 mL of distilled water saturated with oxygen are added.
  • CAA [(Abs extract 120 min - Abs control 120 min) / (Abs control 0 min - Abs control 120 min)] * 100
  • the filtered liquid extracts of Couroupita guianensis obtained according to the extraction procedure, have an oxidation inhibitory activity concentration of between 35% and 60% at 250 ppm.
  • a PBS buffer pH 7.4 is prepared (8.0 g NaCl, 0.2 g KH 2 PO 4 , 1.15 g Na 2 HPO 4 , 0.2 g KCl, 0.2 g sodium azide).
  • the TEAC reagent 38.4 mg ABTS ** 7 mM, 6.62 mg potassium persulfate 2.45 mM in 10 mL PBS buffer) is prepared. It is stirred for 16 hours in the absence of light. The absorbance is read at 734 years. The reagent is diluted until it has an absorbance close to 0.7. 20 ⁇ L of the antioxidant extract is added to 2 mL of the reagent.
  • the filtered liquid extracts of Couroupita guianensis obtained according to the extraction procedure, have a capacity of inhibition of the ABTS * * radical equivalent to between 0.400 to 0.800 mM of Trolox at a concentration of 100 ppm. This implies a weight / weight ratio of the dry extract of Couroupita guianensis / Trolox from 1/1 to 1/2 (g / g).
  • the antioxidant extract is diluted in ethanol, 2.5 mL of phosphate buffer pH 0.6 0.2 M and 2.5 mL of 1% potassium ferrocyanide are added to 1 mL thereof. Incubate at 50 ° C for 20 minutes. 2.5 mL of 10% trichloroacetic acid are added. 2.5 mL of the supernatant is pipetted and mixed with 2.5 mL of distilled water and 0.5 mL of 0.1% ferric chloride. The absorbance at 700 nm is measured and the results are compared with a straight standard of ascorbic acid or vitamin C (0.1-1 mM).
  • the filtered liquid extracts of Couroupita guianensis obtained according to the extraction procedure, have at a concentration of 100 ppm an iron reducing power of between 20% to 40% and an equivalence between 0.200 mM to 0.400 mM in ascorbic acid or vitamin C This implies a weight / weight ratio of the dry extract of Couroupita guianensis / ascorbic acid or vitamin C between 1 / 0.35 to 1 / 0.70 (g / g).
  • the dried extract of Couroupita guianensis obtained is subjected to a new continuous solid-liquid extractive process, by maceration with stirring, with a liquid ratio: 99 g / g solid, at room temperature and protected from light, for 16 days with a 1: 1 butylene glycol / water mixture.
  • the liquid extract of Couroupita guianensis obtained is filtered, using porous plate No. 3, and the final product, the filtered liquid extract of
  • Couroupita guianensis is stored refrigerated at 4 0 C and protected from light to prevent its alteration.
  • the phenolic content of the extract is determined by comparing the absorbance with a straight gallic acid standard (0-100 ppm).
  • the extract obtained according to the procedure described in this example 1 has a phenolic content of 32%.
  • the IC50 or concentration that inhibits 50% of the DPPH * radical was measured.
  • Oxidation inhibitory activity in an emulsified medium based on the oxidation inhibition of an emulsion of ⁇ -carotene (pro-vitamin A) and linoleic acid [G. J.
  • a solution of ⁇ -carotene (2 mg in 10 mL of chloroform) is prepared. 1 mL of this solution is added in a tube with 20 mg of linoleic acid and 200 mg of the Tween 40 emulsifier. It is homogenized. Chloroform is removed in vacuo. 50 mL of distilled water saturated with oxygen are added. Stir at 6000 rpm to form the emulsion.
  • the absorbances at 470 nm of the sample of the antioxidant extract (200 ⁇ L of the extract in 5 mL of emulsion) and of the control (200 ⁇ L of absolute ethanol in 5 mL of emulsion) are measured after 2 hours at 50 0 C. a blank following the same procedure but without adding ⁇ -carotene.
  • the antioxidant activity coefficient is measured (CAA,%), which is the oxidation ratio of ⁇ -carotene, or pro-vitamin A, in the presence and absence of antioxidant.
  • CAA [(Abs extract 120 min - Abs control 120 min) / (Abs control 0 min - Abs control 120 min)] * 100
  • the extract obtained according to the procedure described in this example 1 has a CAA of 58% at 250 ppm concentration.
  • a PBS buffer pH 7.4 (8.0 g NaCl, 0.2 g KH 2 PO 4 , 1.15 g Na 2 HPO 4 , 0.2 g KCl, 0.2 g sodium azide) is prepared.
  • the TEAC reagent 38.4 mg ABTS * * 7 mM, 6.62 mg potassium persulfate 2.45 mM in 10 mL of PBS buffer) is prepared. It is stirred for 16 hours in the absence of light. The absorbance at 734 nm is read. The reagent is diluted until it has an absorbance close to 0.7.
  • the extract obtained according to the procedure described in this example 1 has an equivalent of 0.733 mM in Trolox at 100 ppm concentration. This implies a weight / weight ratio of the dry extract of Couroupita guianensis / Trolox of 1 / 1.84 (g / g).
  • the antioxidant extract is diluted in ethanol, 2.5 mL of phosphate buffer pH 0.6 0.2 M and 2.5 mL of 1% potassium ferrocyanide are added to 1 mL thereof. Incubate at 50 0 C for 20 minutes. 2.5 mL of 10% trichloroacetic acid are added. 2.5 mL of the supernatant is pipetted and mixed with 2.5 mL of distilled water and 0.5 mL of 0.1% ferric chloride. The absorbance at 700 nm is measured and the results are compared with a straight standard of ascorbic acid or vitamin C (0.1-1 mM).
  • the extract obtained according to the procedure described in this example 1 has at 100 ppm a 32% reducing power equivalent to 0.316 mM in ascorbic acid or vitamin C. This implies a weight / weight ratio of the dry extract of Couroupita guianensis / ascorbic acid or vitamin C of 1 / 0.56 (g / g).
  • the dried leaves of Couroupita guianensis, ground and sieved at a particle size of approximately 0.6 mm, are subjected to a continuous solid-liquid extractive process, with a liquid: solid ratio of 30 g / g, using a soxhlet, with a 1: 1 ethanol / water mixture, at the reflux temperature of the solvent (85-95 0 C), for 12 days and protected from light.
  • the solvent of the extract obtained is evaporated to dryness in vacuo. The yield of extraction is 36% (g / g).
  • the dried extract of Couroupita guianensis obtained is subjected to a new continuous solid-liquid extractive process, by maceration with stirring, with a liquid ratio: 99 g / g solid, at room temperature and protected from light, for 16 days with a 1: 1 propylene glycol / water mixture.
  • the liquid extract of Couroupita guianensis obtained is filtered, using porous plate No. 3, and the final product, the filtered liquid extract of Couroupita guianensis, is stored refrigerated at 4 0 C and protected from light to prevent its alteration.
  • the IC 50 or concentration that inhibits 50% of the DPPH * radical was measured.
  • Oxidation inhibitory activity in emulsified medium based on the oxidation inhibition of an emulsion of ⁇ -carotene and linoleic acid [G. /. Marco, op. cited (1968)].
  • a solution of ⁇ -carotene (2 mg in 10 mL of chloroform) is prepared. 1 mL of this solution is added in a tube with 20 mg of linoleic acid and 200 mg of the Tween 40 emulsifier. It is homogenized. Chloroform is removed in vacuo. 50 mL of distilled water saturated with oxygen are added. Stir at 6000 rpm to form the emulsion.
  • the absorbances at 470 nm of the sample of the antioxidant extract (200 ⁇ L of the extract in 5 mL of emulsion) and of the control (200 ⁇ L of absolute ethanol in 5 mL of emulsion) are measured after 2 hours at 50 0 C. a blank following the same procedure but without adding ⁇ -carotene.
  • the coefficient of antioxidant activity (CAA,%) which is the oxidation ratio of ⁇ -carotene in the presence and absence of antioxidant, is measured.
  • CAA [(Abs extract 120 min - Abs control 120 min) / (Abs control 0 min - Abs control 120 min)] * 100
  • the extract obtained according to the procedure described in this example 2 has a CAA of 40% at 250 ppm concentration.
  • a PBS buffer pH 7.4 (8.0 g NaCl, 0.2 g KH 2 PO 4 , 1.15 g Na 2 HPO 4 , 0.2 g is prepared KCl, 0.2 g sodium azide).
  • the TEAC reagent 38.4 mg ABTS ** 7 mM, 6.62 mg potassium persulfate 2.45 mM in 10 mL of PBS buffer) is prepared. It is stirred for 16 hours in the absence of light. The absorbance at 734 nm is read. The reagent is diluted until it has an absorbance close to 0.7.
  • the extract obtained according to the procedure described in this example 2 has an equivalence of 0.525 mM in Trolox at 100 ppm concentration. This implies a weight / weight ratio of the dry extract of Couroupita guianensis / Trolox of 1 / 1.32 (g / g).
  • the antioxidant extract is diluted in ethanol, 2.5 mL of pH 6.60.2 M phosphate buffer and 2.5 mL of 1% potassium ferrocyanide are added to 1 mL thereof. Incubate at 50 0 C for 20 minutes. 2.5 mL of 10% trichloroacetic acid are added. 2.5 mL of the supernatant is pipetted and mixed with 2.5 mL of distilled water and 0.5 mL of 0.1% ferric chloride. The absorbance at 700 nm is measured and the results are compared with a straight standard of ascorbic acid or vitamin C (0.1-1 mM).
  • the extract obtained according to the procedure described in this example 2 has at 100 ppm a 23% reducing power equivalent to 0.316 mM in ascorbic acid or vitamin C. This implies a weight / weight ratio of the dry extract of Couroupita guianensis / ascorbic acid or Vitamin C of 1 / 0.40 (g / g).

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Abstract

Extract of Couroupita guianenis leaves, method for obtaining same and cosmetic use thereof as an antioxidant/antiradical/UV (ultraviolet) filter. Extracts obtained from dry, ground and sieved leaves, extraction with water and/or alcohols, evaporation of this solvent, subsequent extraction with water and/or glycols of the dry extract of Couroupita guianenis and filtration; that can be used for care of the skin and/or hair, for example as antioxidants or radical inhibitors, as agents for combatting ageing or solar protectors.

Description

DESCRIPCIÓN DESCRIPTION
Extracto de las hojas de Couroupita guianensis, procedimiento de obtención y su uso cosmético como antioxidante/antirradicalario/filtro UV.Couroupita guianensis leaf extract, procedure for obtaining and its cosmetic use as antioxidant / anti-radical / UV filter.
