Method development in inverse modeling applied to supercritical fluid extraction of lipids

Modeling of the supercritical fluid extraction of solid materials is an important aspect in order to understand and predict the process. A comparison of two empirical models, two semi-empirical models and two mechanistic models is performed using calibration of single experiments. It is concluded that the best fit is obtained using a simple empirical expression. Furthermore, single calibrations did not generate reliable parameters with physical meaning and a methodology is proposed for inverse modeling with complete calibration using several experiments. The experimental dataset contained 29 extractions of lipids from crushed linseeds with varying temperatures, pressures and flow rates. A general rate model and a proposed extension of the hot ball model were evaluated for this purpose. The methodology includes data acquisition, model structure estimation, model calibration and a cross-validation. In general, it was found that the solubility model of Sovová outperformed the other evaluated correlations, and for the general rate model the Toth partition isotherm was also found in the top model structures. However, no generalization could be made regarding the correlations describing the Nernst diffusion layer and diffusivity

Comparative Life Cycle Assessment Study of Green Extraction Processes to Obtain Antioxidants from Rosemary Leaves

Trabajo presentado al 10th International Symposium on Supercritical Fluids celebrado en San Francisco (US) del 13 al 16 de mayo de 2012.Rosemary is one of the most appreciated natural sources for bioactive compounds with different activities such as antioxidant, antimicrobial or anticarcinogenic. Antioxidant activity has been associated to some of its components, among them, phenolic diterpenes such as carnosic acid, carnosol and rosmarinic acid. A careful selection of the extraction process together with the optimization of the extraction conditions, are of high importance to obtain rosemary extracts with high bioactivity. Among the different extraction processes, supercritical fluid extraction (SFE) and pressurized hot water extraction (PHWE) have demonstrated to be the most selective and environmental friendly techniques. However, extracts obtained by these processes usually require a drying step (freeze or hot drying step) which is both energy and time consuming. In this work, a new process combining PHWE and powder formation on-line (water extraction and particle formation on-line process, WEPO®) has been developed to obtain dry antioxidant powder from rosemary leaves in one step. In this process, parameters related to the extraction efficiency and selectivity (water flow rate and temperature) as well as parameters involving spray stability and powder formation have been considered at the same time. The obtained extracts have been evaluated in terms of their antioxidant activity using the DPPH method. Finally, in order to assess the viability and environmental impact of the new process, a comparison with other green processes used for antioxidant extraction from rosemary leaves such as SFE and PHWE (both followed by a freeze drying step) has been performed in terms of Life Cycle Assessment (LCA). Moreover, a sensitivity analysis of the LCA has been carried out to study the different environmental impact between the processes whether they are employed in different countries.This work was supported by AGL2008-05108-C03-01 (Ministerio de Educación y Ciencia), CSD2007-00063 FUN-CFOOD (Programa CONSOLIDER-INGENIO 2010), and S-0505/AGR/000153 (Comunidad Autónoma de Madrid) projects. C.T. acknowledges the Swedish Research Council for Environment, Agricultural Sciences and Spatial Planning (FORMAS, 2006-1346); the Swedish Research Council (VR, 2006-4084); and the Swedish Foundation for Strategic Research (SSF, 2005:0073/13, RMA08-0044). M.C-P. thanks MICINN for her “Juan de la Cierva” contract.Peer Reviewe

Aglycone specificity of Thermotoga neapolitana β-glucosidase 1A modified by mutagenesis, leading to increased catalytic efficiency in quercetin-3-glucoside hydrolysis

