Development and validation of an indirect competitive enzyme-linked immunosorbent assay for monitoring quinoxaline-2-carboxylic acid in the edible tissues of animals

The feed drug additive carbadox is a suspected carcinogen and mutagen. To monitor effectively residues of carbadox in the edible tissues of food-producing animals, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) to detect quinoxaline-2-carboxylic acid, the marker residue of carbadox, was developed. Several haptens were synthesised and conjugated to the carrier protein. Nine female New Zealand white rabbits were immunised with the immunising conjugates to produce polyclonal antibodies according to the designed schemes of immunisation. The highly specific antibody that was very sensitive to N-butylquinoxaline-2-carboxamide with an IC₅₀ value of 7.75?µg?l^?1 was selected for the development of an ic-ELISA. The standard curves based on the N-butylquinoxaline-2-carboxamide matrix calibration ranged from 0.2 to 51.2?µg?l^?1. The decision limit and detection capability of the ic-ELISA were 0.60 and 0.83?µg?kg^?1 for liver and 0.68 and 0.79?µg?kg^?1 for muscle of swine, respectively. The recoveries were 57–108% with coefficients of variation of less than 20% when the quinoxaline-2-carboxylic acid was spiked into liver and muscle with the concentrations of 1.0–20.0?µg?kg^?1. Excellent correlations between the results of the ic-ELISA and an HPLC method (r?=?0.9956???0.9969) were observed for incurred tissues. These results suggest that the ic-ELISA is a sensitive, accurate and low-cost method that would be a useful tool for screening residues of carbadox in the edible tissues of food-producing animals.     

Development and Validation of a Sensitive Indirect Competitive Enzyme-Linked Immunosorbent Assay for the Screening of Florfenicol and Thiamphenicol in Edible Animal Tissue and Feed

To monitor the illegal use of florfenicol (FF) and thiamphenicol (TAP) in edible animal tissue and feed, a sensitive monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) has been developed with simple sample preparation and cleanup. The obtained monoclonal antibody (5F4) that has isotype IgG1 showed an IC₅₀ value of 0.21 μg L^?1 for FF and 0.35 μg L^?1 for TAP, respectively. It did not exhibit measurable cross-reactivity with other antibiotics. The limits of detection (LODs) for FF and TAP in a muscle matrix ranged from 0.07 to 0.14 μg kg^?1 and in a feed matrix ranged from 2.9 to 5.2 μg kg^?1. The recoveries were 72.8 to 113.4 % with a coefficient of variation of less than 15 %. Good correlation between the ELISA and HPLC-MS/MS results in the tissues tested demonstrated the reliability of our ic-ELISA. This ELISA is a useful tool for screening FF and TAP in edible animal tissue and feed.     

Development of an Enzyme-Linked Immunosorbent Assay for the Detection of Difenoconazole Residues in Fruits and Vegetables

An indirect, competitive enzyme-linked immunosorbent assay (ic-ELISA) for the detection of difenoconazole was developed. Two haptens were designed and successfully synthesized. Hapten 1 had a particular moiety of difenoconazole, while hapten 2 had its full structure. The polyclonal antibodies against hapten-protein conjugates were prepared by immunizing rabbits. After optimization of the conditions, the detection limit (IC₁₅) and sensitivity (IC₅₀) were 4.58 and 29.10 μg L^?1, respectively. The cross-reactivities of the antibody with 11 triazole fungicides were all less than 0.1%, which showed that the antibody had excellent specificity. The recoveries of difenoconazole from the spiked samples ranged from 89.70 to 102.31% with good accuracy. The matrix effect was easily removed using a simple, rapid, and efficient extraction method on fruits and vegetables. The detection limit was all 229 μg kg^?1 in fruits and vegetables. To validate the ic-ELISA, samples were spiked with difenoconazole at three different concentrations and simultaneously analyzed using high-performance liquid chromatography (HPLC). The results showed a good correlation between the ic-ELISA and HPLC data (R ² = 0.9970). As a result, the developed immunosorbent assay is suitable for the quantitative determination of difenoconazole in fruits and vegetables.     

