Democratization of Copper Analysis in Grape Must Following a Polymer-Based Lab-on-a-Chip Approach.

Quality control in the food industry is of the upmost importance from the food safety, organoleptic and commercial viewpoints. Accordingly, the development of in situ, rapid, and costless analytical tools is a valuable task in which we are working. Regarding this point, the copper content of grape must has to be determined by wineries along the wine production process. For this purpose, grape must samples are sent to laboratories where the copper content is measured usually by flame atomic absorption spectrometry or by inductively coupled plasma mass spectrometry. We herein propose a straightforward, rapid, and inexpensive methodology based both on a film-shaped colorimetric polymer sensor and a smartphone method that at the same time can be used by unskilled personnel. The sensory polymer films change their color upon dipping them on the grape must, and the color evolution is analyzed using the digital color parameters of a picture taken to the film with a smartphone. Furthermore, the analytical procedure is automatically carried out by a smartphone app. The limit of detection of copper of the polymer sensor is 0.08 ppm. Following this approach, 18 production samples coming from the French Groupe ICV company were studied. The copper content of the samples was analyzed by the usual procedure carried out by the company (flame atomic absorption spectrometry) and by the method proposed in this work, ranging this content from 0.41 to 6.08 ppm. The statistical study showed that the results of both methods are fully consistent, showing the validity of the proposed method for the determination of copper in grape must within the frame of wine production wineries and industries.

Smart sensory polymer for straightforward Zn(II) detection in pet food samples.

We report on an innovative method to measure the Zn(II) concentration in commercial pet food samples, both wet and dry food. It is based on a colorimetric sensory polymer prepared from commercial monomers and 0.5 % of a synthetic monomer having a quinoline sensory core (N-(8-(2-azidoacetamido)quinolin-5-yl)methacrylamide). We obtained the sensory polymer as crosslinked films by thermally initiated bulk radical polymerization of the monomers of 100 µm thickness, which we punched into Ø6 mm sensory discs. The immersion of the discs in water solutions containing Zn(II) turned the fluorescence on, allowing for the titration of this cation using the G parameter of a digital picture taken to the discs. The limits of detection and quantification were 29 and 87 µg/L, respectively. Furthermore, we measured the concentration of Zn(II) even in the presence of other cations, detecting no significant interferences. Thus, in a further step, we obtained the concentration of Zn(II) from 15 commercial pet food samples, ranging from 19 to 198 mg/kg, following a simple extraction procedure and contacting the extractant with our sensory discs. These results were contrasted with that obtained by ICP-MS as a reference method.

Easy Nitrite Analysis of Processed Meat with Colorimetric Polymer Sensors and a Smartphone App.

We have developed an in situ methodology for determining nitrite concentration in processed meats that can also be used by unskilled personnel. It is based on a colorimetric film-shaped sensory polymer that changes its color upon contacting the meat and a mobile app that automatically calculates the manufacturing and residual nitrite concentration by only taking digital photographs of sensory films and analyzing digital color parameters. The film-shaped polymer sensor detects nitrite anions by an azo-coupling reaction, since they activate this reaction between two of the four monomers that the copolymer is based on. The sensory polymer is complemented with an app, which analyzes the color in two different digital color spaces (RGB and HSV) and performs a set of 32 data fittings representing the concentration of nitrite versus eight different variables, finally providing the nitrite concentration of the test samples using the best fitting curve. The calculated concentration of nitrite correlates with a validated method (ISO 2918: 1975) usually used to determine nitrite, and no statistically significant difference between these methods and our proposed one has been found in our study (26 meat samples, 8 prepared, and 18 commercial). Our method represents a great advance in terms of analysis time, simplicity, and orientation to use by average citizens.

The role of polymeric chains as a protective environment for improving the stability and efficiency of fluorogenic peptide substrates.

We have faced the preparation of fully water-soluble fluorescent peptide substrate with long-term environmental stability (in solution more than 35 weeks) and, accordingly, with stable results in the use of this probe in determining the activity of enzymes. We have achieved this goal by preparing a co-polymer of the commercial N-vinyl-2-pyrrolidone (99.5% mol) and a fluorescent substrate for trypsin activity determination having a vinylic group (0.5%). The activity of trypsin has been measured in water solutions of this polymer over time, contrasted against the activity of both the commercial substrate Z-L-Arg-7-amido-4-methylcoumarin hydrochloride and its monomeric derivative, prepared ad-hoc. Initially, the activity of the sensory polymer was 74.53 ± 1.72 nmol/min/mg of enzyme, while that of the commercial substrate was 20.44 ± 0.65 nmol/min/mg of enzyme, the former maintained stable along weeks and the latter with a deep decay to zero in three weeks. The 'protection' effect exerted by the polymer chain has been studied by solvation studies by UV-Vis spectroscopy, steady-state & time resolved fluorescence, thermogravimetry and isothermal titration calorimetry.

