Antioxidants Release from Solvent-Cast PLA Film: Investigation of PLA Antioxidant-Active Packaging

The release study of natural antioxidants, i.e., ascorbyl palmitate and α-tocopherol, and synthetic phenolic antioxidants including butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), propyl gallate, and tert-butylhydroquinone (TBHQ) from polylactic acid (PLA) film into food simulants were accomplished. PLA antioxidant films were placed in contact with 95, 50, and 10 % ethanol at 40 and 20 °C. Released antioxidants were regularly measured by high-performance liquid chromatography system for 60 days. Ascorbyl palmitate was completely degraded during film preparation and is not a suitable antioxidant for PLA antioxidant-active packaging. The diffusion coefficient (D) and partition coefficient (K) of antioxidants were calculated based on obtained data. Diffusion of the antioxidants from PLA to the simulants showed a Fick’s behavior with the diffusion coefficient (D) value between 10^?9 and 10^?11 cm² s^?1 with 0–100 % of release. Faster and higher release of antioxidants occurred at 40 °C according to Arrhenius law. At 40 °C, TBHQ in 95 % ethanol decomposed to 2-tert-butyl-1,4-benzoquinone and other quinone derivatives, and α-tocopherol in 50 % ethanol decomposed to some unknown neoformed compounds. Antioxidants molecular weight, Log P, simulant polarity, and temperature were the most influencing factors on antioxidants release rate from PLA films in contact with food simulants. The D and K coefficients of studied antioxidants from PLA in three food simulants and two temperatures can be used to create PLA antioxidant-active packaging to continually control the oxidation reactions in diverse foodstuffs to ensure higher food qualities. The PLA antioxidant-active packaging approach also permits to reduce the amounts of directly added antioxidants in foods to provide safer foods.     

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.     

Determination of Chlorothalonil Residue in Cabbage by a Modified QuEChERS-Based Extraction and Gas Chromatography–Mass Spectrometry

Being susceptible to any matrix with pH >5, taking cabbage as an example, the low recovery of chlorothalonil residues adsorbed onto the cabbage matrix was almost completely improved by extracting with 1/1 (v/v) acetonitrile (containing 5 % acetic acid)/toluene. Under the optimized conditions, the recoveries of chlorothalonil in cabbage fortified at three concentrations of 0.5 to 10 mg kg^?1 were 71–93 % with relative standard deviations (RSDs) lower than 6 %. The limit of detection (LOD) and the limit of quantification (LOQ) of the gas chromatography–mass spectrometry (GC–MS) method for chlorothalonil were 0.05 and 0.5 mg kg^?1, respectively, which were much lower than the maximum residue limits (MRLs). The proposed analytical method demonstrated a potential for its application to monitor for chlorothalonil and to help assure food safety, especially base-sensitive-pesticide analysis.     

Applicability of Pressurized Liquid Extraction for Melamine Analysis in Pet Foods with High-Performance Liquid Chromatography with Diode Array Detection

A pressurized liquid extraction (PLE) method was developed for melamine analysis in pet foods. The PLE method which utilized an accelerated solvent extraction (ASE®) system was also compared with sonication and polytron extraction methods. The parameters for the optimized PLE method were temperature (75?°C for wet pet food, 125?°C for dry pet food), pressure (1,500 psi), static time (10 min), flush volume (40%), purge time (1 min), and number of cycles (1). Recoveries obtained by PLE method were significantly higher (P?≤?0.05) than those of sonication and polytron methods for dry pet food samples. For the analysis of adulterated pet foods, PLE resulted in the highest melamine content followed by sonication and polytron. Using PLE, samples fortified with melamine at 2.5 and 100 mg kg^?1 resulted in recoveries ranging from 55% to 90% for wet samples and from 90% to 116% for dry samples. Low recovery rate from wet samples at low spike level (2.5 mg kg^?1) may have been caused by co-aggregation of polysaccharide and protein with melamine due to low pH during solid-phase extraction cleanup. Limit of detection and limit of quantification values were 0.5 (mg kg^?1) and 1.0 (mg kg^?1) for dry samples. Overall, PLE had the best extraction efficiency compared to sonication and polytron, proving PLE to be a useful tool for melamine analysis of pet foods.     

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.     

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.     

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.     

