Novel Procedure for Lager Beer Clarification and Stabilization Using Sequential Enzymatic,Centrifugal, Regenerable PVPP and Crossflow Microfiltration Processing

In this work, the crossflow microfiltration (CFMF) performance of different lots of lager beer, produced in a pilot scale at the Italian Brewing Research Centre (CERB, Perugia, Italy), was assessed in a bench-top plant, equipped with a 0.8-μm ceramic tubular membrane module, under constant crossflow velocity of 6 m s^?1, transmembrane pressure difference of 3.74 bar, temperature of ~10 °C, and periodic CO₂ backflushing. By feeding different beer samples (i.e., as such, precentrifuged (C), or pretreated with a commercial enzyme preparation to degrade the original arabinoxylans and β-glucans and then centrifuged (EC) to minimize the fouling contribution of yeast cells, aggregates, and polysaccharides), it was possible to increase the average permeation flux (expressed as mean value?±?standard deviation) from 112?±?13 to 199?±?17 or 330?±?22 L m^?2 h^?1, respectively. Only when using the EC-pretreated beer specimens, the permeate turbidity at 20 °C approached the limiting one (<0.6 EBC unit) recommended by the European Brewery Convention standards. As expected, the permeate chill haze at 0 °C was generally higher than the above haze target. By submitting EC-pretreated beer seeded with 0.5 g L^?1 of regenerable polyvinylpolypyrrolidone (PVPP) to CFMF, it was possible to reduce the initial total polyphenol content by 30 % and permeate chill haze to 0.60?±?0.01 EBC unit, but the average permeation flux fell to 84?±?4 L m^?2 h^?1. By performing sequentially EC pretreatments, PVPP stabilization, cartridge filtration, and CFMF, it was possible not only to re-enhance the average permeation flux at about 230 L m^?2 h^?1 near to those achievable with DE filters, but also to obtain a chill haze-free permeate ready for aseptic packaging.     

Beer Clarification by Novel Ceramic Hollow‐Fiber Membranes: Effect of Pore Size on Product Quality

In this work, the crossflow microfiltration performance of rough beer samples was assessed using ceramic hollow‐fiber (HF) membrane modules with a nominal pore size ranging from 0.2 to 1.4 μm. Under constant operating conditions (that is, transmembrane pressure difference, TMP = 2.35 bar; feed superficial velocity, v_S = 2.5 m/s; temperature, T = 10 °C), quite small steady‐state permeation fluxes (J^*) of 32 or 37 L/m²/h were achieved using the 0.2‐ or 0.5‐μm symmetric membrane modules. Both permeates exhibited turbidity <1 EBC unit, but a significant reduction in density, viscosity, color, extract, and foam half‐life with respect to their corresponding retentates. The 0.8‐μm asymmetric membrane module might be selected, its corresponding permeate having quite a good turbidity and medium reduction in the aforementioned beer quality parameters. Moreover, it exhibited J^* values of the same order of magnitude of those claimed for the polyethersulfone HF membrane modules currently commercialized. The 1.4‐μm asymmetric membrane module yielded quite a high steady‐state permeation flux (196 ± 38 L/m²/h), and a minimum decline in permeate quality parameters, except for the high levels of turbidity at room temperature and chill haze. In the circumstances, such a membrane module might be regarded as a real valid alternative to conventional powder filters on condition that the resulting permeate were submitted to a final finishing step using 0.45‐ or 0.65‐μm microbially rated membrane cartridges prior to aseptic bottling. A novel combined beer clarification process was thus outlined.     

