WO2021053259A1 - Method for extracting, from olive leaf, oleuropein in a phenolic fraction; functional food composition; and use - Google Patents
Method for extracting, from olive leaf, oleuropein in a phenolic fraction; functional food composition; and use Download PDFInfo
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- WO2021053259A1 WO2021053259A1 PCT/ES2020/070564 ES2020070564W WO2021053259A1 WO 2021053259 A1 WO2021053259 A1 WO 2021053259A1 ES 2020070564 W ES2020070564 W ES 2020070564W WO 2021053259 A1 WO2021053259 A1 WO 2021053259A1
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- B—PERFORMING OPERATIONS; TRANSPORTING
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- B01D11/02—Solvent extraction of solids
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/63—Oleaceae (Olive family), e.g. jasmine, lilac or ash tree
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- the present invention is applicable in the food, cosmetic and pharmaceutical industries.
- a functional food consisting of polyphenols and sugars
- in the cosmetic industry in anti-aging creams for its ability to capture free radicals
- in the pharmaceutical industry in drugs against oxidative stress or for the prevention of cardiovascular and degenerative diseases such as cancer and Alzheimer's.
- Patent documents such as WO 2000038541 A1 describe a healthy diet in which food has been enriched with antioxidants, mainly phenolic compounds from the olive tree.
- the main phenolic compound in the fruit of the olive tree and the leaf is oleuropein, with concentrations of 2 to 134 mg g _1 of dry matter. Numerous studies demonstrate the significant biological activity of oleuropein. It is an important antioxidant and antibacterial, the main responsible for the anti-inflammatory effect of olive leaf extracts, shows antiproliferative activity on breast and liver cell lines, acts against diabetes, prevents obesity since it has shown hypocholesterolemic effects in rats, and protects against osteoporosis and osteoarthritis. In addition, it helps prevent neurodegenerative diseases such as Parkinson's and Alzheimer's due to its neuroprotective properties and to minimize the progression of pulmonary fibrosis.
- JP 2016079127 A describes an antioxidant composed of dihydroquercetin and olive leaf extract containing oleuropein and hydroxytyrosol, with application as a food additive.
- WO 2008108817 A1 a healthy nutritional supplement is described where the bitter taste of oleuropein is masked by complexing with ci the extrins aided by glycerin monoesters.
- oleuropein and its main metabolites including oleuropein aglycone, hydroxytyrosol and elenolic acid are useful for treating obesity, as well as for inhibiting HIV infectivity.
- Patent ES 2288123 B1 protects a functional food composition based on olive and citrus extracts for the prevention or treatment of problems related to excess cholesterol and triglycerides in the blood.
- the extractions are carried out by hydroalcoholic mixtures.
- NADES natural deep eutectic solvents
- NADES based on glycerol and glycine that is, on non-sugar compounds, have been used to extract polyphenols from olive leaves (Athanasiadis, V. et al. Waste Biomass Valor., 2018, 9, 1985-1992); the use of sugars as an extractant is not suggested in the document.
- the role of methyl b-cyclodextrin as an adjuvant in the extraction of polyphenols from the olive leaf with deep eutectic solvents has also been described (Athanasiadis, V. Biomass Convers.
- the problem of the technique is to achieve a simpler extraction procedure for oleuropein from the olive leaf and avoiding toxic solvents.
- the solution proposed by the present invention is a process that replaces typical organic solvents with sugars or honey, acting as NADES.
- the present invention is a process for the olive leaf extraction of a phenolic fraction containing oleuropein, which comprises heating a mixture comprising powdered dried olive leaf in a deep eutectic solvent of edible natural sugars and water, and filtering to obtain a precipitate and an aqueous phase where the aqueous phase contains the phenolic fraction with the oleuropein.
- pulverized dry leaf is understood as the dry leaf subjected to a pulverization process in a mill with a particle size of 0.05 to 1.0 mm.
- Another aspect of the present invention is a process consisting of the above steps. Yet another aspect is that the mixture consists of powdered dried olive leaf and a deep eutectic solvent.
- said sugars that form NADES are glucose, fructose and sucrose.
- the heating occurs at a temperature between 30 and 120 ° C, preferably between 50% and 100 ° C because above there is a certain degree of decomposition of oleuropein and below 50 ° C the yield is lower, and even more preferably 80 ° C.
- glucose, fructose and sucrose are in a molar ratio in water of 1: 1: 1: 11.
- a highly preferable aspect of the present invention is that said deep eutectic solvent constituted by edible natural sugars is honey.
- the heating should be at a temperature between 35 and 50 ° C so as not to denature the honey, preferably 40 ° C.
- the mixture of sugars will mask the bitterness of oleuropein in the food product in which it is incorporated.
- heating occurs for a time between 20 and 240 min, preferably between 50 and 90 min to avoid some decomposition of oleuropein especially at high times and temperatures, and even more preferably 60 min.
