Alejandro Ruiz-Rodriguez

Recent trends in the advanced analysis of bioactive fatty acids.

The consumption of dietary fats have been long associated to chronic diseases such as obesity, diabetes, cancer, arthritis, asthma, and cardiovascular disease; although some controversy still exists in the role of dietary fats in human health, certain fats have demonstrated their positive effect in the modulation of abnormal fatty acid and eicosanoid metabolism, both of them associated to chronic diseases. Among the different fats, some fatty acids can be used as functional ingredients such as alpha-linolenic acid (ALA), arachidonic acid (AA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), gamma-linolenic acid (GLA), stearidonic acid (STA) and conjugated linoleic acid (CLA), among others. The present review is focused on recent developments in FAs analysis, covering sample preparation methods such as extraction, fractionation and derivatization as well as new advances in chromatographic methods such as GC and HPLC. Special attention is paid to trans fatty acids due its increasing interest for the food industry.

Optimization of countercurrent supercritical fluid extraction conditions for spirits fractionation

Abstract Optimization of the countercurrent supercritical fluid extraction (CC-SFE) conditions to obtain distilled alcoholic drinks extracts with low alcoholic content is presented. The optimization procedure is based on the use of an experimental design and the subsequent statistical study of the data obtained. Experiments have been performed with brandy, using a countercurrent supercritical fluid extraction (CC-SFE) system on a pilot-plant scale. The beverage is brought directly into contact with the carbon dioxide current in a packed column, and the extracts are recovered in two different fractionation cells, where depressurization occurs. The main variables that influence countercurrent extraction efficiency have been studied, such as sample flow rate, solvent flow rate (that is solvent-to-feed ratio) and level of sample introduction into the extraction column. Statistical data analysis indicates the variables that clearly influence the process and also the most convenient parameter values from which to obtain a high-value concentrated extract with a rich brandy aroma and low ethanol content.

Isolation of brandy aroma by countercurrent supercritical fluid extraction

Optimization of the countercurrent supercritical fluid extraction (CC-SFE) conditions to obtain high quality brandy aroma extracts is presented. The main variables that influence CC extraction selectivity and efficiency have been studied, such as extraction pressure and temperature and sample flow rate (related to the solvent-to-feed ratio). A rotatable central composite experimental design is used to optimize the combination of these experimental. Experiments have been performed with brandy using a CC-SFE system at pilot plant scale. The beverage is put directly in contact with the carbon dioxide in a packed column and the extracts are recovered in two different fractionation cells, where depressurization occurs. For each experiment, two extracted fractions and a raffinate are obtained and its aroma characterized by gas chromatography. A statistical study of the data obtained is performed including analysis of variance (ANOVA), fitting of a regression model and response surface study. The obtained results allow to know the variables that clearly influence the process and also show the interest of CC-SFE as an useful technique to obtain high-value concentrated brandy aroma extracts.

Pressurized water extraction of β‐glucan enriched fractions with bile acids‐binding capacities obtained from edible mushrooms

A pressurized water extraction (PWE) method was developed in order to extract β‐glucans with bile acids‐binding capacities from cultivated mushrooms (Agaricus bisporus, Lentinula edodes, and Pleurotus ostreatus) to be used as supplements to design novel foods with hypocholesterolemic properties. Extraction yields were higher in individual than sequential extractions being the optimal extraction parameters: 200°C, 5 cycles of 5 min each at 10.3 MPa. The crude polysaccharide (PSC) fractions, isolated from the PWE extracts contained mainly β‐glucans (including chitooligosaccharides deriving from chitin hydrolysis), α‐glucans, and other PSCs (hetero‐/proteo‐glucans) depending on the extraction temperature and mushroom strain considered. The observed bile acids‐binding capacities of some extracts were similar to a β‐glucan enriched fraction obtained from cereals.

Meat-based functional foods for dietary equilibrium omega-6/omega-3.

Nutritionists encourage improving the diet by combining meat products with fish or other sea-related foods, in order to equilibrate the omega-6/omega-3 ratio. Strong scientific evidence supports the beneficial health effects of a balanced omega-6/omega-3 PUFA (poly unsaturated fatty acids) diets. In the present work, the scientific bases of new functional meat products with both a balanced omega-6/omega-3 ratio and a synergic combination of antioxidants are discussed. The aim is to contribute to the dietary equilibrium omega-6/omega-3 and to increase the antioxidant intake. Conventional meat products supplemented with a specific fatty acids and antioxidants combination led to functional foods with healthier nutritional parameters.

Study on the 3-hydroxy-3-methyl-glutaryl CoA reductase inhibitory properties of Agaricus bisporus and extraction of bioactive fractions using pressurised solvent technologies.

BACKGROUND Agaricus bisporus mushrooms were able to lower cholesterol levels in hypercholesterolaemic rats and it was suggested that dietary fibre might inhibit cholesterol absorption. However, A. bisporus extracts were also able to inhibit the 3-hydroxy-3-methyl-glutaryl CoA reductase (HMGCR, the key enzyme in the cholesterol biosynthetic pathway) and this might also contribute to the observed lowering of cholesterol levels in serum. RESULTS The methanol-water extracts obtained from A. bisporus were able to inhibit up to 60% the HMGCR activity using an in vitro assay. The HMGCR inhibitory capacities depended on cultivation conditions, strains, etc. The potential inhibitors were not statins, they might be β-glucans able to scavenge the substrate and impair the enzymatic reaction. They were present during all mushroom developmental stages and similarly distributed through all the tissues including the parts discarded as a by-product. Accelerated solvent extractions using 1:1 ethanol-water as pressurised solvent (10.7 MPa, 25°C, five cycles of 5 min) were more effective in the extraction of the HMGCiR inhibitor(s) than supercritical fluid extractions (9 MPa, 40°C) using CO2 with 10% ethanol. CONCLUSION A mushroom cultivation and two extraction procedures were optimised to obtain fractions from A. bisporus with high HMGCR inhibitory activities to design novel ingredients for hypocholesterolaemic functional foodstuffs.

