A Short Extraction Time of Polysaccharides from Fenugreek (<Emphasis Type="Italic">Trigonella foencem graecum</Emphasis>) Seed Using Continuous Ultrasound Acoustic Cavitation: Process Optimization,Characterization and Biological Activities

In this study, the effective variables involved in ultrasound-assisted extraction (UAE) of fenugreek (Trigonella foenum graecum) seeds polysaccharides (FSPs) were optimized through Box–Behnken response surface design. The maximum yield of FSPs was 33.49% under the modified optimum conditions of an ultrasound power of 120 W, an irradiation time of 22 min and a liquid-to-solid ratio (L/S) of 30:1 mL g^?1. The predicted yield was in a very good agreement with the experimental yield of 33.41%. This value was higher than the FSPs yield obtained using conventional extraction (CE) method for 180 min at the same extraction temperature and L/S ratio. The primary chemical and structural characteristics were investigated by UV, FT-IR, and GC-MS. FSPs found to be a heteropolysaccharide consisted of galactose (38.18%), glucose (3.71%), mannose (46.13%), rhamnose (1.02%), and arabinose (0.83%). Furthermore, the FSPs exhibited considerable scavenging activity against 1,1-Diphenyl-2-Picrylhydrazyl free radicals and ferric reducing antioxidant power and reducing power, in a concentration-dependent mode in vitro. Our results suggested that UAE technique gave a higher yield of FSPs with a shorter extraction time than the CE and FSPs can also be used as promising resources of natural agent for functional foods and medicinal industries.     

Simultaneous Detection of Azodicarbonamide and the Metabolic Product Semicarbazide in Flour by Capillary Electrophoresis

In the present work, capillary electrophoresis (CE) was used for the first time for the simultaneous analysis of azodicarbonamide (ADA) and semicarbazide (SEM), and the capillary electrophoresis separation conditions, extraction agents, and derivatization conditions were investigated. In 20 mmol L^?1 sodium tetraborate, 30 mmol L^?1 β-cyclodextrin (β-CD), 17 % isopropanol (v/v), and 25 mmol L^?1 sodium dodecyl sulfate (SDS) running buffer, ADA and SEM previously derivatized with 9-fluorenylmethyl chloroformate (FMOC) were separated in less than 25 min with good sensitivity. The linear ranges were 8.3?×?10^?4～6.6?×?10^?2 mmol L^?1 and 1.9?×?10^?3～3.4?×?10^?2 mmol L^?1, and detection limits (S/N?=?10) were 0.5 and 0.15 mg kg^?1 for ADA and SEM, respectively. The proposed method was successfully applied for the simultaneous analysis of ADA and SEM in five flour samples with satisfactory recovery data from 88.0 to 93.0 % for ADA and 98.0 to 106.0 % for SEM, indicating the valuable potential application of this method for food analysis.     

Thermal transition and thermo-physical properties of potato (<Emphasis Type="Italic">Solanum tuberosum L</Emphasis>.) var. Russet brown

Potatoes are an important food in many regions of the world and are commonly used in a variety of food products. Thermal transition and thermo-physical properties of potatoes are important in order to design efficient food processes and select appropriate storage conditions. In this study, we determined the thermal transitions and thermophysical properties of raw and blanched/par-fried potato for a temperature range of ??32 to 21.1 °C. Using differential scanning calorimetry, we found an initial freezing point (T_f) at ??1.8?±?0.1 °C, an onset of melting (T_m^′) at ??9.9?±?0.2 °C and an unfreezable water content (X^′_w) for maximally freeze-concentrated raw potato at 0.21 kg water/kg potato. Corresponding values for blanched/par-fried potatoes were ??0.9?±?0.1 °C, ??11.0?±?0.2 °C and 0.18 kg water/kg potato. Results show that an increase in solids content decreased T_f of both raw and blanched potatoes. We modelled the relationship between them using the Chen model. The apparent specific heat (C_app) increased around T_f to 31.7?±?1.13 kJ/kg K for raw potato and 26.7?±?0.62 kJ/kg K for blanched/par-fried potato. For frozen raw potato at ??32 °C, thermal diffusivity (α) was 0.89?±?0.01?×?10?^?6 m²/s and thermal conductivity (k), 1.82?±?0.14 W/m K, respectively. These values were higher for frozen raw potato than for the unfrozen raw potato (0.15?±?0.01?×?10?^?6 m²/s and 0.56?±?0.08 W/m K, respectively at 21.1 °C). The apparent density (ρ) of frozen raw potato (992?±?4.00 kg/m³ at ??32 °C) was less than that for unfrozen raw potato (1053?±?4.00 kg/m³ at 21.1 °C), and a similar trend was obtained for blanched/par-fried potato (993?±?2.00 kg/m³ at ??32 °C and 1188?±?7.00 kg/m³ at 21.1 °C, respectively). This study established a correlation between thermo-physical properties and temperature. Findings may be used to inform the design and optimization of freezing processes and frozen storage for potato products.     

