Estudio del 3,4-dihidroxifenilglicol (DHFG): fuentes, obtención y propiedades antiinflamatoria y fitorreguladora

Programa de Doctorado en Biotecnología, Ingeniería y Tecnología QuímicaLínea de Investigación: Biotecnología en la Agricultura, el Medioambiente, la Industria y la AlimentaciónClave Programa: DBICódigo Línea: 109La cuenca de los países del Mediterráneo en su conjunto forma parte de los mayores productores y exportadores de aceite de oliva a nivel mundial, y se han enfrentado durante décadas al exigente reto que supone la gestión de los subproductos que se generan en la industria olivícola. A lo largo de la historia, este sector se ha ido modernizando, buscando siempre calidad y bajos costes en su producción e indagando en la mejora de la incidencia que estos subproductos generan en la salud humana, animal y medio ambiental. Originalmente, la producción de aceite de oliva se llevaba a cabo a través de un sistema discontinuo que fue desplazado por los más recientes sistemas continuos, ya sean de dos o tres fases. En España, el decanter de dos fases, donde se genera aceite de oliva y alperujo se ha establecido como prioritario frente al de tres fases en el que además del aceite se generan orujo y alpechín. Además, los grandes volúmenes de salmueras que se producen en la industria de la aceituna de mesa es otro gran desafío a la hora su gestión. Por otro lado, existe un subproducto que hasta ahora era destinado principalmente a la quema y genera millones de toneladas como son las hojas y restos de poda del olivo. La importancia de la gestión de los subproductos no es solo por motivos medio ambientales, sino que también hay que tener en cuenta que la principal riqueza de éstos radica en que poseen la mayor parte de los componentes más bioactivos de la aceituna. Hay que resaltar, entre otros muchos, los compuestos fenólicos como el hidroxitirosol, el tirosol y la oleuropeína, que son objeto de estudio por sus innumerables propiedades como la de ser extraordinarios antioxidantes, antifúngicos, anticancerosos, antiagregantes plaquetarios, antiinflamatorios y un largo etc. Sin embargo, uno de los más recientemente descubiertos en los subproductos y por ello menos estudiado es el 3,4- dihidroxifenilglicol (DHFG), que sólo o en sinergia con el HT se está convirtiendo en el fenol simple con mayor potencial en farmacología, cosmética, alimentación y en el mundo agrario como un magnífico fitorregulador del crecimiento de cultivos. Uno de los objetivos del presente trabajo fue la búsqueda de fuentes de DHFG accesibles, económicas y sostenibles a partir de los subproductos de la industria de la aceituna. En esta búsqueda se han estudiado efluentes líquidos generados en el almacenamiento del alperujo que, al ser enriquecidos con aguas procedentes de la almazara, han aumentado su contenido en DHFG. Así mismo, con la finalidad de posibilitar la obtención de una fuente líquida rica en DHFG a partir de los tratamientos térmicos, que actualmente se están realizando para el aprovechamiento del alperujo, se ha hecho uso de los disolventes eutécticos profundos (DES). La combinación de estos disolventes con tratamientos térmicos no solo es muy efectiva para la obtención de DHFG, sino que además disminuye drásticamente la temperatura en el proceso, permitiendo una considerable reducción en el consumo energético. Así mimo, otro recurso para la obtención de fenoles estudiado son las aguas de conservación de las aceitunas de mesa en tres líquidos de conservación (salmuera, salmuera con ácido acético y agua acidificada). Los nuevos efluentes líquidos basados en el uso del ácido acético en la industria de las aceitunas de mesa facilitan la solubilización y extracción de HT y de DHFG. A partir de la purificación de estos fenoles se ha determinado la actividad antiinflamatoria in vitro mediante la evaluación de citoquinas proinflamatorias, para poder hacer uso de ellos en la industria farmacéutica como potenciales candidatos para combatir enfermedades de origen inflamatorio. Por otro lado, y desde un punto de vista agronómico el DHFG suma al conjunto de sus actividades biológicas la de ser un excelente fitorregulador que es capaz de potenciar la germinación y crecimiento de cultivos como el trigo, y la de frenar o mantener bajo control la de las malas hierbas a muy bajas concentraciones. El uso en agricultura de un compuesto con estas características supondría la eliminación de los riesgos que suponen el uso de los herbicidas sintéticos para los ecosistemas adyacentes a campos destinados a cultivos, así como al hombre. Posteriormente, y con el objeto de poder llevar a nivel industrial la obtención del DHFG para su uso tanto en agricultura como en alimentación o en farmacia, se diseñó un nuevo procedimiento a partir de las distintas fuentes que dio lugar a un novedoso sistema. Éste permite la extracción y purificación de un extracto rico en DHFG con baja concentración de HT. Este extracto se obtiene sólo por procedimientos físicos y económicos y permite la extracción del DHFG con un porcentaje de HT inferior al 10%, lo cual posibilita su uso agrícola. Como resultado de todas las investigaciones llevadas a cabo en este trabajo de tesis se ha conseguido detectar nuevas fuentes del DHFG a partir de los subproductos de las industrias del aceite de oliva y de la aceituna de mesa, así como un sistema sencillo de extracción y purificación de este fenol. Lo que puede permitir aportar beneficios importantes en desarrollar procesos que sean respetuosos con el entorno y económicamente viables. Todo esto ha permitido obtener extractos purificados para la verificación de propiedades poco estudiadas del DHFG como son la antiiflamatoria y fitorreguladora, así como hallar en este compuesto un producto comercial natural no tóxico, no contaminante, que no genera residuos y que es inocuo para la salud humana, animal y para el medio ambiente.Universidad Pablo de Olavide de Sevilla. Escuela de Doctorad