Preparation of a Broadly Specific Monoclonal Antibody-Based Indirect Competitive ELISA for the Detection of Benzodiazepines in Edible Animal Tissues and Feed

Incorrect use of benzodiazepines could result in serious health problems. To monitor the illegal use of benzodiazepine compounds in animals, a group-specific monoclonal antibody (mAb) was prepared in this study. The obtained 3D7 mAb, which is an IgG1 isotype mAb, displayed an IC₅₀ value of 8.9 ng mL^?1 for diazepam and exhibited cross-reaction for diazepam (100 %), nitrazepam (49 %), nordiazepam (140 %), temazepam (32 %), oxazepam (17 %), estazolam (7.5 %), and alprazolam (2.4 %). Based on this mAb, for the first time in this study, an optimized indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) protocol that did not require complicated sample preparation and clean-up was developed. The detection limits of this ic-ELISA for benzodiazepines ranged from 1.2 to 3.3 μg kg^?1 in muscle matrix and from 25.2 to 55.4 μg kg^?1 in feed matrix. The recoveries ranged from 70.9 to 111.3 % with coefficients of variation below 15.0 %. Good correlations (r?>?0.9494) between the results of the ic-ELISA and high-performance liquid chromatography were also observed. This simple method reduced the time required for sample preparation, ensured greater throughput, and met the requirements for benzodiazepine residue analyses. In conclusion, the proposed method is a sensitive and rapid multi-residue technique that offers a cost-effective alternative to current published procedures without any concession in the ability to detect benzodiazepine sedative misuse.     

Rapid Detection of Four Organophosphorous and Neonicotinoid Toxicants Using Bi-enzyme Tracer Competitive Enzyme-Linked Immunosorbent Assay

A bi-enzyme tracer direct competitive enzyme-linked immunosorbent assay (dc-ELISA) based on two generic antibodies was developed. The effects of several physicochemical factors, such as heterologous antigen–horseradish peroxidase (HRP), methanol concentration, ionic strength, and pH value, were optimized to obtain satisfactory sensitivity of the assay. Under the optimized conditions, the 50 % inhibition concentration (IC₅₀) value for parathion, parathion-methyl, imidaclothiz, and imidacloprid was 57.28?±?2.73, 169.82?±?5.64, 52.48?±?3.46, and 53.08?±?2.05 μg L^?1, with a limit of detection (LOD, IC₁₀) of 0.56, 3.16, 0.62, and 0.51 μg L^?1, respectively. There was no obvious cross-reactivity (CR) with most of the neonicotinoids and organophosphorus pesticides. The recoveries of parathion, parathion-methyl, imidaclothiz, and imidacloprid in environmental and agricultural samples, including river water, soil, and cabbage, ranged from 82.54 to 116.29 %. The relative standard deviation (RSD) ranged from 1.59 to 8.09 %. The ELISA results were confirmed by gas chromatography (GC) with a high correlation coefficient of 0.9882. These results showed that the bi-enzyme tracer competitive enzyme-linked immunosorbent assay could be used as a sensitive tool for monitoring parathion, parathion-methyl, imidaclothiz, and imidacloprid in environmental samples and agricultural products.     

Development of an ELISA and Immunochromatographic Assay for Tetracycline,Oxytetracycline, and Chlortetracycline Residues in Milk and Honey Based on the Class-Specific Monoclonal Antibody

A sensitive class-specific monoclonal antibody against tetracyclines (TCs) was generated and used to develop an enzyme-linked immunosorbent assay (ELISA) and an immunochromatographic assay for TC, oxytetracycline (OTC), and chlortetracycline (CTC) detection in milk and honey samples. The dynamic range of detection for TC in ELISA was 0.26–2.00 μg L^?1 with an IC₅₀ of 0.72 μg L^?1. The IC₅₀ value of OTC and CTC was 3.2 and 6.4 μg L^?1, respectively. The recovery of TC, OTC, and CTC in milk samples was 82–102, 91–105, and 90–101 %, respectively, and 88–101, 89–101, and 89–95 % in honey samples, respectively. In the immunochromatographic assay, the cutoff values for TC, OTC, and CTC were 15, 15, and 50 μg L^?1 in milk, respectively, and 40, 40, and 40 μg L^?1 in honey, respectively. The results revealed that ELISA and the immunochromatographic assay can be applied for the rapid and sensitive detection of TC, OTC, and CTC in milk and honey samples.     