Biocomposite Films Based on κ-Carrageenan/Locust Bean Gum Blends and Clays: Physical and Antimicrobial Properties

The aims of this work were to evaluate the physical and antimicrobial properties of biodegradable films composed of mixtures of κ-carrageenan (κ-car) and locust bean gum (LBG) when organically modified clay Cloisite 30B (C30B) was dispersed in the biopolymer matrix. Film-forming solutions were prepared by adding C30B (ranging from 0 to 16 wt.%) into the κ-car/LBG solution (40/60 wt.%) with 0.3 % (w/v) of glycerol. Barrier properties (water vapour permeability, P _vapour; CO₂ and O₂ permeabilities), mechanical properties (tensile strength, TS, and elongation-at-break, EB) and thermal stability of the resulting films were determined and related with the incorporation of C30B. Also, X-ray diffraction (XRD) was done in order to investigate the effect of C30B in film structure. Antimicrobial effects of these films against Listeria monocytogenes, Escherichia coli and Salmonella enterica were also evaluated. The increase of clay concentration causes a decrease of P _vapour (from 5.34?×?10^?11 to 3.19?×?10^?11 g (m s?Pa)^?1) and an increase of the CO₂ permeability (from 2.26?×?10^?14 to 2.91?×?10^?14 g (m s?Pa)^?1) and did not changed significantly the O₂ permeability for films with 0 and 16 wt.% C30B, respectively. Films with 16 wt.% clay exhibited the highest values of TS (33.82 MPa) and EB (29.82 %). XRD patterns of the films indicated that a degree of exfoliation is attained depending on clay concentration. κ-car/LBG–C30B films exhibited an inhibitory effect only against L. monocytogenes. κ-car/LBG–C30B composite films are a promising alternative to synthetic films in order to improve the shelf life and safety of food products.     

Automated Sequential Injection Method for Determination of Caffeine in Coffee Drinks

An automated sequential injection analysis spectrophotometric assay for the determination of purine alkaloids in coffee drinks was developed. The sample was treated with a carrez reagent for matrix suppression followed by filtration; subsequently, alkaloids were separated from organic acids using a short C18 monolithic column (10 × 4.6 mm). The flow rate of the separation step was 10 μL s^?1 with 10% v/v of methanol as the mobile phase. The sum of alkaloids evaluated as caffeine was detected at 274 nm. The influence of the main parameters affecting the quantification of purine alkaloids was optimized. One sample analysis lasted 15 min when aspirated in triplicate. The linear range was 1–15 mg L^?1, and the determination coefficient (r ²) was 0.9969. The limit of detection and limit of quantitation were 0.128 and 0.425 mg L^?1, respectively. The repeatability evaluated as the relative standard deviation (RSD) was 3.58% (n = 12, 10 mg L^?1). Under optimal conditions, the method was successfully applied to determine purine alkaloids in different real samples including soluble coffee, coffee from an espresso machine, and brewed coffee drinks.     

Use of a Core-Shell Column for the Development of a Green LC Method for Thiol Determination in Fresh Fruits Following Derivatization with Methyl Propiolate

A novel HPLC method has been established for the determination of thiols in fruit samples, introducing, for the first time, methyl propiolate as an advantageous precolumn derivatization reagent for cysteine (CYS), glutathione (GSH), and N-acetylcysteine (NAC). The formed derivatives were detected at 285 nm, following isocratic separation on a core-shell column (Ascentis Express C18, 50?×?2.1 mm i.d., 2.7 μm) with a mobile phase of 15 mmol L^?1 (ΝΗ₄)₂ΗPO₄/H₃PO₄ (pH?=?2.2)/methanol (92:8?v/v), containing 1 mmol L^?1 EDTA, at a flow rate of 0.2 mL min^?1. Derivatization parameters were optimized including pH and concentration of buffering medium, amount concentration of methyl propiolate, derivatization time, and temperature, by the univariate approach. Under optimal conditions, the developed analytical scheme offers a total analysis time of less than 10 min, limits of detection in the range 0.1–0.5 μmol L^?1, and satisfactory linearity up to 100 μmol L^?1 for all analytes. The method proved also to be equally selective and robust. Endogenous thiols were determined in melon, watermelon, and avocado, using the standards addition approach, after minimal sample preparation, with no use of organic solvents. The accuracy was evaluated by recovery experiments resulting in the range of 86.4–118.5 %.     