Antiviral Properties of Supercritical CO2 Extracts from Oregano and Sage

The antiviral properties of supercritical CO2 extracts obtained from oregano and sage were evaluated against the herpes simplex virus type 1 at different stages during virus infection. All of the extracts tested presented a moderate extracellular direct virucidal activity, although a pre-treatment of Vero cells, with 10 μg/mL of sage extracts before virus addition, inhibited 70% of the virus infection. Moreover, supercritical extracts of sage and oregano were able to significantly inhibit the in vitro virus replication, showing IC50 values of 1.88 and 5.33 μg/mL, respectively. Carvacrol and thymol could be pointed out as the compounds responsible for the antiviral activity found in oregano supercritical extracts; meanwhile, borneol, camphor, and 1,8-cineole could be proposed as antiviral compounds in supercritical sage extracts. Results demonstrated that supercritical extraction was an appropriate technique to obtain antiviral extracts from oregano and sage.

Vaccinium meridionale Swartz Supercritical CO2 Extraction: Effect of Process Conditions and Scaling Up

Vaccinium meridionale Swartz (Mortiño or Colombian blueberry) is one of the Vaccinium species abundantly found across the Colombian mountains, which are characterized by high contents of polyphenolic compounds (anthocyanins and flavonoids). The supercritical fluid extraction (SFE) of Vaccinium species has mainly focused on the study of V. myrtillus L. (blueberry). In this work, the SFE of Mortiño fruit from Colombia was studied in a small-scale extraction cell (273 cm3) and different extraction pressures (20 and 30 MPa) and temperatures (313 and 343 K) were investigated. Then, process scaling-up to a larger extraction cell (1350 cm3) was analyzed using well-known semi-empirical engineering approaches. The Broken and Intact Cell (BIC) model was adjusted to represent the kinetic behavior of the low-scale extraction and to simulate the large-scale conditions. Extraction yields obtained were in the range 0.1%–3.2%. Most of the Mortiño solutes are readily accessible and, thus, 92% of the extractable material was recovered in around 30 min. The constant CO2 residence time criterion produced excellent results regarding the small-scale kinetic curve according to the BIC model, and this conclusion was experimentally validated in large-scale kinetic experiments.

Extraction of bioactive compounds against cardiovascular diseases from Lentinula edodes using a sequential extraction method

Three extraction methods were sequentially combined to obtain fractions from Lentinula edodes (shiitake mushrooms) containing bioactive compounds against cardiovascular diseases (CVDs). Fruiting bodies were first extracted with plain water, obtained residue was then submitted to supercritical fluid extraction (SFE) and remaining residue submitted to hot water extraction. Sequential design allowed reutilization of the nonextracted material as raw material for the successive extractions increasing extraction yields and separating interesting compounds. Obtained fractions contained different amounts of ß‐glucans, chitins, eritadenine, lenthionine, ergosterol, proteins/peptides and phenolic compounds conferring them different bioactivities. Water soluble fractions showed high antioxidant activities (ABTS+• and DPPH• scavenging capacity and reducing power), they were also able to inhibit one of the main enzymes involved in hypertension (angiotensin‐I converting enzyme) and the key enzyme of cholesterol metabolism (3‐hydroxy‐3‐methylglutaryl coenzyme A reductase). The latter inhibitory activity was also noticed in SFE extracts although ergosterol and other lipid‐like molecules were isolated. Dietary fibers were separated in the third extraction. Therefore, with this sequential extraction procedure bioactive compounds against CVDs can be selectively separated from a single batch of shiitake powder.

Vaccinium meridionale Swartz extracts and their addition in beef burgers as antioxidant ingredient

BACKGROUND Vaccinium meridionale Swartz (mortiño) constitutes a source of bioactive phytochemicals, but reports related to its efficient and green production are scarce. In this study, pressurized liquid extraction (PLE) and ultrasound-assisted extraction of mortiño were compared. Total phenolic content (TPC) and antioxidant capacity (ABTS•+ ) were determined. Beef burgers with 20 g kg-1 of mortiño (MM) or its PLE extract (ME) were manufactured. Lipid oxidation (TBARS) and instrumental color changes were measured after refrigerated storage. RESULTS High TPC (up to 72 g gallic acid equivalent kg-1 extract) was determined in mortiño extracts, which was positively correlated with antioxidant activity. TBARS values of beef burgers containing either MM or ME did not change after refrigerated storage, whereas lipid oxidation of control burgers increased significantly. The color of burgers with added MM or ME was different (lower b* and a* values) from that of control burgers. However, the evolution of color after storage was similar between control and ME samples. CONCLUSION Mortiño extracts with high TPC can be obtained by PLE. Both mortiño and its PLE extract are able to control lipid oxidation of beef burgers, but the extract is preferred from the color quality point of view.