Recent progress in promoting the bioavailability of polyphenols in plant-based foods.

Polyphenols are important constituents of plant-based foods, exhibiting a range of beneficial effects. However, many phenolic compounds have low bioavailability because of their low water solubility, chemical instability, food matrix effects, and interactions with other nutrients. This article reviews various methods of improving the bioavailability of polyphenols in plant-based foods, including fermentation, natural deep eutectic solvents, encapsulation technologies, co-crystallization and amorphous solid dispersion systems, and exosome complexes. Several innovative technologies have recently been deployed to improve the bioavailability of phenolic compounds. These technologies may be utilized to increase the healthiness of plant-based foods. Further research is required to better understand the mechanisms of action of these novel approaches and their potential to be used in food production.

Direct Determination of Peroxide Explosives on Polycarbazole/Gold Nanoparticle-Modified Glassy Carbon Sensor Electrodes Imprinted for Molecular Recognition of TATP and HMTD.

Since peroxide-based explosives (PBEs) lack reactive functional groups, they cannot be determined directly by most detection methods and are often detected indirectly by converting them to H2O2. However, H2O2 may originate from many sources, causing false positives in PBE detection. Here, we developed a novel electrochemical sensor for the direct sensitive and selective determination of PBEs such as triacetone triperoxide (TATP) and hexamethylene triperoxide diamine (HMTD) using electrochemical modification of the glassy carbon (GC) electrode with PBE-memory polycarbazole (PCz) films decorated with gold nanoparticles (AuNPs) by cyclic voltammetry (CV). The prepared electrodes were named TATP-memory-GC/PCz/AuNPs (used for TATP determination) and HMTD-memory-GC/PCz/AuNPs (used for HMTD detection). The calibration lines of TATP and HMTD were found in the concentration range of 0.1-1.0 mg L-1 using the net current intensities of differential pulse voltammetry (DPV) versus analyte concentrations. The limit of detection (LOD) commonly found was 15 µg L-1 for TATP and HMTD. The sensor electrodes could separately determine intact TATP and HMTD in the presence of nitro-aromatic, nitramine, and nitrate ester energetic materials. The proposed electrochemical sensing method was not interfered by electroactive substances such as paracetamol, caffeine, acetylsalicylic acid, aspartame, d-glucose, and detergent (containing perborate and percarbonate) used as camouflage materials for PBEs. This is the first molecularly imprinted polymeric electrode for PBEs accomplishing such low LODs, and the DPV method was statistically validated in contaminated clay soil samples against the GC-MS method for TATP and a spectrophotometric method for HMTD using t- and F-tests.

A novel flow injection amperometric method for sensitive determination of total antioxidant capacity at cupric-neocuproine complex modified MWCNT glassy carbon electrode.

A novel amperometric method is presented for the determination of total antioxidant capacity in flow injection analysis (FIA) system using copper(II)-neocuproine complex modified on Nafion-functionalized multi-walled carbon nanotube-glassy carbon electrode ([Cu(Ncp)22+]/Nf@f-MWCNT/GCE). Cyclic voltammetric studies showed that the modified electrode exhibits a very well-formed reversible redox couple for Cu(II)-/Cu(I)-complex. In addition, the [Cu(Ncp)22+]/[Cu(Ncp)2+] redox pair shows very good electrocatalytic activity towards the oxidation of polyphenolic compounds (PPhCs) such as trolox, catechin, and quercetin due to the enhancement of the anodic peak current of the redox couple in the presence of these analytes. This electrocatalytic oxidation current at the [Cu(Ncp)22+]/Nf@f-MWCNT/GCE was used for flow injection (FI) amperometric determination of PPhCs. FI amperometric-time curves recorded under optimized conditions (applied potential: + 0.6 V vs. Ag/AgCl/KCl(0.10 M), flow rate: 2 mL/min) showed that the proposed electrode had a wide linear range (LR) with a very low detection limit (LOD) for PPhCs. LR and LOD were 0.5-800 and 0.2 µM for trolox, respectively and 0.50-250 and 0.14 µM, respectively, for both quercetin and catechin. This sensitive method was successfully applied to the amperometric measurement of total antioxidant capacity (TAC) of some herbal teas, giving compatible results with the spectrophotometric CUPRAC method. The proposed method gave higher rank to fast-reacting antioxidants; it was equally precise but had a wider linear range and lower LOD than the spectrophotometric CUPRAC assay (e.g., LOD for ascorbic acid and gallic acid were 0.07 and 0.08 µM, respectively), and similar electroanalytical methods using the CUPRAC reagent.

