ES2352403B1 - SAXIFRAGA SPATHULARIS EXTRACTS, PROCEDURE FOR OBTAINING AND ITS COSMETIC, PHARMACEUTICAL AND / OR FOOD USE AS ANTIOXIDANTS / ANTIRRADICALARIES / UV FILTERS (ULTRAVIOLET). - Google Patents
SAXIFRAGA SPATHULARIS EXTRACTS, PROCEDURE FOR OBTAINING AND ITS COSMETIC, PHARMACEUTICAL AND / OR FOOD USE AS ANTIOXIDANTS / ANTIRRADICALARIES / UV FILTERS (ULTRAVIOLET). Download PDFInfo
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- ES2352403B1 ES2352403B1 ES200901653A ES200901653A ES2352403B1 ES 2352403 B1 ES2352403 B1 ES 2352403B1 ES 200901653 A ES200901653 A ES 200901653A ES 200901653 A ES200901653 A ES 200901653A ES 2352403 B1 ES2352403 B1 ES 2352403B1
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- saxifraga
- spathularis
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- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 claims 1
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Classifications
-
- 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
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Engineering & Computer Science (AREA)
- Botany (AREA)
- Mycology (AREA)
- Epidemiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Medical Informatics (AREA)
- Alternative & Traditional Medicine (AREA)
- Medicinal Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Dermatology (AREA)
- Birds (AREA)
- Cosmetics (AREA)
Abstract
Extractos de Saxifraga spathularis, procedimiento de obtención y su uso cosmético, farmacéutico y/o alimentario como antioxidantes/antirradicalarios/filtros UV (ultravioleta). Extractos obtenidos a partir de hojas, raíces, tallos y flores secos y/o congelados, molidos y tamizados, extracción con agua, y/o alcoholes, evaporación de este disolvente, posterior extracción con agua y/o glicoles del extracto seco y filtración; y su uso cosmético, farmacéutico y/o alimentario, 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.Extracts of Saxifraga spathularis, procedure for obtaining and its cosmetic, pharmaceutical and / or food use as antioxidants / anti-radicals / UV filters (ultraviolet). Extracts obtained from leaves, roots, stems and dried and / or frozen flowers, ground and sieved, extraction with water, and / or alcohols, evaporation of this solvent, subsequent extraction with water and / or glycols of the dry extract and filtration; and its cosmetic, pharmaceutical and / or food use, for skin and / or hair care, for example as antioxidants or radical inhibitors, as agents for combating aging or sunscreens.
Description
Extractos de Saxifraga spathularis, procedimiento de obtención y su uso cosmético, farmacéutico y/o alimentario como antioxidantes/antirradicalarios/filtros UV (ultravioleta). Extracts of Saxifraga spathularis, procedure for obtaining and its cosmetic, pharmaceutical and / or food use as antioxidants / anti-radiation / UV filters (ultraviolet).
Objeto de la invención Object of the invention
La presente invención se engloba dentro del campo técnico de los extractos vegetales de Saxifraga spathularis,y su obtención para su uso cosmético, farmacéutico y/o alimentario como antioxidantes, antirradicalarios y filtros UV. The present invention falls within the technical field of plant extracts of Saxifraga spathularis, and its preparation for cosmetic, pharmaceutical and / or food use as antioxidants, anti-radicals and UV filters.
Antecedentes de la invención Background of the invention
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: 1O• 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. At 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: 1O • 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.
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 catalasa. Therefore, since the ERO / RLO are species that are generated continuously as products of the cellular use of O2, biological strategies have been developed in aerobic organisms 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 enough 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 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), βcaroteno (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 al., Journal of Agricultural 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 fl avonoids 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 al., Journal of Agricultural 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.
Además, si los antioxidantes naturales absorben radiación ultravioleta pueden ser empleados como filtros solares [EPO781544B1]. 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. In addition, if natural antioxidants absorb ultraviolet radiation they can be used as solar filters [EPO781544B1]. 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.
Saxifraga spathularis (Plumbaginaceae) es una planta endémica del noroeste de la Península Ibérica (España y Portugal) e Irlanda. Se ha reportado su uso interno como antiinflamatoria y analgésica para tratamientos veterinarios Saxifraga spathularis (Plumbaginaceae) is an endemic plant in the northwest of the Iberian Peninsula (Spain and Portugal) and Ireland. Its internal use as an anti-inflammatory and analgesic for veterinary treatments has been reported.