Background: The thermostable beta-glucosidase (TnBgl1A) from Thermotoga neapolitana is a promising biocatalyst for hydrolysis of glucosylated flavonoids and can be coupled to extraction methods using pressurized hot water. Hydrolysis has however been shown to be dependent on the position of the glucosylation on the flavonoid, and e. g. quercetin-3-glucoside (Q3) was hydrolysed slowly. A set of mutants of TnBgl1A were thus created to analyse the influence on the kinetic parameters using the model substrate para-nitrophenyl-beta-D-glucopyranoside (pNPGlc), and screened for hydrolysis of Q3. Results: Structural analysis pinpointed an area in the active site pocket with non-conserved residues between specificity groups in glycoside hydrolase family 1 (GH1). Three residues in this area located on beta-strand 5 (F219, N221, and G222) close to sugar binding sub-site +2 were selected for mutagenesis and amplified in a protocol that introduced a few spontaneous mutations. Eight mutants (four triple: F219L/P165L/M278I, N221S/P165L/M278I, G222Q/P165L/M278I, G222Q/V203M/K214R, two double: F219L/K214R, N221S/P342L and two single: G222M and N221S) were produced in E. coli, and purified to apparent homogeneity. Thermostability, measured as T-m by differential scanning calorimetry (101.9 degrees C for wt), was kept in the mutated variants and significant decrease (Delta T of 5 -10 degrees C) was only observed for the triple mutants. The exchanged residue(s) in the respective mutant resulted in variations in K-M and turnover. The K-M-value was only changed in variants mutated at position 221 (N221S) and was in all cases monitored as a 2-3 x increase for pNPGlc, while the K-M decreased a corresponding extent for Q3. Turnover was only significantly changed using pNPGlc, and was decreased 2-3 x in variants mutated at position 222, while the single, double and triple mutated variants carrying a mutation at position 221 (N221S) increased turnover up to 3.5 x compared to the wild type. Modelling showed that the mutation at position 221, may alter the position of N291 resulting in increased hydrogen bonding of Q3 (at a position corresponding to the +1 subsite) which may explain the decrease in K-M for this substrate. Conclusion: These results show that residues at the +2 subsite are interesting targets for mutagenesis and mutations at these positions can directly or indirectly affect both K-M and turnover. An affinity change, leading to a decreased K-M, can be explained by an altered position of N291, while the changes in turnover are more difficult to explain and may be the result of smaller conformational changes in the active site

Pressurised hot water extraction in continuous flow mode for thermolabile compounds: extraction of polyphenols in red onions

Abstract Extraction and analysis of labile compounds in complex sample matrices, such as plants, is often a big analytical challenge. In this work, the use of a "green and clean" pressurised hot water extraction (PHWE) approach performed in continuous flow mode is explored. Experimental data for extraction and degradation kinetics of selected compounds were utilised to develop a continuous flow extraction (CFE) method targeting thermolabile polyphenols in red onions, with detection by high-performance liquid chromatography (HPLC)-diode array detection (DAD)-mass spectrometry (MS). Water containing ethanol and formic acid was used as extraction solvent. Method performance was focused on extraction yield with minimal analyte degradation. By adjusting the flow rate of the extraction solvent, degradation effects were minimised, and complete extraction could be achieved within 60 min. The CFE extraction yields of the polyphenols investigated were 80-90 % of the theoretically calculated quantitative yields and were significantly higher than the yields obtained by conventional methanol extraction and static batch extraction (70-79 and 58-67 % of the theoretical yields, respectively). The precision of the developed method was lower than 8 % expressed as relative standard deviation

Composition and physicochemical properties of dried berry pomace

[EN] BACKGROUND Berry pomace is a valuable but little used by-product of juice manufacturing. When processed to a stable fruit powder, the composition differs from that of the whole fruit. To facilitate application in foods, a detailed knowledge of its composition and physicochemical properties is essential. RESULTS Blackcurrant, redcurrant, chokeberry, rowanberry and gooseberry were selected for analysis. All pomace powders had a high fibre content (> 550 g kg(-1)) and a fat content of up to 200 g kg(-1). Despite identical milling conditions, the particle sizes of the pomace powders varied. This can be traced back to seed content and brittleness, which also becomes apparent with respect to surface characteristics. Blackcurrant pomace powder differed from other varieties in terms of its low water-binding capacity (3.2 g g(-1)) and a moderate moisture uptake, whereas chokeberry pomace powder showed the highest polyphenol content and rowanberry pomace powder was rich in flavonols. CONCLUSION The results obtained in the present study provide a comprehensive overview of the properties of berry pomace powder and allow conclusions to be made regarding their applicability for use in complex food systems. (c) 2018 Society of Chemical IndustryThe research project was approved during the second SUSFOOD ERA-Net call (www.susfood-era.net).The funding of the project, assured through the national partner organizations, is gratefully acknowledged: Federal Ministry of Education and Research via PTJ in Germany (grant 031B0004), INIA in Spain and FORMAS in Sweden.Reibner, AM.; Al Hamimi, S.; Quiles Chuliá, MD.; Schmidt, C.; Struck, S.; Hernando Hernando, MI.; Turner, C.... (2018). Composition and physicochemical properties of dried berry pomace. Journal of the Science of Food and Agriculture. 99(3):1284-1293. https://doi.org/10.1002/jsfa.930212841293993Elleuch, M., Bedigian, D., Roiseux, O., Besbes, S., Blecker, C., & Attia, H. (2011). 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Structure dependent antioxidant capacity of phlorotannins from Icelandic Fucus vesiculosus by UHPLC-DAD-ECD-QTOFMS