- a preferable aspect of the invention comprises a dilution with water prior to the filtration step to improve the performance of the filtrate.
- Filtering can be done under vacuum on filter paper with a pore size between 2.5-25 pm. It is taken up by the liquid fraction, which can then be extracted with an organic phase, preferably ethyl acetate, and concentrated on a rotary evaporator.
- organic phase preferably ethyl acetate
- the present invention describes the extraction of oleuropein from the olive leaf by a method that respects the environment within the framework of green chemistry, in which the extractant formed by a mixture of sugars or honey allows a functional food to be obtained directly.
- the main advantage of the process of the invention is that it avoids the use of conventional organic solvents, which are toxic and involve purification steps that complicate the process.
- Oleuropein as a phenolic compound with significant bioactivity, is a powerful antioxidant, it helps preserve food, preventing rancidity, loss of color and deterioration of aroma and flavor; Furthermore, it has interesting anti-inflammatory, anti-atherogenic, anti-cancer, hypolipidemic and hypoglycemic pharmacological properties.
- the composition obtained can help defend against oxidative damage in cells, preventing cardiovascular and neurodegenerative diseases and cancer.
- a highly preferable aspect of the invention is a functional food composition
- a functional food composition comprising a phenolic extract with oleuropein in a percentage of between 80% - 85% by weight with respect to the weight of the total phenolic extract.
- composition of the invention as an ingredient in food products such as candies or gummies, carbonated drinks, juices, bakery products, energy or dairy products.
- composition of the invention as a dietary supplement, more preferably still an antioxidant and anti-aging dietary supplement.
- honey enriched in polyphenols from the olive tree could be used in the manufacture of anti-aging creams due to its ability to capture free radicals; and in the pharmaceutical industry, the polyphenol content would allow its use against oxidative stress and for the prevention of cardiovascular and degenerative diseases such as cancer and Alzheimer's.
- compositions in the prevention or treatment of a tumor, neurodegenerative, infectious, inflammatory or cardiovascular disease, or any other for which oleuropein or phenolic extract of the olive leaf be beneficial.
- Figure 1 shows the chromatogram obtained after the extraction of the phenolic fraction of the olive leaf with NADES according to Example 1. It shows the presence of oleuropein with a purity of 85% accompanied by hydroxyoleuropein, ligustroside and two oleuropein isomers ( deduced from UHPLC-MS), as minor compounds. The values are multiplied by a factor of 1000.
- the extract quantified by HPLC contained 90 mg of a phenolic fraction, of which 85% is oleuropein.
- Example 2 Ingredients:
- the extract quantified by HPLC contained 45 mg of a phenolic fraction, of which oleuropein constitutes 83%.
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Abstract
The present invention relates to a method for extracting, from olive leaf, a phenolic fraction containing oleuropein, which comprises: heating a mixture of crushed dry olive leaf and a deep eutectic solvent of edible natural sugars and water; and filtering to obtain a precipitate and an aqueous phase, wherein the aqueous phase contains the phenolic fraction with the oleuropein. The invention also relates to a functional food composition with an oleuropein content of 80-85% by weight with respect to the weight of the total phenolic extract, and to the use thereof in the food, cosmetic or pharmaceutical industry.
Description
PROCEDIMIENTO PARA LA EXTRACCIÓN EN HOJA DE OLIVO DE OLEUROPEÍNA EN UNA FRACCIÓN FENÓLICA, COMPOSICIÓN ALIMENTARIA PROCEDURE FOR THE EXTRACTION IN OLIVE LEAF OF OLEUROPEINE IN A PHENOLIC FRACTION, FOOD COMPOSITION
FUNCIONAL Y USO FUNCTIONAL AND USE
DESCRIPCIÓN DESCRIPTION
Sector de la técnica Technical sector
La presente invención es de aplicación en la industria de alimentación, cosmética y farmacéutica. En la industria de alimentación como alimento funcional formado por polifenoles y azúcares; en la industria cosmética en cremas antienvejecimiento por su capacidad de captación de radicales libres; y en la industria farmacéutica en medicamentos contra el estrés oxidativo o para la prevención de enfermedades cardiovasculares y degenerativas como cáncer y Alzhéimer. The present invention is applicable in the food, cosmetic and pharmaceutical industries. In the food industry as a functional food consisting of polyphenols and sugars; in the cosmetic industry in anti-aging creams for its ability to capture free radicals; and in the pharmaceutical industry in drugs against oxidative stress or for the prevention of cardiovascular and degenerative diseases such as cancer and Alzheimer's.