A principios del siglo XX Mulliken investigó la reactividad fotolumínica demostrando que durante el proceso de peroxidación y fotoexcitación se formaba un estado excitado del oxígeno (oxígeno singlete: '0* 2). El desarrollo de estas investigaciones llevó a la introducción del término de especies reactivas de oxígeno (ERO) y de radicales libres de oxígeno (RLO) en el campo de la biología y a la demostración de que estas especies radicalarias eran las responsables de los efectos tóxicos del oxígeno. Por lo tanto, dado que los ERO/RLO son especies que se generan de forma continuada como productos de la utilización celular del O2, en los organismos aerobios se han desarrollado estrategias biológicas para desactivarlos, como el sistema enzimático superóxido dismutasa, glutatión peroxidasa o catalasaAt the beginning of the twentieth century Mulliken investigated photoluminal reactivity by demonstrating that during the peroxidation and photoexcitation process an excited state of oxygen was formed (singlet oxygen: '0 * 2 ). The development of these investigations led to the introduction of the term reactive oxygen species (ERO) and oxygen free radicals (RLO) in the field of biology and to the demonstration that these radical species were responsible for the toxic effects of oxygen. Therefore, since the ERO / RLO are species that are generated continuously as products of the cellular use of O 2 , in aerobic organisms biological strategies have been developed to deactivate them, such as the enzyme system superoxide dismutase, glutathione peroxidase or catalase
Con frecuencia, estos mecanismos básicos de defensa no son suficientes para eliminar todas las especies radicalarias generadas de forma endógena (respiración celular y otros procesos fisiológicos) o las de origen exógeno (radiaciones, polución ambiental, fármacos, tóxicos) que desde el entorno inciden en el organismo. La pérdida de equilibrio entre el nivel de prooxidación y el tono antioxidante de un órgano o tejido puede producir daño celular y, secundariamente, condicionar la aparición de estados de enfermedad. Se define el estrés oxidativo como la situación de daño celular que resulta cuando la generación y/o incidencia exógena de RLO supera la capacidad de los diferentes mecanismos fisiológicos del organismo para prevenir o interceptar su acumulación.Frequently, these basic defense mechanisms are not sufficient to eliminate all radical species generated endogenously (cellular respiration and other physiological processes) or those of exogenous origin (radiation, environmental pollution, drugs, toxic) that affect the environment from the organism. The loss of balance between the level of prooxidation and the antioxidant tone of an organ or tissue can cause cellular damage and, secondarily, condition the onset of disease states. Oxidative stress is defined as the situation of cellular damage that results when the generation and / or exogenous incidence of RLO exceeds the capacity of the different physiological mechanisms of the organism to prevent or intercept its accumulation.
Además de los mecanismos de defensa enzimáticos a lo largo de la evolución biológica, los organismos han desarrollado la capacidad de sintetizar moléculas con actividad antioxidante: ubiquinona, glutatión, ceruloplasmina, transferina y ácido úrico en mamíferos; y compuestos como ácido ascórbico (vitamina C), α-tocoferol (vitamina E), β- caroteno (pro-vitamina A), diversos flavonoides y otros compuestos fenólicos en el caso de los vegetales. Así, es bien conocida la relación entre el contenido fenólico de los extractos vegetales y su actividad antioxidante [Y. S. Velioglu et ai, Journal of Agricultura and Food Chemistry; 1998, 46 (10), 4113-4117].In addition to enzymatic defense mechanisms throughout biological evolution, organisms have developed the ability to synthesize molecules with antioxidant activity: ubiquinone, glutathione, ceruloplasmin, transferin and uric acid in mammals; and compounds such as ascorbic acid (vitamin C), α-tocopherol (vitamin E), β-carotene (pro-vitamin A), various flavonoids and other phenolic compounds in the case of vegetables. Thus, the relationship between the phenolic content of plant extracts and their antioxidant activity is well known [Y. S. Velioglu et ai, Journal of Agriculture and Food Chemistry; 1998, 46 (10), 4113-4117].
Los antioxidantes naturales además de proteger al organismo del daño producido por los radicales libres, responsables de las enfermedades degenerativas como el cáncer, arterioesclerosis, artritis y procesos neurodegenerativos y de envejecimiento; pueden presentar actividad antibactericida, antivírica, antimutagénica, antiúlcera o anticarlogénica.Natural antioxidants in addition to protecting the body from damage caused by free radicals, responsible for degenerative diseases such as cancer, arteriosclerosis, arthritis and neurodegenerative and aging processes; they can present antibactericidal, antiviral, antimutagenic, antiulcer or anticarlogenic activity.
HOJA DE SUSTITUCIÓN (REGLA 26) Además, si los antioxidantes naturales absorben radiación ultravioleta pueden ser empleados como filtros solares [EP0781544B1]. La radiación UV (ultravioleta) puede ser clasificada en UV-C (longitud de onda menor de 280 nm); UV-B (longitud de onda entre 280 nm y 320 nm) y UV-A (longitud de onda entre 320 nm y 400 nm). De estas radiaciones ultravioletas, la más letal es la radiación UV-C, aunque la mayoría de la misma es absorbida por el ozono. Por ello, las que pueden tener mayor influencia sobre la piel son las radiaciones UV-A y UV-B.SUBSTITUTE SHEET (RULE 26) In addition, if natural antioxidants absorb ultraviolet radiation they can be used as sunscreens [EP0781544B1]. UV (ultraviolet) radiation can be classified in UV-C (wavelength less than 280 nm); UV-B (wavelength between 280 nm and 320 nm) and UV-A (wavelength between 320 nm and 400 nm). Of these ultraviolet radiation, the most lethal is UV-C radiation, although most of it is absorbed by ozone. Therefore, those that may have the greatest influence on the skin are UV-A and UV-B radiation.
Estado de la técnica Couroupita guianensis (Lecitidaceaé) es un árbol original de la alta selva tropical que se extiende desde el Amazonas hasta Venezuela. También se encuentra en Panamá y Costa Rica y en la India y Sri Lanka.State of the art Couroupita guianensis (Lecitidaceaé) is an original tree of the high tropical rainforest that extends from the Amazon to Venezuela. It is also found in Panama and Costa Rica and in India and Sri Lanka.
Couroupita guianensis es una planta utilizada en la India para el tratamiento de enfermedades de la piel [a) A. S. R. Anjaneyulu et al, Indian Journal ofChemistry, Section B; 1998, 37B(4), 382-386. b) The useful plants of India; Publications & InformationCouroupita guianensis is a plant used in India for the treatment of skin diseases [a) A. S. R. Anjaneyulu et al, Indian Journal of Chemistry, Section B; 1998, 37B (4), 382-386. b) The useful plants of India; Publications & Information
Directorate; CSIR, New Delhi, 1986, 144. c) J. Anjaria et al., Nature Helas, A glossary of selected indigenous medicinal plants of India; SRISTI Innovations, 1997, 23].Directorate; CSIR, New Delhi, 1986, 144. c) J. Anjaria et al., Nature Helas, A glossary of selected indigenous medicinal plants of India; SRISTI Innovations, 1997, 23].
Se ha determinado actividad antimicrobiana, antibacteriana y antibiótica de extractos de hojas, tallos, flores, frutos, raíces y corteza de Couroupita guianensis y actividad antifúngica de extractos de tallos, corteza y flores [a) S. J. Vahanwala et al., Indian Drugs;It has been determined antimicrobial, antibacterial and antibiotic activity of extracts of leaves, stems, flowers, fruits, roots and bark of Couroupita guianensis and antifungal activity of extracts of stems, bark and flowers [a) S. J. Vahanwala et al., Indian Drugs;
2000, 37, 343-345. b) M. R. Khan et al, Journal ofherbs, spices & medicinal plants; 2003,2000, 37, 343-345. b) M. R. Khan et al, Journal ofherbs, spices & medicinal plants; 2003,
10 (3), 95-108. c) M. Poonkothai et al., Plant Archives; 2005, 5 (1), 93-95].10 (3), 95-108. c) M. Poonkothai et al., Plant Archives; 2005, 5 (1), 93-95].
Por otra parte, se ha estudiado la actividad antilarvaria de extractos de flores y frutos de Couroupita guianensis [S. T. Desal et al., Indian Drug; 2003, 40 (8), 484-486]. Asimismo, se ha determinado la actividad analgésica y antiinflamatoria de extractos de flores y corteza de Couroupita guianensis [M. Geetha et al, Journal of Natural Remedies; 2004, 4 (1), 52-55].On the other hand, the antilarvaria activity of flower and fruit extracts of Couroupita guianensis [S. has been studied. T. Desal et al., Indian Drug; 2003, 40 (8), 484-486]. Likewise, the analgesic and anti-inflammatory activity of flowers and bark extracts of Couroupita guianensis [M. Geetha et al, Journal of Natural Remedies; 2004, 4 (1), 52-55].
Por otra parte, hay estudios sobre el efecto de extractos de corteza y flores deOn the other hand, there are studies on the effect of bark and flower extracts of
Couroupita guianensis en el ciclo reproductivo en ratas [M. Geetha et al, Journal of Natural Remedies; 2005, 5 (2), 121-125] y estudios sobre el efecto de extractos de semillas de Couroupita guianensis en la reproducción de peces [G. R. Dave et al, Indian BotanicalCouroupita guianensis in the reproductive cycle in rats [M. Geetha et al, Journal of Natural Remedies; 2005, 5 (2), 121-125] and studies on the effect of Couroupita guianensis seed extracts on fish reproduction [G. R. Dave et al, Indian Botanical
Contactor; 1991, 8 (3), 99-106].Contactor; 1991, 8 (3), 99-106].
También, se ha determinado la actividad analgésica y antiinflamatoria de dos extractos, butanólico y acuoso, de las hojas de Couroupita guianensis obtenidos tras un fraccionamiento entre disolventes orgánicos de diferente polaridad (metanol, hexano, diclorometano y acetona) [Λ. M. Martínez Moran; Estudio químico de la Couropita guianensis y de la Ilex guayosa y nuevas aplicaciones sintéticas de las l-(3- feniletil)isoquinolinas y de las 2-bencilidén-l-indanonas: síntesis total de benzofluorenos y de benzofuronaftalenos, Tesis doctoral, 1999],Also, the analgesic and anti-inflammatory activity of two extracts, butanolic and aqueous, of the leaves of Couroupita guianensis obtained after a fractionation between organic solvents of different polarity (methanol, hexane, dichloromethane and acetone) [Λ. M. Martínez Moran; Chemical study of Couropita guianensis and Ilex guayosa and new synthetic applications of l- (3- phenylethyl) isoquinolines and 2-benzylidene-l-indanones: total synthesis of benzofluorenes and benzofuronaphthalenes, PhD Thesis, 1999],
Por último, hay bastantes estudios de actividad de los alcaloides indólicos, de tipo indolo[2,l-6]quinazolo-6,12-dionas, aislados de los frutos de Couroupita guianensis, y en concreto del tryphantrin y sus derivados. Estos alcaloides presentan actividad antibacteriana, antituberculosa [a) L. A. Mitscher et al., Puré & Applied Chemistry; 1998, 70 (2), 365-371. b) WO951380], antimalárica [US2006160827 Al] y actividad antiinflamatoria, por actuar como inhibidores de prostaglandinas y leucotrienos [H. Danz et al., Planta Medica; 2001, 67, 411-416].Finally, there are quite a few studies of the activity of indole alkaloids, of the indole type [2, 1-6] quinazolo-6,12-diones, isolated from the fruits of Couroupita guianensis, and in particular of tryphantrin and its derivatives. These alkaloids have antibacterial, antituberculous activity [a) L. A. Mitscher et al., Puré & Applied Chemistry; 1998, 70 (2), 365-371. b) WO951380], antimalarial [US2006160827 Al] and anti-inflammatory activity, for acting as inhibitors of prostaglandins and leukotrienes [H. Danz et al., Planta Medica; 2001, 67, 411-416].
Hay diversos estudios sobre la composición fitoquímica de Couroupita guianensis.There are several studies on the phytochemical composition of Couroupita guianensis.