Determination of Benzalkonium Homologues and Didecyldimethylammonium in Powdered and Liquid Milk for Infants by Hydrophilic Interaction Liquid Chromatography–Mass Spectrometry

In this study, a hydrophilic interaction liquid chromatography–mass spectrometry (HILIC-MS/MS) method for the determination of benzalkonium (BAC) homologues and didecyldimethylammonium (DDAC) was developed. A satisfactory chromatographic separation of BAC homologues and DDAC was achieved using, as mobile phase, acetonitrile–aqueous 50 mM ammonium formate (pH 3.2) (93?+?7 v/v) at 0.3 mL min^?1. The elution order of BAC homologues was from benzyldimethylhexadecylammonium chloride (C₁₆-BAC) to benzyldimethyldecylammonium chloride (C₁₀-BAC), the exact opposite with respect to separation using reversed liquid chromatography. The instrumental method was successfully applied to powdered and liquid milk for infants (about 50 samples). From powdered milk samples, BAC and DDAC were extracted using 5 % formic acid in methanol for 60 min at 60 °C in an ultrasonic bath; after dilution with water and 5 % NH₄OH solution, a purification step using a weak cationic exchange column was performed. Satisfactory limit of detections (LODs) were achieved, below 1.0 μg kg^?1 and 0.05 μg L^?1 for powdered and liquid milk for infants, respectively. No sample was free of BAC homologues and DDAC, and in one powdered milk sample, the contamination level exceeded 500 μg kg^?1, the maximum level recommended by the Standing Committee on the Food Chain and Animal Health for food and feed.     

Turbidity potentials of single long‐chain fatty acids and gelatinised starch in synthetic lautering wort

The turbidity potentials of long‐chain fatty acids as well as gelatinised starch were investigated in synthetic lautering wort to determine their contribution to turbidity according to concentration. Synthetic wort proved to be a standardised and reproducible model solution. Concentration series of gelatinised starch and single long‐chain fatty acids as pure substances showed high correlations between concentrations and turbidity levels of turbidity 11° and turbidity 90° measurements. Gelatinised barley starch has the turbidity potential of 0.04 EBC/(mg L^?1) on turbidity 11° levels. Regarding long‐chain fatty acids, saturated fatty acids showed distinctly higher turbidity potentials on turbidity 11° (≥0.32 EBC/(mg L^?1) and turbidity 90° (≥0.15 EBC/(mg L^?1) than unsaturated fatty acids. The turbidity potential of the saturated fatty acids corresponded with their chain lengths, with stearic acid (18:0) showing the highest turbidity potential. However, the impact on turbidity is primarily determined by concentration and not by turbidity potential.     

Dextrin as the main turbidity components in wort produced from major malting barley cultivars of Jiangsu province in China

As wort is the sweet starter liquid for beer, its quality needs to be strictly controlled. It is generally accepted that wort turbidity is of importance in terms of the brewing process and the resulting beer quality. This study investigated the major ingredients of wort turbidity using the malting barley cultivar Dan'er, grown in Jiangsu province, China. It was found that dextrins of low molecular weight constituted the vast majority of the polysaccharides causing wort turbidity. To solve the turbidity problem with the Dan'er malt, the present study supplemented 80 U g^?1 of β‐amylase, and as an activator, an extra 50 mg  L^?1 of Ca²⁺ to the malting process of Dan'er barley. The turbidity of the resulting congress wort of the Dan'er malt declined to <5.0 EBC units, which then met the quality guidelines of the brewery. The results of the present study may also help in developing new turbidity detection methods and yield breeding clues for quality improvement of the barley cultivars in the Jiangsu province of China. Copyright © 2016 The Institute of Brewing & Distilling     

Micellar Electrokinetic Chromatography Method for Determination of the Ten Water-Soluble Vitamins in Food Supplements