Rapid screening of unground cocoa beans based on their content of bioactive compounds by NIR spectroscopy

Near Infrared (NIR) spectroscopy was evaluated as a fast and easy method for identifying the most interesting cocoa genotypes according to chemical composition, including bioactive compound contents, in more than 80 samples of cocoa beans from the Mexican Germplasm Bank, which were harvested over three years. Clear dif ferences in chemical composition were detected in fermented and dried samples among genotypes. The contents in fat, protein, total sugars, total phenols, phenolic compounds and theobromine were determined for both whole cocoa beans and ground cotyledon samples; and calibration models were developed from the spectra of intact beans, nibs and ground cotyledons. In general, the calibration models obtained for cotyledon composition from the spectra of cocoa nibs and ground beans were better than those obtained from the spectra of intact beans. Fat content showed better calibration statistic values from the spectra of nibs and ground cotyledon (r2 = 0.70). Bioactive compounds, such as theobromine (r2 = 0.77), total sugars (r2 = 0.74), total phenols (r2 = 0.66) and derivatives of epicatechin (r2 = 0.88), together with fat (r2 = 0.70), protein (r2 = 0.64) and husk content (r2 = 0.82), were well-predicted using NIR spectroscopy in intact beans, cocoa nibs and/or ground cotyledon. The potential of NIRS technology was confirmed to support germplasm banks and breeding programs for the rapid identification of interesting genotypes based on their contents in bioactive compounds

The Effect of the Extra Virgin Olive Oil Minor Phenolic Compound 3′,4′-Dihydroxyphenylglycol in Experimental Diabetic Kidney Disease

The aim of this study was to analyze the possible nephroprotective effect of 3’,4’-dihydroxyphenylglycol (DHPG), a polyphenolic compound of extra virgin olive oil (EVOO), on renal lesions in an experimental model of type 1 diabetes. Rats were distributed as follows: healthy normoglycemic rats (NDR), diabetic rats treated with saline (DR), and DR treated with 0.5 mg/kg/day or 1 mg/kg/day of DHPG. DR showed a significantly higher serum and renal oxidative and nitrosative stress profile than NDR, as well as reduced prostacyclin production and renal damage (defined as urinary protein excretion, reduced creatinine clearance, increased glomerular volume, and increased glomerulosclerosis index). DHPG reduced the oxidative and nitrosative stress and increased prostacyclin production (a 59.2% reduction in DR and 34.7–7.8% reduction in DHPG-treated rats), as well as 38–56% reduction in urinary protein excretion and 22–46% reduction in glomerular morphological parameters (after the treatment with 0.5 or 1 mg/kg/day, respectively). Conclusions: DHPG administration to type 1-like diabetic rats exerts a nephroprotective effect probably due to the sum of its antioxidant (Pearson’s coefficient 0.68–0.74), antinitrosative (Pearson’s coefficient 0.83), and prostacyclin production regulator (Pearson’s coefficient 0.75) effects.This study was supported, in part, by the Consejería de Salud. Junta de Andalucía (Spain), Proyectos de Investigación en Salud [Regional Ministry of Health. Junta de Andalucía (Spain), Health Research Projects] (PI-0129-2017). Partial funding for open access charge: Universidad de Málag

Neuroprotective Effect of 3′,4′-Dihydroxyphenylglycol in Type-1-like Diabetic Rats—Influence of the Hydroxytyrosol/3′,4′-dihydroxyphenylglycol Ratio