Steady- and Unsteady-State Determination of the Water Vapor Permeance (<Emphasis Type="Italic">WVP</Emphasis>) of Polyethylene Film to Estimate the Moisture Gain of Packed Dry Mango

The water vapor permeance (WVP; g m^?2 d^?1 Pa^?1) of packaging films quantifying the water vapor transfer rate between foods and its surroundings is usually determined in units operating under steady-state conditions that do not necessarily reflect food handling scenarios. This study evaluated the determination of the WVP of a polyethylene (PE) film by steady-state method ASTM F1249-06 using a permeability cell and unsteady-state method ASTM E96/E96M in which 102 vacuum-sealed PE bags containing silica gel were stored (37.8 °C, 75% relative humidity) and weighed over 25 days. Average steady-state WVP (2.935 ± 0.365 × 10^?3, n = 4) fell within the 95% quantiles of unsteady-state WVP values (1.818–3.183 × 10^?3, n = 2142). Moisture uptake of dehydrated mango stored at 37.8 °C and 75% relative humidity was predicted with WVP values obtained by both methods. Predictions were validated by monitoring over 25 days the weight gain of 100 PE bags with dry mango. Experimental moisture averages during storage fell within one standard deviation of predictions using the unsteady-state WVP (R ² = 0.974). The same was observed only until day 15 for predictions obtained with the steady-state WVP. Calculations for days 20–25 overestimated the moisture uptake by 6.0–7.2%, resulting in registered R ² = 0.924. The unsteady-state WVP determination is low-cost, uses large numbers of film samples, and allowed more accurate predictions of dry mango moisture uptake. Knowledge of the moisture uptake controlled by the film WVP is essential when predicting the safety and quality changes limiting the shelf-life of foods.     

A mathematical model for moisture movement during continous and intermittent drying of <Emphasis Type="Italic">Eucalyptus saligna</Emphasis>

A modified diffusion-based mathematical model is proposed to describe the moisture movement during continuous and intermittent drying of Eucalyptus saligna. This model includes the temperature change, the surface drying coefficient (β _n ) and 2 diffusion coefficients [from green to FSP (D _f ) and from FSP to dry condition (D _o )] as important parameters. The final model expression obtained was M?=?exp (??25 β _n ² D _t /l²) with the β _n used was 1.5807 kg m^?2 s^?1, the D _f was 2.26?×?10^?11 m² s^?1, and the D _o was 5.85?×?10^?12 m² s^?1. The range of temperature change between heating and non-heating phases in the intermittent drying regimes was from 24.9 to 31.8 °C. The R² values obtained when the model was fitted into the drying data of different intermittent regimes ranged from 71.5 to 85.9%. The R² value was 87.4% when the model was fitted into continuous trial data. The high values of R² indicate that the model can be used to understand the moisture reduction both in intermittent and continuous regimes.     

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.     

Smart NMR Method of Measurement of Moisture Content of Vegetables During Microwave Vacuum Drying