Polymer film as starch azure container for the easy diastase activity determination in honey.

A new original application for a polyacrylic film based on the monomers 2-(dimethylamino)ethyl methacrylate (NNDA), 2-hydroxyethyl acrylate (2HEA) and methylmethacrylate (MMA) as a starch azure container has been set up for a simple determination of honey diastase activity. The proposed method is based on the correlation of reducing sugars generated during the enzymatic process with the Schade reference assay. The polyacrylic film is charged with starch azure acting as a container for this substance; thus, the starch does not interfere in the measurement of reducing sugars, so that the diastase activity is easily calculated. The method has been contrasted with Schade method, showing good correlation and differences under 0.4% between methods in some honey samples. The polyacrylic film has great potential for the routine honey diastase activity assessment in small laboratories, dramatically reducing analysis time and cost.

A simple one-pot determination of both total phenolic content and antioxidant activity of honey by polymer chemosensors.

We have developed a new method for the rapid (2 h) and inexpensive (materials cost < 0.02 €/sample) "2-in-1" determination of the total phenolic content (TPC) and the antioxidant activity (AOX) in honey samples. The method is based on hydrophilic colorimetric films with diazonium groups, which react with phenols rendering highly colored azo groups. The TPC of the sample is correlated to its trolox equivalent antioxidant capacity (TEAC). The intensity of the color allows us to determine both TPC and TEAC of the sample by the analysis of a picture taken with a smartphone that is analysed by the use of the color-definition-parameters (RGB). The controlled light conditions and the systematic use of the same camera avoid the periodical calibration of the system improving the efficiency of the method. Thus, it is a simple method carried out by non-specialized personnel and it involves much lower money and time investment compared to traditional methods.

Why is the Sensory Response of Organic Probes within a Polymer Film Different in Solution and in the Solid-State? Evidence and Application to the Detection of Amino Acids in Human Chronic Wounds.

We anchored a colourimetric probe, comprising a complex containing copper (Cu(II)) and a dye, to a polymer matrix obtaining film-shaped chemosensors with induced selectivity toward glycine. This sensory material is exploited in the selectivity detection of glycine in complex mixtures of amino acids mimicking elastin, collagen and epidermis, and also in following the protease activity in a beefsteak and chronic human wounds. We use the term inducing because the probe in solution is not selective toward any amino acid and we get selectivity toward glycine using the solid-state. Overall, we found that the chemical behaviour of a chemical probe can be entirely changed by changing its chemical environment. Regarding its behaviour in solution, this change has been achieved by isolating the probe by anchoring the motifs in a polymer matrix, in an amorphous state, avoiding the interaction of one sensory motif with another. Moreover, this selectivity change can be further tuned because of the effectiveness of the transport of targets both by the physical nature of the interface of the polymer matrix/solution, where the target chemicals are dissolved, for instance, and inside the matrix where the recognition takes place. The interest in chronic human wounds is related to the fact that our methods are rapid and inexpensive, and also considering that the protease activity can correlate with the evolution of chronic wounds.

Potassium-Ion-Selective Fluorescent Sensors To Detect Cereulide, the Emetic Toxin of B.†cereus, in Food Samples and HeLa Cells.

We report the development of new chemical probes for cereulide, a toxic metabolite produced by specific strains of Bacillus cereus, through displacement of potassium cations from a preformed specific complex and a subsequent change in the fluorescence emission. For this purpose, we designed fluorescent probes for potassium cations that were suitable for displacement assays with cereulide from organic extracts. The fluorescence detection of natural cereulide in rice samples was achieved by using synthetic cereulide as a reference and a potassium fluorescent reporter, and this was found to be useful as a portable and fast method for the in situ detection of cereulide in food extracts. To study the fate of cereulide in live cells, we designed a procedure that was suitable for live-cell microscopy imaging of HeLa cells by comparing the cellular location of the potassium fluorogenic probe, which stained intracellular endolysosomes, in the absence and presence of cereulide; we concluded that in the presence of cereulide, the fluorescence of the probe was decreased because of complexation of the potassium ions by cereulide.