The separation and determination of the ten water-soluble vitamins by using capillary electrophoresis in the micellar electrokinetic chromatography in a single run are proposed. The method uses low toxicity and cost solvent (ethanol) as modifier of background electrolyte (BGE) attending to the Green Chemistry principles. The electrophoretic method uses 10.0 % (v/v) ethanol, 2.0 % (w/v) SDS, 0.02 mol?L^?1 borate at pH 8.70 as BGE. The standard and real sample solutions were injected in the eletrophoretic system by hydrodynamic injection under pressure of 0.80 psi for 8 s, and the separation was carried out in a fused silica capillary under a potential of 28 kV at 25 °C; the analytical signals were monitored at 214 nm. The analytical method is precise (r.s.d.?<?6 %), accurate (better than 9 %), selective, sensitive, robust, simple, and presents high analytical frequency as ten water-soluble vitamins were separated in only 18 min, with migration times of 5.75?±?0.02, 6.81?±?0.02, 8.13?±?0.04, 8.80?±?0.07, 8.98?±?0.06, 11.10?±?0.08, 11.34?±?0.05, 13.85?±?0.15, 14.82?±?0.04, and 17.85?±?0.30 min. Detection and quantification limits of 0.34, 0.32, 0.27, 0.20, 2.50, 4.98, 4.92, 0.30, 0.86 and 0.28 mg?L^?1 and 1.02, 0.97, 0.83, 0.62, 7.56, 15.09, 14.91, 0.90, 2.59 and 0.83 mg?L^?1, for vitamins PP (nicotinamide), B₁₂ (cyanocobalamin), B₂ (riboflavin), B₆ (pyridoxine), B₈ (biotin), C (ascorbic acid), B₅ (pantothenic acid), B₃ (nicotinic acid), B₁ (thiamine), and B₉ (folic acid), respectively. Excellent recoveries (intra and inter-day) were obtained and, when the method was applied to food supplement analyses the results were in agreement with the conventional HPLC methods.     

Simultaneous Determination of Cyadox and Its Metabolites in Chicken Tissues by LC-MS/MS

A specific and sensitive LC-MS/MS method was firstly established for the simultaneous extraction and determination of cyadox and its three main metabolites—1,4-bisdesoxycyadox, 4-desoxycyadox, and quinoxaline-2-carboxylic acid—in chicken muscle, liver, kidney, and fat tissues. Samples were subjected to extraction using ethyl acetate and followed by acetonitrile–chloroform (1:4, v/v) and further purified by Oasis mixed mode anion exchange SPE cartridge. Analysis was performed on a C₁₈ column by detection with MS in multiple-reaction monitoring mode. A gradient elution program with 0.1 % formic acid solution, acetonitrile, and 1 % formic acid (adjusted to pH 8 with ammonia) was performed at a flow rate of 0.2 mL min^?1. The correlation coefficients (r) for each calibration curve are higher than 0.99 within the experimental concentration range. The recoveries of the four target analytes at three spiking levels of 2.5, 25 and 250 μg kg^?1 were between 74.5 and 93.8 %, with relative standard deviations less than 12 %. The decision limits (CCαs) of the four analytes in chicken edible tissues ranged from 0.3 to 1.5 μg kg^?1, and the detection capabilities (CCβs) were below 2.3 μg kg^?1. The developed method demonstrated a satisfactory applicability in incurred chicken tissue samples.     

Antioxidant Activity and Determination of Phenolic Compounds from <Emphasis Type="Italic">Eugenia involucrata</Emphasis> DC. Fruits by UHPLC-MS/MS

Fruits have been the focus of several studies aimed at finding new antioxidant sources for protection against the damage caused by reactive species. In this study, the antioxidant activity and the presence of phenolic compounds in all parts (peel, pulp, and seeds) of Eugenia involucrata DC. fruits were evaluated. DPPH^·, ABTS^·+, and ORAC methods were used to determine the antioxidant activity, and an UHPLC-MS/MS method was developed for determining the phenolic compounds (gallic, chlorogenic, ferulic, p-coumaric and ellagic acids, quercetin, and myricetin). In the determination of both antioxidant activity and phenolic composition, the efficiency of solvents with different polarities—methanol/H₂O (80:20, v/v), ethanol/H₂O (80:20, v/v), methanol/acidified water with phosphoric acid pH 3.00 (80:20, v/v), and ethyl acetate—for the extraction of the phenolic compounds, was also evaluated. All parts of E. involucrata fruits showed antioxidant activity, in the range of 36.68 ± 1.44 to 873.87 ± 18.24 μmol TE g^?1, being the highest values found in the seeds and peel when more polar extraction solvents were used. Six, five, and three phenolic compounds were identified and quantified in the pulp, peel, and seeds, respectively, with the highest abundance as p-coumaric acid (14 ± 2 mg kg^?1) in the pulp, quercetin (47 ± 5 mg kg^?1) in the peel, and gallic acid (74 ± 4 mg kg^?1) in the seeds, also when more polar solvents were used. Although antioxidant activity methods suggested that the peel and seeds have more antioxidant potential, a wider variety of compounds were determined in the pulp.     

