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Synergies between compounds in food ingredients have been commonly obtained by the mixing of extracts produced individually. A different approach, based on the coextraction of plants and a new theoretical framework to identify synergistic and antagonistic effects in the coextracts, has been proposed in this study for the first time. The Pressurized Liquid Extraction (PLE) of rosemary and marjoram with parsley at several plant ratios was carried out and the antioxidant activity (ABTS and cellular assays) and phenolic compounds of the extracts were evaluated. Synergistic effects on the antioxidant activity of the extracts were identified from all the plant combinations. The HPLC-PAD analysis of the samples did not show a relation between the concentration of the main phenolic compounds in the coextracts and the observed antioxidant synergistic effects. The proposed theoretical framework may be successfully used for the determination of synergies in the bioactivity of extracts obtained by coextraction.

Los insectos comestibles destacan como una fuente alternativa de proteínas. No obstante, durante el procesado de algunas especies se genera igualmente una fracción grasa relevante, como es el caso de las larvas de Hermetia illucens (mosca soldado negra), siendo un coproducto de gran interés para alimentación animal y humana. Por tanto, estudiar la influencia del procesado tecnológico sobre esta fracción grasa es clave para garantizar su calidad y uso en diversos fines. En el presente estudio, se llevó a cabo la evaluación del efecto de distintas formas de procesado de las larvas de H. illucens sobre la composición, calidad y bioactividad de la grasa obtenida. En concreto, se evaluaron dos formas de sacrificio (congelación o escaldado), seguido de dos formas de secado (horno o liofilización) y, de dos formas de desgrasado (prensado o extracción con fluidos supercríticos, SFE). Atendiendo al rendimiento de grasa, para todas las combinaciones de tratamientos, fue preferible el procedimiento de SFE, consiguiéndose el menor rendimiento al combinar escaldado-liofilización-prensa. De manera general, el perfil de ácidos grasos no sufrió cambios relevantes en función del procesado. No obstante, sí se evidenció una evidente lipólisis, con la consecuente generación de acidez libre, en función del tratamiento. Así, la aplicación de métodos no térmicos de congelación junto con liofilización dio lugar a la mayor degradación de la grasa en forma de ácidos grasos libres. Sin embargo, esta acidez fue minoritaria al aplicar métodos térmicos, como sacrificio por escaldado y secado por horno. Por otro lado, la combinación de congelación-liofilización-prensado produjo el mejor estado oxidativo inicial. De igual manera, el proceso de liofilización-prensado produjo las grasas más estables durante su almacenamiento a lo largo de 6 meses. En cuanto a la bioactividad de la grasa, se encontró una relación significativa entre la estabilidad oxidativa y la actividad antioxidante de la grasa, su

El interés por la investigación de las propiedades beneficiosas para la salud que presentan las plantas ha aumentado considerablemente debido a los numerosos compuestos bioactivos que poseen relacionados con actividades antioxidantes, anti-inflamatorias o antibacterianas, entre otras. En este trabajo, ha sido objeto de estudio la obtención de extractos enriquecidos en compuestos bioactivos de semillas de apio (Apium graveolens) para su potencial uso como ingrediente de un alimento o en la formulación de un nutraceútico con propiedades beneficiosas para la salud. Los extractos fueron obtenidos con diferentes disolventes mediante la técnica de extracción sólido-líquido asistida con ultrasonidos y se determinó de ellos tanto la composición química como las actividades biológicas antioxidante, anti-inflamatoria y antibacteriana frente a una cepa específica de Helicobacter pylori (H. pylori). Los resultados mostraron que la composición química de los extractos variaba en función de la polaridad del disolvente empleado y, por tanto, también su actividad biológica. Los extractos etanólicos fueron los que mayor contenido en compuestos fenólicos presentaron, así como en compuestos de la familia de las luteolinas y apigeninas, potentes antioxidantes, mientras que los extractos de hexano mayor contenido en compuestos del aceite esencial. Respecto a las actividades biológicas, el extracto etanólico presentó la mayor actividad antioxidante, con el menor valor del índice EC50, los extractos acuosos y etanólicos la mayor actividad anti-inflamatoria, mientras que los extractos de etanol y hexano fueron los que mostraron mayor actividad antibacteriana frente al H. pylori. En base a los resultados obtenidos, el extracto etanólico de las semillas de apio podría ser un ingrediente muy interesante para incorporar en un nutracéutico o un alimento con potenciales efectos beneficiosos para la salud en personas con problemas gástricos derivados de la infección por H. pylori como posible com