Micronutrients (B,Co, Cu,Fe, Mn,Mo, and Zn) content in made tea (Camellia sinensis L.) and tea infusion with health prospect: A critical review

Tea (Camellia sinensis L.) is a perennial acidophilic crop, and known to be a nonalcoholic stimulating beverage that is most widely consumed after water. The aim of this review paper is to provide a detailed documentation of selected micronutrient contents, viz. boron (B), cobalt (Co), copper (Cu), iron (Fe), manganese (Mn), molybdenum (Mo), and zinc (Zn) in made tea and tea infusion. Available data from the literature were used to calculate human health aspect associated with the consumption of tea infusion. A wide range of micronutrients reported in both made tea and tea infusion could be the major sources of micronutrients for human. The content of B, Co, Cu, Fe, Mn, Mo, and Zn in made tea are ranged from 3.04 to 58.44 μg g^?1, below detectable limit (BDL) to 122.4 μg g^?1, BDL to 602 μg g^?1, 0.275 to 13,040 μg g^?1, 0.004 to 15,866 μg g^?1, 0.04 to 570.80 μg g^?1 and 0.01 to 1120 μg g^?1, respectively. Only 3.2 μg L^?1 to 7.25 mg L^?1, 0.01 μg L^?1 to 7 mg L^?1, 3.80 μg L^?1 to 6.13 mg L^?1, 135.59 μg L^?1 ?11.05 mg L^?1, 0.05 μg L^?1 to 1980.34 mg L^?1, 0.012 to 3.78 μg L^?1, and 1.12 μg L^?1 to 2.32 μg L^?1 of B, Co, Cu, Fe, Mn, Mo, and Zn, respectively, are found in tea infusion which are lower than the prescribed limit of micronutrients in drinking water by World Health Organization. Furthermore, micronutrient contents in tea infusion depend on infusion procedure as well as on the instrument used for analysis. The proportion of micronutrients found in different tea types are 1.0–88.9% for B, 10–60% for Co, 2.0–97.8% for Cu, 67.8–89.9% for Fe, 71.0–87.4% for Mn, 13.3–34% for Mo, and 34.9–83% for Zn. From the results, it can also be concluded that consumption of three cups of tea infusion per day does not have any adverse effect on human health with respect to the referred micronutrients rather got beneficial effects to human.     

Simultaneous Determination of Aflatoxin B1 and Aflatoxin M1 in Food Matrices by Enzyme-Linked Immunosorbent Assay

Aflatoxins are a class of highly toxic chemical contaminants occurring in food. Consumption of aflatoxin-contaminated food can lead to harmful effects on human health. A rapid and reliable analysis of aflatoxins in food is crucial. In this study, we generated a broad–specific monoclonal antibody (MAb) 3 C10 against aflatoxin B₁ (AFB₁). The MAb 3 C10 binds specifically to AFB₁ and AFM₁ and has a IC₅₀ of 0.13 μg L^?1 for AFB₁ and 0.16 μg L^?1 for AFM₁. Furthermore, the MAb showed high cross-reactivity to AFB₂, AFG₁, and AFG₂. To enable simultaneous AFB₁ and AFM₁ detection in different food matrices, an indirect competitive enzyme-linked immunosorbent assay (icELISA) based on MAb 3 C10 has been developed and optimized. In addition, the extraction methods of different food matrices (peanut, corn, soybean, wheat flour, rice, soy sauce, vinegar, wine, raw milk, pure milk, skimmed milk, and yogurt) were established. The average recovery ranged from 73 to 121 %, with relative standard deviation values less than 15 %. The limit of detection was 0.52?+?0.36 μg kg^?1 (mean?+?3SD) for AFB₁ in eight agricultural products and 0.031?+?0.015 μg kg^?1 (mean?+?3SD) for AFM₁ in four dairy products. The sensitivity of icELISA was below the limit set by the European Commission for aflatoxin detection in different food matrices and similar to LC–MS/MS method. We demonstrate a rapid, simple, and reliable method for simultaneous screening of AFB₁ and AFM₁ in different food matrices.     