Antecedentes de la invención Background of the invention
Los hábitos alimentarios en la cuenca mediterránea abundantes en frutas, verduras y aceite de oliva virgen extra, son ricos en polifenoles. Esta es la principal razón de la baja incidencia de varias patologías degenerativas; por ejemplo, la reducción del riesgo cardiovascular y el descenso del riesgo de desarrollar tumores. Documentos de patente como la WO 2000038541 A1 describen una dieta saludable en la que los alimentos han sido enriquecidos con antioxidantes, principalmente compuestos fenólicos del olivo.The eating habits in the Mediterranean basin abundant in fruits, vegetables and extra virgin olive oil, are rich in polyphenols. This is the main reason for the low incidence of various degenerative pathologies; for example, the reduction of cardiovascular risk and the decrease of the risk of developing tumors. Patent documents such as WO 2000038541 A1 describe a healthy diet in which food has been enriched with antioxidants, mainly phenolic compounds from the olive tree.
El compuesto fenólico mayoritario en el fruto del olivo y la hoja es la oleuropeína, con concentraciones de 2 a 134 mg g_1 de materia seca. Numerosos estudios demuestran la importante actividad biológica de la oleuropeína. Es un importante antioxidante y antibacteriano, el principal responsable del efecto antinflamatorio de los extractos de hoja de olivo, muestra actividad antiproliferativa sobre líneas celulares de mama y de hígado, actúa contra la diabetes, previene la obesidad ya que ha demostrado efectos hipocolesterolémicos en ratas, y protege contra la osteoporosis y la osteoartritis. Además, ayuda a prevenir enfermedades neurodegenerativas como el Parkinson y Alzhéimer por sus propiedades neuroprotectoras y a minimizar el avance de la fibrosis pulmonar. The main phenolic compound in the fruit of the olive tree and the leaf is oleuropein, with concentrations of 2 to 134 mg g _1 of dry matter. Numerous studies demonstrate the significant biological activity of oleuropein. It is an important antioxidant and antibacterial, the main responsible for the anti-inflammatory effect of olive leaf extracts, shows antiproliferative activity on breast and liver cell lines, acts against diabetes, prevents obesity since it has shown hypocholesterolemic effects in rats, and protects against osteoporosis and osteoarthritis. In addition, it helps prevent neurodegenerative diseases such as Parkinson's and Alzheimer's due to its neuroprotective properties and to minimize the progression of pulmonary fibrosis.
En los últimos años se han aplicado tecnologías de alta presión, fluido supercrítico o destilación molecular para extraer estos componentes e incluirlos en matrices alimentarias. El interés en los alimentos funcionales, alimentos enriquecidos y
suplementos alimenticios actuales es ofrecer efectos protectores frente a enfermedades de elevada incidencia, como obesidad, trastornos cardiovasculares y cáncer, entre otras. In recent years, high pressure, supercritical fluid or molecular distillation technologies have been applied to extract these components and include them in food matrices. Interest in functional foods, fortified foods, and Current nutritional supplements is to offer protective effects against diseases of high incidence, such as obesity, cardiovascular disorders and cancer, among others.
Así, la solicitud WO 2017032722 A1 describe una bebida de un extracto de hojas de olivo con derivados secoiridoides obtenido por calentamiento en agua y aditivos enzimáticos; y la JP 2016079127 A, describe un antioxidante compuesto de dihidroquercetina y extracto de hoja de olivo que contiene oleuropeína e hidroxitirosol, con aplicación como aditivo alimentario. Thus, the application WO 2017032722 A1 describes a drink made from an extract of olive leaves with secoiridoid derivatives obtained by heating in water and enzymatic additives; and JP 2016079127 A, describes an antioxidant composed of dihydroquercetin and olive leaf extract containing oleuropein and hydroxytyrosol, with application as a food additive.
En la WO 2008108817 A1 se describe un suplemento nutricional saludable donde el sabor amargo de la oleuropeína se enmascara al formar complejo con ci el od extrinas ayudado con monoésteres de glicerina. De acuerdo con US 20090061031 A1 , la oleuropeína y sus principales metabolitos, incluyendo el aglicón de la oleuropeína, el hidroxitirosol y el ácido elenólico son útiles para tratar la obesidad, así como para inhibir la infectividad del VIH. In WO 2008108817 A1 a healthy nutritional supplement is described where the bitter taste of oleuropein is masked by complexing with ci the extrins aided by glycerin monoesters. According to US 20090061031 A1, oleuropein and its main metabolites, including oleuropein aglycone, hydroxytyrosol and elenolic acid are useful for treating obesity, as well as for inhibiting HIV infectivity.
La patente ES 2288123 B1 protege una composición alimentaria funcional a base de extractos de olivo y cítricos para la prevención o tratamiento de problemas relacionados con el exceso de colesterol y triglicéridos en sangre. Las extracciones se llevan a cabo por mezclas hidroalcohólicas. Patent ES 2288123 B1 protects a functional food composition based on olive and citrus extracts for the prevention or treatment of problems related to excess cholesterol and triglycerides in the blood. The extractions are carried out by hydroalcoholic mixtures.