Stabilization of Al(III) solutions by complexation with cacodylic acid: speciation and binding features.

Aluminium ions are believed to play a role in a number of neurological and skeletal disorders in the human body. The study of the biological processes and molecular mechanisms that underlie these pathological disorders is rendered a difficult task due to the wide variety of complex species that result from the hydrolysis of Al(3+) ions. In addition, this ion displays a pronounced tendency to precipitate as a hydroxide, so certain complexing agents should be envisaged to stabilize Al(III) solutions in near physiological conditions. In this work, we show that the common buffer cacodylic acid (dimethylarsinic acid, HCac) interacts with Al(III) to give stable complexes, even at pH 7. After preliminary analyses of the speciation of the metal ion and also of the ligand, a systematic study of the formation of different Al/Cac complexes at different pH values has been conducted. UV-Vis titrations, mass spectrometry NMR measurements and DTF calculations were performed to enlighten the details of the speciation and stoichiometry of Al/Cac complexes. The results altogether show that Al/Cac dimer complexes prevail, but monomer and trimer forms are also present. Interestingly, it was found that cacodylate promotes the formation of such relatively simple complexes, even under conditions where the polymeric form, Al13O4(OH)24(7+), should predominate. The results obtained can help to shed some light into the reactivity of aluminium ions in biological environments.

Chemical speciation of MeHg+ and Hg2+ in aqueous solution and HEK cells nuclei by means of DNA interacting fluorogenic probes.

Selected new fluorogenic probes that interact in different ways with Hg2+ and MeHg+ have been prepared and used for the chemical speciation of both cations in aqueous solution as well as in HEK293 cells. The best selective speciation of Hg2+ and MeHg+ has been achieved by in vitro approaches based on fluorogenic probes supported in cultured cells, due to the particular sensitivity of the HEK293 cells to permeation by Hg2+, MeHg+ and the fluorogenic probes. In particular, MeHg+ was selectively detected in cell nuclei by probe JG45.

Acrylic Polymers with Pendant Phenylboronic Acid Moieties as "Turn-Off" and "Turn-On" Fluorescence Solid Sensors for Detection of Dopamine, Glucose, and Fructose in Water.

We report herein a fluorescence polymer membrane as a film-shaped solid sensory kit for the detection and quantification in water of saccharides, namely, fructose and glucose, and dopamine. The sensory motifs are phenylboronic acids, which are chemically incorporated in the polymer network in the radically initiated bulk polymerization process. The sensory membrane is fluorescent. The interaction of the sensory motifs with dopamine "turn-off" the fluorescence due to a dynamic quenching, while stable complexes are formed with saccharides giving rise to a fluorescence "turn-on". The variation of the fluorescence intensity and the wavelength of the maxima permitted the titration of the species with a detection limit of 3-4 × 10-4 M. The hydrophilic membrane allowed for the detection in water in spite of the lack of solubility in this medium of the sensory phenylboronic acid derivative monomer.

Forced solid-state interactions for the selective "turn-on" fluorescence sensing of aluminum ions in water using a sensory polymer substrate.

Selective and sensitive solid sensory substrates for detecting Al(III) in pure water are reported. The material is a flexible polymer film that can be handled and exhibits gel behavior and membrane performance. The film features a chemically anchored salicylaldehyde benzoylhydrazone derivative as an aluminum ion fluorescence sensor. A novel procedure for measuring Al(III) at the ppb level using a single solution drop in 20 min was developed. In this procedure, a drop was allowed to enter the hydrophilic material for 15 min before a 5 min drying period. The process forced the Al(III) to interact with the sensory motifs within the membrane before measuring the fluorescence of the system. The limit of detection of Al(III) was 22 ppm. Furthermore, a water-soluble sensory polymer containing the same sensory motifs was developed with a limit of detection of Al(III) of 1.5 ppb, which was significantly lower than the Environmental Protection Agency recommendations for drinking water.

Selective and sensitive detection of aluminium ions in water via fluorescence "turn-on" with both solid and water soluble sensory polymer substrates.