The interrelated effect of different slaughtering, drying and defatting methods of black soldier fly larvae (BSFL) on the lipid composition and properties of the fat was studied. Blanching and freezing were compared as slaughtering methods, oven or freeze-drying as drying methods, and mechanical pressing or supercritical fluid extraction (SFE) as defatting methods. The different modes of slaughtering, drying, and defatting, along with both binary and ternary interactions caused significant effects on processes yields, lipid composition, moisture content and thermal properties. Thus, considering the defatting degree and the yield in total valued products (defatted meal plus fat), the combination of blanching, freeze-drying plus mechanical pressing was the worst option (51.2% and 87.5%, respectively). In contrast, the other combinations demonstrated better and comparable efficiency, although SFE is preferable for defatting (83.2% and 96.9%, respectively). The content of major fatty acids (lauric, palmitic and myristic acids) was significantly affected by the BSFL treatments, although with unsignificant impact on the total saturated fatty acids content. To preserve the integrity of the fat, the combination of blanching and oven-drying was preferred, as non-thermal methods of slaughtering and drying caused intense lipolysis, releasing free fatty acids (FFA) in the range of 18.6-23.5%. To achieve the lowest moisture content in the fats (≤0.1%), oven-drying with mechanical pressing were desired, regardless of the slaughtering method; while values > 1% were reached for freezing, freeze-drying and SFE. Both differences in FFA and moisture contents caused different thermal behaviors in the samples. Specially, the melting temperature was lower for samples with higher FFA and moisture contents, with a notable difference when freezing, freeze-drying and SFE were combined (14.5 °C vs 30.6 °C, as the mean value for the rest of samples). The different modes of processing did not a

[Simple Summary] In addition to the potential of edible insects as an alternative source of proteins, the production of insect meals generates a significant amount of fat that is also of interest for food and feed. The consideration of the impact of processing factors on the quality of insect fats, such as oxidative stability, is indispensable to fully realize their value as a co-product. Therefore, this study aimed to understand how the different successive modes of processing of Hermetia illucens larvae (black soldier fly larvae, BSFL), such as slaughtering (by blanching or freezing), drying (by oven-drying or freeze-drying) and defatting (by mechanical pressing or supercritical CO2 SFE), as well as their interactions, affect the oxidation of the resulting fat and defatted meal. The preferred procedure would be a combination of freezing with freeze-drying and mechanical pressing, especially the combination of freeze-drying and mechanical pressing, for the best oxidative quality during storage. Such oxidative stability was strongly related to the antioxidant activity of the fats, which was conditioned by the processing methods used for BSFL. Therefore, this study provides guidance on selecting the best combinations of slaughtering, drying, and defatting processes for BSFL to obtain fats with the best oxidative quality., The interrelation effect of slaughtering, drying, and defatting methods of BSFL on the oxidative quality of the derived fat was studied. Blanching and freezing were compared as slaughtering methods, followed by oven or freeze-drying for drying and mechanical pressing or SFE for defatting. The oxidative state and stability of the extracted fat and defatted meals were monitored immediately after their production, using peroxide value (PV) and Rancimat test, and over 24 weeks of storage. Slaughtering and drying methods had an independent effect on PV, with freezing and freeze-drying being the best methods. Mechanical pressing and SFE were comparable and superior to conventional hexane defatting. Interactions were observed between slaughtering and defatting, drying and defatting, and between all three factors. Generally, freeze-drying combined with any of the slaughtering and defatting methods resulted in the lowest PVs, with mechanical pressing being preferred. Freeze-drying plus mechanical pressing also produced the most stable fats during storage according to the evolution of PV, while the combination of blanching and SFE produced the least stable. A significant correlation was found between the PV at 24 weeks and the antioxidant activity of the fats. Contrary to storage assays, in accelerated Rancimat assays, freeze-dried samples were the least stable, which was partially attributed to the significant correlation with the acid values of the samples. Defatted meals followed a similar pattern to the extracted fat, except for worse oxidation for SFE defatting. Therefore, the different processing methods of slaughtering, drying, and defatting of BSFL differently affect lipid oxidation, with interactions between such successive steps.