Todos estos métodos de extracción de la oleuropeína requieren de disolventes orgánicos (MeOH/H20, EtOH/H20 o MeCN/H20) que son tóxicos y hay que eliminar en la elaboración de un producto alimenticio. All these oleuropein extraction methods require organic solvents (MeOH / H 2 0, EtOH / H 2 0 or MeCN / H 2 0) which are toxic and must be eliminated in the preparation of a food product.
Frente a este inconveniente, los disolventes eutécticos profundos naturales (NADES) evitan la necesidad de reducir o eliminar el extractante. Los NADES son de gran interés, ya que son una alternativa verde frente a los disolventes convencionales tóxicos. Faced with this drawback, natural deep eutectic solvents (NADES) avoid the need to reduce or eliminate the extractant. NADES are of great interest, as they are a green alternative to toxic conventional solvents.
NADES basados en glicerol y glicina, es decir en compuestos no azúcares, han sido usados para extraer polifenoles de las hojas de olivo (Athanasiadis, V. et al. Waste Biomass Valor., 2018, 9, 1985-1992); en el documento no se sugiere el uso de azúcares como extractante. También se ha descrito el papel de la metil b-ciclodextrina como coadyuvante en la extracción de polifenoles de la hoja del olivo con disolventes eutécticos profundos (Athanasiadis, V. Biomass Convers. Biorefin., 2018, 8, 345-355), y el uso de disolventes eutécticos profundos basados en colina en la extracción asistida por microondas de compuestos fenólicos de la hoja de olivo (Alañón, M.E. et al. Arab. J. Chem., 2018, en prensa, https://doj.Org/10.1016/j.arabjc.2Q18.Q1.Q03); donde junto con colina como aceptor de enlace de hidrógeno, se utiliza ácido acético, ácido oxálico, ácido
tartárico, butano-1,4-diol, etilenglicol, xilitol, propano-1,2-diol, maltosa o urea como donador de enlace de hidrógeno. NADES based on glycerol and glycine, that is, on non-sugar compounds, have been used to extract polyphenols from olive leaves (Athanasiadis, V. et al. Waste Biomass Valor., 2018, 9, 1985-1992); the use of sugars as an extractant is not suggested in the document. The role of methyl b-cyclodextrin as an adjuvant in the extraction of polyphenols from the olive leaf with deep eutectic solvents has also been described (Athanasiadis, V. Biomass Convers. Biorefin., 2018, 8, 345-355), and the use of choline-based deep eutectic solvents in microwave-assisted extraction of phenolic compounds from olive leaf (Alañón, ME et al. Arab. J. Chem., 2018, in press, https://doj.Org/10.1016 /j.arabjc.2Q18.Q1.Q03); where together with choline as a hydrogen bond acceptor, acetic acid, oxalic acid, acid tartaric, butane-1,4-diol, ethylene glycol, xylitol, propane-1,2-diol, maltose or urea as a hydrogen bond donor.
En resumen, no hay antecedentes en la técnica del uso de azúcares como únicos componentes de un NADES para extraer polifenoles de la hoja de olivo. In summary, there is no prior art to the use of sugars as the sole components of a NADES to extract polyphenols from olive leaf.
El problema de la técnica es conseguir un procedimiento de extracción de oleuropeína de la hoja del olivo más sencillo y evitando disolventes tóxicos. La solución que propone la presente invención es un procedimiento que sustituye los disolventes orgánicos típicos por azúcares o miel actuando como NADES. The problem of the technique is to achieve a simpler extraction procedure for oleuropein from the olive leaf and avoiding toxic solvents. The solution proposed by the present invention is a process that replaces typical organic solvents with sugars or honey, acting as NADES.
Descripción de la invención Description of the invention
La presente invención es un procedimiento para la extracción en hoja de olivo de una fracción fenólica que contiene oleuropeína, que comprende calentar una mezcla que comprende hoja seca de olivo pulverizada en un disolvente eutéctico profundo de azúcares naturales comestibles y agua, y filtrar para obtener un precipitado y una fase acuosa donde la fase acuosa contiene la fracción fenólica con la oleuropeína. The present invention is a process for the olive leaf extraction of a phenolic fraction containing oleuropein, which comprises heating a mixture comprising powdered dried olive leaf in a deep eutectic solvent of edible natural sugars and water, and filtering to obtain a precipitate and an aqueous phase where the aqueous phase contains the phenolic fraction with the oleuropein.
En el ámbito de la presente invención se entiende por hoja seca “pulverizada” la hoja seca sometida a un proceso de pulverización en molinillo con tamaño de partícula de 0.05 a 1.0 mm. Within the scope of the present invention, "pulverized" dry leaf is understood as the dry leaf subjected to a pulverization process in a mill with a particle size of 0.05 to 1.0 mm.
