Katherina Fernández

An enzymatic extraction of proanthocyanidins from País grape seeds and skins.

Proanthocyanidins (PAs) from the skins and seeds of País grapes were obtained by enzymatic extraction by using three enzymes (pectinase, cellulase and tannase) and an enzymatic blend to increase the phenol concentrations and reduce the PA molecular size. The total phenol concentrations (as indicated by Folin-Ciocalteu reagent), mean degree of polymerisation (mDP), galloylation percentage (%G) and structural proportion (phloroglucinolysis) were analysed, in addition to the extracts capacity to inhibit angiotensin I-converting enzyme (ACE) activity. On grape skins, pectinase had the greatest effective on the release of total phenols, to 0.01g/ml solid/liquid (S/L) and 1% enzyme/substrate (E/S). On grape seed, the three enzymes were effective in increasing the phenolic extraction (p<0.05). The effects of enzymes on the mDP and %G of the extracts were related to their enzymatic activity. All the extracts inhibited ACE, but ACE inhibition was thought to be improved by the increased number of terminal units in the seed samples.

Startup and oxygen concentration effects in a continuous granular mixed flow autotrophic nitrogen removal reactor.

The startup and performance of the completely autotrophic nitrogen removal over nitrite (CANON) process was tested in a continuously fed granular bubble column reactor (BCR) with two different aeration strategies: controlling the oxygen volumetric flow and oxygen concentration. During the startup with the control of oxygen volumetric flow, the air volume was adjusted to 60mL/h and the CANON reactor had volumetric N loadings ranging from 7.35 to 100.90mgN/Ld with 36-71% total nitrogen removal and high instability. In the second stage, the reactor was operated at oxygen concentrations of 0.6, 0.4 and 0.2mg/L. The best condition was 0.2 mgO2/L with a total nitrogen removal of 75.36% with a CANON reactor activity of 0.1149gN/gVVSd and high stability. The feasibility and effectiveness of CANON processes with oxygen control was demonstrated, showing an alternative design tool for efficiently removing nitrogen species.

Simulated digestion of proanthocyanidins in grape skin and seed extracts and the effects of digestion on the angiotensin I-converting enzyme (ACE) inhibitory activity.

This study investigated the effect of in vitro gastrointestinal digestion on the stability and composition of flavan-3-ols from red grape skin and seed extracts (raw and purified, which are high in proanthocyanidins (PAs)). In addition, the effects of digestion on the angiotensin I-converting enzyme (ACE) inhibitory activities of these extracts were evaluated. The extracts were digested with a mixture of pepsin-HCl for 2 h, followed by a 2 h incubation with pancreatin and bile salts including a cellulose dialysis tubing (molecular weight cut-off 12 kDa) at 37°C with shaking in the dark and under N2. Under gastric conditions, the mean degree of polymerisation (mDP) of seed extracts, raw (mDP≈6, p<0.05), and purified (mDP≈10, p<0.05) was stable. The mDP of the raw skin extracts increased from 19 to 25 towards the end of the digestion. The PAs were significantly degraded (up to 80%) during the pancreatic digestion, yielding low-molecular-weight compounds that diffused into the serum-available fraction (mDP≈2). The overall mass transfer coefficient (K) of the seed extracts was 10(-7) m(2)/s. After simulated gastrointestinal digestion, over 80% of ACE inhibition by raw seed and skin extracts was preserved. However, the purified seed and skin extracts lost their ability to inhibit ACE after intestinal digestion.

Factorial design optimization and characterization of poly-lactic acid (PLA) nanoparticle formation for the delivery of grape extracts

Proanthocyanidins (PAs) extracted from grapes have several bioactive properties, giving them potential medical uses. However, PAs are unstable in the digestive tract and must be stabilized to allow oral administration, which can be accomplished by nanoencapsulation. In this study, PAs extracted from grape seed and skin were stabilized with poly-d,l-lactide (PLA) polymer by the emulsion-evaporation method. An experimental Box-Behnken design was implemented, evaluating the influence of three factors: sonication time (30-360s) for the emulsion formation, loading of grape extracts (5-20%) and concentration of stabilizing agent (polyvinyl alcohol, PVA: 1-3%). The process was optimized to achieve higher encapsulation efficiency (EE=82.7%) and a smaller size (256 nm). The nanoparticles (NPs) were physically analyzed by TEM, FT-IR, TGA and DTG to characterize the nanoencapsulation process. In vitro release studies, through stomach and intestinal simulation, showed a sustained release of PAs from PLA-NPs.

EFFECT OF THE BENCH SCALE EXTRACTION CONDITIONS ON Pinus radiata BARK EXTRACT YIELD, ANTIOXIDANT PROPERTIES AND COMPOSITION

Many pine bark extracts have antioxidant properties. This work examined bench scale extraction on the yield, the composition and antioxidant properties of Pinus radiata. Acetone solvent (98.7% w/w at 120 °C) yielded maximum phenols concentration, tannins concentration, anti-radical activity (DPPH) and reducing activity (FRAP), although its extraction yield was the lowest. When the organic solvent in the mixture decreased, the chelating activity increased (ICA) and antiradical and reducing activities decreased. The extract largest yield was obtained with water at 120oC, but the antioxidant properties were relatively poor. Taxifolin (dihydroquercetin) and catechin were the main extract components detected by RP-HPLC. The best extraction conditions for high extractive yield and antioxidant properties was achieved with ethanol/water at 120 °C.

Comparison of phenolic extracts obtained of Pinus radiata bark from pulp and paper industry and sawmill industry

Pinus radiata barks obtained from tress of different ages, as subproduct of pulp and paper (trees less than 10 years) and sawmill (trees between 20 to 25 years) industries, were used to produce extracts containing phenolic compounds. A factorial design 23 was used to evaluate the influence of the variables temperature (25 – 35 oC), solvent type (acetone - ethanol) and extraction time (1-12 h). The extracts were compared in their extraction yield (%), total phenols (by FolinCiocalteau), and radical scavenging activity (by DPPH). The extract obtained from old trunks presented a higher extraction yield than from young trees. The highest yield value was 2.56%, which was obtained using acetone as solvent for 12 h and 35°C. The highest concentration of phenol (5.84±0.18 g CE g extract-1), and scavenging activity (IP=86.1±4.4%) were also obtained for this type of extract. The extraction duration was the variable that most influenced the parameters studied. The bark’s radical scavenging power was greater than BHT (40%) and slightly lower than ascorbic acid (92%), common commercial antioxidants.

Technical and economical optimization of a full-scale poultry manure treatment process: total ammonia nitrogen balance.

ABSTRACT A full-scale process for the treatment of 80 tons per day of poultry manure was designed and optimized. A total ammonia nitrogen (TAN) balance was performed at steady state, considering the stoichiometry and the kinetic data from the anaerobic digestion and the anaerobic ammonia oxidation. The equipment, reactor design, investment costs, and operational costs were considered. The volume and cost objective functions optimized the process in terms of three variables: the water recycle ratio, the protein conversion during AD, and the TAN conversion in the process. The processes were compared with and without water recycle; savings of 70% and 43% in the annual fresh water consumption and the heating costs, respectively, were achieved. The optimal process complies with the Chilean environmental legislation limit of 0.05 g total nitrogen/L.