Microwave drying is usually combined with vacuum environment in conjunction with hot air flow to draw the moisture rapidly. The moisture content of the vegetables undergoing drying is hard to measure online. This research designed a microwave vacuum drying (MVD)-low-field nuclear magnetic resonance (NMR) smart device and investigated the feasibility of NMR method for online measurement of state of moisture during MVD. The relation between the signal amplitude (A ₂) and the true moisture content (M ₁) of six kinds of vegetables (mushroom, carrot, potato, lotus, edamame, vegetable corn) was fitted to estimate if NMR can measure the M ₁ of vegetables directly. Results showed that A ₂ and M ₁ of different fresh vegetables had no single empirical mathematical model to fit. However, for each kind of these vegetables, the A ₂ and corresponding M ₁ in different MVD stages showed a significant linear relationship. The predicted moisture content (M ₂) of mushroom: M ₂ = 5.25351 × 10^?4 A ₂ ? 0.34042, R = 0.996; carrot: M ₂ = 5.78756 × 10^?4 A ₂ ? 0.14108, R = 0.998; potato: M ₂ = 3.10019 × 10^?4 A ₂ ? 0.10612, R = 0.991; lotus: M ₂ = 2.32415 × 10^?4 A ₂ ? 0.01573, R = 0.998; edamame: M ₂ = 3.13310 × 10^?4 A ₂ ? 0.4198, R = 0.996; vegetable corn: M ₂ = 1.69461 × 10^?4 A ₂ ? 0.09063, R = 0.995. The linear models between M ₂ and A ₂ were able to estimate the end point (M ₁ < 8%) of MVD with a high accuracy (P > 0.950).     

Characterization of Biodegradable Films Based on <Emphasis Type="Italic">Salvia hispanica</Emphasis> L. Protein and Mucilage

Biodegradable films of chia by-products (mucilage and protein-rich fraction (PF)) incorporated with clove essential oil (CEO) were obtained and characterized. The effects of polymer concentration (PC; 1.0–3.0 %, w/v) and CEO concentration (0.1–1.0 %, v/v) were evaluated as well as the pH (7–10), using a 2³ factorial design with four central points. The films exhibited moisture values between 11.6 and 52.1 % (d.b.), which decreased (p?<?0.05) with increasing PC and CEO. The thickness of the films increased (p?<?0.05) with increasing PC. PC and pH influenced (p?<?0.05) the lightness (L) and variation in color between red and green (a). The orientation of the color to yellow-blue hues (b) decreased significantly (p?<?0.05) with increasing PC. Transparency was significantly lower and higher (p?<?0.05) than PC and CEO, respectively. The film surface morphology was evaluated using atomic force miscrocope images, and thermogravimetric analysis (TGA) was performed to study the thermal stability of the films. The displacement and tensile strength were significantly lower (p?<?0.05) at higher concentrations of CEO, this variable being the only one with a significant effect. The chemical composition of the films was confirmed utilizing Fourier transform infrared (FTIR) spectroscopy. The proportion of CEO added to the films had a significant influence on antimicrobial activity, inhibiting the growth of both Escherichia coli and Staphylococcus aureus.     

Development and characterization of reconstituted hydrogel from <Emphasis Type="Italic">Aloe vera</Emphasis> (<Emphasis Type="Italic">Aloe barbadensis</Emphasis> Miller) powder

Structural and rheological characterization of reconstituted hydrogels developed from A. vera non-fibrous alcohol insoluble residue (NFAIR) powder using different methods [viz., shaking (S), heating-shaking (HS), and heating (H)] and concentrations (viz., 0.2–1.6 %, w/v) was carried out. Functional group distribution by FTIR spectroscopy and Congo red (CR) method revealed the presence of acetylated acemannan in A. vera powder. Dynamic oscillation studies of A. vera (NFAIR) fluids at all concentrations of 0.2–1.6 %, w/v, showed gel strength in the order of H > HS > S method. However, in H method, increase in concentration from 0.2 to 1.6 %, w/v showed the conformational transition from semi-diluted solution to weak gel nature. Rheological models described the effect of heating temperatures (HT); 30–90 °C, and times (Ht); 15–60 min on viscoelastic behavior in reconstituted A. vera fluids. The reconstituted A. vera hydrogel prepared with a concentration of 1.6 %, w/v using 50 °C (HT) and 30 min (Ht) condition showed a good agreement with the Power law (storage modulus, G′) and Weak gel model (complex modulus, G*) fitted data (R² > 0.94) resulting higher viscoelastic moduli intercepts; G′₀ (71.5 Pa s ^n′), G″₀ (33.5 Pa s ^n″), lower slopes; n′ (0.22), n″ (0.06), higher network strength (A _F , 121.3 Pa s^1/z ) and number of network (z, 5.3) values. The obtained results suggested that heating at 50 °C/30 min can develop aqueous weak gel networks of A. vera with enhanced gel strength which may be utilized as a novel gelling agent for wide variety of targeted applications in food and pharmaceutical sectors.     