Otro aspecto de la presente invención es un procedimiento que consiste en las etapas anteriores. Otro aspecto más es que la mezcla consista en hoja seca de olivo pulverizada y un disolvente eutéctico profundo. Another aspect of the present invention is a process consisting of the above steps. Yet another aspect is that the mixture consists of powdered dried olive leaf and a deep eutectic solvent.
Otro aspecto preferible de la invención es que dichos azúcares que forman el NADES son glucosa, fructosa y sacarosa. En este caso, el calentamiento se produce a una temperatura de entre 30 y 120°C, preferiblemente entre 50% y 100°C porque por encima existe cierto grado de descomposición de la oleuropeína y por debajo de 50°C el rendimiento es menor, y aún más preferiblemente de 80°C. Another preferable aspect of the invention is that said sugars that form NADES are glucose, fructose and sucrose. In this case, the heating occurs at a temperature between 30 and 120 ° C, preferably between 50% and 100 ° C because above there is a certain degree of decomposition of oleuropein and below 50 ° C the yield is lower, and even more preferably 80 ° C.
En un aspecto más preferible aún la glucosa, fructosa y sacarosa están en una relación molar en agua de 1:1 :1 :11. In an even more preferable aspect the glucose, fructose and sucrose are in a molar ratio in water of 1: 1: 1: 11.
Un aspecto muy preferible de la presente invención es que dicho disolvente eutéctico profundo constituido por azúcares naturales comestibles sea miel. En este caso, el calentamiento debe ser a una temperatura de entre 35 y 50°C para no desnaturalizar la miel, preferiblemente de 40°C.
La mezcla de azúcares va a enmascarar el amargor propio de la oleuropeína en el producto alimentario en que se incorpore. A highly preferable aspect of the present invention is that said deep eutectic solvent constituted by edible natural sugars is honey. In this case, the heating should be at a temperature between 35 and 50 ° C so as not to denature the honey, preferably 40 ° C. The mixture of sugars will mask the bitterness of oleuropein in the food product in which it is incorporated.
Otro aspecto preferible de la invención es que dicho calentamiento se produce durante un tiempo entre 20 y 240 min, preferiblemente entre 50 y 90 min para evitar cierta descomposición de la oleuropeína especialmente a tiempos y temperaturas altos, y aún más preferiblemente 60 min. Another preferable aspect of the invention is that said heating occurs for a time between 20 and 240 min, preferably between 50 and 90 min to avoid some decomposition of oleuropein especially at high times and temperatures, and even more preferably 60 min.
Un aspecto preferible de la invención comprende una dilución con agua previa a la etapa de filtración para mejorar el rendimiento del filtrado. A preferable aspect of the invention comprises a dilution with water prior to the filtration step to improve the performance of the filtrate.
El filtrado puede ser en vacío sobre filtro de papel con un tamaño de poro de entre 2,5 - 25 pm. La toma la fracción líquida, que seguidamente se puede extraer con una fase orgánica, preferiblemente acetato de etilo, y concentrarse en rotavapor. Filtering can be done under vacuum on filter paper with a pore size between 2.5-25 pm. It is taken up by the liquid fraction, which can then be extracted with an organic phase, preferably ethyl acetate, and concentrated on a rotary evaporator.
En la presente invención se describe la extracción de oleuropeína de la hoja de olivo por un método respetuoso con el medio ambiente enmarcado dentro de la química verde, en que el extractante formado por una mezcla de azúcares o por miel permite obtener directamente un alimento funcional. The present invention describes the extraction of oleuropein from the olive leaf by a method that respects the environment within the framework of green chemistry, in which the extractant formed by a mixture of sugars or honey allows a functional food to be obtained directly.
La ventaja principal del procedimiento de la invención es que evita el uso de disolventes orgánicos convencionales, que resultan tóxicos e implican etapas de purificación que complican el procedimiento. The main advantage of the process of the invention is that it avoids the use of conventional organic solvents, which are toxic and involve purification steps that complicate the process.
La oleuropeína, en tanto que compuesto fenólico con importante bioactividad resulta un potente antioxidante, ayuda a preservar alimentos, evitando el enranciamiento, la pérdida de color y el deterioro de aroma y sabor; además, presenta interesantes propiedades farmacológicas antinflamatorias, antiaterogénicas, anti-cancer, hipolipidémicas e hipoglicémicas. Oleuropein, as a phenolic compound with significant bioactivity, is a powerful antioxidant, it helps preserve food, preventing rancidity, loss of color and deterioration of aroma and flavor; Furthermore, it has interesting anti-inflammatory, anti-atherogenic, anti-cancer, hypolipidemic and hypoglycemic pharmacological properties.
La composición obtenida puede ayudar a la defensa del daño oxidativo en las células, previniendo enfermedades cardiovasculares, neurodegenerativas y cáncer. The composition obtained can help defend against oxidative damage in cells, preventing cardiovascular and neurodegenerative diseases and cancer.