Se han aislado de los frutos de Couroupita guianensis los ácidos cítrico, málico e isocítrico [a) E. K. Nelson et al., Journal of the American Chemical Society; 1937, 59, 2499-2500. b) T. R. Soderstrom, American Journal of Botany; 1962, 49(8), 850-855], ácidos clorogénicos [P. V. Pontes et al, Journal of the Science of Food and Agricultura; 2002, 82(10), 1177-1181], los esteróles estigmaesterol y camfesterol; y los alcaloides indólicos tryptanthrina, couropitina A, couropitina B o indirrubina, índigo e isatina [a) J. Bergman et al, Tetrahedron; 1985, 41(14), 2879-2881. b) J. Bergman et al, Tetrahedron Letters; 1977, 30, 2625-2626. c)A. K. Sen et al, Tetrahedron Letters; 1974, 7, 609-610. c) A. L Dyakonov et al, Chemistry of Natural Compounds; 1997, 33 (3), 221-267].Citrus, malic and isocitric acids have been isolated from the fruits of Couroupita guianensis [a) E. K. Nelson et al., Journal of the American Chemical Society; 1937, 59, 2499-2500. b) T. R. Soderstrom, American Journal of Botany; 1962, 49 (8), 850-855], chlorogenic acids [P. V. Pontes et al, Journal of the Science of Food and Agriculture; 2002, 82 (10), 1177-1181], stigmaesterol and camfesterol sterols; and the tryptanthrine, couropitin A, couropitin B or indirubin, indigo and isatin [alkaline] alkaloids [a) J. Bergman et al. 1985, 41 (14), 2879-2881. b) J. Bergman et al, Tetrahedron Letters; 1977, 30, 2625-2626. AC. K. Sen et al, Tetrahedron Letters; 1974, 7, 609-610. c) A. L Dyakonov et al, Chemistry of Natural Compounds; 1997, 33 (3), 221-267].
Hay estudios sobre el contenido en ácidos grasos, proteínas, terpenos, esteróles, y aminoácidos de las semillas de Couroupita guianensis. [a) E. H. A. Andrade et al, Journal of Food Composition and Analysis; 1999, 12(1), 37-51. b) R. C. A. Lago et al, Acta Amazónica; 1987, Volunte Date 1986, 16-17(1), 369-376. c) G. R. Dave et al, Fette, Seifen, Anstrichmittel; 1985, 87(3), 111-112. d) N. S. Bai, BuIl Central Research Inst. Univ. Travancor, Trivandrum; 1954, 3, 114-117. e) W. N. Zuo et al, Journal of Agricultural and Food Chemistry; 1996, 44(5), 1206-1210]. Se ha determinado la presencia de taninos, de terpenos, esteroides, ketosteroides y esteróles en la composición química de la corteza [a) A. S. R. Anjaneyulu, op. cited (1998). b) L. R. Row et al, Current Science; 1966, 35(6), 146-147]. Se ha determinado la presencia de antocianinas, flavonoides, ácidos cinámicos y compuestos de naturaleza lipídica, en los estigmas [F. W. Martin, American Journal of Botany; 1969, 56(9), 1023-1027].There are studies on the content of fatty acids, proteins, terpenes, sterols, and amino acids of the seeds of Couroupita guianensis. [a) EHA Andrade et al, Journal of Food Composition and Analysis; 1999, 12 (1), 37-51. b) RCA Lago et al, Amazon Act; 1987, Volunte Date 1986, 16-17 (1), 369-376. c) GR Dave et al, Fette, Seifen, Anstrichmittel; 1985, 87 (3), 111-112. d) NS Bai, BuIl Central Research Inst. Univ. Travancor, Trivandrum; 1954, 3, 114-117. e) WN Zuo et al, Journal of Agricultural and Food Chemistry; 1996, 44 (5), 1206-1210]. The presence of tannins, terpenes, steroids, ketosteroids and sterols in the chemical composition of the cortex [a) ASR Anjaneyulu, op. cited (1998). b) LR Row et al, Current Science; 1966, 35 (6), 146-147]. The presence of anthocyanins, flavonoids, cinnamic acids and compounds of a lipid nature in stigmas has been determined [FW Martin, American Journal of Botany; 1969, 56 (9), 1023-1027].
De las flores de Couroupita guianensis se han identificado, entre los componentes volátiles, linalool, eugenol, nerol, geraniol y (£,£>farnesol [a) E. H. A. Andrade et al, Journal ofEssential OiI Research; 2000, 12(2), 163-166. b) K. C. Wong et al, Journal of Essential OiI Research; 1995, 7(2), 225-227. c) Y. S. Lewis et al, Perfumery and Essential OiI Record; 1969, 60(1), 23-24. d) J. T. Knudsen et al, Biotropica; 1996, 28, 42-6O]. Asimismo, se han aislado de las flores de Couroupita guianensis un hidrocarburo alifático y estigmasterol [J. B. Rane et al, Indian Journal of Pharmaceutical Sciences; 1994, 56, 72-73].Among the flowers of Couroupita guianensis, among the volatile components, linalool, eugenol, nerol, geraniol and (£, £> farnesol [a) E. H. A. Andrade et al, Journal of Essential OiI Research; 2000, 12 (2), 163-166. b) K. C. Wong et al, Journal of Essential OiI Research; 1995, 7 (2), 225-227. c) Y. S. Lewis et al, Perfumery and Essential OiI Record; 1969, 60 (1), 23-24. d) J. T. Knudsen et al, Biotropica; 1996, 28, 42-6O]. Likewise, an aliphatic hydrocarbon and stigmasterol have been isolated from the flowers of Couroupita guianensis [J. B. Rane et al, Indian Journal of Pharmaceutical Sciences; 1994, 56, 72-73].
De las hojas de Couroupita guianensis se han aislado el éster triperpénico β-amirin palmitato [A. A. Eknath et al, Indian Drugs; 2002, 39(4), 213-216], el kaemferol-3-0- neohesperidósido, y dos ácidos hidroxicinámicos, el ácido cafeico y un derivado del mismo, el ácido rosmarínico [A. M. Martínez Moran, op. cited (1999)].From the leaves of Couroupita guianensis, the β-amirin palmitate triperpenic ester [A. A. Eknath et al, Indian Drugs; 2002, 39 (4), 213-216], kaemferol-3-0-neohesperidósido, and two hydroxycinnamic acids, caffeic acid and a derivative thereof, rosmarinic acid [A. M. Martínez Moran, op. cited (1999)].
DescripciónDescription
La presente invención describe extractos de las hojas de Couroupita guianenis, su procedimiento de obtención y su uso cosmético como antioxidantes/antirradicalarios/filtros UV (ultravioleta).The present invention describes extracts of Couroupita guianenis leaves, their method of obtaining and their cosmetic use as antioxidants / anti-radicals / UV filters (ultraviolet).
Se describe, entre otras cosas, un nuevo procedimiento para la obtención de extractos de Couroupita guianensis a partir de las hojas, que permite emplear disolventes inocuos, como son agua, glicoles y otros alcoholes, de bajo coste y operación segura para obtener un producto estable, fácil de manejar, de adicionar a diferentes productos y que puede ser empleado como ingrediente de la industria cosmética. También describe dichos extractos de Couroupita guianensis, obtenidos por el procedimiento descrito, es decir, los propios extractos de Couroupita guianensis, caracterizados por presentar, entre otros, elevado contenido fenólico y relevantes propiedades antioxidantes, antirradicalarias y de protección UV, que hace que puedan ser utilizados para el cuidado de la piel y/o el cabello, por ejemplo como antioxidantes o inhibidores de radicales, como agentes para combatir el envejecimiento o protectores solares. Los extractos presentan frecuentemente mayor actividad antioxidante que antioxidantes sintéticos como el BHT (butirohidroxitolueno), el BHA (butirohidroxianisol) o Trolox (equivalente soluble del α-tocoferol o vitamina E). La práctica de la presente invención implica: Procedimiento de obtención i) Preparación de las hojas de Couroupita guianensis:It describes, among other things, a new procedure for obtaining extracts of Couroupita guianensis from the leaves, which allows the use of harmless solvents, such as water, glycols and other alcohols, of low cost and safe operation to obtain a stable product , easy to handle, to add to different products and that can be used as an ingredient of the cosmetic industry. It also describes said extracts of Couroupita guianensis, obtained by the described procedure, that is, the extracts of Couroupita guianensis, characterized by presenting, among others, high phenolic content and relevant antioxidant, anti-radical and UV protection properties, which makes them possible to be used for skin and / or hair care, for example as antioxidants or radical inhibitors, as agents to combat aging or sunscreens. Extracts frequently have higher antioxidant activity than synthetic antioxidants such as BHT (butyrohydroxytoluene), BHA (butyrohydroxyanisole) or Trolox (soluble equivalent of α-tocopherol or vitamin E). The practice of the present invention involves: Procedure for obtaining i) Preparation of Couroupita guianensis leaves:
Se emplean las hojas secas de Couroupita guianensis molidas y tamizadas. Se someten a un molido, preferentemente en un molino de aspas, y se tamiza, preferentemente a tamaño de partícula de 0,6 mm. aproximadamente. ii) Extracción de las hojas de Couroupita guianensis con disolventes: Las hojas secas, molidas y tamizadas de Couroupita guianensis se someten a un proceso de extracción sólido-líquido en continuo con disolventes, preferentemente se realiza una extracción por soxhleL Se emplean relaciones líquido:sólido elevadas, preferentemente de 30 g/g.The dried leaves of ground and sifted Couroupita guianensis are used. They are subjected to a grind, preferably in a blade mill, and screened, preferably at a particle size of 0.6 mm. approximately. ii) Extraction of the Couroupita guianensis sheets with solvents: The dried, ground and sieved sheets of Couroupita guianensis are subjected to a continuous solid-liquid extraction process with solvents, preferably a soxhle extraction is performed Liquid: solid ratios are used high, preferably 30 g / g.
Como disolvente a extractar se emplean agua y/o disolventes hidroalcohólicos y/o alcohólicos. Se elige ventajosamente alcoholes Ci a C4 y, preferentemente, etanol o metanol. Entre ellos, se elige ventajosamente una mezcla etanol/agua 1:1.Water and / or hydroalcoholic and / or alcoholic solvents are used as solvent to be extracted. Ci to C 4 alcohols and, preferably, ethanol or methanol are advantageously chosen. Among them, a 1: 1 ethanol / water mixture is advantageously chosen.
La temperatura de la extracción es la de reflujo del disolvente o mezcla de disolventes de extracción y está en el rango de 60-1200C y, preferentemente, entre 85-95 0C. El tiempo de extracción está en el intervalo de 3 horas a 21 días. El proceso extractivo se realiza protegido de la luz para evitar posibles alteraciones. El rendimiento del proceso extractivo oscila entre el 15% y el 40% (g/g). iii) Obtención de los extractos secos o extractos acuosos de Couroupita guianensis: Los extractos acuosos, alcohólicos o hidroalcohólicos obtenidos de las hojas de Couroupita guianensis se someten a evaporación a vacío para eliminar el disolvente y obtener los extractos secos de Couroupita guianensis.The extraction temperature is the reflux of the solvent or mixture of extraction solvents and is in the range of 60-120 0 C and preferably between 85-95 0 C. The extraction time is in the range of 3 hours to 21 days The extractive process is protected from light to avoid possible alterations. The yield of the extractive process ranges from 15% to 40% (g / g). iii) Obtaining dry extracts or aqueous extracts of Couroupita guianensis: Aqueous, alcoholic or hydroalcoholic extracts obtained from the leaves of Couroupita guianensis are subjected to vacuum evaporation to remove the solvent and obtain dry extracts of Couroupita guianensis.