Brown algae are rich in polyphenolic compounds, phlorotannins, which have been found to possess high in vitro antioxidant capacity, especially DPPH radical scavenging activity, due to the high number of hydroxyl groups. Whereas, the overall antioxidant capacity of brown algae extracts has been widely studied, the antioxidant capacity of individual phlorotannins has been rarely explored. The aim of this study was to determine the structure dependant antioxidant capacity of phlorotannins from Icelandic brown algae, Fucus vesiculosus. The antioxidant capacity of individual phlorotannins was determined by an on-line method using liquid chromatography and an electrochemical detector followed by quadrupole Time of Flight mass spectrometry (UHPLC-DAD-ECD-QTOFMS). Tentative structural elucidation of 13 phlorotannin isomers from EAF was obtained by LC-DAD-QTOFMS, ranging from 374 to 870 Da. On-line determination of antioxidant capacity of the individual phlorotannins generally showed that low molecular phlorotannins exhibited higher antioxidant capacity and that the capacity decreased with polymerisation

Adding Value to Fruit Processing Waste: Innovative Ways to Incorporate Fibers from Berry Pomace in Baked and Extruded Cereal-based Foods A SUSFOOD Project

This article communicates the set-up of BERRYPOM, a European research project established in the second call of the SUStainable FOOD Production and Consumption (SUSFOOD) network. The project deals with the by-product from berry processing, which is frequently recycled as animal feed, composted or utilized for biogas production. With BERRYPOM it is proposed to analyze the value of berry pomace, to optimize the recovery of bioactive compounds from pomace material, and to incorporate processed berry pomace in cereal-based foods to take advantage of nutritional benefits that originate from its fiber and the content of bioactive substances. Additionally, extraction methods will be evaluated to obtain products rich in phytochemicals, and the influence of OPEN ACCESS Foods 2015, 4 691 processing steps on the antioxidant capacity of pomace will be analyzed. The fiber extracts will then also be utilized in different cereal-based foods and extruded products. As project outcome we expect a substantial increase of knowledge concerning fiber and phytochemicals extraction from berry pomace, its suitability for enhancing nutritional and sensory properties of cereal-based foods, and its effects on the sustainability of the food chain.Rohm, H.; Brennan, C.; Turner, C.; Guenther, E.; Campbell, G.; Hernando Hernando, MI.; Struck, S.... (2015). Adding Value to Fruit Processing Waste: Innovative Ways to Incorporate Fibers from Berry Pomace in Baked and Extruded Cereal-based Foods A SUSFOOD Project. Foods. 4(4):690-697. doi:10.3390/foods4040690S6906974

Characterization of carotenoids in Rhodothermus marinus

Rhodothermus marinus, a marine aerobic thermophile, was first isolated from an intertidal hot spring in Iceland. In recent years, the R. marinus strain PRI 493 has been genetically modified, which opens up possibilities for targeted metabolic engineering of the species, such as of the carotenoid biosynthetic pathway. In this study, the carotenoids of the R. marinus type‐strain DSM 4252T, strain DSM 4253, and strain PRI 493 were characterized. Bioreactor cultivations were used for pressurized liquid extraction and analyzed by ultra‐high performance supercritical fluid chromatography with diode array and quadropole time‐of‐flight mass spectrometry detection (UHPSFC‐DAD‐QTOF/MS). Salinixanthin, a carotenoid originally found in Salinibacter ruber and previously detected in strain DSM 4253, was identified in all three R. marinus strains, both in the hydroxylated and nonhydroxylated form. Furthermore, an additional and structurally distinct carotenoid was detected in the three strains. MS/MS fragmentation implied that the mass difference between salinixanthin and the novel carotenoid structure corresponded to the absence of a 4‐keto group on the ß‐ionone ring. The study confirmed the lack of carotenoids for the strain SB‐71 (ΔtrpBΔpurAcrtBI’::trpB) in which genes encoding two enzymes of the proposed pathway are partially deleted. Moreover, antioxidant capacity was detected in extracts of all the examined R. marinus strains and found to be 2–4 times lower for the knock‐out strain SB‐71. A gene cluster with 11 genes in two operons in the R. marinusDSM 4252T genome was identified and analyzed, in which several genes were matched with carotenoid biosynthetic pathway genes in other organisms.Vetenskapsrådet, Grant/Award Number: 622-2010-333; Svenska Forskningsrådet Formas, Grant/Award Number: 239-2013-971 and 942-2015-1952; Seventh Framework Programme, Grant/Award Number: 311932Peer Reviewe