The mealworm (Tenebrio molitor Linnaeus 1758) is gaining importance as one of the most popular edible insects. Studies focusing on its bioactivities are increasing, although alternative forms of consumption other than the whole insect or flour, such as bioactive non-protein extracts, remain underexplored. Furthermore, the incidence of metabolic syndrome-related pathologies keeps increasing, hence the importance of seeking novel natural sources for reducing the impact of certain risk factors. The aim was to study the potential of a non-protein mealworm extract on metabolic syndrome-related pathologies, obtained with ethanol:water (1:1, v/v) by ultrasound-assisted extraction. We characterized the extract by gas-chromatography mass-spectrometry and assessed its hypolipidemic potential, its ability to scavenger free radicals, to attenuate the inflammatory response in microglial cells, to affect mitochondrial respiration and to enhance insulin sensitivity in mouse hepatocytes. The extract contained fatty acids, monoglycerides, amino acids, certain acids and sugars. The mealworm extract caused a 30% pancreatic lipase inhibition, 80% DPPH· scavenging activity and 55.9% reduction in the bioaccessibility of cholesterol (p = 0.009). The extract was effective in decreasing iNOS levels, increasing basal, maximal and ATP coupled respiration as well as enhancing insulin-mediated AKT phosphorylation at low insulin concentrations (p < 0.05). The potential of a non-protein bioactive mealworm extract against metabolic syndrome-related pathologies is shown, although further studies are needed to elucidate the mechanisms and relationship with compounds., [Simple Summary] Among edible insects, mealworm has gained attention due to its proven beneficial effects; however, mealworm products other than flour, such as concentrated forms or extracts different to proteins, are still rare. This novel food source can have a positive impact on multiple diseases such as via the metabolic syndrome. Our objective was to describe the composition of a mealworm extract and study its effect on metabolic syndrome-related pathologies. The extract interfered with the absorption of cholesterol and partially blocked the digestion of fats. It also reduced the inflammatory response, enhanced the sensitivity to insulin and showed strong antioxidant potential. Mealworm extract might be an interesting candidate for ameliorating the risk factors associated with the metabolic syndrome.

Nowadays, there is great interest in the discovery of food compounds that might inhibit gut microbial TMA production from its methylamine precursors. In this work, an innovative novel screening strategy capable of rapidly determining the differences in the metabolic response of Klebsiella pneumoniae, a bacteria producing TMA under aerobic conditions, to a library of extracts obtained from food and natural sources was developed. The proposed high-throughput screening (HTS) method combines resazurin reduction assay in 384-well plates and Gaussian Processes as a machine learning tool for data processing, allowing for a fast, cheap and highly standardized evaluation of any interfering effect of a given compound or extract on the microbial metabolism sustained by L-carnitine utilization. As a proof-of-concept of this strategy, a pilot screening of 39 extracts and 6 pure compounds was performed to search for potential candidates that could inhibit in vitro TMA formation from L-carnitine. Among all the extracts tested, three of them were selected as candidates to interfere with TMA formation. Subsequent in vitro assays confirmed the potential of oregano and red thyme hexane extracts (at 1 mg mL−1) to inhibit TMA formation in bacterial lysates. In such in vitro assay, the red thyme extract exerted comparable effects on TMA reduction (∼40%) as 7.5 mM meldonium (∼50% TMA decrease), a reported L-carnitine analogue. Our results show that metabolic activity could be used as a proxy of the capacity to produce TMA under controlled culture conditions using L-carnitine to sustain metabolism.