De modo que un aspecto muy preferible de la invención es una composición alimentaria funcional que comprende un extracto fenólico con oleuropeína en un porcentaje de entre 80% - 85% en peso con respecto al peso del extracto fenólico total. Thus, a highly preferable aspect of the invention is a functional food composition comprising a phenolic extract with oleuropein in a percentage of between 80% - 85% by weight with respect to the weight of the total phenolic extract.
Otro aspecto preferible más es el uso de la composición de la invención como ingrediente en productos alimentarios como caramelos o gominolas, bebidas carbonatadas, zumos, productos de panadería, productos energéticos o lácteos.
Otro aspecto preferible es el uso de la composición de la invención como complemento dietético, más preferiblemente aún un complemento dietético antioxidante y antienvejecimiento. Yet another preferable aspect is the use of the composition of the invention as an ingredient in food products such as candies or gummies, carbonated drinks, juices, bakery products, energy or dairy products. Another preferable aspect is the use of the composition of the invention as a dietary supplement, more preferably still an antioxidant and anti-aging dietary supplement.
En la industria cosmética la miel enriquecida en polifenoles del olivo podría utilizarse en la fabricación de cremas antienvejecimiento por su capacidad de captación de radicales libres; y en la industria farmacéutica el contenido en polifenoles permitiría su utilización frente al estrés oxidativo y para la prevención de enfermedades cardiovasculares y degenerativas como cáncer y Alzheimer. In the cosmetic industry, honey enriched in polyphenols from the olive tree could be used in the manufacture of anti-aging creams due to its ability to capture free radicals; and in the pharmaceutical industry, the polyphenol content would allow its use against oxidative stress and for the prevention of cardiovascular and degenerative diseases such as cancer and Alzheimer's.
De forma que otro aspecto preferible de la invención es el uso de la composición en la prevención o tratamiento de una enfermedad tumoral, neurodegenerativa, infecciosa, inflamatoria o cardiovascular, o cualquier otra para la que la oleuropeína o el extracto fenólico de la hoja del olivo resulte beneficiosa. So another preferable aspect of the invention is the use of the composition in the prevention or treatment of a tumor, neurodegenerative, infectious, inflammatory or cardiovascular disease, or any other for which oleuropein or phenolic extract of the olive leaf be beneficial.
Descripción de las figuras Description of the figures
La figura 1 muestra el cromatograma obtenido después de la extracción de la fracción fenólica de la hoja de olivo con el NADES según el Ejemplo 1. Muestra la presencia de oleuropeína con una pureza del 85% acompañado de hidroxioleuropeína, ligustrósido y dos isómeros de oleuropeína (deducido de UHPLC-MS), como compuestos minoritarios. Los valores se encuentran multiplicados por un factor de 1000. Figure 1 shows the chromatogram obtained after the extraction of the phenolic fraction of the olive leaf with NADES according to Example 1. It shows the presence of oleuropein with a purity of 85% accompanied by hydroxyoleuropein, ligustroside and two oleuropein isomers ( deduced from UHPLC-MS), as minor compounds. The values are multiplied by a factor of 1000.
Ejemplos Ejemplo 1 : Examples Example 1:
Ingredientes: Ingredients:
Agua . 27,2 g Water . 27.2 g
Glucosa . 2,0 g Glucose 2.0 g
Fructosa . 2,0 g Fructose 2.0 g
Sacarosa . 3,8 g Sucrose 3.8 g
Hoja de olivo seca y pulverizada . 10,0 g Dried and powdered olive leaf. 10.0 g
Se calentaron 10 g de hoja seca de olivo pulverizada con NADES (10 g, 2,0 g de glucosa, 2,0 g de fructosa, 3,8 g de sacarosa, 2,2 g de agua) a 80 °C durante una hora. Pasado
ese tiempo se diluyó con 25 g de agua y se filtró en embudo Büchner a vacío a través de filtro de papel. Se extrajo entonces la fracción líquida obtenida con 25 ml_ de acetato de etilo y seguidamente se concentró esa fracción líquida en rotavapor. 10 g of dried olive leaf sprayed with NADES (10 g, 2.0 g of glucose, 2.0 g of fructose, 3.8 g of sucrose, 2.2 g of water) were heated at 80 ° C during a hour. Last This time it was diluted with 25 g of water and filtered in a Büchner funnel under vacuum through a paper filter. The liquid fraction obtained was then extracted with 25 ml_ of ethyl acetate and then this liquid fraction was concentrated in a rotary evaporator.
El 1H RMN del extracto muestra que se obtuvo oleuropeína como producto mayoritario. El cromatograma obtenido de la fracción fenólica se muestra en la Figura 1. The 1 H NMR of the extract shows that oleuropein was obtained as the major product. The chromatogram obtained from the phenolic fraction is shown in Figure 1.