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.     

The Use of Microwave and Pulsed Electric Field as a Pretreatment Step in Ultrasonic Extraction of Polyphenols from Defatted Hemp Seed Cake (Cannabis sativa) Using Response Surface Methodology

The use of microwave and pulsed electric field (PEF) processing to optimize polyphenol extraction from defatted hemp seed cake in mixed solvent of methanol, acetone and water (MAW, 7:7:6 v/v/v) was investigated using the Box-Behnken response surface method. A variables combination for microwave processing of time, microwave power and liquid to solid (L:S) ratio and for PEF (ethanol concentration, time, frequency and voltage) were used in the investigation. Following microwave or PEF treatments, the polyphenols were extracted from the samples under ultrasound with fixed variables (200 W of ultrasonic power, water bath of 70 °C and 20 min of extraction time). The measured responses were total phenolics (TP), total flavonoids (TF), DPPH˙ scavenging activity and ferric reducing/antioxidant power (FRAP). The optimum variables combination for microwave processing (5 min treatment time, L:S ratio of 6, 700 W power and volume 30 mL) and PEF-assisted extraction (voltage (30 V), frequency (30 Hz), ethanol concentration (10 %) and time (10 s)) resulted in maximum yields in all measured responses. The results suggest that microwave processing and PEF can be integrated in processing defatted hemp seed cake to enhance polyphenol extraction and maximize the yield.     

Development and Optimization of a SERS Method for On-site Determination of Nitrite in Foods and Water

A rapid, sensitive, and selective detection method of nitrite in foods and water was established by surface-enhanced Raman spectroscopy (SERS) based on the diazo reaction of nitrite with p-nitroaniline and 1-naphthylamine in acidic solution. The azo dye (4-(4-nitrophenyldiazenyl)naphthalene-1-aminium, NNA), which was derived from the diazo reaction, was determined by SERS using citrate-coated silver nanoparticles (AgNPs) as SERS substrates. The concentrations of nitrite in samples were finally calculated from the intensities of SERS signals generated by NNA in the testing solutions. By using the present method combined with a portable miniature Raman spectrometer, on-site determination of nitrite could be performed easily and efficiently. The effects of several experimental parameters on the intensity of SERS signals, such as the volume of AgNP solution and the mixing time of AgNPs and NNA, were investigated. The linear range of the method was 0.1–10.0 mg L^?1. The limits of detection (LODs) were 0.01, 0.03, and 0.05 mg L^?1 at 720, 1,459, and 1,609 cm^?1, respectively. The method was applied to the determination of nitrite in real food and water samples, with recoveries in the range of 86.9–103.4 % and relative standard deviations (RSDs) less than 9.64 %.     

Extraction of Antioxidants from Aloe vera Leaf Gel: a Response Surface Methodology Study

Extraction of antioxidants from aloe gel powder with water–ethanol solvents was studied using response surface methodology. The independent variables were solvent composition, temperature, extraction time, and the liquid-to-solid (L/S) ratio. The concentrations of potentially antioxidant compounds (aloin A and B, aloesin, total phenolics, and total carbohydrates) and a few indicators of antioxidant capacity (DPPH, FRAP, CUPRAC) served as the response functions. It was found that aloe gel contains both phenolic and non-phenolic antioxidants, and it was supposed that the phenolic fraction consists almost exclusively of chromones and anthrones. Different approaches to the optimization of extraction procedures are discussed, and here, the maximum recovery yield of antioxidants is achieved with 34 % ethanol at 60 °С and a L/S ratio of 46 in 1 h. The use of 90 % ethanol results in a higher antioxidant capacity of the product, but also results in a much lower extraction yield, decreasing the overall productivity of the process.     