Colour Changes in Brandy Spirits Induced by Light-Emitting Diode Irradiation and Different Temperature Levels

One aged brandy was subjected to an artificial visible light source from light-emitting diodes (2?×?10³ lx) and under dark conditions. The temperature was also controlled at 25, 35, and 45 °C (±2 °C); the samples were monitored for colour changes; and selected chemical parameters were at 0, 10, 20, 30, and 40 days. Colorimetric indices, CIELab parameters and spectra were also determined with UV–Vis spectrometry. Visible light modified the colour of brandy and an important colour decrease was observed, showing maximum values (ΔE _ab*) at 25 °C, under the studied experimental conditions. On the contrary, at higher temperatures, significant colour degradation was not revealed. Non-colour parameters were not significantly affected by both irradiation and storage temperatures (p?>?0.05). Kinetic parameters including rate constant and activation energy were calculated. Based on Arrhenius model, activation energies of a*, browning index and h _ab* at darkness were 12.2, 55.1, and 21.6 kJ mol^?1 respectively.     

Stimulation of Saccharomyces cerevisiae Cultures by Pulsed Electric Fields

The effects of stimulation of Saccharomyces cerevisiae cells in an aqueous suspension by pulsed electric field (PEF) with electric field strength E?=?20–2,000 V cm^?1 and effective PEF treatment time t _PEF?=?10^?5–1 s were investigated. At relatively high electric field strengths (E?>?1,000 V cm^?1) and moderate times of PEF treatment (t _PEF?>?100 μs), the extraction of ionic components from yeast was observed, which can be related to electroporation of cell membranes. Petri dishes counting revealed dependency of the colony sizes on the time of preliminary fermentation t _f and power consumption W. The “logarithmic” and “saturated” types of electrostimulation were distinguished. At “logarithmic” electrostimulation (10^?7 J mL^?1?W??1 J mL^?1), the viability of yeast cells increased with the increase of power consumption and was higher for longer fermentation (t _f ?=?24 h). However, at “saturated” electrostimulation (10^?1 J mL^?1?W?1 J mL^?1), the viability of yeast cells was noticeably higher for t _f ?=?1 h than for t _f ?=?24 h. The impact of preliminary fermentation time and PEF protocol on biological activity of cells and consumption of nutrients was also discussed.     

Speciation Determination of Selenium in Seafood by High-Performance Ion-Exchange Chromatography-Hydride Generation-Atomic Fluorescence Spectrometry

A high-performance ion-exchange chromatography coupled with hydride generation-atomic fluorescence spectrometry (HPIEC-HG-AFS) method was developed for simultaneous speciation of selenium in seafood. Three selenium species including of selenocystine (Se-Cys), selenome-thionine (Se-Met), and selenite Se(IV) were separated on an anion-exchange column (PRP-X100) with eluent of 30 mM NH₄H₂PO₄ and methanol (39:1, v/v) in 10 min at the flow rate of 1.0 mL min^?1. Variables affecting the HG-AFS detection of selenium species were optimized. The optimum conditions found were the following: reducing agent, 2.0 % of KBH₄, and 5.0 % of HCl; lamp current, 90 mA; photomultiplier tube voltage, 280 V; flow rate of carrier gas, 300 mL min^?1; and shielding gas, 800 mL min^?1. Under the optimized conditions, the good linearity of calibration curves (R ²?>?0.999) between signal of fluorescence and concentration of selenium species was obtained in the range of detection limits (DLs), 80 μg L^?1, and the DLs of Se-Cys, Se-Met, Se(IV) were 1.66, 0.990, 1.10 μg L^?1, respectively. The repeatability of the method, expressed as relative standard deviation, was less than 5.0 % (n?=?10), and the average recoveries for spiked test were from 87.3 to 103 % for three analytes in real seafood samples. The developed HPIEC-HG-AFS method was successfully applied for the speciation of selenium in seafood samples.     