The aim of this study was to assess the possible neuroprotective effect of 3′,4′-dihydroxyphenylglycol (DHPG), a polyphenol from extra virgin olive oil (EVOO), in an experimental model of diabetes and whether this effect is modified by the presence of another EVOO polyphenol, hydroxytyrosol (HT). The neuroprotective effect was assessed in a hypoxia–reoxygenation model in brain slices and by quantifying retinal nerve cells. The animals were distributed as follows: (1) normoglycemic rats (NDR), (2) diabetic rats (DR), (3) DR treated with HT (5 mg/kg/day p.o.), (4) DR treated with DHPG (0.5 mg/kg/day), or (5) with 1 mg/kg/day, (6) DR treated with HT plus DHPG 0.5 mg/kg/day, or (7) HT plus 1 mg/kg/day p.o. DHPG. Diabetic animals presented higher levels of oxidative stress variables and lower numbers of neuronal cells in retinal tissue. The administration of DHPG or HT reduced most of the oxidative stress variables and brain lactate dehydrogenase efflux (LDH) as an indirect index of cellular death and also reduced the loss of retinal cells. The association of DHPG+HT in the same proportions, as found in EVOO, improved the neuroprotective and antioxidant effects of both polyphenols.This study was supported, in part, by the Consejería de Salud. Junta de Andalucía (Spain), Proyectos de Investigación en Salud [Regional Ministry of Health. Junta de Andalucía (Spain), Health Research Projects] (PI-0129-2017). Partial funding for open access charge: Universidad de Málaga

An Acid-Free Alternative to Pectin Production from the Cell Walls of Olive Oil Waste and Different Fruits Using Choline Chloride

The pectin from the cell walls of olive waste (alperujo) and apple, orange and strawberry fruits was extracted using choline chloride (ChCl) and the yield and chemical and structural compositions were compared to pectin extracted using citric acid (CA) and ammonium oxalate/oxalic acid (AOOA). According to the results, the alperujo pectin extracted using ChCl from alcohol-insoluble residue (AIR) showed a higher yield (2.20–2.88% on the basis of dry weight of AIR) than using CA (0.65–1.22%) but lower than using AOOA (3.92–5.42%). For fruit pectin, the highest yield was obtained using CA (8.81–16%), followed by AOOA (5.4–6.63%), although for apple pectin, ChCl gave a similar yield (5.36%) to AOOA. The uronic acid contents in all ChCl pectins (45.9–70.6% dry basis AIR) were higher or similar to that of the other extracting agents (30.6–65.2%), although a lower level of neutral sugar side chains was detected, with a lower degree of branching and degree of methylation. The NMR and FT-IR spectroscopy of the pectin isolated using ChCl confirmed its slightly different structural composition with respect to CA and AOOA pectin. Therefore, depending on the source material and functionality, pectin isolated using ChCl could be an acid-free alternative to pectin production

An Acid-Free Alternative to Pectin Production from the Cell Walls of Olive Oil Waste and Different Fruits Using Choline Chloride

The pectin from the cell walls of olive waste (alperujo) and apple, orange and strawberry fruits was extracted using choline chloride (ChCl) and the yield and chemical and structural compositions were compared to pectin extracted using citric acid (CA) and ammonium oxalate/oxalic acid (AOOA). According to the results, the alperujo pectin extracted using ChCl from alcohol-insoluble residue (AIR) showed a higher yield (2.20–2.88% on the basis of dry weight of AIR) than using CA (0.65–1.22%) but lower than using AOOA (3.92–5.42%). For fruit pectin, the highest yield was obtained using CA (8.81–16%), followed by AOOA (5.4–6.63%), although for apple pectin, ChCl gave a similar yield (5.36%) to AOOA. The uronic acid contents in all ChCl pectins (45.9–70.6% dry basis AIR) were higher or similar to that of the other extracting agents (30.6–65.2%), although a lower level of neutral sugar side chains was detected, with a lower degree of branching and degree of methylation. The NMR and FT-IR spectroscopy of the pectin isolated using ChCl confirmed its slightly different structural composition with respect to CA and AOOA pectin. Therefore, depending on the source material and functionality, pectin isolated using ChCl could be an acid-free alternative to pectin production

Extraction of polyphenols associated with pectin from olive waste (alperujo) with choline chloride