(E. Blanco et al., El Caurel, las plantas y sus habitantes. Estudio etnobotánico de la Sierra del Caurel (Lugo): la importancia de las plantas para nuestros antepasados; Fundación Caixa Galicia, A Coruña, 1996, 203) y su uso tópico para el tratamiento de forúnculos (M. P. González-Hernández et al., A proceedings of the XXVI International Horticultural Congress, Toronto, 2002, 63-75). (E. Blanco et al., El Caurel, plants and their inhabitants. Ethnobotanical study of the Sierra del Caurel (Lugo): the importance of plants for our ancestors; Caixa Galicia Foundation, A Coruña, 1996, 203) and its topical use for the treatment of boils (MP González-Hernández et al., A proceedings of the XXVI International Horticultural Congress, Toronto, 2002, 63-75).
Descripción de la invención Description of the invention
La presente invención describe extractos de las hojas, flores, raíces y tallos de Saxifraga spathularis su procedimiento de obtención y su uso cosmético, farmacéutico y/o alimentario como antioxidantes/antirradicalarios/filtros UV (ultravioleta). The present invention describes extracts of the leaves, flowers, roots and stems of Saxifraga spathularis, its method of obtaining and its cosmetic, pharmaceutical and / or food use as antioxidants / anti-radical / UV filters (ultraviolet).
Se describe, entre otras cosas, un nuevo procedimiento para la obtención de extractos de Saxifraga spathularis a partir de las hojas, flores, raíces y tallos, 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 Saxifraga spathularis obtenidos por el procedimiento descrito, 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) o el BHA (butirohidroxianisol). It describes, among other things, a new procedure for obtaining extracts of Saxifraga spathularis from the leaves, flowers, roots and stems, which allows the use of harmless solvents, such as water, glycols and other alcohols, of low cost and operation safe 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 Saxifraga spathularis obtained by the described procedure, characterized by presenting, among others, high phenolic content and relevant antioxidant, anti-radical and UV protection properties, which makes them suitable for skin care and / or hair, 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) or BHA (butyrohydroxyanisole).
La práctica de la presente invención implica: The practice of the present invention involves:
Procedimiento de obtención Obtaining procedure
i) Preparación de las hojas, flores, raíces y tallos de Saxifraga spathularis: i) Preparation of the leaves, flowers, roots and stems of Saxifraga spathularis:
Se emplean las hojas, flores, raíces y tallos secos o congelados de Saxifraga spathularis 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. The dried and frozen leaves, flowers, roots and stems of ground and sifted Saxifraga spathularis 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) Extracción de hojas, flores, raíces y tallos de Saxifraga spathularis con disolventes: Las hojas secas o congeladas molidas y tamizadas se someten a un proceso de extracción sólido-líquido en continuo ii) Extraction of leaves, flowers, roots and stems of Saxifraga spathularis with solvents: The dried or frozen leaves ground and sieved are subjected to a continuous solid-liquid extraction process
con disolventes. with solvents.
Se emplean relaciones líquido:sólido elevadas, preferentemente de 30 g/g. High liquid: solid ratios are used, preferably 30 g / g.
Como disolvente a extractar se emplean agua y/o disolventes hidroalcohólicos y/o alcohólicos. Se elige ventaWater and / or hydroalcoholic and / or alcoholic solvents are used as solvent to be extracted. Sale is chosen
josamente alcoholes C1 aC4 y, preferentemente, etanol o metanol. Entre ellos, se elige ventajosamente una mezcla etanol/agua 1:1. especially C1 aC4 alcohols and, preferably, ethanol or methanol. 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 60-120ºC y, preferentemente, entre 85-95ºC. The temperature of the extraction is the reflux of the solvent or mixture of extraction solvents and is in the range of 60-120 ° C and, preferably, between 85-95 ° C.
El tiempo de extracción está en el intervalo de1a3 horas. The extraction time is in the range of 1-3 hours.
El proceso extractivo se realiza protegido de la luz para evitar posibles alteraciones. The extractive process is protected from light to avoid possible alterations.