Direct Electrochemical Determination of Peroxide-Type Explosives Using Well-Dispersed Multi-Walled Carbon Nanotubes/Polyethyleneimine-Modified Glassy Carbon Electrodes.

The sensitive and selective determination of peroxide-based explosives (PBEs) in the field/on site is an important analytical challenge. Most methods claiming to detect PBEs are indirect, actually detecting their decomposition product, H2O2. Here, we present an electrochemical sensor for direct detection of organic peroxide explosives, that is, triacetone triperoxide (TATP) and hexamethylenetriperoxide diamine (HMTD), using well-dispersed multiwalled carbon nanotubes/polyethyleneimine (MWCNTs/PEI)-modified glassy carbon (GC) electrode, namely, GC/MWCNTs/PEI electrode. This is the first use of the conductive polyelectrolyte PEI as an electrode modifier for pristine PBE sensing. The potential range, scan rate, solvent selection, and supporting electrolyte concentration were optimized for PBEs. As a distinct advantage over other similar methods, our sensor electrode responded to intact TATP solutions in neutral medium, meaning that TATP did not interact with acids/bases that would transform it into H2O2. Calibration curves were linear in the range of 10-200 mg L-1 for TATP and 25-200 mg L-1 for HMTD. Using differential pulse voltammetry, detection limits of 1.5 mg L-1 TATP and 3.0 mg L-1 HMTD were obtained from direct electrochemical reduction in 80/20% (v/v) H2O-acetone solvent medium. Electroactive camouflage materials such as passenger belongings (e.g., sweetener, detergent, sugar, and paracetamol-caffeine-based analgesic drugs), common ions, and other explosives were shown not to interfere with the proposed method. The nonresponsive behavior of our electrode to H2O2 prevents "false positives" from other peroxide materials of everyday use. This electrochemical sensor could also detect other nitro-explosives at different potentials and was statistically validated against standard GC-MS and spectrophotometric methods.

A Simple Determination of Trinitrotoluene (TNT) Based on Fluorescence Quenching of Rhodamine 110 with FRET Mechanism.

Sensitive and selective detection of nitroaromatic explosives is an important issue in regard to human health, environment, public security and military issues. In this study, a simple and sensitive fluorescence quenching - based assay utilizing Rhodamine 110 as fluorophore probe was developed for the determination of trinitrotoluene (TNT). This sensitive fluorometric method could measure the decrease in fluorescence of Rhodamine 110 (λex = 490 nm, λem = 521 nm) owing to the primary amine groups of Rhodamine 110 (different from other rhodamines) capable of donor-acceptor interaction with TNT. The resulting TNT-amine complex can strongly quench the fluorescence emission of Rhodamine 110 by fluorescence resonance energy transfer (FRET) which occurs as the excited Rhodamine 110 fluorophore (donor) transfers its energy to TNT (acceptor) by non-radiative dipole-dipole interaction. Fluorescence quenching varied linearly with TNT concentration, with LOD and the LOQ of 0.71 and 2.38 mg L- 1 TNT, respectively. Similar explosives, common soil ions, and possible camouflage materials were found not to interfere with the proposed method, offering significant advantages with its easy methodology, low-cost, sensitivity, and rapidity of analysis. FRET mechanism based on dye donor-TNT acceptor interaction.

Ethylenediamine-bound magnetite nanoparticles as dual function colorimetric sensor having charge transfer and nanozyme activity for TNT and tetryl detection.