Validation of a HS-SPME-GC Method for Determining Higher Fatty Esters and Oak Lactones in White Rums

Higher fatty esters and oak lactones are the main components of white rum aroma and furthermore, they have an important sensorial impact in these distilled alcoholic beverages. A method for analyzing these volatile compounds was validated. It involves a separation and concentration step using headspace solid-phase microextraction (HS-SPME) and determination by capillary gas chromatography using flame ionization detection. The method showed a good within-day (RSD?<?3 %) and between-day precision (RSD?<?5 %). The calibration curves were linear at the tested ranges (R?>?0.99) and the limits of detection and quantification were 0.001–0.018 and 0.003–0.054 mg L^?1 (12 %?v/v alcohol), respectively. Good recoveries were obtained (98.6–100.3 %). The method is suitable for the quality control of higher fatty esters and oak lactones in white rums.     

A Combination of Chemical and Ultrasonication Treatments to Reduce Campylobacter jejuni on Raw Poultry

Cost-effective and acceptable decontamination technologies that can be readily applied to the broiler-processing industry have been proposed as a strategy to reduce pathogenic organisms on carcasses and improve food safety. The objective of this study was to determine the potential of combining chemical and ultrasonication processes to reduce Campylobacter, total viable counts (TVCs) and total Enterobacteriaceae counts (TECs) on broiler carcasses. Drumsticks were inoculated with Campylobacter jejuni and immersed in 12 % (w/v) trisodium phosphate (TSP), 2 % (w/v) citric acid (CA) or 5 % (w/v) capric acid sodium salt (CP) solutions for 1 min, while ultrasonication was performed at 40, 60 or 80 kHz. In addition, chemicals were administered in sequential combination (TSP?+?CA, TSP?+?CP or CA?+?CP) while ultrasonication was performed at the frequencies mentioned. The sequential treatment of TSP and CP with ultrasonication at 80 kHz achieved the greatest reductions for C. jejuni (4.5–4.6 log₁₀ colony forming units (CFU)/cm²) and TVC (1.9 log₁₀ CFU/cm²). Enterobacteriaceae were susceptible to the sequential application of CA and CP and ultrasonication at 80 kHz (2 log₁₀ CFU/cm²). This study demonstrated that combining chemical decontaminants with ultrasonication can significantly (p?相似文献   

Determination of Eugenol in Aquatic Products by Dispersive Solid-Phase Extraction and Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry

A novel procedure, dispersive solid-phase extraction coupled with ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), was developed for the determination of eugenol in aquatic products (shrimp, crab, and carp). Aquatic products were extracted with acetonitrile and primarily purified by dispersive solid-phase extraction with graphitized carbon black as absorbent. The pretreated acetonitrile extract was detected by UHPLC-MS/MS. UHPLC was carried out on Dikma Endeavorsil C₁₈ (30 mm × 2.1 mm, 1.8 μm) column eluted by methanol and water (80:20 v/v) at a rate of 0.30 mL min^?1. Tandem mass spectrometry was performed by electrospray ionization in negative ion mode to identify and quantify eugenol during multiple reaction monitoring. Under optimized analytical conditions, the matrix-matched spiked calibration sample demonstrates good linearity between 5.0 and 500.0 μg kg^?1 with a linear regression coefficient of 0.9996. The average recovery of eugenol from aquatic products is 95.3–103.4% at spiked levels between 5 and 50 μg kg^?1 with a relative standard deviation (n = 6) less than 5.4%. The limits of detection and quantification for eugenol were calculated to be 1.47 and 4.91 μg kg^?1, respectively. In comparison with those reported, the proposed method has advantages in low detection limit, high recovery, and short analysis time, meeting the requirements for the determination of trace eugenol residue in aquatic products.     