Plant extracts are being studied in the development of novel edible food coatings. The antioxidant and antimicrobial compounds that naturally occur in some plants are the key substances that contribute to preserving food quality. Besides the plant material, the method utilized to produce the extract influences its chemical and preservative characteristics. In general, hydrodistillation produce plant products (essential oils) that are well recognized for containing high concentration of antioxidant or antimicrobial volatile compounds. Supercritical fluid technology produces high quality bioactive extracts with higher yield but lower concentration of volatile compounds, in comparison with hydrodistillation. In this work, six different natural extracts of five different plants were produced by hydrodistillation and supercritical fluid extraction, and their antioxidant and antimicrobial activities were compared. The most active extracts were used to produce chitosan-based edible coatings with the aim of assessing the effect of essential oils versus supercritical extracts on the preservation of strawberries.

Supercritical anti-solvent precipitation (SAS) using carbon dioxide is a novel technique that can be used to produce powdered ingredients in small size particles, facilitating their incorporation into food matrices. In this work, the SAS precipitation of a licorice root ethanolic extract was studied. SAS assays were carried out at 15–20 MPa, 308.15 and 313.15 K, and two different concentrations (9.6 and 14.2 mg/ml) of the ethanolic licorice extract. In the range of conditions investigated, SAS pressure and temperature did not affect significantly the precipitation yield, but phytochemicals recovery was higher with the lower licorice extract concentration. Moreover, the fractionation of licorice bioactives (liquiritin, liquiritigenin, isoliquiritigenin, glabridin and glycyrrhizic acid) was assessed, together with the content of total phenolic compounds and antioxidant activity of the powders and oleoresin by-products obtained. In this respect, precipitates and oleoresins presented significant differences in the concentration of some licorice bioactives, and higher antioxidant activity was observed in precipitates. Additionally, significant effect of pressure, temperature and licorice extract concentration on the morphology and particle size of precipitates was observed, recovering smaller and more regular particles at 15–20 MPa, 313.15 K and 9.6 mg/ml licorice extract concentration, attaining satisfactory yield and antioxidant activity.

Heather (Calluna vulgaris L.) and marigold (Calendula officinalis L.) are two rich sources of bioactive pentacyclic triterpenes. The supercritical carbon dioxide (SCCO2) extraction of these two plants was explored at a pressure range of 25–50 MPa, 50 °C, with or without fractionation, different extraction times (1.5–4.0 h) and using ethanol as a co-solvent (0 and 10% w/w). In order to determine potential synergisms, a combined extraction (heather + marigold 50:50) was also studied. In general, higher extraction yields were achieved when the co-solvent was added. Higher concentrations of total triterpenic acids were obtained in heather extracts, specially using ethanol. The co-solvent did not increase the terpene concentration in marigold extracts. For the combined extraction, an antagonist effect in the triterpene concentration was observed in absence of a co-solvent, whereas a synergistic effect was exhibited in its presence, especially for ursolic acid. In general, the extracts showed a certain anti-inflammatory effect, although a straight correlation with the analyzed triterpenic acids concentration was not exhibited. Moreover, the combined extraction exposed a similar anti-inflammatory activity in comparison with the individual plant extracts.

Licorice (Glycyrrhiza glabra L.) is a plant used widely in herbal medicines due to their several biological potentials. The supercritical extraction of licorice roots was investigated to assess the antioxidant and antimicrobial activity of the extracts. Extraction conditions were pressures from 15 to 40 MPa, 313.15 and 333.15 K, and ethanol cosolvent in the range of 0 to 20% mass. In the case of high-pressure extractions using pure carbon dioxide (CO2) fractionation of the supercritical extract was accomplished in a two-cell decompression system. Fractionation was carried out with the aim to examine the potential separation of the antioxidant and antimicrobial licorice compounds and thus increase the bioactive properties of the fractions obtained in each separation cell. Main licorice bioactive compounds, liquiritin, liquiritigenin, glycyrrhizin, isoliquiritigenin and glabridin, were identified by HPLC and quantified using standards. Extracts obtained with supercritical CO2 and ethanol cosolvent contain the higher amounts of phenolic compounds and also the higher antioxidant activity but exhibit low or even no antimicrobial activity. Using pure CO2 at high pressure coupled with the on-line fractionation of the extract, two samples were obtained which showed, respectively, lower phenolic compounds content and good antimicrobial capacity (first fraction) and higher phenolic compounds content and antioxidant capacity (second fraction). Thus, the advantages of supercritical on-line fractionation are demonstrated in the extraction of Licorice roots.