El agua se elimina mediante evaporación a vacío, preferentemente mediante liofilización. Si, para la obtención de los extractos líquidos de la etapa iv), se emplean como disolventes de extracción agua o hidroglicoles, en algún caso puede eliminarse a vacío sólo el disolvente alcohólico de la etapa ii) y no eliminar el agua, obteniéndose extractos acuosos de Couroupita guianensis. iv) Obtención de los extractos líquidos de Couroupita guianensis: Los extractos secos o los extractos acuosos de Couroupita guianensis obtenidos en la etapa iii), se someten a un nuevo proceso extractivo sólido-líquido en continuo con disolventes, preferentemente mediante maceración con agitación.Water is removed by evaporation under vacuum, preferably by lyophilization. If, in order to obtain the liquid extracts of stage iv), water or hydroglycols are used as extraction solvents, in some cases only the alcoholic solvent of stage ii) can be removed under vacuum and the water cannot be removed, obtaining aqueous extracts of Couroupita guianensis. iv) Obtaining liquid extracts of Couroupita guianensis: Dry extracts or aqueous extracts of Couroupita guianensis obtained in stage iii), are subjected to a new solid-liquid extractive process in continuous with solvents, preferably by maceration with stirring.
Se emplean relaciones líquido:sólido elevadas, preferentemente de 99 g/g. Como disolvente a extractar se emplea agua y/o disoluciones hidroglicólicas y/o glicólicas. Se elige ventajosamente glicoles Cz a Ce. Entre estos glicoles se prefiere utilizar propilenglicol y bútilenglicol. Entre ellos, se elige ventajosamente una mezcla butilenglicol/agua 1:1. La temperatura de extracción es la temperatura ambiente, 200C aproximadamente.High liquid: solid ratios are used, preferably 99 g / g. As solvent to be extracted, water and / or hydro-glycol and / or glycol solutions are used. Cz to Ce glycols are advantageously chosen. Among these glycols, it is preferred to use propylene glycol and butylene glycol. Among them, a 1: 1 butylene glycol / water mixture is advantageously chosen. The extraction temperature is room temperature, approximately 20 0 C.
El tiempo de extracción es prolongado, de 3 a 21 días.Extraction time is prolonged, from 3 to 21 days.
El proceso extractivo se realiza protegido de la luz para evitar posibles alteraciones, v) Obtención de los extractos líquidos filtrados de Couroupita guianensis: Los sólidos de los extractos líquidos de Couroupita guianensis, obtenidos en la etapa iv), se separan por filtración, preferentemente mediante placa porosa del n° 3 ó 4, o papel de filtro de gramaje 73 g/m2, obteniéndose los extractos líquidos filtrados de Couroupita guianensis.The extractive process is carried out protected from light to avoid possible alterations, v) Obtaining the filtered liquid extracts of Couroupita guianensis: The solids of the liquid extracts of Couroupita guianensis, obtained in stage iv), are separated by filtration, preferably by filtration porous plate No. 3 or 4, or weight filter paper 73 g / m 2 , obtaining filtered liquid extracts of Couroupita guianensis.
Estos extractos se almacenan refrigerados, preferentemente a 4 0C, y protegidos de la luz para evitar posibles alteraciones. Caracterización de los extractos líquidos filtrados de Couroupita guianensis i) Determinación de la absorción UV (ultravioleta) de los extractos líquidos filtrados de Couroupita guianensis:These extracts are stored refrigerated, preferably at 4 0 C, and protected from light to avoid possible alterations. Characterization of filtered liquid extracts of Couroupita guianensis i) Determination of UV (ultraviolet) absorption of filtered liquid extracts of Couroupita guianensis:
La radiación UV (ultravioleta) se clasifica en UV-C (longitud de onda menor de 280 nm); UV-B (longitud de onda entre 280 nm y 320 nm) y UV-A (longitud de onda entre 320 nm y 400 nm), por ello, si un producto absorbe radiación entre 250 a 400 nm puede ser empleado como filtro solar.UV (ultraviolet) radiation is classified as UV-C (wavelength less than 280 nm); UV-B (wavelength between 280 nm and 320 nm) and UV-A (wavelength between 320 nm and 400 nm), therefore, if a product absorbs radiation between 250 to 400 nm it can be used as a sunscreen.
Se mide la absorbencia (Abs) de los extractos líquidos filtrados de Couroupita guianensis, obtenidos según el procedimiento de extracción, a la concentración de 100 ppm en etanol entre las longitudes de onda de 250 a 400 nm frente a un blanco de etanol. Todos presentan máximos de absorción en la región UV (ultravioleta). Así a la longitud de onda de 250 nm presentan Abs > 1,000; a 280 nm presentan Abs > 0,800; a 320 nm presentan Abs > 0,300 y a 400 nm presentan Abs > 0, 100. ii) Determinación del contenido fenólico de los extractos líquidos filtrados de Couroupita guianensis: Los compuestos fenólicos presentan una elevada actividad antioxidante [Y. S. Velioglu, op. cited (1998)]. Por ello, se determina el contenido fenólico de los extractos líquidos de Couroupita guianensis según el método de Folin Ciocalteu's [A. Escarpa et al., Analytica Chinaca Acta; 2001, 427(1), 119-127]. Se mide la absorbancia a 765 nm de una disolución de 0,5 mL del extracto antioxidante, 3,75 mL de agua destilada, 0,25 mL del reactivo de Folin Ciocalteau's (1:1 con agua destilada) y 0,5 mL de Na2CO3 al 10 %; después de 1 hora a temperatura ambiente, frente a un blanco sin extracto. Se determina el contenido fenólico del extracto comparando la absorbencia con un recta patrón de ácido gálico (0-100 ppm). Los extractos líquidos filtrados de Couroupita guianensis, obtenidos según el procedimiento de extracción, presentan, según el procedimiento descrito, un alto contenido fenólico, entre 25-35%. iii) Determinación de la actividad captadora del radical OC, a-difenil-β-picrilhidracilo (DPPVt) y de su equivalencia en BHT y BHA de los extractos líquidos filtrados de Couroupita guianensis:The absorbency (Abs) of the filtered liquid extracts of Couroupita guianensis, obtained according to the extraction procedure, is measured at the concentration of 100 ppm in ethanol between the wavelengths of 250 to 400 nm against an ethanol blank. All have maximum absorption in the UV (ultraviolet) region. Thus at the wavelength of 250 nm they present Abs>1,000; at 280 nm they present Abs>0.800; at 320 nm they present Abs> 0.300 and at 400 nm they present Abs> 0.100. ii) Determination of the phenolic content of the filtered liquid extracts of Couroupita guianensis: The phenolic compounds have a high antioxidant activity [YS Velioglu, op. cited (1998)]. Therefore, the phenolic content of the liquid extracts of Couroupita guianensis is determined according to the Folin Ciocalteu's method [A. Escarpa et al., Analytica Chinaca Acta; 2001, 427 (1), 119-127]. The absorbance at 765 nm of a solution of 0.5 mL of the antioxidant extract, 3.75 mL of distilled water, 0.25 mL of the reagent is measured Ciocalteau's Folin (1: 1 with distilled water) and 0.5 mL of 10% Na 2 CO 3 ; after 1 hour at room temperature, against a white without extract. The phenolic content of the extract is determined by comparing the absorbency with a straight standard of gallic acid (0-100 ppm). The filtered liquid extracts of Couroupita guianensis, obtained according to the extraction procedure, have, according to the described procedure, a high phenolic content, between 25-35%. iii) Determination of the activity of the OC radical, a-diphenyl-β-picrylhydracil (DPPVt) and its equivalence in BHT and BHA of the filtered liquid extracts of Couroupita guianensis:
Para medir la actividad captadora del radical α,α-difenil-β-picrilhidracilo (DPPH*) //.To measure the activity of the radical α, α-diphenyl-β-picrylhydracil (DPPH * ) //.
Parejo et ai, Journal ofAgricultural and Food Chemistry; 2002, 50, 6882-6890], se mide la disminución de la absorbancia a 515 nm, después de 16 minutos, de una disolución de 2 mL de DPPH* 6 10"5 M en metanol y 50 μL del extracto antioxidante. El porcentaje de inhibición se calcula según la siguiente fórmula:Parejo et ai, Journal of Agricultural and Food Chemistry; 2002, 50, 6882-6890], the decrease in absorbance at 515 nm, after 16 minutes, of a 2 mL solution of DPPH * 6 10 "5 M in methanol and 50 µL of the antioxidant extract is measured. of inhibition is calculated according to the following formula:
PI (Porcentaje de Inhibición) PI(%): [(Abs t=0min-Abs t=16 min)/Abs t=0min)]*100PI (Percent Inhibition) PI (%): [(Abs t = 0min-Abs t = 16 min) / Abs t = 0min)] * 100
Se mide la IC50 o concentración que inhibe el 50% del radical DPPH*. Los extractos líquidos filtrados de Couroupita guianensis, obtenidos según el procedimiento de extracción, presentan una IC50 entre 200 a 400 ppm. La IC50 del BHA, medida según el mismo procedimiento, es de 240 ppm y la del BHT de 2790 ppm. iv) Determinación de la actividad inhibitoria de la oxidación en medio emulsionado, basada en la inhibición de la oxidación de una emulsión de β-caroteno y ácido linoleico, y de su equivalencia en BHA, de los extractos líquidos filtrados de Couroupita guianensis:The IC 50 or concentration that inhibits 50% of the DPPH * radical is measured. The filtered liquid extracts of Couroupita guianensis, obtained according to the extraction procedure, have an IC 50 between 200 and 400 ppm. The IC 50 of the BHA, measured according to the same procedure, is 240 ppm and that of the BHT of 2790 ppm. iv) Determination of oxidation inhibitory activity in an emulsified medium, based on the inhibition of the oxidation of an emulsion of β-carotene and linoleic acid, and its equivalence in BHA, of the filtered liquid extracts of Couroupita guianensis:
Para medir la actividad inhibitoria de la oxidación en medio emulsionado, basada en la inhibición de la oxidación de una emulsión de β-caroteno (pro-vitamina A) y ácido linoleico [G. /. Marco, Journal American OiI Chemists' Society; 1968, 45, 594-8], se prepara una disolución de β-caroteno (2 mg en 10 mL de cloroformo). Se añade 1 mL de esta disolución en un tubo con 20 mg de ácido linoleico y 200 mg del emulgente Tween 40. Se homogeniza. Se elimina el cloroformo a vacío. Se añaden 50 mL de agua destilada saturada de oxígeno. Se agita a 6000 rpm para formar la emulsión. Se miden las absorbancias a 470 nm de la muestra del extracto antioxidante (200 μL del extracto en 5 mL de emulsión) y del control (200 μL de etanol absoluto en 5 mL de emulsión) después de 2 horas a 500C. Se hace un blanco siguiendo el mismo procedimiento pero sin añadir β- caroteno. Se mide el coeficiente de actividad antioxidante (CAA, %), que es la relación de oxidación del β-caroteno en presencia y ausencia de antioxidante.To measure oxidation inhibitory activity in an emulsified medium, based on the oxidation inhibition of an emulsion of β-carotene (pro-vitamin A) and linoleic acid [G. /. Marco, Journal American OiI Chemists'Society; 1968, 45, 594-8], a solution of β-carotene (2 mg in 10 mL of chloroform) is prepared. 1 mL of this solution is added in a tube with 20 mg of linoleic acid and 200 mg of the Tween 40 emulsifier. It is homogenized. Chloroform is removed in vacuo. 50 mL of distilled water saturated with oxygen are added. Stir at 6000 rpm to form the emulsion. The absorbances at 470 nm of the sample of the antioxidant extract are measured (200 μL of the extract in 5 mL of emulsion) and control (200 µL of absolute ethanol in 5 mL of emulsion) after 2 hours at 50 0 C. A blank is made following the same procedure but without adding β-carotene. The coefficient of antioxidant activity (CAA,%), which is the oxidation ratio of β-carotene in the presence and absence of antioxidant, is measured.