Development of an Aqueous Polyethylene Glycol-Based Extraction and Recovery Method for Almond (<Emphasis Type="Italic">Prunus armeniaca L</Emphasis>.) Protein

A novel method for protein extraction from sweet almonds with aqueous polyethylene glycol (PEG) as solvent and recovery from the extraction solution was developed. The extraction yields of different solvents, such as sodium hydroxide, sodium chloride, PEG 200, PEG 400, and PEG 600 aqueous solutions, were investigated and PEG 200 showed the highest extraction efficiency. The PEG-based microwave-assisted extraction (MAE) parameters were then optimized using response surface methodology. Under optimum condition, PEG 200 concentration of 25 % (w/w), liquid to solid ratio of 22 mL g^?1, microwave power of 120 W, extraction temperature of 45 °C, and extraction time of 4 min, the average extraction yield was 93.75?±?3.15 %. Subsequently, the almond protein was recovered from the extraction solution containing PEG with an isoelectric point-ethanol synergy precipitation protocol. The combined technique integrated the speed of isoelectric point precipitation with the completeness of alcohol precipitation. The recovery of almond protein was 98.81 % with a time of 3–5 min. The proposed PEG-based MAE and synergy precipitation protocol provide a rapid and effective method for almond protein extraction and recovery and have the potential to be used for other plant proteins.     

An approach for extraction,purification, characterization and quantitation of acylated-anthocyanins from <Emphasis Type="Italic">Nitraria tangutorun</Emphasis> Bobr. fruit

In the present study, a systematic approach for extraction, purification and analysis of acylated-anthocyanins from Nitraria tangutorun Bobr. fruit was explored. Six acylated-anthocyanins in N. tangutorun fruit were identified by HPLC-MS/MS, and a rapid and efficient HPLC-DAD method was developed to analyze the acylated-anthocyanins. Ultrasonic-assisted extraction conditions of acylated-anthocyanins were optimized using response surface methodology, extraction at 70 °C for 32 min using 70% methanol solution (0.1% HCl, v/v) rendered an extract with 80.37?±?2.66 mg/100 g of cyanidin-3-O-(trans-p-coumaroyl)-diglucoside and 97.88?±?4.06 mg/100 g of total acylated-anthocyanins. Nine macroporous resins were investigated for preliminary purification of acylated-anthocyanins. According to the static/dynamic adsorption and desorption tests, XDA-6 macroporous resin exhibited the maximum potential for preparing acylated-anthocyanins. The purity of cyanidin-3-O-(trans-p-coumaroyl)-diglucoside (43.30 mg/g) in purified acylated-anthocyanins was 201.89 times of that of the extract (0.21 mg/g), and the purity of total acylated-anthocyanins increased from 0.36 to 56.44 mg/g. Besides, the stability (t _1/2) of cyanidin-3-O-(trans-p-coumaroyl)-diglucoside and total acylated-anthocyanins increased by more than five-fold after purification using XDA-6. The established methods of analysis, extraction and purification of acylated-anthocyanins were hopefully utilized in food industry.     

Quantification and Discrimination of Viable and Dead <Emphasis Type="Italic">Escherichia coli</Emphasis> O157:H7 Cells from Chicken Without Enrichment by Ethidium Bromide Monoazide Real-time Loop-Mediated Isothermal Amplification