A reusable, low-cost, and convenient ethylenediamine (EDA)-bound magnetite nanoparticles (MNPs)-based colorimetric sensor has been developed for dual function colorimetric determination of nitroaromatic explosives such as TNT and tetryl. Colorimetric detection of analytes may occur through two independent routes: (1) nano-Fe3O4- EDA- NH2 as σ-donor may interact with the σ- and π-acceptor aromatic-poly(NO2) groups to produce a colored charge-transfer (CT) complex; (2) nano-Fe3O4-EDA-NH2 as a Fenton-type nanozyme may generate reactive species that comprise hydroxyl radicals (•OH) with H2O2 to oxidize 3,3',5,5'-tetramethylbenzidine (TMB) to a blue-colored diimine (oxTMB-TMB) CT complex, where this color is bleached with TNT/tetryl because of donor-acceptor interactions between the explosive -NO2 groups and the -NH2 group of Fe3O4-EDA nanoparticles of restricted nanozyme activity. Both methods can quantify TNT well below the EPA recommended TNT residential screening level in soil, LOD being in the micromolar range. As EDA was covalently bound to MNPs, the same sensor can be separately reused six times for TNT and eight times for tetryl determination, using method (1). Common metal ions, anions, energetic materials, several camouflage materials, and soil components such as humates did not interfere with the nanosensor performance for TNT and tetryl. The combination of charge-transfer and nanozyme ability of Fe3O4- EDA-NH2 nanoparticles may bring a new approach to dual function colorimetric sensor design. To the best of our knowledge, this is the first dual function colorimetric sensor for TNT and tetryl using the same nanoparticles as sensing elements in two different detection systems involving either formation or bleaching of colored species. The proposed colorimetric sensor can determine nitroaromatic explosives in two different ways: method-1 for TNT and tetryl sensing with EDA-MNPs relies on the donor-acceptor interaction between the electron-deficient nitroaromatics and electron-rich amine groups covalently functionalized on MNPs to produce an absorbance at 512 nm. In method-2, EDA-MNPs having nanozyme activity react with H2O2 to form reactive species that can oxidize TMB to its blue-colored charge-transfer (CT) complex, where TNT and tetryl addition may partially inhibit the nanozyme activity of EDA-MNPs and cause color bleaching (decrement of 650 nm absorbance) by disrupting the CT complex formed from TMB. This is the first dual function colorimetric sensor for nitro explosives uniquely combining charge-transfer and nanozyme ability of EDA-Fe3O4 nanoparticles in the same nano-sensor.

A colorimetric probe based on 4-mercaptophenol and thioglycolic acid-functionalized gold nanoparticles for determination of phytic acid and Fe(III) ions.

As a first of its kind, we developed a highly sensitive colorimetric nanoprobe for phytic acid (PA) and Fe(III) ion detection based on 4-mercaptophenol (4MP) and thioglycolic acid (TGA)-functionalized gold nanoparticles {AuNPs@(4MP-TGA)}. AuNPs were easily derivatized by 4MP and TGA through -SH binding to gold. Fe(III) ions possibly are bound first to the phenolate groups of 4MP-AuNPs, and further coordinated several nanoparticles via the carboxylate groups of TGA-AuNPs to cause aggregation, resulting in a red-to-purple color change and a bathochromic shift in the SPR absorption band of the nanoprobe. With the addition of PA to the AuNPs@(4MP-TGA)-Fe(III) system, the aggregated particles were released due to strong complex formation between Fe(III) and PA, resulting in a restoration of the color (purple-to-red) and of the SPR band to the original 520 nm wavelength maximum. Thus, the 650-nm absorption is attenuated and the 520-nm band is enhanced upon PA-Fe(III) chelation. This means that the absorption ratio A650/A520 is an indication of Fe(III) whereas the reverse ratio A520/A650 of the PA content of complex samples. The limits of detection (LOD) of the AuNPs@(4MP-TGA) were 1.0 µM for Fe(III) ions and 0.15 µM for PA. Phytic acid extracted from bean grains was determined with the proposed probe, yielding good recoveries. In addition, common metal ions, anions, and several biomolecules did not show an adverse effect on the nanoprobe performance for ferric ions and phytate. The developed method was statistically validated against a LC-MS/MS literature method. Graphical abstract Mercaptophenolate (4MP)- and thioglycolic acid (TGA)-functionalized gold nanoparticles were prepared as nanoprobes to detect Fe(III) ions through nanoparticle aggregation accompanied by red-to-purple color shift. The same nanoprobe determined phytic acid in food through disaggregation of Fe(III)-aggregated nanoparticles by strong Fe(III)-phytate chelation and restoration of solution color from purple to red.