A solid substrate comprised of a cross-linked polymer network is shaped as a film with gel-like behaviour and is used to detect aluminium ions in water; concurrently, a water soluble sensory polymer synthesised towards the same purpose is also discussed. The detection in both systems was achieved via fluorescence "turn-on". The limits of detection for Al(III) were 1.6 and 25ppb for the former and latter materials, respectively; these levels are significantly lower than the EPA recommendations for drinking water.

Microwave dielectric relaxation spectroscopy study of alkan-1-ol/alkylbenzoate binary solvents.

The structure and dynamics of alkan-1-ol/alkylbenzoate binary mixtures have been studied by microwave dielectric relaxation spectroscopy in the 200 MHz to 20 GHz frequency range. The binary mixtures of methanol, ethanol, propan-1-ol, butan-1-ol, and pentan-1-ol with methyl, ethyl, propyl, and butyl benzoates were studied at 298.15 K. The relaxational response of the pure alcohols, pure esters, and their binary mixtures over the full composition range is properly described by the Havriliak-Negami model. The alcohol content, alcohol length, and alkyl side-chain effects on the relaxational properties have been studied for these mixtures over the whole composition range. From the experimental readings, the effective and the corrective Kirkwood and Bruggeman correlation factors have been calculated. The data gathered have been interpreted in terms of the alkyl side-chain effect and their reliance on the mixture composition.

An organic/inorganic hybrid membrane as a solid "turn-on" fluorescent chemosensor for coenzyme A (CoA), cysteine (Cys), and glutathione (GSH) in aqueous media.

The preparation of a fluorogenic sensory material for the detection of biomolecules is described. Strategic functionalisation and copolymerisation of a water insoluble organic sensory molecule with hydrophilic comonomers yielded a crosslinked, water-swellable, easy-to-manipulate solid system for water "dip-in" fluorogenic coenzyme A, cysteine, and glutathione detection by means of host-guest interactions. The sensory material was a membrane with gel-like behaviour, which exhibits a change in fluorescence behaviour upon swelling with a water solution of the target molecules. The membrane follows a "turn-on" pattern, which permits the titration of the abovementioned biomolecules. In this way, the water insoluble sensing motif can be exploited in aqueous media. The sensory motif within the membrane is a chemically anchored piperazinedione-derivative with a weakly bound Hg(II). The response is caused by the displacement of the cation from the membrane due to a stronger complexation with the biomolecules, thus releasing the fluorescent sensory moieties within the membrane.

Methacrylate copolymers with pendant piperazinedione-sensing motifs as fluorescent chemosensory materials for the detection of Cr(VI) in aqueous media.

A fluorogenic sensory film, or dense membrane, capable of detecting Cr(VI), Fe(III), and Hg(II) in water was prepared. The film was prepared by a bulk radical polymerization of different comonomers, one of which contained a piperazinedione motif as sensory fluorophore. The film exhibited gel-like behavior and was highly tractable, even after being swollen in water. The sensing conditions were chosen to overcome interference from iron and mercury cations, giving rise to a material with a detection limit of 1 ppb for Cr(VI).

A turn-on fluorogenic probe for detection of MDMA from ecstasy tablets.

We report a fluorogenic probe that is able to discriminate a range of primary or secondary biogenic amines and their natural or synthetic mimics, in water or buffer, by means of the turn-on transient generation of green fluorescence, with high quantum yields and low detection limits, thus making the system suitable for the detection of abuse drugs, such as MDMA, from ecstasy tablets.

Phosphine and thiophene cyclopalladated complexes: hydrolysis reactions in strong acidic media.

The mechanisms for the hydrolysis of organopalladium complexes [Pd(CNN)R]BF(4) (R=P(OPh)(3), PPh(3), and SC(4)H(8)) were investigated at 25 °C by using UV/Vis absorbance measurements in 10 % v/v ethanol/water mixtures containing different sulphuric acid concentrations in the 1.3-11.7 M range. In all cases, a biphasic behavior was observed with rate constants k(1obs), which corresponds to the initial step of the hydrolysis reaction, and k(2obs), where k(1obs)>k(2obs). The plots of k(1obs) and k(2obs) versus sulfuric acid concentration suggest a change in the reaction mechanism. The change with respect to the k(1obs) value corresponds to 35 %, 2 %, and 99 % of the protonated complexes for R=PPh(3), P(OPh)(3), and SC(4)H(8), respectively. Regarding k(2obs), the change occurred in all cases at about 6.5 M H(2)SO(4) and matched up with the results reported for the hydrolysis of the 2-acetylpyridinephenylhydrazone (CNN) ligand. By using the excess acidity method, the mechanisms were elucidated by carefully looking at the variation of k(i),(obs) (i=1,2) versus cH+. The rate-determining constants, k(0,A-1), k(0,A-2), and k(0,A-SE2) were evaluated in all cases. The R=P(OPh)(3) complex was most reactive due to its π-acid character, which favors the rupture of the trans nitrogen-palladium bond in the A-2 mechanism and also that of the pyridine nitrogen-palladium bond in the A-1 mechanism. The organometallic bond exerts no effect on the relative basicity of the complexes, which are strongly reliant on the substituent.