El rendimiento del proceso extractivo oscila entre el 25% y el 30% (g/g). The yield of the extractive process ranges from 25% to 30% (g / g).
iii) Obtención de los extractos secos o extractos acuosos de Saxifraga spathularis: iii) Obtaining dry extracts or aqueous extracts of Saxifraga spathularis:
Los extractos acuosos, alcohólicos o hidroalcohólicos obtenidos se someten a evaporación a vacío para eliminar el disolvente y obtener los extractos secos de Saxifraga spathularis. The aqueous, alcoholic or hydroalcoholic extracts obtained are subjected to vacuum evaporation to remove the solvent and obtain the dried extracts of Saxifraga spathularis.
El agua se elimina mediante evaporación a vacío, preferentemente mediante liofilización. Water is removed by evaporation under vacuum, preferably by lyophilization.
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 Saxifraga spathularis. 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 from Saxifraga spathularis.
iv) Obtención de los extractos líquidos de Saxifraga spathularis: iv) Obtaining the liquid extracts of Saxifraga spathularis:
Los extractos secos o los extractos acuosos de Saxifraga spathularis obtenidos en la etapa iii), se someten a un nuevo proceso extractivo sólido-líquido en continuo con disolventes, preferentemente mediante maceración con agiDry extracts or aqueous extracts of Saxifraga spathularis obtained in step iii), are subjected to a new solid-liquid extractive process in continuous with solvents, preferably by maceration with agi
tación. Lotion
Se emplean relaciones líquido:sólido elevadas, preferentemente de 99 g/g. High liquid: solid ratios are used, preferably 99 g / g.
Como disolvente a extractar se emplea agua y/o disoluciones hidroglicólicas y/o glicólicas. Se elige ventajosaAs solvent to be extracted, water and / or hydro-glycol and / or glycol solutions are used. It is chosen advantageously
mente glicoles C2 aC6. Entre estos glicoles se prefiere utilizar propilenglicol y butilenglicol. Entre ellos, se elige ventajosamente una mezcla butilenglicol/agua 1:1. mind glycols C2 BC6. Among these glycols, propylene glycol and butylene glycol are preferred. Among them, a 1: 1 butylene glycol / water mixture is advantageously chosen.
La temperatura de extracción es la temperatura ambiente, 20ºC aproximadamente. The extraction temperature is room temperature, approximately 20 ° C.
El tiempo de extracción es de 1 a 7 días. The extraction time is 1 to 7 days.
El proceso extractivo se realiza protegido de la luz para evitar posibles alteraciones. The extractive process is protected from light to avoid possible alterations.
v) Obtención de los extractos líquidos filtrados de Saxifraga spathularis: v) Obtaining the fi ltered liquid extracts of Saxifraga spathularis:
Los sólidos de los extractos líquidos obtenidos en la etapa iv), se separan por filtración, preferentemente mediante placa porosa del nº 3 ó 4, o papel de filtro de gramaje 73 g/m2, obteniéndose los extractos líquidos filtrados de Saxifraga spathularis. The solids of the liquid extracts obtained in step iv), are separated by filtration, preferably by porous plate of No. 3 or 4, or weight filter paper 73 g / m2, obtaining the fi ltered liquid extracts of Saxifraga spathularis.
Estos extractos se almacenan refrigerados, preferentemente a 4ºC, y protegidos de la luz para evitar posibles alteraciones. These extracts are stored refrigerated, preferably at 4 ° C, and protected from light to avoid possible alterations.
Caracterización de los extractos líquidos filtrados de Saxifraga spathularis Characterization of fi ltered liquid extracts of Saxifraga spathularis
i) Determinación de la absorción UV (ultravioleta) de los extractos líquidos filtrados de Saxifraga spathularis: i) Determination of the UV (ultraviolet) absorption of the fi ltered liquid extracts of Saxifraga spathularis:
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 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), therefore, if a product absorbs radiation between 250 to 400 nm it can be used as a solar fi lter.
Se mide la absorbancia (Abs) de los extractos líquidos filtrados de Saxifraga spathularis obtenidos según el procedimiento de extracción, a la concentración de 100 ppm en agua destilada entre las longitudes de onda de 250 a 400 nm frente a un blanco de agua destilada. Todos presentan máximos de absorción en la región UV (ultravioleta). The absorbance (Abs) of the fi ltered liquid extracts of Saxifraga spathularis obtained according to the extraction procedure is measured at the concentration of 100 ppm in distilled water between the wavelengths of 250 to 400 nm against a blank of distilled water. All have maximum absorption in the UV (ultraviolet) region.