Preparation of Artificial Antigen and Development of Indirect Competitive ELISA Based on Chicken IgY for the Detection of Acid Orange II in Food Samples

To evaluate the feasibility of specific egg yolk antibodies (IgY) technology for the detection of hazardous chemical residues in foodstuffs, IgY were generated to detect acid orange II (AO2) residues. Bovine serum albumin (BSA) was made to bind with AO2 by succinic anhydride (SA), and the conjugate was used to immunize the laying chickens. PEG-6000 precipitation was used to extract IgY antibodies. The titer of anti-AO2 IgY attained the peak (1:12,800) after fifth booster immunization. Checkerboard titration showed that 1:640 dilution of anti-AO2 IgY could approximately give an optical density (OD) 1.0 at 5 μg/mL AO2-OVA coating concentration. The anti-AO2 IgY based indirect competitive ELISA (ic-ELISA) showed that the IC₅₀ value of anti-AO2 IgY was 10.72 μg/mL and regression curve equation was y?=?24.41?×?+75.11 (R ²?=?0.946). The ic-ELISA showed less cross-reactivity in range between 0.09 and 21.07 %. Recoveries from dried bean curd, sausage, and chilli powder samples were from 78.80 to 152.90 %, with relative standard deviation lower than 19.13 %. The study suggests that generated anti-AO2 IgY antibodies could be used in routine screening analysis of AO2 residues in food samples.     

Development of an Indirect Competitive ELISA for Analysis of Alternariol in Bread and Bran Samples

Alternariol (AOH) is one of the major mycotoxins produced by various species of Alternaria fungi. Natural occurrences of AOH have been reported in various foods, including fruits; processed fruit products such as apple juice, tomato products; wheat and other grains; oilseeds and products thereof, such as sunflower seeds, oilseed rape meal, and flax seed/linseed; and pecans. In this study, AOH-specific polyclonal antibodies were generated and developed an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) for monitoring AOH in bread and bran samples. The assay was very sensitive with a limit of detection (LOD) of 2.4?±?0.6 ng/g and a half maximal inhibitory concentration (IC₅₀) of 15.2?±?2.6 ng/g in bread and a LOD of 8.4?±?1.2 ng/g and IC₅₀ of 52.8?±?10.8 ng/g in bran extract. The assay was very specific to AOH and showed no cross-reactivity to alternariol monomethyl ether, altertoxin, altenuene, tentoxin, or tenuazonic acid. The effect of organic solvents on the assay was tested. The ELISA system tolerated methanol and acetonitrile as co-solvents at up to 5% content without significant loss of IC₅₀ value. Recoveries in all cases were greater than 75%, and the results using this method were comparable to those obtained from mass spectrometry methods. We conclude that this method is suitable for rapid detection of AOH in bread and bran samples, without expensive analytical equipment or time-consuming sample preparation.     

Overlapping of Second Order Scattering and Frequency Double Scattering Spectra Method and Resonance Rayleigh Scattering Method for the Determination of 6-Benzyladenine in Bean Sprout