CAA = [(Abs extract 120 min - Abs control 120 min)/(Abs control 0 min - Abs control 120 min)]*100CAA = [(Abs extract 120 min - Abs control 120 min) / (Abs control 0 min - Abs control 120 min)] * 100
Los extractos líquidos filtrados de Couroupita guianensis, obtenidos según el procedimiento de extracción, presentan a 250 ppm de concentración actividad inhibitoria de la oxidación de entre el 35% al 60%. Los valores de CAA para el BHA a 250 ppm, según el mismo procedimiento, fueron de 80%. v) Determinación de la actividad inhibitoria del radical ABTS** y de su equivalente en Trolox, de los extractos líquidos filtrados de Couroupita guianensis: Para medir la capacidad de inhibición del radical ABTS** y determinar su equivalente en Trolox (equivalente soluble de la vitamina E o α-tocoferol) [R. Re et al, Free Radical Biology and Medicine; 1999, 26, 1231-1237], se prepara un tampón de PBS pH 7,4 (8,0 g NaCl, 0,2 g KH2PO4 , 1,15 g Na2HPO4, 0,2 g KCl, 0,2 g azida sódica). Se prepara el reactivo de TEAC (38,4 mg ABTS** 7 mM, 6,62 mg persulfato potásico 2,45 mM en 10 mL de tampón PBS). Se agita durante 16 horas en ausencia de luz. Se lee la absorbancia a 734 nía Se diluye el reactivo hasta que presente una absorbancia próxima a 0,7. Se añaden 20 μL del extracto antioxidante a 2 mL del reactivo. Se calienta a 30 0C y se mide la absorbancia 734 nm a los 10 minutos. Se hace un control con etanol y un blanco con el tampón PBS. Se compara la absorbancia con los valores de una recta patrón con Trolox (0,1 a 1 mM en agua destilada).The filtered liquid extracts of Couroupita guianensis, obtained according to the extraction procedure, have an oxidation inhibitory activity concentration of between 35% and 60% at 250 ppm. The CAA values for the BHA at 250 ppm, according to the same procedure, were 80%. v) Determination of the inhibitory activity of the ABTS radical ** and its equivalent in Trolox, of the filtered liquid extracts of Couroupita guianensis: To measure the inhibition capacity of the ABTS * * radical and determine its equivalent in Trolox (soluble equivalent of the vitamin E or α-tocopherol) [R. Re et al, Free Radical Biology and Medicine; 1999, 26, 1231-1237], a PBS buffer pH 7.4 is prepared (8.0 g NaCl, 0.2 g KH 2 PO 4 , 1.15 g Na 2 HPO 4 , 0.2 g KCl, 0.2 g sodium azide). The TEAC reagent (38.4 mg ABTS ** 7 mM, 6.62 mg potassium persulfate 2.45 mM in 10 mL PBS buffer) is prepared. It is stirred for 16 hours in the absence of light. The absorbance is read at 734 years. The reagent is diluted until it has an absorbance close to 0.7. 20 μL of the antioxidant extract is added to 2 mL of the reagent. Heat at 30 0 C and the absorbance measured at 734 nm after 10 minutes. A control is made with ethanol and a blank with the PBS buffer. The absorbance is compared with the values of a standard line with Trolox (0.1 to 1 mM in distilled water).
Los extractos líquidos filtrados de Couroupita guianensis, obtenidos según el procedimiento de extracción, presentan a una concentración de 100 ppm una capacidad de inhibición del radical ABTS** equivalente a entre 0,400 a 0,800 mM de Trolox. Esto implica una relación peso/peso del extracto seco de Couroupita guianensis /Trolox de 1/1 a 1/2 (g/g). v) Determinación de la actividad antioxidante o de reducción del hierro y de su equivalencia en ácido ascórbico o vitamina C de los extractos líquidos filtrados de Couroupita guianensis: Se determina el poder reductor del hierro o actividad antioxidante de los extractos líquidos de Couroupita guianensis según el método de Oyaizu [Af. Oyaizu, J apáñese Journal of Nutrition; 1986, 44, 307-315]. El extracto antioxidante se diluye en etanol, a 1 mL del mismo se le añaden 2,5 mL de tampón fosfato pH 6,6 0,2 M y 2,5 mL de ferrocianuro potásico al 1%. Se incuba a 50 °C durante 20 minutos. Se añaden 2,5 mL de ácido tricloroacético al 10%. Se pipetean 2,5 mL del sobrenadante y se mezclan con 2,5 mL de agua destilada y 0,5 mL de cloruro férrico al 0,1%. Se mide la absorbencia a 700 nm y se comparan los resultados con un a recta patrón de ácido ascórbico o vitamina C (0,1- 1 mM). Los extractos líquidos filtrados de Couroupita guianensis, obtenidos según el procedimiento de extracción, presentan a una concentración de 100 ppm un poder reductor del hierro de entre el 20% al 40% y una equivalencia entre 0,200 mM a 0,400 mM en ácido ascórbico o vitamina C. Esto implica una relación peso/peso del extracto seco de Couroupita guianensis/áddo ascórbico o vitamina C entre 1/0,35 a 1/0,70 (g/g).The filtered liquid extracts of Couroupita guianensis, obtained according to the extraction procedure, have a capacity of inhibition of the ABTS * * radical equivalent to between 0.400 to 0.800 mM of Trolox at a concentration of 100 ppm. This implies a weight / weight ratio of the dry extract of Couroupita guianensis / Trolox from 1/1 to 1/2 (g / g). v) Determination of the antioxidant or reduction activity of iron and its equivalence in ascorbic acid or vitamin C of the filtered liquid extracts of Couroupita guianensis: The reducing power of iron or antioxidant activity of liquid extracts of Couroupita guianensis is determined according to the method of Oyaizu [Af. Oyaizu, J bathe Journal of Nutrition; 1986, 44, 307-315]. The antioxidant extract is diluted in ethanol, 2.5 mL of phosphate buffer pH 0.6 0.2 M and 2.5 mL of 1% potassium ferrocyanide are added to 1 mL thereof. Incubate at 50 ° C for 20 minutes. 2.5 mL of 10% trichloroacetic acid are added. 2.5 mL of the supernatant is pipetted and mixed with 2.5 mL of distilled water and 0.5 mL of 0.1% ferric chloride. The absorbance at 700 nm is measured and the results are compared with a straight standard of ascorbic acid or vitamin C (0.1-1 mM). The filtered liquid extracts of Couroupita guianensis, obtained according to the extraction procedure, have at a concentration of 100 ppm an iron reducing power of between 20% to 40% and an equivalence between 0.200 mM to 0.400 mM in ascorbic acid or vitamin C This implies a weight / weight ratio of the dry extract of Couroupita guianensis / ascorbic acid or vitamin C between 1 / 0.35 to 1 / 0.70 (g / g).
Ejemplo 1Example 1
Preparación del extracto hidrobutilenglicólico de Couroupita guianensis. Las hojas secas de Couroupita guianensis, molidas y tamizadas a tamaño de partícula de 0,6 mm aproximadamente, se someten a un proceso extractivo sólido-líquido en continuo, con una relación líquido: sólido de 30 g/g, mediante un soxhlet, con una mezcla etanol/agua 1:1, a la temperatura de reflujo del disolvente (85-95 0C), durante 12 días y protegido de la luz. Se evapora hasta sequedad, a vacío, el disolvente del extracto obtenido. El rendimiento de la extracción es del 36% (g/g).Preparation of the hydrobutylene glycol extract of Couroupita guianensis. The dried leaves of Couroupita guianensis, ground and sieved at a particle size of approximately 0.6 mm, are subjected to a continuous solid-liquid extractive process, with a liquid: solid ratio of 30 g / g, using a soxhlet, with a 1: 1 ethanol / water mixture, at the reflux temperature of the solvent (85-95 0 C), for 12 days and protected from light. The solvent of the extract obtained is evaporated to dryness in vacuo. The yield of extraction is 36% (g / g).
El extracto seco de Couroupita guianensis obtenido se somete a un nuevo proceso extractivo sólido-líquido en continuo, mediante maceración con agitación, con una relación líquido: sólido de 99 g/g, a temperatura ambiente y protegido de la luz, durante 16 días con una mezcla butilenglicol/agua 1:1. El extracto líquido de Couroupita guianensis obtenido se filtra, mediante placa porosa del n° 3, y el producto final, el extracto líquido filtrado deThe dried extract of Couroupita guianensis obtained is subjected to a new continuous solid-liquid extractive process, by maceration with stirring, with a liquid ratio: 99 g / g solid, at room temperature and protected from light, for 16 days with a 1: 1 butylene glycol / water mixture. The liquid extract of Couroupita guianensis obtained is filtered, using porous plate No. 3, and the final product, the filtered liquid extract of
Couroupita guianensis, se almacena refrigerado a 4 0C y protegido de la luz para evitar su alteración.Couroupita guianensis, is stored refrigerated at 4 0 C and protected from light to prevent its alteration.
- Se midió la absorbancia de este extracto a la concentración de 100 ppm en etanol a las longitudes de onda de 250 nm (Abs = 1,234), 280 nm (Abs = 1,005), 3^0 nm (Abs = 0,380) y 400 nm (Abs = 0,145) frente a un blanco de etanol. - Contenido fenólico según el método de Folin Ciocalteu's [A. Escarpa, op. cited (2001)]. Se mide la absorbencia a 765 nm de una disolución de 0,5 mL del extracto antioxidante, 3,75 mL de agua destilada, 0,25 mL del reactivo de Folin Ciocalteau's (1: 1 con agua destilada) y 0,5 mL de Na2CÜ3 al 10 %; después de 1 hora a temperatura ambiente, frente a un blanco sin extracto. Se determina el contenido fenólico del extracto comparando la absorbancia con un recta patrón de ácido gálico (0-100 ppm).- The absorbance of this extract was measured at the concentration of 100 ppm in ethanol at the wavelengths of 250 nm (Abs = 1,234), 280 nm (Abs = 1,005), 3 ^ 0 nm (Abs = 0,380) and 400 nm (Abs = 0.145) against an ethanol blank. - Phenolic content according to the method of Folin Ciocalteu's [A. Escarp, op. cited (2001)]. The absorbance at 765 nm of a solution of 0.5 mL of the antioxidant extract, 3.75 mL of distilled water, 0.25 mL of Folin Ciocalteau's reagent (1: 1 with distilled water) and 0.5 mL of 10% Na 2 CÜ3; after 1 hour at room temperature, against a white without extract. The phenolic content of the extract is determined by comparing the absorbance with a straight gallic acid standard (0-100 ppm).
El extracto obtenido según el procedimiento descrito en este ejemplo 1 presenta un contenido fenólico del 32%.The extract obtained according to the procedure described in this example 1 has a phenolic content of 32%.