Los extractos de Saxifraga spathularis a la longitud de onda de 280 nm presentan Abs > 0,700; a 320 nm presentan Abs > 0,250 y a 400 nm presentan Abs > 0,050. Saxifraga spathularis extracts at the wavelength of 280 nm have Abs> 0.700; at 320 nm they present Abs> 0.250 and at 400 nm they present Abs> 0.050.
ii) Determinación del contenido fenólico de los extractos líquidos filtrados de Saxifraga spathularis: ii) Determination of the phenolic content of the fi ltered liquid extracts of Saxifraga spathularis:
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 Saxifraga spathularis según el método de Folin Ciocalteu’s Phenolic compounds have a high antioxidant activity [Y. S. Velioglu, op. cited (1998)]. Therefore, the phenolic content of the liquid extracts of Saxifraga spathularis is determined according to the Folin Ciocalteu’s method
[A. Escarpa et al., Analytica Chimica 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 absorbancia con un recta patrón de ácido gálico (0-100 ppm). [TO. Escarpa et al., Analytica Chimica 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 Folin Ciocalteau's reagent (1: 1 with distilled water) and 0.5 mL of 10% Na2CO3; 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).
Los extractos líquidos filtrados de Saxifraga spathularis obtenidos según el procedimiento de extracción, presentan, según el procedimiento descrito, un alto contenido fenólico, entre 20-35%. The fi ltered liquid extracts of Saxifraga spathularis obtained according to the extraction procedure, have, according to the described procedure, a high phenolic content, between 20-35%.
iii) Determinación de la actividad captadora del radical α,α-difenil-β-picrilhidracilo (DPPH•) y de su equivalencia en BHT y BHA de los extractos líquidos filtrados de Saxifraga spathularis: iii) Determination of the capture activity of the α, α-diphenyl-β-picrylhydracil (DPPH •) radical and its equivalence in BHT and BHA of the fi ltered liquid extracts of Saxifraga spathularis:
Para medir la actividad captadora del radical α,α-difenil-β-picrilhidracilo (DPPH•)[I. Parejo et al., Journal of Agricultural 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• 610−5 M en metanol y 50 μL del extracto antioxidante. El porcentaje de inhibición se calcula según la siguiente fórmula: To measure the activity of the radical α, α-diphenyl-β-picrylhydracil (DPPH •) [I. Parejo et al., 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 • 610-5 M in methanol and 50 µL of the antioxidant extract is measured. The percentage of inhibition is calculated according to the following formula:
PI (Porcentaje de Inhibición) PI(%): [(Abs t=0 min-Abs t=16 min)/Abs t=0 min)]*100 PI (Percent Inhibition) PI (%): [(Abs t = 0 min-Abs t = 16 min) / Abs t = 0 min)] * 100
Se mide la IC50 o concentración que inhibe el 50% del radical DPPH• . The IC50 or concentration that inhibits 50% of the DPPH radical is measured.
Los extractos líquidos filtrados de Saxifraga spathularis, obtenidos según el procedimiento de extracción, presentan una IC50 entre 300 a 350 ppm. The fi ltered liquid extracts of Saxifraga spathularis, obtained according to the extraction procedure, have an IC50 between 300 to 350 ppm.
La IC50 del BHA, medida según el mismo procedimiento, es de 240 ppm y la del BHT de 2790 ppm. The IC50 of the BHA, measured according to the same procedure, is 240 ppm and that of the BHT of 2790 ppm.
iv) Determinación de la actividad antioxidante o de reducción del hierro (FRAP) y de su equivalencia en ácido ascórbico o vitamina C de los extractos líquidos filtrados Saxifraga spathularis: iv) Determination of the antioxidant or iron reduction activity (FRAP) and its equivalence in ascorbic acid or vitamin C of the leaked liquid extracts Saxifraga spathularis:
Se determina el poder reductor del hierro o actividad antioxidante de los extractos líquidos de Saxifraga spathularis según el método FRAP [Benzie IFF et al. Anal Biochem 1996; 239: 70-76]. Se mide la capacidad de reducir el complejo de Fe(III)/tripiridiltriazine y las actividades reductivas de los extractos se expresan como equivalentes nM de ácido ascórbico (AscAE),/g de extracto seco. A 0,1 mL del extracto antioxidante se le añaden 3 mL del reactivo FRAP se deja a temperatura ambiente durante 6 minutos. Se mide la absorbancia a 593 nm y se comparan los resultados con una recta patrón de ácido ascórbico o vitamina C (0,1-1 mM). The reducing power of iron or antioxidant activity of the liquid extracts of Saxifraga spathularis is determined according to the FRAP method [Benzie IFF et al. Anal Biochem 1996; 239: 70-76]. The ability to reduce the Fe (III) / tripyridyltriazine complex is measured and the reductive activities of the extracts are expressed as nM equivalents of ascorbic acid (AscAE), / g of dry extract. To 0.1 mL of the antioxidant extract is added 3 mL of the FRAP reagent is left at room temperature for 6 minutes. The absorbance at 593 nm is measured and the results are compared with a straight standard of ascorbic acid or vitamin C (0.1-1 mM).