In this work, second order scattering (SOS), frequency double scattering (FDS), the overlapping method of the two (SFOM), and resonance Rayleigh scattering (RRS) method had been developed for sensitive determination of trace 6-benzyladenine (BA) in bean sprout samples. In pH 1.6 HCl-NaAc medium, Pd(II) reacted with BA to form a 1:1 chelate complex, and then, the complex further self-aggregated into nanoparticles [Pd(II)-BA]_n. This resulted in a remarkable enhancement of resonance Rayleigh-scattering (RRS), second-order scattering (SOS), and frequency-double scattering (FDS) spectra. The maximum wavelengths were located at 312 nm (RRS), 632 nm (SOS), and 323 nm (FDS), respectively. The increments of scattering intensities ΔI were directly proportional to the concentration of BA in certain ranges. The detection limits were 7.0 nmol L^?1 (0.79 μg/kg, RRS), 10.3 nmol L^?1 (1.16 μg/kg, SOS), 39.4 nmol L^?1 (4.44 μg/kg, FDS), and 8.6 nmol L^?1 (0.96 μg/kg, SFOM). In addition, the optimum conditions of the reaction, and the effects of coexisting substances were investigated. The results showed that these methods exhibited a high selectivity. The reaction mechanism and the reasons for the enhancement of scattering were also discussed. Moreover, the feasibility for the SFOM method was illustrated in this paper. The proposed method had been successfully applied to determine a trace amount of BA in bean-sprout samples with the recoveries of 97.0–103.0 %.     

Immunoassay based on monoclonal antibodies versus LC-MS: deoxynivalenol in wheat and flour in Southern Brazil

An indirect competitive enzyme-linked immunosorbent assay (ELISA) method using a monoclonal antibody for deoxynivalenol (DON) detection in wheat and flour was standardised and validated (detection limit?=?177.1?µg?kg^?1) and its performance was compared with LC-MS, quantification limit?=140?µg?kg^?1). DON recovery ranged from 88.7% to 122.6% for wheat grain and from 70.6% to 139.3% for flour. Among the 38 wheat samples evaluated, DON was detected in 29 samples (76.3%) by ic-ELISA (281.6–12?291.4?µg?kg^?1) and in 22 samples (57.9%) by LC-MS (155.3–9906.9?µg?kg^?1). The 0.93 correlation coefficient between ic-ELISA and LC-MS data in 19 positive DON wheat samples demonstrated the reliability and efficiency of ic-ELISA. Results indicated that standardised ic-ELISA was suitable for DON screening in wheat samples and the need for continuous monitoring of mycotoxin levels in foodstuffs.     

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 %.     

Preconcentration and Simultaneous Analysis of Benzimidazole Anthelmintics in Milk Samples by Ultrasound-Assisted Surfactant-Enhanced Emulsification Microextraction and High-Performance Liquid Chromatography

The proposed ultrasound-assisted surfactant-enhanced emulsification microextraction (UASEME) coupled with high-performance liquid chromatography-photodiode array detection (HPLC-PDA) has been developed for the preconcentration and simultaneous analysis of five benzimidazole anthelmintics. Dichloromethane (extraction solvent) and Triton X-114 (emulsifier) was used for extraction of the target analytes. The parameters affecting the extraction efficiency were investigated and optimized. Under the optimum conditions, linearity was in the range from 10 to 150 μg?L^?1 with good coefficients of determination (R ²) higher than 0.994. Preconcentration factors were obtained up to 60, corresponding to limits of detection range of 1.8???3.6 μg?L^?1. Intra-day (n?=?5) and inter-day (n?=?4?×?3) precisions were obtained with relative standard deviation of retention time and peak area below 0.8 and 9.2 %, respectively. Good recoveries for the spiked target anthelmintics at different concentrations (e.g., 20, 50, and 100 μg?L^?1) of milk samples were obtained in 72.5–113.5 %. The results demonstrated that the proposed UASEME-HPLC-PDA can be used as an alternative powerful method for the simultaneous determination of the target analytes in milk samples.     

Simple and fast spectrophotometric determination of low levels of thiabendazole residues in fruit and vegetables after pre-concentration with ionic liquid phase microextraction