El extracto cuantificado por HPLC contuvo 90 mg de una fracción fenólica, de la cual el 85% es oleuropeína. The extract quantified by HPLC contained 90 mg of a phenolic fraction, of which 85% is oleuropein.
Ejemplo 2: Ingredientes: Example 2: Ingredients:
Agua . 25,0 g Water . 25.0 g
Miel . 10,0 g Honey . 10.0 g
Hoja de olivo seca y pulverizada . 10,0 g Dried and powdered olive leaf. 10.0 g
Se calentaron 10 g de hoja seca de olivo pulverizado con miel natural (10 g) a 40 °C durante una hora. Pasado ese tiempo se diluyó con 25 g de agua y se filtró en embudo Büchner a vacío a través filtro de papel. 10 g of dried olive leaf sprayed with natural honey (10 g) were heated at 40 ° C for one hour. After that time, it was diluted with 25 g of water and filtered in a Büchner funnel under vacuum through a paper filter.
El extracto cuantificado por HPLC contuvo 45 mg de una fracción fenólica, de la cual la oleuropeína constituye el 83%.
The extract quantified by HPLC contained 45 mg of a phenolic fraction, of which oleuropein constitutes 83%.
Claims
1. Procedimiento para la extracción en hoja de olivo de una fracción fenólica que contiene oleuropeína, caracterizado por que dicho procedimiento comprende: calentar una mezcla que comprende hoja seca de olivo pulverizada y un disolvente, en que dicho disolvente es un disolvente eutéctico profundo que consiste en azúcares naturales comestibles y agua, y filtrar para obtener un precipitado y una fase acuosa, en que dicha fase acuosa contiene la fracción fenólica y la oleuropeína. 1. Procedure for the olive leaf extraction of a phenolic fraction containing oleuropein, characterized in that said procedure comprises: heating a mixture comprising dried powdered olive leaf and a solvent, in which said solvent is a deep eutectic solvent consisting of in edible natural sugars and water, and filter to obtain a precipitate and an aqueous phase, in which said aqueous phase contains the phenolic fraction and oleuropein.
2. Un procedimiento según la reivindicación 1, caracterizado por que dicho calentamiento se produce a una temperatura de entre 35°C y 120°C. 2. A process according to claim 1, characterized in that said heating occurs at a temperature between 35 ° C and 120 ° C.
3. Un procedimiento según la reivindicación 2, caracterizado por que dicha temperatura es de entre 50°C y 100°C 3. A process according to claim 2, characterized in that said temperature is between 50 ° C and 100 ° C
4. Un procedimiento según la reivindicación 3, caracterizado por que dicha temperatura es de 80°C. 4. A process according to claim 3, characterized in that said temperature is 80 ° C.
5. Un procedimiento según cualquiera de las reivindicaciones 1 a 4, caracterizado por que dichos azúcares son glucosa, fructosa y sacarosa.5. A process according to any of claims 1 to 4, characterized in that said sugars are glucose, fructose and sucrose.
6. Un procedimiento según la reivindicación 5, caracterizado por que dichas glucosa, fructosa y sacarosa están en una relación molar en agua de 1 :1 :1 :11. 6. A process according to claim 5, characterized in that said glucose, fructose and sucrose are in a molar ratio in water of 1: 1: 1: 11.
7. Un procedimiento según la reivindicación 1, caracterizado por que dicho disolvente eutéctico profundo constituido por azúcares naturales comestibles es miel. 7. A process according to claim 1, characterized in that said deep eutectic solvent constituted by edible natural sugars is honey.
8. Un procedimiento según la reivindicación 7, caracterizado por que dicho calentamiento se produce a una temperatura de entre 35 y 50°C. 8. A process according to claim 7, characterized in that said heating occurs at a temperature between 35 and 50 ° C.
9. Un procedimiento según la reivindicación 8, caracterizado por que dicha temperatura de 40°C. 9. A process according to claim 8, characterized in that said temperature of 40 ° C.
10. Un procedimiento según cualquiera de las reivindicaciones anteriores, caracterizado por que dicho calentamiento se produce durante un tiempo entre 20 y 240 min. 10. A process according to any of the preceding claims, characterized in that said heating occurs for a time between 20 and 240 min.
11. Un procedimiento según la reivindicación 10, caracterizado por que dicho tiempo de calentamiento es entre 50 y 90 min. 11. A method according to claim 10, characterized in that said heating time is between 50 and 90 min.
12. Un procedimiento según la reivindicación 11, caracterizado por que dicho tiempo de calentamiento son 60 min.
12. A method according to claim 11, characterized in that said heating time is 60 min.
13. Composición alimentaria funcional caracterizada por que comprende un extracto fenólico con oleuropeína, en que dicha oleuropeína está presente en una proporción de 80% - 85% en peso con respecto al peso del extracto fenólico total. 13. Functional food composition characterized in that it comprises a phenolic extract with oleuropein, in which said oleuropein is present in a proportion of 80% -85% by weight with respect to the weight of the total phenolic extract.