En los últimos años, ha aumentado el uso de CO2 supercrítico (SCCO2) como antisolvente para la precipitación de compuestos bioactivos a partir de extractos vegetales. Este proceso de precipitación supercrítica antisolvente (SAS) permite emplear bajas temperaturas de procesamiento y evita el uso de disolventes orgánicos tóxicos. El objetivo de este trabajo, consistió en el fraccionamiento de compuestos bioactivos de romero (Rosmarinus oficinalis) mediante procesos SAS, evaluando las condiciones de extracción y el diseño de diferentes celdas de precipitación en las que se modificaron las conexiones de entrada y salida tanto del extracto como del SCCO2. El fraccionamiento de los compuestos se realizó a diferentes condiciones de presión (10-20 MPa), temperatura (40-60 ºC) y flujo másico de SCCO2 (50-60 g/min) durante 60 min, partiendo de una solución etanólica obtenida por ultrasonidos (UAE). De todas las fracciones obtenidas se determinó el contenido de compuestos fenólicos totales (TPC), el contenido en los principales compuestos antioxidantes del romero, ácido carnósico(CA) y ácido rosmarínico (RA), así como la actividad antioxidante. Además, la morfología de las partículas y la distribución de tamaño de partícula se determinaron mediante microscopia electrónica de barrido (SEM) y difracción láser respectivamente. Los resultados obtenidos por medio de la precipitación SAS en las diferentes condiciones mostraron una recuperación del extracto inicial entre un 80-90%. En las dos fracciones obtenidas, extracto seco en la celda de precipitación y fracción etanólica en el separador, los rendimientos fueron de 45-69% y 18-44%, respectivamente, dependiendo de las condiciones de precipitación. En el análisis de las fracciones se observó que el RA se concentró en la celda de precipitación hasta más del doble de la concentración inicial y el CA en el separador hasta casi el triple de la concentración inicial. El contenido en TPC fue superior en todos los casos al contenido del

La especie Cochlospermun angolensis (borututu) es un árbol nativo de las regiones tropicales de América Latina y África, ampliamente utilizado en la medicina tradicional. La raíz de borututu presenta una alta cantidad de compuestos fenólicos, sin embargo, su extracción se ha llevado a cabo únicamente mediante métodos tradicionales, utilizando principalmente agua como disolvente. En este trabajo se llevó a cabo la extracción de compuestos fenólicos mediante líquidos presurizados (PLE), a partir de la raíz y de las flores de borututu. Se estudiaron cuatro disolventes (agua, etanol, acetato de etilo y agua/etanol 50:50) y se aplicaron temperaturas entre 50 y 200 ºC. Como cabía esperar, el rendimiento de extracción aumentó con la temperatura y al aumentar la polaridad del disolvente (rendimiento del 57,3 % con agua a partir de la raíz y próximo al 50 % a partir de las flores). El rendimiento obtenido de las flores con acetato de etilo fue aproximadamente 2,5 veces superior al de la raíz, sugiriendo la presencia de una mayor cantidad de compuestos apolares en esta materia prima. Del mismo modo, la concentración de compuestos fenólicos (Folin-Ciocalteu) y la capacidad antioxidante (ensayo ABTS) aumentaron con la polaridad del disolvente. No obstante, la mayor concentración de estos compuestos y la mayor capacidad antioxidante de los extractos de la raíz se obtuvieron con la mezcla etanol/agua (374 mg GAE/g y 4,2 mmol Trolox/g).

Calendula officinalis, commonly known as marigold, has been used in traditional medicine for its anti-inflammatory and antiseptic virtues, and against gastric spasms. Here, we had obtained marigold extracts through supercritical CO2 extraction (SFE) and through ultrasonic assisted extraction (UAE), and we had characterized their bioactivity in the framework of cancer. We have found that marigold UAE extracts have a higher content of phenolic and flavonoid than marigold SFE extract, as well as a higher antioxidant activity. On the contrary, marigold SFE extract has a stronger cytotoxic and antiproliferative activity against pancreatic tumor-derived cells, being independent to its antioxidant effect. These results suggest that marigold SFE could be considered for further studies as a potential nutritional supplement which could improve current cancer therapies.