Solubility determination as an alternative to migration measurements

Solubility values for six UV stabilisers (Cyasorb UV-1164, Tinuvin P, Tinuvin 234, Tinuvin 326, Tinuvin 327 and Tinuvin 1577) and five antioxidants (Irgafos 168, Irganox 1010, Irganox 3114, Irganox 3790 and Irganox 565) were determined in all the liquid food simulants (3% (m/V) acetic acid–water mixture, 10% (V/V), 20% (V/V), 50% (V/V) ethanol–water mixture and vegetable oil) proposed in European Union Regulation No. 10/2011/EC, as well as in fruit juice and cola drink. The applied method was obtained by modification of the method for the determination of water solubility as described in OECD guideline Test No. 105. By using ultrasonication and shorter equilibration time, the time demand of the solubility determinations were decreased notably. Solubility values proved to be lower than the specific migration limits (as specified in 10/2011/EC) at 25°C for almost all target compounds in food simulants A, B, C and D1 as well as in fruit juice and cola drink. The exceptions were Tinuvin P and Irganox 3790 in simulant D1. The solubility in food simulant D2 was higher than 1000 µg ml^–1 for all target compounds. These results show that the solubility of some additives in food simulants can be so low that it makes migration studies for certain additive–food simulant pairs dispensable.     

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.     

Stable Isotope Ratios and Aroma Profile Changes Induced Due to Innovative Wine Dealcoholisation Approaches

The high ethanol level in wine has become an important issue for all the main wine producing countries. Several techniques are available to the wine industry to reduce the ethanol content; among them, the membrane contactors are certainly one of the newest. Very few studies on the effect of this practice on the wine quality and aroma profile and on the stable isotopes composition are available. A pilot and industrial plant equipped with the membrane contactor system were used in the study in the dealcoholisation on several white and red wines. Significant changes for several classes of aroma compounds in both pilot- and industrial-scale experiments were observed, even though these changes were not always in perfect agreement with the sensory evaluation carried out. Finally, modifications on the δ¹⁸O of up to 1?‰ for 2 %?v/v and of up to 4?‰ for 8 %?v/v ethanol removal were encountered. An increase of δ¹³C of ethanol of up to 1.1?‰ for 2 % and of up to 2.3?‰ for 4 % of dealcoholisation rate was also observed. Dealcoholisation via membrane contactor seemed to affect the overall wine composition (aroma and flavour), even though the main concern resided on the alteration of the isotopic composition which could be linked to product authenticity issues.     

Screening of Phenolic Antioxidants in Edible Oils by HPTLC-DPPH Assay and MS Confirmation

The cytotoxicity of phenolic antioxidants had attracted marked attention, posing serious challenges to food safety. This paper presented a screening method for two major phenolic antioxidants (butylated hydroxytoluene and tert-butylhydroquinone) added in edible oils. To specifically visualize the targeted compounds after developing with toluene/ethyl acetate/methanol 8:1:1 (v/v/v) to 70 mm solvent front, the plate was subjected to a standardized 1,1-diphenyl-2-picrylhydrazyl assay. In addition to synoptical eye inspection, accurate quantification was realized by modified densitometric measurements: fluorescence mode, excitation wavelength 530 nm (D₂ and W lamp) without optical filter, which offered satisfactory sensitivity (8.5–17.5 mg/kg) and acceptable linearity (R²?>?0.999 within 50–200 ng/zone). Moreover, the established method was validated with edible oil samples, against EU Directive 2006/52/EC. Apart from that, the unambiguous confirmation of positive results was conveniently achieved by TLC-MS interface-mediated mass spectrometry. Featuring the merits of screening conception, the proposed method not only reached the goal of accurate quantification and conclusive identification of multi-phenolic antioxidants, but also excellently balanced the simplicity, detectability, and throughput of the screening workup. Therefore, it might be an attractive alternative to conventional methods.     

Optimization of Extraction Parameters of Total Phenolics from <Emphasis Type="Italic">Annona crassiflora</Emphasis> Mart. (Araticum) Fruits Using Response Surface Methodology

In this study, response surface methodology was used to optimize the extraction temperature (25–75 °C) and ethanol concentration (0–70 %, ethanol/water, v/v) to maximize the extraction of total phenolic compounds (TPC) from araticum pulp. The efficiency of the extraction process was monitored over time, and equilibrium conditions were reached between 60–90 min. A second-order polynomial model was adequately fit to the experimental data with an adjusted R ² of 0.9793 (p < 0.0001) showing that the model could efficiently predict the TPC content. Optimum extraction conditions were ethanol concentration of 46 % (v/v), extraction temperature of 75 °C and extraction time of 90 min. Under the optimum conditions, the araticum pulp showed high TPC content (4.67 g GAE/100 g dw) and also high antioxidant activity in the different assays used (46.56 μg/mL, 683.65 μmol TE/g and 1593.72 μmol TE/g for DPPH IC₅₀, TEAC and T-ORAC_FL, respectively). From our extraction procedure, we successfully recovered a significantly higher amount of TPC compared to other studies in the literature to date (1.5–22-fold higher). Furthermore, TPC and antioxidant activity were present in the fruit in levels that are difficult to find in other common fruits. These results expose a potential approach for improving human health through consumption of araticum fruit.     