10 Páginas.-- 6 Figuras.-- 1 TablaThe main by-product from olive oil extraction (alperujo) was extracted with hot water, citric acid, natural deep eutectic solvent (choline chloride: citric acid), and only choline chloride. The purified extracts were composed of macromolecular complexes constituting polyphenols associated with pectin. The extracts were structurally characterized by FT-IR and solid-NMR spectroscopy and an in vitro test revealed distinct antioxidant and antiproliferative activity, depending on the extracting agents. The choline chloride-extracted complex contained the highest amount of polyphenols among the examined agents, which exhibited a strong antioxidant activity and significant antiproliferative capacity. However, the complex extracted by hot water showed the highest antiproliferative capacity in vitro against the colon carcinoma Caco-2 cell line. In this finding, choline chloride could be used as a novel, green and promising alternative to the conventional extracting agent for the production of complexes that combine the antioxidant activity of phenolic compounds and the physiological effects of pectic polysaccharides.A. Bermúdez-Oria received funding from Council of Economic Transformation, Industry, Knowledge and Universities of the Junta de Andalucía, co-financed by the European Social Fund. Researcher doctor (DOC_00166). M.L Castejón received funding from Ministerio de Universidades of Spain under grant Margarita Salas (COD 19982). This work was supported by the Junta de Andalucia, Spain (Poyect PY18-616).Peer reviewe

Isolation and structural determination of cis- and trans-p-coumaroyl-secologanoside (comselogoside) from olive oil waste (alperujo). Photoisomerization with ultraviolet irradiation and antioxidant activities

8 Páginas.-- 4 Figuras.-- 2 Tablasp-Coumaroyl-6́-secologanoside (comselogoside) is a secoiridoid identified in large amounts in olive fruits, although no studies in vitro or in vivo of comselogoside have been reported. This work focuses on the recovery and purification of this compound from olive mill waste (alperujo). The successive isolation on Amberlite XAD-16 and Sephadex LH-20 resins, allowed a comselogoside extract with 80-85% of purity. A photoisomerization of the vinyl-double bond in the p-coumaroyl moiety occurred when the extract was exposed to ultraviolet radiation and a mixture of the trans and cis-isomers was obtained. Both isomers were characterized using NMR, mass spectroscopy, and UV spectrometry. The J (coupling constant) of the protons on the C7 and C8 on the unsaturated chain were found to be the difference between cis (12.8 Hz) and trans- (15.9 Hz) comselogoside. Cis-isomer exhibited lower radical-scavenging activity than trans, although a synergistic effect occurred when the cis-isomer was supplement by the trans-isomer.This study was funded by Project PY18-616 funded by the regional government of Andalucía, Junta de Andalucía, Consejería de Economía, Conocimiento, Empresas y Universidad, Andalucía, Spain, A. Bermúdez-Oria received funding from Council of Economic Transformation, Industry, Knowledge and Universities of the Junta de Andalucía, co-financed by the European Social Fund. Researcher doctor (DOC_00166). M.L Castejón received funding from Ministerio de Universidades of Spain under grant Margarita Salas (COD 19982).Peer reviewe

New Liquid Source of Antioxidant Phenolic Compounds in the Olive Oil Industry: Alperujo Water

The current management of alperujo as the main solid by-product from the two-phase olive oil extraction system has led to the appearance of a new liquid effluent that until now was treated together with the alperujo itself. The composition and antioxidant properties of its bioactive components at different depths of the pond were studied using colorimetric and HPLC with UV and MS detectors, DPPH, reducing power and rancimat. The concentration of suspended solids varied between 1.71 and 8.49 g/L, total fat was between 0.74 and 1.47 g/L, and total phenols were found between 3.74 and 4.11 g/L, which included hydroxytyrosol, 3,4-dihydroxyphenylglycol and tyrosol as the main phenols. Two types of extracts were obtained through two industrial systems with ethyl acetate and by chromotography, with an average content in total sugars of 2.1% and 3.16%, total phenols of 17.9% and 28.6% and hydroxytyrosol of 51.5 and 79.0 mg/g of extract, respectively. The activity presented by the chromatographic extract was higher in terms of free radical sequestering capacity, reducing power and the inhibition of lipid oxidation. Obtaining bioactive extracts would improve the formulation of food with natural components and at the same time would be the first step in a biorefinery to improve the management of the new effluent

Use of 3,4-dihydroxyphenylglycol (DHPG) as a plant growth regulator

The present invention relates to the use of the compound 3,4-dihydroxyphenylglycol (DHPG), compositions containing same, and methods for using said compositions as effective plant growth regulators for the treatment of crops and weedsPeer reviewedConsejo Superior de Investigaciones Científicas (España), Tecnofood ID Soluciones S.L.A1 Solicitud de patente con informe sobre el estado de la técnic