[Background] Pancreatic cancer is one of the most aggressive and mortal cancers. Although several drugs have been proposed for its treatment, it remains resistant and new alternatives are needed. In this context, plants and their derivatives constitute a relevant source of bioactive components which might efficiently inhibit tumor cell progression., [Methods] In this study, we have analyzed the potential anti-carcinogenic effect of different Asteraceae (Achillea millefolium and Calendula officinalis) and Lamiaceae (Melissa officinalis and Origanum majorana) plant extracts obtained by different green technologies (Supercritical CO2 Extraction –SFE- and Ultrasonic Assisted Extraction –UAE-) to identify efficient plant extracts against human pancreatic cancer cells that could constitute the basis of novel treatment approaches., [Results] Asteraceae extracts showed better results as antitumoral agents than Lamiaceae by inducing cytotoxicity and inhibiting cell transformation, and SFE extracts were most efficient than UAE extracts. In addition, SFE derived plant extracts from Achillea millefolium and Calendula officinalis displayed synergism with the chemotherapeutic 5-Fluororacil., [Conclusion] These results show how Yarrow and Marigold SFE-derived extracts can inhibit pancreatic cancer cell growth, and could be proposed for a comprehensive study to determine the molecular mechanisms involved in their bioactivity with the final aim to propose them as potential adjuvants in pancreatic cancer therapy.

The efficient production of saponin-rich extracts is of increasing interest due to the bioactive properties that have being demonstrated for these compounds. However, saponins have a poor bioavailability. In this respect, the knowledge about the bioaccessibility of saponins as a first step before bioavailability has been scarcely explored. In this study, the production of ultrasound-assisted extracts of saponins from edible seeds (quinoa, soybean, red lentil, fenugreek and lupin) was carried out with ethanol, ethanol:water or water. Extraction yield, total saponin (TSC), fat and total phenolics content (TPC) were determined. Then, the bioaccessibility of saponins after the in vitro gastrointestinal digestion of the extracts was determined and the effect of TPC and fat in the extracts on bioaccessibility was evaluated. The highest saponin-rich extracts were obtained by ethanol, being fenugreek and red lentil the richest extracts (12% and 10%, respectively). Saponins from ethanol:water extracts displayed variable bioaccessibility (from 13% for fenugreek to 83% for lentil), but a bioaccessibility closer to 100% was reached for all ethanol extracts. Correlation studies showed that TPC of the extracts negatively affected the bioaccessibility of saponins, whereas fat of the extracts enhanced this parameter. As summary, ultrasound-assisted extraction is shown as an efficient method for obtaining saponin-rich extracts from edible seeds, being ethanol the most advantageous solvent due to the richness of saponins and the successful bioaccessibility from these extracts, likely caused by the co-extracted fat with ethanol. Regardless of the extracts, phenolic compounds or fat may hinder or enhance the bioaccessibility of saponins, respectively. Additionally, an adequate balance between saponins to lipids has shown to be relevant on such an effect.

Asteraceae (Achillea millefolium and Calendula officinalis) and Lamiaceae (Melissa officinalis and Origanum majorana) extracts were obtained by applying two sequential extraction processes: supercritical fluid extraction with carbon dioxide, followed by ultrasonic assisted extraction using green solvents (ethanol and ethanol:water 50:50). The extracts were analyzed in terms of the total content of phenolic compounds and the content of flavonoids; the volatile oil composition of supercritical extracts was analyzed by gas chromatography and the antioxidant capacity and cell toxicity was determined. Lamiaceae plant extracts presented higher content of phenolics (and flavonoids) than Asteraceae extracts. Regardless of the species studied, the supercritical extracts presented the lowest antioxidant activity and the ethanol:water extracts offered the largest, following the order Origanum majorana > Melissa officinalis ≈ Achillea millefolium > Calendula officinalis. However, concerning the effect on cell toxicity, Asteraceae (especially Achillea millefolium) supercritical extracts were significantly more efficient despite being the less active as an antioxidant agent. These results indicate that the effect on cell viability is not related to the antioxidant activity of the extracts.