Electrochemical Determination of TNT, DNT, RDX, and HMX with Gold Nanoparticles/Poly(Carbazole-Aniline) Film-Modified Glassy Carbon Sensor Electrodes Imprinted for Molecular Recognition of Nitroaromatics and Nitramines.

Since nitroaromatic- and nitramine-type energetic materials, mostly arising from military activities, are persistent pollutants in soil and groundwater, on-site sensing of these hazardous chemicals has gained importance. A novel electrochemical sensor was designed for detecting nitroaromatic- and nitramine-type energetic materials, relying on gold nanoparticles (Aunano), modified glassy carbon (GC) electrode coated with nitro-energetic memory-poly(carbazole-aniline) copolymer (Cz- co-ANI) film (e.g., TNT memory-GC/P(Cz- co-ANI)-Aunano modified electrode). Current was recorded against concentration to build the calibration curves that were found to be linear within the range of 100-1000 µg L-1 for 2,4,6-trinitrotoluene (TNT) and 2,4-dinitrotoluene (DNT): 50-1000 µg L-1 for 1,3,5-trinitro-1,3,5-triazacyclohexane (RDX) and octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX). The corresponding limits of detection were 25 µg L-1 for TNT, 30 µg L-1 for DNT, and 10 µg L-1 for both RDX and HMX, using nitro-energetic memory-GC/P(Cz- co-ANI)-Aunano electrodes. These electrodes were used separately, and specific determinations were made in various mixtures of nitro-energetic materials. The developed method could be efficiently used in electroanalyzing nitroaromatics and nitramines in military explosives (i.e., comp B, octol, and comp A5). The sensor electrodes were specific for the tested nitro-energetic compounds and did not respond to paracetamol-caffeine-based analgesic drug, acetylsalicylic acid (aspirin), sweetener, and sugar that can be used as camouflage materials in passenger belongings. The developed method was statistically validated against the standard LC-MS reference method in contaminated clay soil samples containing TNT and RDX explosives.

Novel Spectroscopic and Electrochemical Sensors and Nanoprobes for the Characterization of Food and Biological Antioxidants.

Since an unbalanced excess of reactive oxygen/nitrogen species (ROS/RNS) causes various diseases, determination of antioxidants that can counter oxidative stress is important in food and biological analyses. Optical/electrochemical nanosensors have attracted attention in antioxidant activity (AOA) assessment because of their increased sensitivity and selectivity. Optical sensors offer advantages such as low cost, flexibility, remote control, speed, miniaturization and on-site/in situ analysis. Electrochemical sensors using noble metal nanoparticles on modified electrodes better catalyze bioelectrochemical reactions. We summarize the design principles of colorimetric sensors and nanoprobes for food antioxidants (including electron-transfer based and ROS/RNS scavenging assays) and important milestones contributed by our laboratory. We present novel sensors and nanoprobes together with their mechanisms and analytical performances. Our colorimetric sensors for AOA measurement made use of cupric-neocuproine and ferric-phenanthroline complexes immobilized on a Nafion membrane. We recently designed an optical oxidant/antioxidant sensor using N,N-dimethyl-p-phenylene diamine (DMPD) as probe, from which ROS produced colored DMPD-quinone cationic radicals electrostatically retained on a Nafion membrane. The attenuation of initial color by antioxidants enabled indirect AOA estimation. The surface plasmon resonance absorption of silver nanoparticles as a result of enlargement of citrate-reduced seed particles by antioxidant addition enabled a linear response of AOA. We determined biothiols with Ellman reagent-derivatized gold nanoparticles.

CUPRAC colorimetric and electroanalytical methods determining antioxidant activity based on prevention of oxidative DNA damage.