The Release of Carotenoids from a Light-Protected Antioxidant Active Packaging Designed to Improve the Stability of Soybean Oil

An antioxidant active packaging was developed by coextruding two layers of high-density polyethylene (HDPE). Bags were made with the film in which the inner layer, designed to be in contact with food, contained marigold flower (Tagetes erecta) extract rich in carotenoids. The outer layer of the film was added with titanium dioxide (TiO₂) to decrease the effect of commercial lighting on the degradation of carotenoids. Four bilayer films were produced: added with TiO₂ and carotenoids, added with carotenoids, added with TiO₂, and with no additives at all (control film). Degradation of color and astaxanthin in the films was delayed by the addition of the TiO₂ when they were stored under commercial lighting at 25 °C. Bags made of these films produced an improvement on the soybean oil stability at 25 °C as a result of a synergic effect of light protection by TiO₂ and carotenoids release. This release was measured as diffusion coefficients of carotenoids from the films toward soybean oil at 10, 25, and 40 °C (2.10–19.26?×?10^?11 cm² s^?1) with activation energy of 53.66 kJ mol^?1. In conclusion, the combination of the two layers of HDPE added with TiO₂ and carotenoids introduced opacity and permitted to extend the active effect of the films in contact with soybean oil. Moreover, the effect of temperature on the diffusion of carotenoids showed that this new active packaging is able to exert its function in conditions of transport, storage, and commercialization of food.     

Development of an Efficient Acid Digestion Procedure Utilizing High-Pressure Asher Technique for the Determination of Iodine and Metallic Elements in Milk Powder

An effective wet digestion procedure for the determination of total iodine contents in milk powder samples was developed utilizing a high-pressure asher technique. The optimized method based on a two-stage digestion procedure. In the first stage, 500 mg samples were digested at 300 °C for 2 h using 15.2 mol L^?1 HNO₃ (5 mL) and 30 % H₂O₂ (3 mL). After the first digestion stage, digestion vessels were allowed to cool down and 1.2 mL of 20 % (w/v) Na₂S₂O₈ solution was added as an additional oxidizing agent to the samples. After that, the vessels were closed, and they were heated at 100 °C for 30 min, resulting in clear and colorless sample solutions. Iodine concentrations in the digested samples were measured with inductively coupled plasma mass spectrometry. The accuracy of the optimized method was confirmed by analyzing milk powder reference material (BCR-151, milk powder). The obtained value for iodine (5.29?±?0.37 mg kg^?1, n?=?6) was in good agreement with the certified value (5.35?±?0.14 mg kg^?1). Furthermore, the results obtained for reference material showed that the developed method can be applied also for the determination of other elements, e.g., copper, iron, and lead in the digested milk powder samples.     

The Cryoprotective Effect of Different Konjac Glucomannan (KGM) Hydrolysates on the Glass Carp (Ctenopharyngodon idella) Myofibrillar During Frozen Storage

The physicochemical properties, Ca²⁺–ATPase activity, and surface hydrophobicity of grass carp myobrillar with different cryoprotectants (0.5 % irradiated degraded KGM, 0.5 % β-glucanase KGM hydrolysate, 8 % sucrose-sorbitol (1:1 w/w)) were investigated. The molecular weight analysis showed that the irradiated degraded KGM and β-glucanase KGM hydrolysate were 1.209?×?10⁴ and 2.093?×?10⁴ Da, respectively. The solubility, Ca²⁺–ATPase activity, and total and reactive sulfhydryl (SH) content of treatments containing KGM hydrolysates were higher than other treatments during frozen storage. Addition of KGM hydrolysates could effectively prevent the increase of surface hydrophobicity. The gel strength and whiteness of surimi showed that the addition of KGM hydrolysates in this experiment resulted in surimi with comparable quality.