Fast and Selective Determination of Ochratoxin A in Wines Using an Optimized and Validated Liquid Chromatographic Method

Determination of ochratoxin A (OTA) in wines requires a cleanup step using solid phase extraction (SPE). Immunoaffinity columns are commonly the columns of choice but due to its high cost, other SPE columns have been assayed without optimal results. The present work reports an optimized and validated liquid chromatographic method for a fast and selective quantification of OTA in wines using C₁₈ columns for cleanup. Chromatographic conditions were optimized using a central composite design, establishing the following optimal conditions: acetonitrile/water/acetic acid (59.5:39.5:1.0 v/v/v) as mobile phase, flow rate of 1.2 mL min^?1, and column temperature of 30 °C. With these conditions, OTA had a retention time (~4 min) up to five times lower than those reported earlier. Regarding validation, calibration data (n?=?8) fitted a linear regression model with a determination coefficient (R ²) of 0.9992. Repeatability (relative standard deviation (RSD)) and intermediate precision (RSD) in matrix showed values of 1.3 % (n?=?6) and 0.8 % (n?=?3), respectively. Recoveries at five levels ranged from 87.2 to 118.9 % (mean RSD of 7.4 %). Fifty-three samples from different origins were analyzed, finding only seven samples (13 %) with quantifiable OTA content (0.15–0.26 μg L^?1). According to the levels found, the contribution of wine consumption to OTA daily intake was at least 58 times lower than the current tolerable daily intake. In view of optimization and validation results as well as its applicability to real samples, this method could be considered a good alternative for routine analysis of OTA in wines.     

Parameters affecting 5-hydroxymethylfurfural exposure from beer

5-Hydroxymethylfurfural (HMF) is generated during food and beverage heating processes and/or storage. Its daily intake, estimated as 4–10 mg day^?1, is several orders of magnitude higher than other process contaminants. Beer can be of relevance to the evaluation of HMF exposure; however, the information concerning its occurrence in different types of beer and during product storage is scarce. Therefore, the major goal of this work was to assess the amounts of HMF in different commercial beers, as well as the impact of storage, to deepen knowledge about the contribution of beer to HMF exposure. Blonde beers presented a mean content of 4.29 ± 1.05 mg L^?1, which was significantly lower (P ≤ 0.05) than those obtained for amber (6.84 ± 0.75 mg L^?1) and dark beers (6.99 ± 0.52 mg L^?1). Additionally, to study kinetic of HMF formation, fresh pilsner beers were stored at 30, 40 and 50°C during 40 days; a zero-order reaction was observed. The dependence of the rate constant on temperature was described by the Arrhenius equation and calculated activation energy was 101.85 kJ mol^?1. Storage can increase drastically HMF content, which means higher exposure for consumers. Thus, beer contribution to HMF exposure should not be neglected, since the intake of 1 L of beer entails a consumption of 4–7 mg of HMF or even more, depending on storage time and temperature.     

Production optimization,purification, and characterization of a novel acid protease from a fusant by Aspergillus oryzae and Aspergillus niger

The production of a novel acid protease was enhanced by 44 % through statistical optimization. The cultural parameters, such as inoculum size, temperature, moisture content, and incubation time, were 8.59 × 10⁵ g^?1 dry koji, 31 °C, 57 %, and 86 h, respectively. This novel acid protease was purified by 17 folds with a recovery yield of 33.56 % and a specific activity of 4,105.49 U mg^?1. Far-UV circular dichroic spectra revealed that this purified protease contained 7.1 % α-helix, 64.1 % β-sheet, and 32 % aperiodic coil. This novel acid protease was active over the temperature range of 35–55 °C with optimum temperature of 40 °C and was stable in the pH range of 2.5–6.5 with optimum pH of 3.5. Mn²⁺ enhanced its activity while Co²⁺ showed inhibitory effect. With casein as substrate, the kinetic parameters of K _m, V _max, energy of activation (E _a), and attenuation index of inactivation velocity by heat inducing (λ) were 0.96 mg mL^?1, 135.14 μmol min^?1 mg^?1, 64.11 kJ mol^?1, and 0.59, respectively.