There is a great importance of monitoring thiabendazole (TBZ) residues in fruits and vegetables to ensure food safety. Therefore, a new ionic liquid (IL) phase microextraction method using IL, 1-butyl-3-methylimidazoliumhexafluorophosphate [C₄mim][PF₆], as extracting solvent is proposed for simple and fast determination of low levels of TBZ in fruits and vegetables by spectrophotometry. The method is based on selective complex formation of TBZ with Cu(II) ions in presence of PF₆^– as counter ion at pH 5.5, and then microextraction of the complex into the fine micro-drops of IL phase. After optimisation of variables affecting microextraction efficiency, the analytical parameters of the method were determined by calibration curves. The method exhibits a linear relationship (0.3–280 μg L^?1), low detection limit (0.1 μg L^?1), good intra- and inter-day precision (2.4–4.5% as RSD_r%, 2.1–5.6% as RSD_R%), good recovery (≥95.1–98.2%) and high sensitivity enhancement factor (150) by solvent-based calibration curve. It allows a detection limit of 0.24 μg L^?1 and a range of 0.8–250 μg L^?1 by the matrix-matched calibration curve. After validation, the method was successfully applied to the determination of TBZ residues with method quantification limits in fruit and vegetables of 2.0 and 2.5 µg kg^?1 with and without adding polyvinylpyrrolidone (PVP-15) solution. Recoveries range from 85.5% to 98.2% after spiking (10, 50 and 100 µg kg^?1, n: 3).     

Application of a New Functionalized Nanoporous Silica for Simultaneous Trace Separation and Determination of Cd(II), Cu(II), Ni(II), and Pb(II) in Food and Agricultural Products

A novel sorbent for simultaneous separation of cadmium, copper, nickel, and lead was prepared by functionalizing SBA-15 nanoporous silica with dithizone. A solid-phase extraction method using the above sorbent was developed to separate and preconcentrate trace amounts of cadmium, copper, nickel, and lead ions from food and agricultural products by flame atomic absorption spectrometry measurements. The optimum experimental conditions such as pH; flow rates; type, concentration, and volume of the eluent; breakthrough volume; and effect of coexisting ions on the separation and determination of these heavy metals were evaluated. The extraction efficiencies for the mentioned heavy metals were greater than 97 %, and the limits of detection were 0.09, 0.16, 0.21, and 0.45 μg?L^?1 for cadmium, copper, nickel, and lead, respectively. The preconcentration factor for simultaneous analysis of the four heavy metals was found to be 100 approximately. The relative standard deviations of the method were <5 % for 10 separate column experiments for the determination of 5.0 μg of cadmium, copper, nickel, and lead ions. The adsorption capacity of the dithizone-SBA-15 was 189 mg?g^?1 for cadmium, 102 mg?g^?1 for copper, 91 mg?g^?1 for nickel, and 208 mg?g^?1 for lead.     

Sensitive and Selective Determination of Riboflavin in Milk and Soymilk Powder by Multi-walled Carbon Nanotubes and Ionic Liquid [BMPi]PF<Subscript>6</Subscript> Modified Electrode

A novel electrochemical method to detect riboflavin was proposed using a multi-walled carbon nanotubes and ionic liquid N-butyl-N-methyl-piperidinium hexafluorophosphate composite film modified glassy carbon electrode (MWNTs-[BMPi]PF₆/GCE). A well-defined CV behavior with a pair of sensitive and well-shaped redox peak was observed, and the response value of riboflavin at MWNTs-[BMPi]PF₆/GCE is much higher than that at MWNTs/GCE in 0.1 mol L^?1 HAc-NaAc buffer solution (pH 4.5). The electrochemical approach based on a sensitive DPV analytical signal exhibits an excellent analytical performance with a wide linear range (2.6 × 10^?8 to 1.3 × 10^?6 mol L^?1) and low detection limit (4.7 × 10^?9 mol L^?1) for the riboflavin. Moreover, the proposed method possesses a potential practical application value and can be employed for the quantitative analysis of trace riboflavin with a recovery of 95.8–102.4 % in food samples such as milk and soymilk powder.     