An unbalanced excess of oxygen/nitrogen species (ROS/RNS) can give oxidative hazard to DNA and other biomacromolecules under oxidative stress conditions. While the 'comet' assay for measuring DNA damage is neither specific nor practical, monitoring oxidative changes on individual DNA bases and other oxidation products needs highly specialized equipment and operators. Thus, we developed a modified CUPRAC (cupric ion reducing antioxidant capacity) colorimetric method to determine the average total damage on DNA produced by Fenton oxidation, taking advantage of the fact that the degradation products of DNA but not the original macromolecule is CUPRAC-responsive. The DNA-protective effects of water-soluble antioxidants were used to devise a novel antioxidant activity assay, considered to be physiologically more realistic than those using artificial probes. Our method, based on the measurement of DNA oxidative products with CUPRAC colorimetry proved to be 2 orders-of-magnitude more sensitive than the widely used TBARS (thiobarbituric acid-reactive substances) colorimetric assay used as reference. Additionally, the DNA damage was electrochemically investigated using pencil graphite electrodes (PGEs) as DNA sensor platform in combination with differential pulse voltammetry (DPV). The interaction of the radical species with DNA in the absence/presence of antioxidants was detected according to the changes in guanine oxidation signal.

Spectrophotometric Determination of Phenolic Antioxidants in the Presence of Thiols and Proteins.

Development of easy, practical, and low-cost spectrophotometric methods is required for the selective determination of phenolic antioxidants in the presence of other similar substances. As electron transfer (ET)-based total antioxidant capacity (TAC) assays generally measure the reducing ability of antioxidant compounds, thiols and phenols cannot be differentiated since they are both responsive to the probe reagent. In this study, three of the most common TAC determination methods, namely cupric ion reducing antioxidant capacity (CUPRAC), 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt/trolox equivalent antioxidant capacity (ABTS/TEAC), and ferric reducing antioxidant power (FRAP), were tested for the assay of phenolics in the presence of selected thiol and protein compounds. Although the FRAP method is almost non-responsive to thiol compounds individually, surprising overoxidations with large positive deviations from additivity were observed when using this method for (phenols + thiols) mixtures. Among the tested TAC methods, CUPRAC gave the most additive results for all studied (phenol + thiol) and (phenol + protein) mixtures with minimal relative error. As ABTS/TEAC and FRAP methods gave small and large deviations, respectively, from additivity of absorbances arising from these components in mixtures, mercury(II) compounds were added to stabilize the thiol components in the form of Hg(II)-thiol complexes so as to enable selective spectrophotometric determination of phenolic components. This error compensation was most efficient for the FRAP method in testing (thiols + phenols) mixtures.

Electrochemical Determination of Food Preservative Nitrite with Gold Nanoparticles/p-Aminothiophenol-Modified Gold Electrode.

Due to the negative impact of nitrate and nitrite on human health, their presence exceeding acceptable levels is not desired in foodstuffs. Thus, nitrite determination at low concentrations is a major challenge in electroanalytical chemistry, which can be achieved by fast, cheap, and safe electrochemical sensors. In this work, the working electrode (Au) was functionalized with p-aminothiophenol (p-ATP) and modified with gold nanoparticles (Au-NPs) to manufacture the final (Au/p-ATP-Aunano) electrode in a two-step procedure. In the first step, p-ATP was electropolymerized on the electrode surface to obtain a polyaminothiophenol (PATP) coating. In the second step, Au/p-ATP-Aunano working electrode was prepared by coating the surface with the use of HAuCl4 solution and cyclic voltammetry. Determination of aqueous nitrite samples was performed with the proposed electrode (Au/p-ATP-Aunano) using square wave voltammetry (SWV) in pH 4 buffer medium. Characteristic peak potential of nitrite samples was 0.76 V, and linear calibration curves of current intensity versus concentration was linear in the range of 0.5-50 mg·L(-1) nitrite with a limit of detection (LOD) of 0.12 mg·L(-1). Alternatively, nitrite in sausage samples could be colorimetrically determined with high sensitivity by means of p-ATP‒modified gold nanoparticles (AuNPs) and naphthylethylene diamine as coupling agents for azo-dye formation due to enhanced charge-transfer interactions with the AuNPs surface. The slopes of the calibration lines in pure NO2(-) solution and in sausage sample solution, to which different concentrations of NO2(-) standards were added, were not significantly different from each other, confirming the robustness and interference tolerance of the method. The proposed voltammetric sensing method was validated against the colorimetric nanosensing method in sausage samples.