- Actividad captadora del radical α,α-difenil-β-picrilhidracilo (DPPH*) //. Parejo, op. cited ( 2002)]. Se mide la disminución de la absorbancia a 515 nm, después de 16 minutos, de una disolución de 2 mL de DPPH* 6 10"5 M en metanol y 50 μL del extracto antioxidante. El porcentaje de inhibición se calcula según la siguiente fórmula:- Activity of the radical α, α-diphenyl-β-picrylhydracil (DPPH * ) //. Even, op. cited (2002)]. The decrease in absorbance at 515 nm is measured, after 16 minutes, of a 2 mL solution of 10 " 5M DPPH * 6 in methanol and 50 µL of the antioxidant extract. The percent inhibition is calculated according to the following formula:
PI (Porcentaje de Inhibición) PI(%): [(Abs t=0min-Abs t=16 min)/Abs t=0min)]*100PI (Percent Inhibition) PI (%): [(Abs t = 0min-Abs t = 16 min) / Abs t = 0min)] * 100
Se midió la IC50 o concentración que inhibe el 50% del radical DPPH*. El extracto obtenido según el procedimiento descrito en este ejemplo 1 presenta una IC50 = 205 ppm. La actividad inhibitoria del BHA, medida según el mismo procedimiento, es menor, pues presenta una IC50 = 240 ppm, al igual que la del BHT con una IC50 = 2790 ppm.The IC50 or concentration that inhibits 50% of the DPPH * radical was measured. The extract obtained according to the procedure described in this example 1 has an IC 50 = 205 ppm. The inhibitory activity of BHA, measured according to the same procedure, is lower, since it has an IC 50 = 240 ppm, just like that of BHT with an IC 50 = 2790 ppm.
- Actividad inhibitoria de la oxidación en medio emulsionado, basada en la inhibición de la oxidación de una emulsión de β-caroteno (pro-vitamina A) y ácido linoleico [G. J.- Oxidation inhibitory activity in an emulsified medium, based on the oxidation inhibition of an emulsion of β-carotene (pro-vitamin A) and linoleic acid [G. J.
Marco, op. cited (1968)]. Se prepara una disolución de β-caroteno (2 mg en 10 mL de cloroformo). Se añade 1 mL de esta disolución en un tubo con 20 mg de ácido linoleico y 200 mg del emulgente Tween 40. Se homogeniza. Se elimina el cloroformo a vacío. Se añaden 50 mL de agua destilada saturada de oxígeno. Se agita a 6000 rpm para formar la emulsión. Se miden las absorbancias a 470 nm de la muestra del extracto antioxidante (200 μL del extracto en 5 mL de emulsión) y del control (200 μL de etanol absoluto en 5 mL de emulsión) después de 2 horas a 50 0C. Se hace un blanco siguiendo el mismo procedimiento pero sin añadir β-caroteno. Se mide el coeficiente de actividad antioxidante (CAA, %), que es la relación de oxidación del β-caroteno, o pro-vitamina A, en presencia y ausencia de antioxidante.Marco, op. cited (1968)]. A solution of β-carotene (2 mg in 10 mL of chloroform) is prepared. 1 mL of this solution is added in a tube with 20 mg of linoleic acid and 200 mg of the Tween 40 emulsifier. It is homogenized. Chloroform is removed in vacuo. 50 mL of distilled water saturated with oxygen are added. Stir at 6000 rpm to form the emulsion. The absorbances at 470 nm of the sample of the antioxidant extract (200 μL of the extract in 5 mL of emulsion) and of the control (200 µL of absolute ethanol in 5 mL of emulsion) are measured after 2 hours at 50 0 C. a blank following the same procedure but without adding β-carotene. The antioxidant activity coefficient is measured (CAA,%), which is the oxidation ratio of β-carotene, or pro-vitamin A, in the presence and absence of antioxidant.
CAA = [(Abs extract 120 min - Abs control 120 min)/(Abs control 0 min - Abs control 120 min)]* 100CAA = [(Abs extract 120 min - Abs control 120 min) / (Abs control 0 min - Abs control 120 min)] * 100
El extracto obtenido según el procedimiento descrito en este ejemplo 1 presenta a 250 ppm de concentración un CAA de 58%. Los valores de CAA para el BHA a 250 ppm, según el mismo procedimiento, fueron de 80%.The extract obtained according to the procedure described in this example 1 has a CAA of 58% at 250 ppm concentration. The CAA values for the BHA at 250 ppm, according to the same procedure, were 80%.
- Actividad capacidad de inhibición del radical ABTS** y determinación de su equivalente en Trolox (equivalente soluble de la vitamina E) [R. Re, op. cited (1999)]. Se prepara un tampón de PBS pH 7,4 (8,0 g NaCl, 0,2 g KH2PO4 , 1,15 g Na2HPO4, 0,2 g KCl, 0,2 g azida sódica). Se prepara el reactivo TEAC (38,4 mg ABTS** 7 mM, 6,62 mg persulfato potásico 2,45 mM en 10 mL de tampón PBS). Se agita durante 16 horas en ausencia de luz. Se lee la absorbancia a 734 nm. Se diluye el reactivo hasta que presente una absorbancia próxima a 0,7. Se añaden 20 μL del extracto antioxidante a 2 mL del reactivo. Se calienta a 300C y se mide la absorbancia 734 nm a los 10 minutos. Se hace un control con etanol y un blanco con el tampón PBS. Se compara la absorbancia con los valores de una recta patrón con Trolox (0,1 a 1 mM en agua destilada).- ABTS ** radical inhibition ability activity and determination of its equivalent in Trolox (soluble equivalent of vitamin E) [R. Re, op. cited (1999)]. A PBS buffer pH 7.4 (8.0 g NaCl, 0.2 g KH 2 PO 4 , 1.15 g Na 2 HPO 4 , 0.2 g KCl, 0.2 g sodium azide) is prepared. The TEAC reagent (38.4 mg ABTS * * 7 mM, 6.62 mg potassium persulfate 2.45 mM in 10 mL of PBS buffer) is prepared. It is stirred for 16 hours in the absence of light. The absorbance at 734 nm is read. The reagent is diluted until it has an absorbance close to 0.7. 20 μL of the antioxidant extract is added to 2 mL of the reagent. Heat at 30 0 C and the absorbance measured at 734 nm after 10 minutes. A control is made with ethanol and a blank with the PBS buffer. The absorbance is compared with the values of a standard line with Trolox (0.1 to 1 mM in distilled water).
El extracto obtenido según el procedimiento descrito en este ejemplo 1 presenta a 100 ppm de concentración una equivalencia de 0,733 mM en Trolox. Esto implica una relación peso/peso del extracto seco de Couroupita guianensis /Trolox de 1/1,84 (g/g).The extract obtained according to the procedure described in this example 1 has an equivalent of 0.733 mM in Trolox at 100 ppm concentration. This implies a weight / weight ratio of the dry extract of Couroupita guianensis / Trolox of 1 / 1.84 (g / g).
- Poder reductor del hierro o actividad antioxidante según el método de Oyaizu y determinación de su equivalencia en ácido ascórbico o vitamina C [M Oyaizu, op. cited (1986)]. El extracto antioxidante se diluye en etanol, a 1 mL del mismo se le añaden 2,5 mL de tampón fosfato pH 6,6 0,2 M y 2,5 mL de ferrocianuro potásico al 1%. Se incuba a 50 0C durante 20 minutos. Se añaden 2,5 mL de ácido tricloroacético al 10%. Se pipetean 2,5 mL del sobrenadante y se mezclan con 2,5 mL de agua destilada y 0,5 mL de cloruro férrico al 0,1%. Se mide la absorbancia a 700 nm y se comparan los resultados con un a recta patrón de ácido ascórbico o vitamina C (0,1- 1 mM).- Reducing power of iron or antioxidant activity according to the Oyaizu method and determination of its equivalence in ascorbic acid or vitamin C [M Oyaizu, op. cited (1986)]. The antioxidant extract is diluted in ethanol, 2.5 mL of phosphate buffer pH 0.6 0.2 M and 2.5 mL of 1% potassium ferrocyanide are added to 1 mL thereof. Incubate at 50 0 C for 20 minutes. 2.5 mL of 10% trichloroacetic acid are added. 2.5 mL of the supernatant is pipetted and mixed with 2.5 mL of distilled water and 0.5 mL of 0.1% ferric chloride. The absorbance at 700 nm is measured and the results are compared with a straight standard of ascorbic acid or vitamin C (0.1-1 mM).
El extracto obtenido según el procedimiento descrito en este ejemplo 1 presenta a 100 ppm un 32% de poder reductor que equivale a 0,316 mM en ácido ascórbico o vitamina C. Esto implica una relación peso/peso del extracto seco de Couroupita guianensis /ácido ascórbico o vitamina C de 1/0,56 (g/g).The extract obtained according to the procedure described in this example 1 has at 100 ppm a 32% reducing power equivalent to 0.316 mM in ascorbic acid or vitamin C. This implies a weight / weight ratio of the dry extract of Couroupita guianensis / ascorbic acid or vitamin C of 1 / 0.56 (g / g).
Ejemplo 2 Preparación del extracto hidropropilenglicólico de Couroupita guianensis.Example 2 Preparation of the hydropropylene glycol extract of Couroupita guianensis.
Las hojas secas de Couroupita guianensis, molidas y tamizadas a tamaño de partícula de 0,6 mm aproximadamente, se someten a un proceso extractivo sólido-líquido en continuo, con una relación líquido: sólido de 30 g/g, mediante un soxhlet, con una mezcla etanol/agua 1:1, a la temperatura de reflujo del disolvente (85-95 0C), durante 12 días y protegido de la luz. Se evapora hasta sequedad, a vacío, el disolvente del extracto obtenido. El rendimiento de la extracción es del 36% (g/g).The dried leaves of Couroupita guianensis, ground and sieved at a particle size of approximately 0.6 mm, are subjected to a continuous solid-liquid extractive process, with a liquid: solid ratio of 30 g / g, using a soxhlet, with a 1: 1 ethanol / water mixture, at the reflux temperature of the solvent (85-95 0 C), for 12 days and protected from light. The solvent of the extract obtained is evaporated to dryness in vacuo. The yield of extraction is 36% (g / g).
El extracto seco de Couroupita guianensis obtenido se somete a un nuevo proceso extractivo sólido-líquido en continuo, mediante maceración con agitación, con una relación líquido: sólido de 99 g/g, a temperatura ambiente y protegido de la luz, durante 16 días con una mezcla propilenglicol/agua 1:1. El extracto líquido de Couroupita guianensis obtenido se filtra, mediante placa porosa del n° 3, y el producto final, el extracto líquido filtrado de Couroupita guianensis, se almacena refrigerado a 40C y protegido de la luz para evitar su alteración.The dried extract of Couroupita guianensis obtained is subjected to a new continuous solid-liquid extractive process, by maceration with stirring, with a liquid ratio: 99 g / g solid, at room temperature and protected from light, for 16 days with a 1: 1 propylene glycol / water mixture. The liquid extract of Couroupita guianensis obtained is filtered, using porous plate No. 3, and the final product, the filtered liquid extract of Couroupita guianensis, is stored refrigerated at 4 0 C and protected from light to prevent its alteration.
- Se midió la absorbancia de este extracto a la concentración de 100 ppm en etanol a las longitudes de onda de 250 nm (Abs = 1,049), 280 nm (Abs = 0,806), 320 nm (Abs = 0,377) y 400 nm (Abs = 0,154) frente a un blanco de etanol.- The absorbance of this extract was measured at the concentration of 100 ppm in ethanol at the wavelengths of 250 nm (Abs = 1,049), 280 nm (Abs = 0.806), 320 nm (Abs = 0.377) and 400 nm (Abs = 0.154) against an ethanol blank.