Use of Multivariate Optimization to Develop Methods for Direct Copper and Lead Determination in Breast Milk by Graphite Furnace Atomic Absorption Spectrometry

A direct method for lead and copper determination in breast milk by graphite furnace atomic absorption spectrometry, using aqueous calibration, was proposed in this study. Samples were diluted with hydroximethylaminomethane 80 %?v/v, pH 8. The dilution determination for Pb and Cu was 1:1 and 1:9, respectively. Fractional factorial (2^4?1) and central composite designs were used to optimize experimental conditions (pyrolysis and atomization temperatures, pyrolysis time, and modifier) using 10 μL samples introduced into a graphite furnace. The methods allowed for copper and lead determination under optimized conditions with an aqueous calibration curve between 0 and 180 μg L^?1 for Cu and 0 and 48 μg L^?1 for Pb. The detection limits were 0.92 μg L^?1 and 6.4 μg L^?1 for Pb and Cu, respectively. Intra and inter-assay studies revealed coefficients of variation of 5.0 and 6.3 %, and 6.4 and 5.5 % for Pb and Cu, respectively. Recovery studies at three concentration levels (three consecutive days, n?=?7/day) presented results between 107 and 109 % for Pb and 102 and 103 % for Cu. Good accuracy was obtained for both metals through recoveries studies using certified reference material (infant formula NIST® 1846). The method determined lead and copper in six samples and the concentrations ranged from 2.90 to 27.9 μg L^?1 for Pb and 384 to 1,212 μg L^?1 for Cu. While copper is an essential element, lead has no nutritional function and is cumulative at low concentrations. Therefore, safe, efficient, and validated methods should be available to determine its concentration in breast milk.     

Large-Volume Sample Staking of Rice Polyphenols Prior to Their Determination by Non-aqueous Capillary Electrophoresis

A rapid non-aqueous capillary electrophoretic (NACE) method for the separation of (?)-epicatechin (EPI), (+)-catechin (?CAT), kaempferol (KAE), quercetin (QUR), naringanin (NAR), ferulic acid (FA), and p-coumaric acid (p-CA) has been developed and applied to the determination of these compounds in different rice varieties. All seven compounds were separated on capillary of 50 μm?×?68 cm (60-cm effective length) using 20 mmol L^?1 borate buffer of pH 9.0 and 5 % acetonitrile in methanol. Large-volume sample stacking (LVSS) technique was optimized and used to preconcentrate non-detectable polyphenols of white polished rice. Rice extracts were concentrated on-line by LVSS prior to separation by non-aqueous capillary electrophoresis. An improvement of 10–55 times in detectability was achieved with injection at 50 mbar for 30 s followed by voltage inversion (?20 kV) for 5 s. Linear calibration range of 1–300 μg L^?1 and 0.01–60 μg L^?1 was observed for NACE and NACE-LVSS method respectively, with the detection limit of 0.33–2.0 and 0.006–0.19 μg L^?1. Good reproducibility with standard deviations of less than 5 % was achieved. Polyphenol contents of different rice varieties were determined using developed method.     

The Effects of Ultrasound Treatment and Nano-zinc Oxide Coating on the Physiological Activities of Fresh-Cut Kiwifruit

This study evaluated the efficacy of ultrasound treatment and nano-zinc oxide (ZnO) coating individually and in combination in preserving the quality of fresh-cut kiwifruit. The nano-ZnO coating solution was prepared by mixing the ZnO nanoparticles in premixed chitosan–acetic acid solution. The fresh-cut kiwifruit were dipped in NaClO solution (50 μL?L^?1 sodium, control), subjected to ultrasound treatment (40 KHz, 350 W, 10 min), or coated with nano-ZnO solution. The fresh-cut kiwifruit samples were also subjected to combined ultrasound treatment and nano-ZnO coating. All of these test samples were stored at 4 °C for 10 days. The effects of these treatments on the quality parameters such as the production of carbon dioxide and ethylene, mass loss, and flesh firmness were investigated. At the end of storage, the combination treatments with ultrasound treatment at 40 KHz with 1.2 g?L^?1 nano-ZnO coating showed lower production of ethylene (1.86 μL?kg^?1?h^?1) and carbon dioxide (10.01 mg?kg^?1?h^?1), water loss (0.46 %), and texture (7.87 N). Hence, it was concluded that the combination of ultrasound treatment with nano-ZnO coating is a promising approach to extend the shelf-life of fresh-cut kiwifruit.