In this study, a rapid and sensitive method of real-time loop-mediated isothermal amplification (Rti-LAMP) assays was developed for quantification and discrimination of viable and heat-killed E. coli O157:H7 cells treated with low concentration of ethidium bromide monoazide (EMA). Four micrograms per milliliter of EMA was chosen as the optimal concentration which did not inhibit DNA amplification derived from viable cells, but significantly increased the Tt values of dead cells in Rti-LAMP assays. When the DNA from 2.0?×?10³ viable CFU of E. coli O157:H7 was subjected to EMA-Rti-LAMP, the resulting Tt value was 17.73 min. In contrast, the DNA from 2.0?×?10³?CFU completely heat destroyed CFU of E. coli O157:H7 did not yield a positive amplification which Tt value was regarded as 60 min. When the DNA from viable plus heat-killed CFU at a ratio of 5:2995 was subjected to EMA-Rti-LAMP, the resulting Tt value was 23.06 min, which was statistically identical (P?<?0.05) to the Tt value of 24.07 min obtained with the DNA from only 5 viable CFU. The results indicate that even though 3.0?×?10³ dead cells yielded a negative amplification setting the Tt value as 60 min, low numbers of viable cells in the presence of much higher numbers of dead cells still yielded a linear plot for enumerating viable CFU from Tt values. Detection of E. coli O157:H7 derived from contaminated chicken samples, the EMA-Rti-LAMP could notably distinguish viable and heat-killed cells from 5.0?×?10¹ to 1.0?×?10⁴?CFU/g without enrichment.     

A Rapid and Visual Single Primer Isothermal Amplification-Based Method for the Detection of <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> in Raw Pork Products

Staphylococcus aureus (S. aureus) is an important food-borne pathogen which poses a severe threat to public health worldwide. Rapid detection of S. aureus with high sensitivity is of particular importance for food safety. In this study, a novel single primer isothermal amplification (SPIA) method was established to detect S. aureus in food, targeting the accessory gene regulator (agr) gene with a DNA/RNA primer. The developed SPIA method has the advantages of visualization and avoiding tedious electrophoresis. In order to confirm the specificity of this method, 7 S. aureus strains and 26 non-S. aureus strains were detected with their pure cultures. The sensitivity and detection limit of S. aureus with artificially inoculated raw pork products by SPIA were evaluated through fluorescence and turbidity by naked eye and the amplification curve, which were 4.3?×?10⁰ CFU/mL and 5.6?×?10⁰ CFU/g, respectively. Compared with the conventional PCR method, the SPIA has 100-fold higher sensitivity and 100-fold lower detection limit. Therefore, the developed SPIA method is a potentially reliable tool for rapid and visual detection of S. aureus in food.     

TLC-Digital Image-Based Fluorometric Analysis of Ergosterol and Chitin Content in Food Grains Artificially Infested with <Emphasis Type="Italic">Aspergillus flavus</Emphasis> and <Emphasis Type="Italic">Fusarium verticillioides</Emphasis>

Novel thin-layer chromatography-digital image-based analytical methods were developed for the quantitation of ergosterol and chitin content in six food matrices (rice, wheat, maize, sorghum, groundnut, and sunflower), artificially infested with Aspergillus flavus (MTCC 6513)/Fusarium verticillioides (MRC 826). For ergosterol, single-step method, based on liquid/liquid extraction, was followed by thin-layer chromatography (TLC). Chitin was solubilized using lithium chloride (5%) in dimethyl acetamide and converted to chitosan using 5 N NaOH and subsequently complexed with calcofluor white dye. The absorption and emission maxima of chitosan-calcofluor complex were recorded at λ 350/230 and 430 nm, respectively. The sensitivity based on the limit of detection (LOD) was found to be 100 ng both for ergosterol and chitin analysis. Based on ergosterol and chitin analysis, groundnut and maize were found to be suitable substrates for A. flavus (p?<?0.013 and p?<?0.01), while sorghum followed by groundnut and sunflower were found to be ideal for F. verticillioides (p?<?0.01 and p?<?0.0001) and rice was established as poor substrate as there was no growth on it up to 12 days of incubation. A strong correlation was found between ergosterol and chitin contents with regression (r ²) values of 0.974 and 0.997 in food grains inoculated with A. flavus and F. verticillioides, respectively, during the period of infection. The authenticity of the two methods developed was further confirmed by applying them to commercial food grains and flours. Thus, ergosterol in combination with chitin analysis could be successfully used as an index of fungal contamination employing TLC-digital-based analytical methods.     