- Contenido fenólico según el método de Folin Ciocalteu's [A. Escarpa, op. cited (2001)]. Se mide la absorbancia a 765 nm de una disolución de 0,5 mL del extracto antioxidante, 3,75 mL de agua destilada, 0,25 mL del reactivo de Folin Ciocalteau's (1:1 con agua destilada) y 0,5 mL de NaaCCb al 10 %; después de 1 hora a temperatura ambiente, frente a un blanco sin extracto. Se determina el contenido fenólico del extracto comparando la absorbancia con un recta patrón de ácido gálico (0-100 ppm). El extracto obtenido según el procedimiento descrito en este ejemplo 2 presenta un contenido fenólico del 26%.- Phenolic content according to the method of Folin Ciocalteu's [A. Escarp, op. cited (2001)]. The absorbance at 765 nm of a solution of 0.5 mL of the antioxidant extract, 3.75 mL of distilled water, 0.25 mL of Folin Ciocalteau's reagent (1: 1 with distilled water) and 0.5 mL of 10% NaaCCb; after 1 hour at room temperature, against a white without extract. The phenolic content of the extract is determined by comparing the absorbance with a straight gallic acid standard (0-100 ppm). The extract obtained according to the procedure described in this example 2 has a phenolic content of 26%.
- Actividad captadora del radical α,α-difenil-β-picrilhidracilo (DPPH*) [I. Parejo, op. cited C 2002)]. Se mide la disminución de la absorbancia a 515 nm después de 16 minutos de una disolución de 2 mL de DPPH" 6 10"5 M en metanol y 50 μL del extracto antioxidante. El porcentaje de inhibición se calcula según la siguiente fórmula:- Activity of the radical α, α-diphenyl-β-picrylhydracil (DPPH * ) [I. Even, op. cited C 2002)]. The absorbance decrease at 515 nm is measured after 16 minutes of a 2 mL solution of DPPH "6 10 " 5 M in methanol and 50 µL of the antioxidant extract. The percentage of inhibition is calculated according to the following formula:
PI (Porcentaje de Inhibición) PI(%): [(Abs t=0min-Abs t=16 min)/Abs t=0min)]*100PI (Percent Inhibition) PI (%): [(Abs t = 0min-Abs t = 16 min) / Abs t = 0min)] * 100
Se midió la IC50 o concentración que inhibe el 50% del radical DPPH*. El extracto obtenido según el procedimiento descrito en este ejemplo 2 presenta una IC50 = 327 ppm. La actividad inhibitoria del BHA, medida según el mismo procedimiento, es mayor pues presenta una IC50 = 240 ppm, y la del BHT, con IC50 = 2790 ppm, menor.The IC 50 or concentration that inhibits 50% of the DPPH * radical was measured. The extract obtained according to the procedure described in this example 2 has an IC 50 = 327 ppm. The inhibitory activity of BHA, measured according to the same procedure, is greater because it has an IC 50 = 240 ppm, and that of BHT, with IC 50 = 2790 ppm, lower.
- Actividad inhibitoria de la oxidación en medio emulsionado, basada en la inhibición de la oxidación de una emulsión de β-caroteno y ácido linoleico [G. /. Marco, op. cited (1968)]. Se prepara una disolución de β-caroteno (2 mg en 10 mL de cloroformo). Se añade 1 mL de esta disolución en un tubo con 20 mg de ácido linoleico y 200 mg del emulgente Tween 40. Se homogeniza. Se elimina el cloroformo a vacío. Se añaden 50 mL de agua destilada saturada de oxígeno. Se agita a 6000 rpm para formar la emulsión. Se miden las absorbancias a 470 nm de la muestra del extracto antioxidante (200 μL del extracto en 5 mL de emulsión) y del control (200 μL de etanol absoluto en 5 mL de emulsión) después de 2 horas a 50 0C. Se hace un blanco siguiendo el mismo procedimiento pero sin añadir β-caroteno. Se mide el coeficiente de actividad antioxidante (CAA, %), que es la relación de oxidación del β-caroteno en presencia y ausencia de antioxidante.- Oxidation inhibitory activity in emulsified medium, based on the oxidation inhibition of an emulsion of β-carotene and linoleic acid [G. /. Marco, op. cited (1968)]. A solution of β-carotene (2 mg in 10 mL of chloroform) is prepared. 1 mL of this solution is added in a tube with 20 mg of linoleic acid and 200 mg of the Tween 40 emulsifier. It is homogenized. Chloroform is removed in vacuo. 50 mL of distilled water saturated with oxygen are added. Stir at 6000 rpm to form the emulsion. The absorbances at 470 nm of the sample of the antioxidant extract (200 μL of the extract in 5 mL of emulsion) and of the control (200 µL of absolute ethanol in 5 mL of emulsion) are measured after 2 hours at 50 0 C. a blank following the same procedure but without adding β-carotene. The coefficient of antioxidant activity (CAA,%), which is the oxidation ratio of β-carotene in the presence and absence of antioxidant, is measured.
CAA = [(Abs extract 120 min - Abs control 120 min)/(Abs control 0 min - Abs control 120 min)]*100CAA = [(Abs extract 120 min - Abs control 120 min) / (Abs control 0 min - Abs control 120 min)] * 100
El extracto obtenido según el procedimiento descrito en este ejemplo 2 presenta a 250 ppm de concentración un CAA del 40%. Los valores de CAA para el BHA a 250 ppm, según el mismo procedimiento, fueron de 80%.The extract obtained according to the procedure described in this example 2 has a CAA of 40% at 250 ppm concentration. The CAA values for the BHA at 250 ppm, according to the same procedure, were 80%.
- Actividad capacidad de inhibición del radical ABTS1+ y determinación de su equivalente en Trolox (equivalente soluble de la vitamina E) [R. Re, op. cited (1999)]. Se prepara un tampón de PBS pH 7,4 (8,0 g NaCl, 0,2 g KH2PO4 , 1,15 g Na2HPO4, 0,2 g KCl, 0,2 g azida sódica). Se prepara el reactivo TEAC (38,4 mg ABTS** 7 mM, 6,62 mg persulfato potásico 2,45 mM en 10 mL de tampón PBS). Se agita durante 16 horas en ausencia de luz. Se lee la absorbancia a 734 nm. Se diluye el reactivo hasta que presente una absorbancia próxima a 0,7. Se añaden 20 μL del extracto antioxidante a 2 mL del reactivo. Se calienta a 30 "C y se mide la absorbancia 734 nm a los 10 minutos. Se hace un control con etanol y un blanco con el tampón PBS. Se compara la absorbancia con los valores de una recta patrón con Trolox (0,1 a 1 mM en agua destilada).- ABTS 1+ radical inhibition ability activity and determination of its equivalent in Trolox (soluble equivalent of vitamin E) [R. Re, op. cited (1999)]. A PBS buffer pH 7.4 (8.0 g NaCl, 0.2 g KH 2 PO 4 , 1.15 g Na 2 HPO 4 , 0.2 g is prepared KCl, 0.2 g sodium azide). The TEAC reagent (38.4 mg ABTS ** 7 mM, 6.62 mg potassium persulfate 2.45 mM in 10 mL of PBS buffer) is prepared. It is stirred for 16 hours in the absence of light. The absorbance at 734 nm is read. The reagent is diluted until it has an absorbance close to 0.7. 20 μL of the antioxidant extract is added to 2 mL of the reagent. It is heated to 30 "C and the absorbance is measured 734 nm at 10 minutes. A control is made with ethanol and a blank with the PBS buffer. The absorbance is compared with the values of a standard line with Trolox (0.1 a 1 mM in distilled water).
El extracto obtenido según el procedimiento descrito en este ejemplo 2 presenta a 100 ppm de concentración una equivalencia de 0,525 mM en Trolox. Esto implica una relación peso/peso del extracto seco de Couroupita guianensis /Trolox de 1/1,32 (g/g).The extract obtained according to the procedure described in this example 2 has an equivalence of 0.525 mM in Trolox at 100 ppm concentration. This implies a weight / weight ratio of the dry extract of Couroupita guianensis / Trolox of 1 / 1.32 (g / g).
- Poder reductor del hierro o actividad antioxidante según el método de Oyaizu [M Oyaizu, op. cited (1986)]. El extracto antioxidante se diluye en etanol, a 1 mL del mismo se le añaden 2,5 mL de tampón fosfato pH 6,60,2 M y 2,5 mL de ferrocianuro potásico al 1%. Se incuba a 50 0C durante 20 minutos. Se añaden 2,5 mL de ácido tricloroacético al 10%. Se pipetean 2,5 mL del sobrenadante y se mezclan con 2,5 mL de agua destilada y 0,5 mL de cloruro férrico al 0,1%. Se mide la absorbancia a 700 nm y se comparan los resultados con un a recta patrón de ácido ascórbico o vitamina C (0,1- 1 mM).- Reducing power of iron or antioxidant activity according to the method of Oyaizu [M Oyaizu, op. cited (1986)]. The antioxidant extract is diluted in ethanol, 2.5 mL of pH 6.60.2 M phosphate buffer and 2.5 mL of 1% potassium ferrocyanide are added to 1 mL thereof. Incubate at 50 0 C for 20 minutes. 2.5 mL of 10% trichloroacetic acid are added. 2.5 mL of the supernatant is pipetted and mixed with 2.5 mL of distilled water and 0.5 mL of 0.1% ferric chloride. The absorbance at 700 nm is measured and the results are compared with a straight standard of ascorbic acid or vitamin C (0.1-1 mM).
El extracto obtenido según el procedimiento descrito en este ejemplo 2 presenta a 100 ppm un 23% de poder reductor que equivale a 0,316 mM en ácido ascórbico o vitamina C. Esto implica una relación peso/peso del extracto seco de Couroupita guianensis/ácido ascórbico o vitamina C de 1/0,40 (g/g). The extract obtained according to the procedure described in this example 2 has at 100 ppm a 23% reducing power equivalent to 0.316 mM in ascorbic acid or vitamin C. This implies a weight / weight ratio of the dry extract of Couroupita guianensis / ascorbic acid or Vitamin C of 1 / 0.40 (g / g).

Claims

REVINDICACIONES REVINDICATIONS
1. Procedimiento de obtención de extractos de Couropita guianensis caracterizado porque comprende las siguientes etapas: i) Secado, molienda y tamización de las hojas de Couropita guianensis. ii) Extracción sólido-líquido en continuo de las hojas secas, molidas y tamizadas de1. Procedure for obtaining extracts of Couropita guianensis characterized in that it comprises the following stages: i) Drying, milling and sieving of the leaves of Couropita guianensis. ii) Continuous solid-liquid extraction of dried, ground and sieved leaves of
Couropita guianensis con disolventes acuosos, alcohólicos o hidroalcohólicos. iii) Evaporación hasta sequedad del disolvente de la extracción de la etapa ii) para la obtención de los extractos secos de Couropita guianensis; o evaporación del disolvente alcohólico de la etapa ii) para la obtención de los extractos acuosos de Couroupita guianensis. iv) Extracción sólido-líquido en continuo de los extractos secos o acuosos de la etapa iii) con disolventes acuosos, glicólicos o hidroglicólicos para la obtención de los extractos líquidos de Couropita guianensis. v) Filtración de los extractos líquidos de la etapa iv) para la obtención de los extractos líquidos filtrados de Couropita guianensis y almacenamiento en frío y ausencia de luz.Couropita guianensis with aqueous, alcoholic or hydroalcoholic solvents. iii) Evaporation to dryness of the solvent from the extraction of stage ii) to obtain the dry extracts of Couropita guianensis; or evaporation of the alcoholic solvent of step ii) to obtain the aqueous extracts of Couroupita guianensis. iv) Continuous solid-liquid extraction of the dry or aqueous extracts from step iii) with aqueous, glycolic or hydro-glycol solvents to obtain the liquid extracts of Couropita guianensis. v) Filtration of the liquid extracts of stage iv) to obtain the filtered liquid extracts of Couropita guianensis and cold storage and absence of light.