Los extractos líquidos filtrados de Saxifraga spathularis obtenidos según el procedimiento de extracción, presentan a una concentración de 100 ppm un poder reductor del hierro equivalente a entre 0,100 mM a 0,150 mM en ácido ascórbico o vitamina C. The fi ltered liquid extracts of Saxifraga spathularis obtained according to the extraction procedure, have at a concentration of 100 ppm an iron reducing power equivalent to between 0,100 mM to 0,150 mM in ascorbic acid or vitamin C.
Ejemplos Examples
Ejemplo 1 Example 1
Preparación del extracto hidrobutilenglicólico de las hojas de Saxifraga spathularis Preparation of the hydrobutylene glycol extract of the leaves of Saxifraga spathularis
Las hojas congeladas con N2 líquido de Saxifraga spathularis, 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, con una mezcla etanol/agua 1:1, a la temperatura de reflujo del disolvente (85-95ºC), durante 3 horas y protegido de la luz. Se evapora hasta sequedad, a vacío, el disolvente del extracto obtenido. El rendimiento de la extracción es del 28% (g/g). The leaves frozen with liquid N2 of Saxifraga spathularis, ground and sieved to 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, with a 1: 1 ethanol / water mixture, at the solvent reflux temperature (85-95 ° C), for 3 hours and protected from light. The solvent of the extract obtained is evaporated to dryness in vacuo. The yield of extraction is 28% (g / g).
El extracto seco de Saxifraga spathularis obtenido se somete a un nuevo proceso extractivo sólido-liquido en continuo, mediante maceración con agitación, con una relación líquido:sólido de 99 g/g, a temperatura ambiente y protegido de la luz, durante 7 días con una mezcla butilenglicol/agua 1:1. El extracto líquido de Saxifraga spathularis obtenido se filtra, mediante placa porosa del nº 3, y el producto final, el extracto líquido filtrado de Saxifraga spathularis,se almacena refrigerado a 4ºC y protegido de la luz para evitar su alteración. The dry extract of Saxifraga spathularis obtained is subjected to a new continuous solid-liquid extractive process, by maceration with stirring, with a liquid: solid ratio of 99 g / g, at room temperature and protected from light, for 7 days with a 1: 1 butylene glycol / water mixture. The liquid extract of Saxifraga spathularis obtained is filtered, using a porous plate of No. 3, and the final product, the filtered liquid extract of Saxifraga spathularis, is stored refrigerated at 4 ° C and protected from light to prevent alteration.
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- Se midió la absorbancia de este extracto a la concentración de 100 ppm en agua destilada a las longitudes de onda de 280 nm (Abs = 0,714), 320 nm (Abs = 0,271) y 400 nm (Abs = 0,053) frente a un blanco de agua destilada. The absorbance of this extract was measured at the concentration of 100 ppm in distilled water at the wavelengths of 280 nm (Abs = 0.714), 320 nm (Abs = 0.271) and 400 nm (Abs = 0.053) against a blank of distilled water.
- --
- 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 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 absorbancia con un recta patrón de ácido gálico (0-100 ppm). 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% Na2CO3; 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 24%. The extract obtained according to the procedure described in this example 1 has a phenolic content of 24%.
- --
- Actividad captadora del radical α,α-difenil-β-picrilhidracilo (DPPH•)[I. 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• 610−5 Men 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-picrilhydracil (DPPH •) [I. Even, op. cited (2002)]. The decrease in absorbance at 515 nm, after 16 minutes, of a 2 mL solution of DPPH • 610-5 Men methanol and 50 µL of the antioxidant extract is measured. The percentage of inhibition is calculated according to the following formula:
PI (Porcentaje de Inhibición) PI(%): [(Abs t=0 min-Abs t=16 min)/Abs t=0 min)]*100 PI (Percent Inhibition) PI (%): [(Abs t = 0 min-Abs t = 16 min) / Abs t = 0 min)] * 100
Se midió la IC50 o concentración que inhibe el 50% del radical DPPH• . The IC50 or concentration that inhibits 50% of the DPPH • radical was measured.