Validated RP-HPLC Method for Simultaneous Determination of Tocopherols and Tocotrienols in Whole Grain Barley Using Matrix Solid-Phase Dispersion

The vitamers of vitamin E such as α-, β-, γ-, and δ-tocotrienol and α-, β-, γ-, and δ- tocopherol are important phytochemical compounds with antioxidant activity and with potential benefits for human health. A high-performance liquid chromatography with fluorescence detection (HPLC-FLD) method was validated for their determination in whole grain barley samples. Tocol extraction was performed by an optimized matrix solid-phase dispersion (MSPD) protocol with neutral alumina (0.5 g) as the dispersion agent and methanol (5 mL) as the elution solvent. The analytical column was an Eclipse XDB C18 column (150?×?4.6 mm, 5 μm) and it was operated at room temperature. Mobile phase was consisted of methanol/acetonitrile/i-propanol (55:40:5?v/v?%) and the elution was isocratic at a flow rate of 0.8 mL/min. Total analysis time was 12 min, and the detection of the tocols was performed with a fluorimetric detector where the excitation and emission wavelengths were set at 295 and 335 nm, respectively. Method validation was performed by means of intra-day (n?=?5) and inter-day accuracy and precision (n?=?8), sensitivity, and linearity. The linear regression coefficient (R ²) was higher than 0.99. The recoveries of the tocols from barley samples with the proposed extraction method were in an acceptable level (74–91 %) where the relative standard deviation ranged from 4.2 to 15.0 %. Limits of detection (LODs) and limits of quantification (LOQs) varied from 0.03 to 0.11 mg kg^?1 and 0.11 to 0.34 mg kg^?1, respectively.     

A Case Study on Optimization of Biomass Flow During Single-Screw Extrusion Cooking Using Genetic Algorithm (GA) and Response Surface Method (RSM)

In the present study, response surface method (RSM) and genetic algorithm (GA) were used to study the effects of process variables like screw speed, rpm (x ₁), L/D ratio (x ₂), barrel temperature (°C; x ₃), and feed mix moisture content (%; x ₄), on flow rate of biomass during single-screw extrusion cooking. A second-order regression equation was developed for flow rate in terms of the process variables. The significance of the process variables based on Pareto chart indicated that screw speed and feed mix moisture content had the most influence followed by L/D ratio and barrel temperature on the flow rate. RSM analysis indicated that a screw speed?>?80 rpm, L/D ratio?>?12, barrel temperature?>?80 °C, and feed mix moisture content?>?20% resulted in maximum flow rate. Increase in screw speed and L/D ratio increased the drag flow and also the path of traverse of the feed mix inside the extruder resulting in more shear. The presence of lipids of about 35% in the biomass feed mix might have induced a lubrication effect and has significantly influenced the flow rate. The second-order regression equations were further used as the objective function for optimization using genetic algorithm. A population of 100 and iterations of 100 have successfully led to convergence the optimum. The maximum and minimum flow rates obtained using GA were 13.19?×?10^?7 m³/s (x ₁?=?139.08 rpm, x ₂?=?15.90, x ₃?=?99.56 °C, and x ₄?=?59.72%) and 0.53?×?10^?7 m³/s (x ₁?=?59.65 rpm, x ₂?=?11.93, x ₃?=?68.98 °C, and x ₄?=?20.04%).     

Application of Bismuth(III)-Entrapped XO Biosensing System for Xanthine Determination in Beverages

A xanthine biosensor was prepared by electrochemical immobilization of xanthine oxidize enzyme onto carbon paste electrode via entrapment of Bi³⁺. After the optimization of experimental parameters, analytical characteristics were investigated. Two linear ranges between 0.02 and 0.06 and 1–7.5 μM with the equation y?=?93.00x?+?0.12 and y?=?1.07x?+?18.03 with the correlation coefficients of R ²?=?0.9951 and R ²?=?0.9931, respectively, were obtained for this biosensing system. RSD value was calculated for 0.04 μM xanthine (n?=?5) and found as 3.84%. LOD and LOQ values were also calculated and revealed as 1.30?×?10^?8 and 4.3?×?10^?8 M, respectively. Then, this biosensor was applied for xanthine detection in real samples. As a sample treatment, only necessary dilutions were made. Four types of beverages including wine, energy drink, peach, and sour cherry juice were used for this purpose. Obtained recovery values demonstrate that this system is applicable for xanthine detection in real samples without needing any laborious sample pretreatment procedures.     