14. Uso de la composición de la reivindicación 13, como ingrediente en un producto alimentario, en que dicho producto alimentario está seleccionado del grupo que consiste en un caramelo o una gominola, una bebida carbonatada, un zumo, un producto de panadería, un producto energético o un lácteo. 14. Use of the composition of claim 13, as an ingredient in a food product, wherein said food product is selected from the group consisting of a caramel or a jelly bean, a carbonated drink, a juice, a bakery product, a product energy or a dairy.
15. Uso según la reivindicación 14, caracterizado por que dicho producto alimentario es un complemento dietético. Use according to claim 14, characterized in that said food product is a dietary supplement.
16. Uso según la reivindicación 15, caracterizado por que dicho complemento dietético es un complemento dietético antioxidante o antienvejecimiento.16. Use according to claim 15, characterized in that said dietary supplement is an antioxidant or anti-aging dietary supplement.
17. Uso de la composición de la reivindicación 13 en la preparación de un medicamento para la prevención o tratamiento de una enfermedad tumoral, neurodegenerativa, infecciosa, inflamatoria o cardiovascular.
17. Use of the composition of claim 13 in the preparation of a medicament for the prevention or treatment of a tumor, neurodegenerative, infectious, inflammatory or cardiovascular disease.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| ES201930815A ES2813774B2 (en) | 2019-09-20 | 2019-09-20 | PROCEDURE FOR THE OLIVE LEAF EXTRACTION OF OLEUROPEIN IN A PHENOLIC FRACTION, FUNCTIONAL FOOD COMPOSITION AND USE |
| ESP201930815 | 2019-09-20 |
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| WO2021053259A1 true WO2021053259A1 (en) | 2021-03-25 |
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| PCT/ES2020/070564 WO2021053259A1 (en) | 2019-09-20 | 2020-09-18 | Method for extracting, from olive leaf, oleuropein in a phenolic fraction; functional food composition; and use |
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| WO (1) | WO2021053259A1 (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008108817A1 (en) * | 2007-03-06 | 2008-09-12 | Ameriden International, Inc. | Nutritional supplement with oleuropein |
| WO2019145586A1 (en) * | 2018-01-24 | 2019-08-01 | Amador Mesa Juan Francisco | Thermal partition for vehicles |
-
2019
- 2019-09-20 ES ES201930815A patent/ES2813774B2/en active Active
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008108817A1 (en) * | 2007-03-06 | 2008-09-12 | Ameriden International, Inc. | Nutritional supplement with oleuropein |
| WO2019145586A1 (en) * | 2018-01-24 | 2019-08-01 | Amador Mesa Juan Francisco | Thermal partition for vehicles |
Non-Patent Citations (4)
| Title |
|---|
| ALANON ET AL.: "CHOLINE CHLORIDE DERIVATIVE-BASED DEEP EUTECTIC LIQUIDS AS NOVEL GREEN ALTERNATIVE SOLVENTS FOR EXTRACTION OF PHENOLIC COMPOUNDS FROM OLIVE LEAF", ARABIAN JOURNAL OF CHEMISTRY, 1 March 2018 (2018-03-01), pages 1 - 17, XP086032937, Retrieved from the Internet <URL:http://doi.org/10.1016/j.arabjc.2018/01/003> [retrieved on 20200123] * |
| DAI, Y. ET AL.: "TAILORING PROPERTIES OF NATURAL DEEP EUTECTIC SOLVENTS WITH WATER TO FACILITATE THEIR APPLICATIONS", FOOD CHEMISTRY, vol. 187, 2015, pages 14 - 19, XP029233810, Retrieved from the Internet <URL:http://dx.doi.org/10-1016/j.foodchem.2015.03.123> [retrieved on 20200123], DOI: 10.1016/j.foodchem.2015.03.123 * |
| GARCÍA A. ET AL.: "EXTRACTION OF PHENOLIC COMPOUNDS FROM VIRGIN OLIVE OIL By DEEP EUTECTIC SOLVENTS (DESs", FOOD CHEMISTRY, vol. 197, October 2016 (2016-10-01), pages 554 - 561, XP029312155, Retrieved from the Internet <URL:http://dx.doi.org/10.2016/jfood.chem.2015.10.131> DOI: 10.1016/j.foodchem.2015.10.131 * |
| ZOIDOU, E. ET AL.: "PREPARATION OF FUNCTIONAL YOGURT ENRICHED WITH OLIVE DERIVED PRODUCTS", YOGURT IN HEALTH AND DISEASE PREVENTION, 2017 * |
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| ES2813774A1 (en) | 2021-03-24 |
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