Biological assays and noncovalent interactions of pyridine-2-carbaldehyde thiosemicarbazonecopper(II) drugs with [poly(dA-dT)](2), [poly(dG-dC)] (2), and calf thymus DNA.

The interaction of the Cu(II) drugs CuL(NO(3)) and CuL'(NO(3)) (HL is pyridine-2-carbaldehyde thiosemicarbazone and HL' is pyridine-2-carbaldehyde 4N-methylthiosemicarbazone, in water named [CuL](+) and [CuL'](+)) with [poly(dA-dT)](2), [poly(dG-dC)](2), and calf thymus (CT) DNA has been probed in aqueous solution at pH 6.0, I = 0.1 M, and T = 25 degrees C by absorbance, fluorescence, circular dichroism, and viscosity measurements. The results reveal that these drugs act as groove binders with [poly(dA-dT)](2), with a site size n = 6-7, whereas they act as external binders with [poly(dG-dC)](2) and/or CT-DNA, thus establishing overall electrostatic interaction with n = 1. The binding constants with [CuL'](+) were slightly larger than with [CuL](+). The title compounds display some cleavage activity in the presence of thiols, bringing about the rupture of the DNA strands by the reactive oxygen species formed by reoxidation of Cu(I) to Cu(II); this feature was not observed in the absence of thiols. Mutagenic assays performed both in the presence and in the absence of S9 mix, probed by the Ames test on TA 98, TA 100, and TA 102, were negative. Weak genotoxic activity was detected for [CuL](+) and [CuL'](+), with a significative dose-response effect for [CuL'](+), which was shown to be more cytotoxic in the Ames test and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell proliferation assays. Methylation of the terminal NH(2) group enhances the antiproliferative activity of the pyridine-2-carbaldehyde thiosemicarbazones.

Hydrolysis mechanisms for the organopalladium complex [Pd(CNN)P(OMe)3]BF4 in sulfuric acid.

The acid-catalyzed hydrolysis of the organopalladium complex [Pd(CNN)P(OMe)3]BF4 species was monitored spectrophotometrically at different sulfuric acid concentrations (3.9 and 11.0 M) in 10% v:v ethanol-water over the 25-45 degrees C temperature range and in 30% and 50% (v/v) ethanol-water at 25 degrees C. Two acidity regions (I and II) could be differentiated. In each of the two regions the kinetic data pairs yielded two different rate constants, k(1obs) and k(2obs), the former being faster. These constants were fitted by an Excess Acidity analysis to different hydrolyses mechanisms: A-1, A-2, and A-SE2. In region I ([H2SO4] < 7.0 M), the k(1obs) values remained constant k(1obs)(av) = 1.6 x 10(-3) s(-1) and the set of k(2obs) values nicely matched an A-SE2 mechanism, yielding a rate-determining constant k(0,ASE2) = 2.4 x 10(-7) M(-1) s(-1). In region II ([H2SO4] > 7.0 M), a switchover was observed from an A-1 mechanism (k(0,A1) = 1.3 x 10(-4) s(-1)) to an A-2 mechanism (k(0,A2) = 3.6 x 10(-3) M(-1) s(-1)). The temperature effect on the rate constants in 10% (v/v) ethanol-water yielded positive DeltaH and negative DeltaS values, except for the A-1 mechanism, where DeltaS adopted positive values throughout. The solvent permittivity effect, epsilonr, revealed that k(1obs)(av) and k(0,A2) dropped with a fall in epsilonr, whereas the k(0,ASE2) value remained unaffected. The set of results deduced is in line with the schemes put forward.