2. Procedimiento de obtención de extractos de Couropita guianensis, según la reivindicación 1, etapa i), caracterizado porque se emplean las hojas secas molidas en un molino de aspas, y se tamizan a tamaño de partícula de 0,6 mm. aproximadamente.2. Procedure for obtaining extracts of Couropita guianensis, according to claim 1, step i), characterized in that the dried leaves ground in a blade mill are used and screened at a particle size of 0.6 mm. approximately.
3. Procedimiento de obtención de extractos de Couropita guianensis, según la reivindicación 1, etapa ii), caracterizado porque el proceso de extracción sólido-líquido en continuo con disolventes se realiza mediante un proceso extractivo por soxhlet; se emplean relaciones líquido:sólido elevadas, preferentemente de 30 g/g; el disolvente es agua, o un alcohol Ci a Gt, preferentemente metanol o etanol, o una mezcla metanol/agua, o etanol/agua; la temperatura de la extracción es la de reflujo del disolvente o mezcla de disolventes de extracción y es^á en el rango de 60-120 0C; el tiempo de extracción está en el intervalo de 3 horas a 21 días; y el proceso extractivo se realiza protegido de la luz.3. Procedure for obtaining Couropita guianensis extracts, according to claim 1, step ii), characterized in that the continuous solid-liquid extraction process with solvents is carried out by means of a soxhlet extraction process; high liquid: solid ratios are used, preferably 30 g / g; the solvent is water, or a Ci to Gt alcohol, preferably methanol or ethanol, or a methanol / water mixture, or ethanol / water; the temperature of the extraction is the reflux of the solvent or mixture of extraction solvents and is in the range of 60-120 0 C; the extraction time is in the range of 3 hours to 21 days; and the extractive process is carried out protected from light.
4. Procedimiento de obtención de extractos de Couropita guianensis, según la reivindicación 3, caracterizado porque el disolvente de extracción es una mezcla etanol/agua 1:1; y la temperatura de la extracción es la de reflujo de la mezcla de disolventes y está en el rango de 85-950C.4. Procedure for obtaining Couropita guianensis extracts according to claim 3, characterized in that the extraction solvent is a 1: 1 ethanol / water mixture; and the extraction temperature is the reflux of the solvent mixture and is in the range of 85-95 0 C.
5. Procedimiento de obtención de extractos de Couropita guianensis, según la reivindicación 1, etapa iii), caracterizado porque la evaporación hasta sequedad del disolvente para la obtención de los extractos secos de Couropita guianensis, se realiza a vacío y el agua se elimina, preferentemente, mediante liofilización. 5. Procedure for obtaining Couropita guianensis extracts, according to claim 1, step iii), characterized in that the evaporation to dryness of the solvent to obtain the dried extracts of Couropita guianensis is carried out under vacuum and the water is preferably removed , by lyophilization.
6. Procedimiento de obtención de extractos de Couropita guianensis, según la reivindicación 1, etapa iii), caracterizado porque la evaporación del disolvente alcohólico para la obtención de los extractos acuosos de Couropita guianensis, se realiza a vacío.6. Procedure for obtaining extracts of Couropita guianensis, according to claim 1, step iii), characterized in that the evaporation of the alcoholic solvent to obtain the aqueous extracts of Couropita guianensis is carried out under vacuum.
7. Procedimiento de obtención de extractos de Couropita guianensis, según la reivindicación 1, etapa iv), caracterizado porque el proceso de extracción sólido-líquido en continuo con disolventes se realiza mediante maceración con agitación; se emplean relaciones líquido: sólido elevadas, preferentemente de 99 g/g; el disolvente es agua, o un glicol C2 a Ce, preferentemente propilenglicol o butilenglicol, o una mezcla propilenglicol/agua, o butilenglicol/agua; la temperatura de extracción es la temperatura ambiente, 20 0C aproximadamente; el tiempo de la extracción es prolongado, de 3 a 21 días y se realiza protegido de la luz.7. Procedure for obtaining Couropita guianensis extracts, according to claim 1, step iv), characterized in that the continuous solid-liquid extraction process with solvents is carried out by maceration with stirring; high liquid: solid ratios are used, preferably 99 g / g; the solvent is water, or a C 2 to Ce glycol, preferably propylene glycol or butylene glycol, or a mixture of propylene glycol / water, or butylene glycol / water; the extraction temperature is room temperature, about 20 0 C; The extraction time is prolonged, from 3 to 21 days and is carried out protected from light.
8. Procedimiento de obtención de extractos de Couropita guianensis, según la reivindicación 7, caracterizado porque el disolvente de extracción es una mezcla butilenglicol /agua 1:1. 8. Method of obtaining extracts of Couropita guianensis, according to claim 7, characterized in that the extraction solvent is a 1: 1 butylene glycol / water mixture.
9. Procedimiento de obtención de extractos de Couropita guianensis, según la reivindicación 1, etapa v), caracterizado porque se filtran los extractos líquidos obtenidos en la etapa iv), preferentemente mediante placa porosa del n° 3 ó 4, o papel de filtro de gramaje 73 g/m2, y los extractos líquidos filtrados de Couropita guianensis se almacenan en frío, preferentemente a 40C, y protegidos de la luz. 9. Procedure for obtaining Couropita guianensis extracts, according to claim 1, step v), characterized in that the liquid extracts obtained in stage iv) are filtered, preferably by porous plate No. 3 or 4, or filter paper of weight 73 g / m 2 , and the filtered liquid extracts of Couropita guianensis are stored cold, preferably at 4 0 C, and protected from light.
10. Extractos secos de Couropita guianensis obtenidos según las revindicaciones 2 a 5.10. Dry extracts of Couropita guianensis obtained according to claims 2 to 5.
11. Extractos líquidos de Couropita guianensis obtenidos según las revindicaciones 10, y 6 a 9.11. Liquid extracts of Couropita guianensis obtained according to claims 10, and 6 to 9.
12. Extractos líquidos filtrados de Couropita guianensis, según la reivindicación 11, caracterizados por presentar absorción ultravioleta entre las longitudes de onda de 250 nm a 400 nm; por poseer un alto contenido fenólico; por su capacidad de inhibición del radical DPPH*; por su capacidad de inhibición de la oxidación del sistema micelar β-caroteno y ácido linoleico; por su capacidad de inhibición del radical ABTS1+; por su capacidad antioxidante o de reducción del hierro.12. Filtered liquid extracts of Couropita guianensis according to claim 11, characterized in that they have ultraviolet absorption between the wavelengths of 250 nm to 400 nm; for having a high phenolic content; for its ability to inhibit the DPPH * radical; for its ability to inhibit oxidation of the micellar system β-carotene and linoleic acid; for its ability to inhibit the radical ABTS 1+ ; for its antioxidant or iron reduction capacity.
13. Extractos líquidos filtrados de Couroupita guianensis, según la reivindicación 12, caracterizados por presentar a la concentración de 100 ppm a las longitudes de onda de 250 nm (Abs > 1,000), 280 nm (Abs > 0,800), 320 nm (Abs > 0,300) y 400 nm (Abs > 0,100); por poseer un contenido fenólico entre 25-35%; por presentar una IC50 entre 200 a 400 ppm de inhibición del radical DPPH*; por presentar un coeficiente de actividad antioxidante (CAA, %) entre el 35% al 60% del sistema micelar β-caroteno y ácido linoleico; por presentar a 100 ppm una capacidad de inhibición del radical ABTS*+ equivalente entre 0,400 a 0,800 mM de Trolox y por su capacidad antioxidante o de reducción del hierro a 100 ppm de entre un 20% al 40% o una equivalencia entre 0,200 mM a 0,400 mM en ácido ascórbico o vitamina C.13. Filtered liquid extracts of Couroupita guianensis according to claim 12, characterized by having a concentration of 100 ppm at wavelengths of 250 nm (Abs> 1,000), 280 nm (Abs> 0.800), 320 nm (Abs> 0.300) and 400 nm (Abs>0.100); for having a phenolic content between 25-35%; for presenting an IC 50 between 200 and 400 ppm of DPPH * radical inhibition; for presenting a coefficient of antioxidant activity (CAA,%) between 35% to 60% of the β-carotene micellar system and linoleic acid; for presenting at 100 ppm a capacity for inhibition of the ABTS * + radical equivalent between 0.400 to 0.800 mM of Trolox and for its antioxidant capacity or reduction of iron at 100 ppm between 20% to 40% or an equivalence between 0.200 mM at 0.400 mM in ascorbic acid or vitamin C.
14. Extracto líquido filtrado de Couroupita guianensis, según la reivindicación 13, caracterizado por presentar a la concentración de 100 ppm a las longitudes de onda de 250 nm (Abs = 1,234), 280 nm (Abs = 1,005), 320 nm (Abs = 0,380) y 400 nm (Abs = 0,145); por poseer un contenido fenólico del 32%; por presentar una IC50 = 205 ppm de inhibición del radical DPPH*; por presentar un coeficiente de actividad antioxidante (CAA, %) del 58% del sistema micelar β-caroteno y ácido linoleico; por presentar a 100 ppm una capacidad de inhibición del radical ABTS** equivalente a 0,733 mM en Trolox y por su capacidad antioxidante o de reducción del hierro a 100 ppm de un 32%, lo que equivale a 0,316 mM en ácido ascórbico o vitamina C. 14. Couroupita guianensis filtered liquid extract according to claim 13, characterized in that it has a concentration of 100 ppm at wavelengths of 250 nm (Abs = 1,234), 280 nm (Abs = 1,005), 320 nm (Abs = 0.380) and 400 nm (Abs = 0.145); for having a phenolic content of 32%; for presenting an IC 50 = 205 ppm of DPPH * radical inhibition; for presenting a coefficient of antioxidant activity (CAA,%) of 58% of the micellar system β-carotene and linoleic acid; for presenting at 100 ppm a capacity for inhibition of the ABTS radical ** equivalent to 0.733 mM in Trolox and for its antioxidant capacity or reduction of iron at 100 ppm of 32%, which is equivalent to 0.316 mM in ascorbic acid or vitamin C .
15. Uso de los extractos de Couroupita guianensis, según las reivindicaciones 10 a 14, como agente cosmético para el cuidado de la piel y/o el cabello, destinado a actuar como agente antienvejecimiento y antioxidante, por prevenir las alteraciones derivadas de la oxidación celular o el estrés oxidativo, y por actuar como inhibidor de radicales y protector contra radiaciones ionizantes y radicales libres. 15. Use of Couroupita guianensis extracts, according to claims 10 to 14, as a cosmetic agent for skin and / or hair care, intended to act as an anti-aging and antioxidant agent, for preventing alterations derived from cellular oxidation or oxidative stress, and for acting as a radical inhibitor and protector against ionizing radiation and free radicals.
16. Uso de los extractos de Couroupita guianensis, según la reivindicaciones 10 a 14, como agente cosmético para el cuidado de la piel y/o el cabello, destinado a actuar como absorbente de la radiación UV (ultravioleta), lo que le aporta propiedades de filtro solar. 16. Use of Couroupita guianensis extracts, according to claims 10 to 14, as a cosmetic agent for skin and / or hair care, intended to act as a UV (ultraviolet) absorber, which gives it properties of sunscreen
PCT/ES2008/000178 2007-03-30 2008-03-27 Extract of the leaves of couroupita guianensis, method for obtaining same and cosmetic use thereof as an antioxidant/antirradical/uv filter WO2008119852A1 (en)

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