Determination of triacetone triperoxide with a N,N-dimethyl-p-phenylenediamine sensor on nafion using Fe3O4 magnetic nanoparticles.

The explosive triacetone triperoxide (TATP) can be easily manufactured from readily accessible reagents and is extremely difficult to detect, owing to the lack of UV absorbance, fluorescence, or facile ionization. The developed method is based on the acidic hydrolysis of TATP into H2O2, pH adjustment to 3.6, and the addition of magnetite nanoparticles (Fe3O4 MNPs) to the medium to produce hydroxyl radicals from H2O2, owing to the peroxidase-like activity of MNPs. The formed radicals converted the N,N-dimethyl-p-phenylenediamine (DMPD) probe to the colored DMPD(+) radical cation, the optical absorbance of which was measured at a wavelength of 554 nm. The molar absorptivity (ε) of the method for TATP was 21.06 × 10(3) L mol(-1) cm(-1). The colored DMPD(+) product in solution could be completely retained on a cation-exchanger Nafion membrane, constituting a colorimetric sensor for TATP and increasing the analytical sensitivity. The proposed method did not respond to a number of hand luggage items like detergent, sweetener, sugar, acetylsalicylic acid (aspirin), and paracetamol-caffeine-based analgesic drugs. On the other hand, TATP could be almost quantitatively recovered from a household detergent and sweetener that can be used as camouflage for the analyte. Neither common soil and groundwater ions (e.g., Ca(2+), Mg(2+), K(+), Cl(-), SO4(2-), and NO3(-)) at 100-fold ratios nor nitro-explosives of trinitrotoluene (TNT), hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), and pentaerythritol tetranitrate (PETN) at 10-fold amounts interfered with the proposed assay. The method was statistically validated against the standard GC/MS reference method.

Determination of total antioxidant capacity of milk by CUPRAC and ABTS methods with separate characterisation of milk protein fractions.

Most milk-applied antioxidant assays in literature are based on the isolation and quantification of individual antioxidative compounds, whereas total antioxidant capacity (TAC) gives a more holistic picture due to cooperative action of antioxidants. Recently, the cupric reducing antioxidant capacity (CUPRAC) method has been modified to measure the antioxidant capacities of thiol-containing proteins, where the classical ammonium acetate buffer - that may otherwise precipitate proteins- was replaced with concentrated urea buffer (able to expose embedded thiol groups of proteins to oxidative attack) adjusted to pH 7.0. Thus, antioxidant capacity of milk was investigated with two competing TAC assays, namely CUPRAC and ABTS (2,2'-azinobis(3-ethylbenzothiazoline-6-sulphonic acid))/persulphate, because only these assays were capable of evaluating protein contribution to the observed TAC value. As milk fat caused turbidity, experiments were carried out with skim milk or defatted milk samples. To determine TAC, modified CUPRAC method was applied to whole milk, separated and redissolved protein fractions, and the remaining liquid phase after necessary operations. Both TAC methods were investigated for their dilution sensitivity and antioxidant power assessment of separate milk fractions such as casein and whey. Proteins like ß-lactoglobulin and casein (but not simple thiols) exhibited enhanced CUPRAC reactivity with surfactant (SDS) addition. Addition of milk protein fractions to whole skim milk produced significant 'negative-biased' deviations (up to -26% relative standard error) from TAC absorbance additivity in the application of the ABTS method, as opposed to that of the CUPRAC method less affected by chemical deviations from Beer's law thereby producing much smaller deviations from additivity (i.e. the property of additivity is valid when the measured TAC of a mixture is equal to the sum of individual antioxidant capacities of its constituents).

4-Aminothiophenol functionalized gold nanoparticle-based colorimetric sensor for the determination of nitramine energetic materials.