El extracto obtenido según el procedimiento descrito en este ejemplo 1 presenta una IC50 = 334 ppm. La IC50 del BHA, medida según el mismo procedimiento, es de 240 ppm y la del BHT de 2790 ppm. The extract obtained according to the procedure described in this example 1 has an IC50 = 334 ppm. The IC50 of the BHA, measured according to the same procedure, is 240 ppm and that of the BHT of 2790 ppm.
- --
- Actividad antioxidante o de reducción del hierro (FRAP) [I.F.F. Benzie, op. cited (1996)]. Se mide la capacidad de reducir el complejo de Fe (III)/tripiridiltriazine y las actividades reductivas de los extractos se expresan como equivalentes nM de ácido ascórbico (AscAE),/g de extracto seco. A 0,1 mL del extracto antioxidante se le añaden 3 mL del reactivo FRAP se deja a temperatura ambiente durante 6 minutos. Se mide la absorbancia a 593 nm y se comparan los resultados con una recta patrón de ácido ascórbico o vitamina C (0,1-1 mM). Antioxidant or iron reduction activity (FRAP) [I.F.F. Benzie, op. cited (1996)]. The ability to reduce the Fe (III) / tripyridyltriazine complex is measured and the reductive activities of the extracts are expressed as nM equivalents of ascorbic acid (AscAE), / g of dry extract. To 0.1 mL of the antioxidant extract is added 3 mL of the FRAP reagent is left at room temperature for 6 minutes. The absorbance at 593 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 una concentración de 100 ppm un poder reductor del hierro equivalente a 0,126 mM en ácido ascórbico o vitamina C. The extract obtained according to the procedure described in this example 1, exhibits at a concentration of 100 ppm an iron reducing power equivalent to 0.126 mM in ascorbic acid or vitamin C.
Claims (18)
- 2. 2.
- Procedimiento de obtención de extractos de Saxifraga spathularis, 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. Method of obtaining extracts of Saxifraga spathularis, according to claim 1, step i), characterized in that the dried leaves ground in a blade mill are used, and they are screened at a particle size of 0.6 mm. approximately.
- 3. 3.
- Procedimiento de obtención de extractos de Saxifraga spathularis, según la reivindicación 1, etapa ii), caracterizado porque el proceso de extracción sólido-líquido en continuo con disolventes; se emplean relaciones líquido:sólido elevadas, preferentemente de 30 g/g; el disolvente es agua, o un alcohol C1 aC4, preferentemente metanol o etanol, Method of obtaining extracts of Saxifraga spathularis, according to claim 1, step ii), characterized in that the process of solid-liquid extraction in continuous with solvents; high liquid: solid ratios are used, preferably 30 g / g; the solvent is water, or a C1 to C4 alcohol, preferably methanol or ethanol,
- 4. Four.
- Procedimiento de obtención de extractos de Saxifraga spathularis, 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 85-95ºC. Method of obtaining extracts of Saxifraga spathularis, 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 ° C.
- 5. 5.
- Procedimiento de obtención de extractos de Saxifraga spathularis, según la reivindicación 1, etapa iii), caracterizado porque la evaporación hasta sequedad del disolvente para la obtención de los extractos secos de Saxifraga spathularis, se realiza a vacío y el agua se elimina, preferentemente, mediante liofilización. Method of obtaining extracts of Saxifraga spathularis, according to claim 1, step iii), characterized in that the evaporation to dryness of the solvent to obtain dry extracts of Saxifraga spathularis is carried out under vacuum and the water is preferably removed by lyophilization.
- 6. 6.
- Procedimiento de obtención de extractos de Saxifraga spathularis, según la reivindicación 1, etapa iii), caracterizado porque la evaporación del disolvente alcohólico para la obtención de los extractos acuosos de Saxifraga spathularis, se realiza a vacío. Method of obtaining extracts of Saxifraga spathularis, according to claim 1, step iii), characterized in that the evaporation of the alcoholic solvent to obtain the aqueous extracts of Saxifraga spathularis is carried out under vacuum.