Application and Optimization of Microwave-Assisted Extraction and Dispersive Liquid–Liquid Microextraction Followed by High-Performance Liquid Chromatography for the Determination of Oleuropein and Hydroxytyrosol in Olive Pomace

In the present study, a new method based on microwave-assisted extraction and dispersive liquid–liquid microextraction (MAE–DLLME) followed by high-performance liquid chromatography (HPLC) was proposed for the separation and determination of oleuropein (Ole) and hydroxytyrosol (HyT) from olive pomace samples. The effective factors in the MAE–DLLME process such as microwave power, extraction time, the type and volume of extraction, and dispersive solvents were studied and optimized with the aid of response surface methodology (RSM) based on a central composite design (CCD) to obtain the best condition for Ole and HyT extraction. At the optimized conditions, parameter values were 220 W microwave power, 12 min extraction time, 60 μL extracting solvent, and 500 μL dispersive solvent. The calibration graphs of the proposed method were linear in the range of 10–500,000 μg L^?1, with the coefficient of determination (R²) higher than 0.99 for Ole and HyT. Repeatability of the method, described as the relative standard deviation (RSD), was 4.12–5.63% (n?=?6). The limits of detection were 35 and 20 μg L^?1 for Ole and HyT, respectively. The recoveries of these compounds in the spiked olive pomace sample were from 93 to 98%. The proposed method, MAE–DLLME–HPLC–UV, was an accurate, rapid, and reliable method when compared with previous methods.     

Saltatory Rolling Circle Amplification (SRCA): a Novel Nucleic Acid Isothermal Amplification Technique Applied for Rapid Detection of <Emphasis Type="Italic">Shigella</Emphasis> Spp. in Vegetable Salad

Shigella spp. are enteric pathogens that pose a serious threat to public health worldwide. A novel saltatory rolling circle amplification (SRCA) assay was developed to detect Shigella spp. in food targeting the ipaH gene. SRCA as an isothermal amplification method requires no expensive thermocycle instrument and could avoid electrophoresis as visualization results was successfully applied for SRCA. In order to confirm the specificity of this assay, 34 strains including 11 strains belonging to different Shigella species and 23 non-Shigella bacteria were detected with pure cultures. The sensitivity of Shigella flexneri by SRCA was evaluated using agarose gel electrophoresis, which was 7.3 × 10¹ fg/μL. In addition, the amplification results were also determined by adding the fluorochrome, SYBR Green I (1 μL of 1000×), allowing naked eye visualization of results, and the sensitivity was 7.3 × 10⁰ fg/μL. Moreover, the sensitivity of PCR was 7.3 × 10² fg/μL, showing that the sensitivity of SRCA by electrophoresis and SYBR Green I fluorescence were 10- and 100-fold higher than that of PCR, respectively. The detection limit of SRCA was also evaluated with artificially inoculated vegetable salad without enrichment, and it was 4.7 × 10² and 4.7 × 10¹ CFU/g by electrophoresis and fluorescence, respectively. The detection limit by PCR was 4.7 × 10³ CFU/g, which was 10- and 100-fold higher than that of SRCA. Therefore, SRCA is a potentially reliable tool for rapid and specific detection of Shigella in food and could be useful in underdeveloped countries with limited resources.     

Comparative inactivation studies of <Emphasis Type="Italic">Aegle marmelos</Emphasis> (<Emphasis Type="Italic">bael</Emphasis>) peroxidase in crude extract of fruit by heat processing and ultrasonication treatment

Bael (Aegle marmelos) is considered as a holy fruit comprised of vast number of phytonutrients. Whole bael tree including all its parts has medicinal significance. Lack of awareness and seasonal nature makes its processing rather challenging. Conventional heat processing may lead to inactivation of quality hampering enzymes such as peroxidase, but at the cost of loss in essential phytonutrients. In the present work it was observed that thermal inactivation of bael peroxidase obeyed first order kinetics with enzyme activation energy of 7.7 kJ mol^?1. Complete inactivation of bael peroxidase was achieved within 11 min at 85?°C while ultrasound treatment attained in lesser time of 4 min at 64.07 W cm^?2 ultrasonic intensity. Loss of marmelosin a well-known phytonutrient in bael fruit was found to be 83.29?% by heat (11 min, 85?°C) and only 50.20?% by ultrasonication (4 min, 64.07 W cm^?2 ultrasonic intensity). Ultrasonication has potential to overcome harmful effects of heat processing with retention of phyto-constituents and hence has promising future in various food processing applications.