The heterocyclic nitramine compounds, hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX), are two most important military-purpose high explosives. Differentiation of RDX and HMX with colorimetric methods of determination has not yet been made because of their similar chemical structures. In this study, a sensitive colorimetric method for the determination of RDX and HMX was proposed on the basis of differential kinetics in the hydrolysis of the two compounds (yielding nitrite as a product) followed by their colorimetric determination using 4-aminothiophenol (4-ATP) modified gold nanoparticles (AuNPs) and naphthylethylene diamine (NED) as coupling agent for azo-dye formation, abbreviated as "4-ATP-AuNP+NED" colorimetric method. After alkaline hydrolysis in a 1 M Na2CO3 + 0.04 M NaOH mixture solution at room temperature, only RDX (but not HMX) was hydrolyzed to give a sufficient colorimetric response in neutralized solution, the molar absorptivity (ε) at 565 nm and the limit of detection (LOD) for RDX being (17.6 ± 1.3) × 10(3) L mol(-1) cm(-1) and 0.55 µg mL(-1), respectively. On the other hand, hot water bath (at 60 °C) hydrolysis enabled both nitramines, RDX and HMX, to give substantial colorimetric responses; i.e., ε and LOD for RDX were (32.8 ± 0.5) × 10(3) L mol(-1)cm(-1) and 0.20 µg mL(-1) and for HMX were (37.1 ± 2.8) × 10(3) L mol(-1)cm(-1) and 0.24 µg mL(-1), respectively. Unlike other AuNP-based nitrite sensors in the literature showing absorbance quenching within a relatively narrow concentration range, the developed sensor operated with an absorbance increase over a wide range of nitrite. Synthetic mixtures of (RDX + HMX) gave additive responses, and the proposed method was statistically validated against HPLC using nitramine mixtures.

Identification and anti-oxidant capacity determination of phenolics and their glycosides in elderflower by on-line HPLC-CUPRAC method.

INTRODUCTION: Development and application of an on-line cupric reducing anti-oxidant capacity (CUPRAC) assay coupled with HPLC for separation and on-line determination of phenolic anti-oxidants in elderflower (Sambucus nigra L.) extracts for their anti-oxidant capacity are significant for evaluating health-beneficial effects. Moreover, this work aimed to assay certain flavonoid glycosides of elderflower that could not be identified/quantified by other similar on-line HPLC methods (i.e. 2,2-diphenyl-1-picrylhdrazyl and 2, 2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid). OBJECTIVE: To identify anti-oxidant constituents in elderflower by HPLC and to evaluate their individual anti-oxidant capacities by on-line HPLC-CUPRAC assay with a post-column derivatisation system. METHODS: The separation and UV detection of polyphenols were performed on a C18 -column using gradient elution with two different mobile phase solutions, that is acetonitrile and 1% glacial acetic acid, with detection at 340 nm. The HPLC-separated anti-oxidant polyphenols in column effluent react with copper(II)-neocuproine in a reaction-coil to reduce the latter to copper(I)-neocuproine (Cu(I)-Nc) chelate having maximum absorption at 450 nm. RESULTS: The detection limits of tested compounds at 450 nm after post-column derivatisation were compared with those of at 340 nm UV-detection without derivatisation. LOD values (µg/mL) of quercetin and its glycosides at 450 nm were lower than those of UV detection at 340 nm. This method was applied successfully to elderflower extract. The flavonol glycosides of quercetin and kaempferol bound to several sugar components (glucose, rhamnose, galactose and rutinose) were identified in the sample. CONCLUSION: The on-line HPLC-CUPRAC method was advantageous over on-line ABTS and DPPH methods for measuring the flavonoid glycosides of elderflower.

Antioxidant/prooxidant effects of α-tocopherol, quercetin and isorhamnetin on linoleic acid peroxidation induced by Cu(II) and H2O2.