- 7. 7.
- Procedimiento de obtención de extractos de Saxifraga spathularis, 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 aC6, preferentemente propilenglicol o butilenglicol, o una mezcla propilenglicol/agua, o butilenglicol/agua; la temperatura de extracción es la temperatura ambiente, 20ºC aproximadamente; el tiempo de la extracción es de 1 a 7 días y se realiza protegido de la luz. Method of obtaining extracts of Saxifraga spathularis, 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 C2 to C6 glycol, preferably propylene glycol or butylene glycol, or a propylene glycol / water mixture, or butylene glycol / water; the extraction temperature is room temperature, approximately 20 ° C; The extraction time is 1 to 7 days and is carried out protected from light.
- 8. 8.
- Procedimiento de obtención de extractos de Saxifraga spathularis, según la reivindicación 7, caracterizado porque el disolvente de extracción es una mezcla butilenglicol/agua 1:1. Method of obtaining extracts of Saxifraga spathularis, according to claim 7, characterized in that the extraction solvent is a 1: 1 butylene glycol / water mixture.
- 9. 9.
- Procedimiento de obtención de extractos de Saxifraga spathularis, según la reivindicación 1, etapa v), caracterizado porque se filtran los extractos líquidos obtenidos en la etapa iv), preferentemente mediante placa porosa del nº 3 ó 4, o papel de de filtro de gramaje 73 g/m2, obteniéndose los extractos líquidos filtrados de Saxifraga spathularis se almacenan en frío, preferentemente a 4ºC, y protegidos de la luz. Method of obtaining extracts of Saxifraga spathularis, according to claim 1, step v), characterized in that the liquid extracts obtained in step iv) are filtered, preferably by porous plate of No. 3 or 4, or weight fi lter paper 73 g / m2, obtaining fi ltered liquid extracts of Saxifraga spathularis are stored cold, preferably at 4 ° C, and protected from light.
- 10. 10.
- Extractos secos de Saxifraga spathularis obtenidos según las reivindicaciones1a5. Dry extracts of Saxifraga spathularis obtained according to claims 1-5.
- 11. eleven.
- Extractos líquidos de Saxifraga spathularis obtenidos según las reivindicaciones1a9. Liquid extracts of Saxifraga spathularis obtained according to claims 1-9.
- 13. 13.
- Extractos líquidos filtrados de Saxifraga spathularis, según la reivindicación 12, caracterizados por presentar a la concentración de 100 ppm a las longitudes de onda de de 280 nm presentan Abs > 0,700; a 320 nm presentan Abs > 0,250 y a 400 nm presentan Abs > 0,050, por poseer un contenido fenólico entre 20-35%; por presentar una IC50 entre 300 a 350 ppm de inhibición del radical DPPH• y por su capacidad antioxidante o de reducción del hierro a 100 ppm equivalente a entre 0,100 mM a 0,150 mM en ácido ascórbico o vitamina C. Filtered liquid extracts of Saxifraga spathularis, according to claim 12, characterized by having a concentration of 100 ppm at wavelengths of 280 nm have Abs> 0.700; at 320 nm they present Abs> 0.250 and at 400 nm they present Abs> 0.050, because they have a phenolic content between 20-35%; for presenting an IC50 between 300 to 350 ppm of inhibition of the DPPH radical • and for its antioxidant capacity or reduction of iron at 100 ppm equivalent to between 0.100 mM to 0.150 mM in ascorbic acid or vitamin C.
- 14. 14.
- Extracto líquido filtrado de las hojas de Saxifraga spathularis, según la reivindicación 13, caracterizado por presentar a la concentración de 100 ppm a las longitudes de onda de 280 nm (Abs = 0,714), 320 nm (Abs = 0,271) y 400 nm (Abs = 0,053); por poseer un contenido fenólico del 24%; por presentar una IC50 = 334 ppm de inhibición del radical DPPH• y por su capacidad antioxidante o de reducción del hierro equivalente a 0,126 mM en ácido ascórbico Liquid extract filtered from the sheets of Saxifraga spathularis, according to claim 13, characterized in that it has a concentration of 100 ppm at wavelengths of 280 nm (Abs = 0.714), 320 nm (Abs = 0.271) and 400 nm (Abs = 0.053); for having a phenolic content of 24%; for presenting an IC50 = 334 ppm of inhibition of the DPPH radical • and for its antioxidant capacity or reduction of iron equivalent to 0.126 mM in ascorbic acid
- 15. fifteen.
- Uso de los extractos de Saxifraga spathularis, 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. Use of the extracts of Saxifraga spathularis, 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. 16.
- Uso de los extractos de Saxifraga spathularis, 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. Use of the extracts of Saxifraga spathularis, according to claims 10 to 14, as a cosmetic agent for skin and / or hair care, intended to act as an absorber of UV (ultraviolet) radiation, which gives it fi lter properties solar.
- 17. 17.
- Uso de los extractos de Saxifraga spathularis, según la reivindicaciones 10 a 14, en la industria farmacéutica y/o alimentaria como antioxidante. Use of the extracts of Saxifraga spathularis, according to claims 10 to 14, in the pharmaceutical and / or food industry as an antioxidant.
- Categoría Category
- Documentos citados Reivindicaciones afectadas Documents cited Claims Affected
- A A To
- JP 10203944 (SHISEIDO CO LTD) 04.08.1998, resumen. JP 10046142 A (SHISEIDO CO LTD) 17.02.1998, resumen. 1-17 1-17 JP 10203944 (SHISEIDO CO LTD) 04.08.1998, summary. JP 10046142 A (SHISEIDO CO LTD) 17.02.1998, summary. 1-17 1-17
- Categoría de los documentos citados X: de particular relevancia Y: de particular relevancia combinado con otro/s de la misma categoría A: refleja el estado de la técnica O: referido a divulgación no escrita P: publicado entre la fecha de prioridad y la de presentación de la solicitud E: documento anterior, pero publicado después de la fecha de presentación de la solicitud Category of the documents cited X: of particular relevance Y: of particular relevance combined with other / s of the same category A: reflects the state of the art O: refers to unwritten disclosure P: published between the priority date and the date of priority submission of the application E: previous document, but published after the date of submission of the application
- El presente informe ha sido realizado • para todas las reivindicaciones • para las reivindicaciones nº: TODAS This report has been produced • for all claims • for claims no: ALL
- Fecha de realización del informe 23.11.2010 Date of realization of the report 23.11.2010
- Examinador A. Amaro Roldán Página 1/4 Examiner A. Amaro Roldán Page 1/4
- Novedad (Art. 6.1 LP 11/1986) Novelty (Art. 6.1 LP 11/1986)
- Reivindicaciones Reivindicaciones 1-17 SI NO Claims Claims 1-17 IF NOT
- Actividad inventiva (Art. 8.1 LP11/1986) Inventive activity (Art. 8.1 LP11 / 1986)
- Reivindicaciones Reivindicaciones 1-17 SI NO Claims Claims 1-17 IF NOT
- Documento Document
- Número Publicación o Identificación Fecha Publicación Publication or Identification Number publication date
- D01 D01
- JP 10203944 (SHISEIDO CO LTD) 04.08.1998 JP 10203944 (SHISEIDO CO LTD) 04.08.1998
- D02 D02
- JP 10046142 A (SHISEIDO CO LTD) 17.02.1998 JP 10046142 A (SHISEIDO CO LTD) 02.17.1998
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| ES200901653A ES2352403B1 (en) | 2009-07-27 | 2009-07-27 | SAXIFRAGA SPATHULARIS EXTRACTS, PROCEDURE FOR OBTAINING AND ITS COSMETIC, PHARMACEUTICAL AND / OR FOOD USE AS ANTIOXIDANTS / ANTIRRADICALARIES / UV FILTERS (ULTRAVIOLET). |
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| ES2420517B1 (en) * | 2012-02-22 | 2014-06-24 | Caroi'line Cosmetica S.L. | Formulations with fibroblast stimulation activity containing extracts of Armeria pubigera and / or Saxifraga spathularis |
| CN113087608B (en) * | 2021-03-08 | 2022-11-11 | 中国科学院西北高原生物研究所 | New diaryl nonane V, VI and VII free radical inhibitor in saxifraga tangutica and separation preparation process and application thereof |
| CN113087606B (en) * | 2021-03-08 | 2022-11-11 | 中国科学院西北高原生物研究所 | New diaryl nonane IV and III free radical inhibitor in saxifraga tangutica and separation preparation process and application thereof |
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| JPH10203944A (en) * | 1997-01-18 | 1998-08-04 | Shiseido Co Ltd | Cosmetic |
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