The peroxidation of linoleic acid (LA) in the presence of copper(II) (Cu(II)) ions alone and with α-tocopherol (α-TocH) was investigated in aerated and incubated emulsions at 37 °C and pH 7. Additionally, the effects of quercetin (QR) and its O-methylated derivative, isorhamnetin (IR), as potential antioxidant protectors were studied in the (Cu(II) + TocH)-induced LA peroxidation system. Cu(II)-induced LA peroxidation followed pseudo-first-order kinetics with respect to primary (hydroperoxides) and secondary (aldehydes- and ketones-like) oxidation products, which were determined by ferric thiocyanate and thiobarbituric acid-reactive substances methods, respectively. As opposed to the concentration-dependent (at 0.6 and 10.0 µM) prooxidative action of α-TocH in the absence of QR and IR, the latter two compounds showed antioxidant effect over TocH. The peroxidation of LA in the presence of Cu(II)-H(2)O(2) combination alone and with TocH, QR and IR were also investigated in aerated and incubated emulsions, where the latter three compounds exhibited antioxidant effects.

Folin-Ciocalteu spectrophotometric assay of ascorbic acid in pharmaceutical tablets and orange juice with pH adjustment and pre-extraction of lanthanum(III)­flavonoid complexes.

BACKGROUND: This study proposes modifications to the conventional Folin-Ciocalteu (FC) spectrophotometric method for individually determining ascorbic acid (AA) in complex matrices in the presence of other phenolics and potential interferents. The conventional FC assay in the aqueous phase,which normally measures total water-soluble phenolics and other antioxidants, has recently been modified by incorporating isobutanol (iso-BuOH) in the solvent mixture for the simultaneous determination of lipophilic and hydrophilic antioxidants in foods. RESULTS: Interference effects of other flavonoids and phenolics to individual AA assays were overcome by preliminary extraction­removal as their La(III) chelates into ethyl acetate (EtAc). The pH of the medium was adjusted to 3.0 in order to prevent conversion of AA into over-oxidation products further beyond the dehydroascorbic acid stage, as encountered in the conventional FC assay carried out at pH 10. This pH does not permit most phenolics to ionise, rendering their oxidation difficult. CONCLUSION: Both methods (conventional and iso-BuOH-modified FC at pH 3, with and without La(III)/EtAc pre-extraction) were applied to the determination of ascorbic acid in pharmaceutical tablets and orange juice, showing good agreement with HPLC results. The proposed spectrophotometric methods with their low cost, simplicity, reliability, versatility and accuracy offer novelty to the determination of AA in complex matrices.

Colorimetric Sensing of Antioxidant Capacity via Auric Acid Reduction Coupled to ABTS Oxidation.

In this study, a simple and sensitive colorimetric assay has been developed for total antioxidant capacity measurement. The assay is based on the absorption measurement of the bluish-green oxidized product (ABTS·+) formed as a result of the oxidation reaction of the chromogenic reagent ABTS (2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) with gold(III). However, in the presence of antioxidants, the ABTS oxidation process is effectively suppressed due to the reduction of gold(III) ions to the zerovalent state forming gold nanoparticles (AuNPs). Relatively lighter colors and a significant decrease in absorbance are observed depending on the total antioxidant capacity. Taking advantage of this situation, qualitative and quantitative total antioxidant capacity (TAC) measurements, with the naked eye and UV-vis spectroscopy, respectively, could be successfully performed. The assay is named "auric reducing antioxidant capacity" (AuRAC) because the gold(III) ion-reducing ability of antioxidants is measured. The AuRAC assay was applied to dietary polyphenols, vitamin C, thiol-type antioxidants, and their synthetic mixtures. Trolox equivalent antioxidant capacity (TEAC) values obtained with the AuRAC assay were found to be compatible with those of the reference CUPRAC (cupric reducing antioxidant capacity) assay. The AuRAC assay was validated through linearity, additivity, precision, and recovery, demonstrating that the assay is reliable and robust. Compared to the simple TAC assays in the literature based on AuNP formation with subsequent surface plasmon resonance (SPR) absorbance measurement, this indirect assay has a smoother linear range starting from lower antioxidant concentrations. This method displays much higher molar absorption coefficients for antioxidant compounds than other conventional single electron transfer (SET) assays because 3-e- reduction of trivalent gold (i.e., Au(III) → Au(0)) produces three chromophore cation radicals (ABTS·+) of the assay reagent. The sensor has been successfully applied to complex matrices, such as tea infusions and pharmaceutical samples. The AuRAC assay stands out with its high molar absorptivity connected to enhanced sensitivity as well as its potential to convert into a paper-based colorimetric sensor.