Use of XRD and SEM/EDX to predict age and sex from fire-affected dental remains.

In fire scenarios, the application and accuracy of traditional odontological methods are often limited. Crystalline studies and elemental profiling have been evaluated for their applicability in determining biological profiles (age and sex) from human dentition, particularly fire- and heat-affected dental remains. Thirty-seven teeth were paired according to tooth type and donor age/sex for the analysis of crown and root surfaces pre- and post-incineration using X-ray diffraction (XRD) and scanning electron microscopy (SEM/EDX). In unburned crowns, carbon (C) content showed a positive correlation with age, whereas phosphorus (P) and calcium (Ca) contents showed a negative correlation with age. In unburned roots, C, P and Ca contents also showed significant changes that were opposite of those observed in the crowns. In relation to sex, females exhibited a higher C ratio than males, whereas males showed significantly higher levels of oxygen (O), P and Ca in unburned roots. Incineration resulted in an increase in the crystallite size that correlated with increasing temperature. No differences in hydroxyapatite (HA) crystallite size were found between age groups; however, unburned teeth from females exhibited a larger crystallite size than did those from males. The challenges of using XRD with a 3D sample were overcome to allow analysis of whole teeth in a nondestructive manner. Further studies may be useful in helping predict the temperature of a fire.

Usefulness of palladium impregnated magnetite nanoparticles for polyphenol determination.

Palladium impregnated magnetite nanoparticles (Pd-Fe3O4NPs) have been synthesized and used as reusable catalyst for the fluorometric determination of polyphenols in wines. The method is based on the decrease of the indocyanine green fluorescence, which is ascribed to its oxidation by dissolved oxygen in the presence of the nanoparticles, and the inhibition of the fluorescence decrease by polyphenols, which is proportional to the polyphenol concentration. The dynamic range of the calibration graph is 0.1-10.0µM gallic acid, which was chosen as model analyte, and the detection limit is 0.02µM. Precision data, expressed as relative standard deviation, ranged between 3.3% and 5.4%. The method was applied to the analysis of several wine samples, obtaining recovery values in the range of 79.7-102.0%. The results obtained were compared with those obtained using the Folin-Ciocalteu and laccase methods, finding that Pd-Fe3O4NPs provide a better selectivity than the first method and show a catalytic behavior similar to that of laccase.

Automatic determination of polyphenols in wines using laccase and terbium oxide nanoparticles.

The analytical usefulness of the combined use of laccase, terbium oxide nanoparticles (Tb4O7NPs) and 8-hydroxypyrene-3-sulphonate trisodium (HPTS) for the determination of polyphenol compounds in wine samples is described. The system is based on the temporal inhibition by polyphenols on the decrease of the HPTS fluorescence in the presence of laccase and on the activating effect of Tb4O7NPs, which increase the reaction rate of the system, shortening analysis times. The method has been developed in a microplate format using an automatic reader, reaching a sample throughput of 35 samples h(-1). The dynamic range of the calibration graph is 0.5-12 µM gallic acid, which was chosen as model analyte, and the detection limit is 0.14 µM. Precision data, expressed as relative standard deviation, ranged between 2.5% and 6.3%. The method was applied to the analysis of several wine samples, obtaining recovery values in the range of 80.0-108.3%.

Application of Tb(4)O(7) nanoparticles for lasalocid and salicylate determination in food analysis.

The usefulness of Tb(4)O(7) nanoparticles (NPs) as analytical reagents using sensitized luminescence as a detection system is described for the first time, and the results obtained are compared with those obtained using Tb(III) ions. Two drugs used in veterinary practice, namely, lasalocid (LAS) and salicylate (SAL), have been chosen as model analytes to carry out this study. The experimental conditions for these systems have been optimized, and their analytical features were obtained. The detection limits obtained for LAS and SAL using Tb(4)O(7) NPs were 1.0 and 4.0 ng mL(-1), respectively, which were comparable to those obtained using Tb(III) ions: 1.8 and 1.0 ng mL(-1), respectively. However, precision data, with relative standard deviation values in the range 2.3-3.8% using the NPs and 3.5-6.5% using Tb(III) ions, were slightly better for LAS with Tb(4)O(7) NPs. The practical analytical usefulness of Tb(4)O(7) NPs as luminescent reagents has been shown by performing the determination of LAS in tap water, feed premix, and egg samples, obtaining recoveries in the range of 80.0-105.0%.

Determination of monensin in milk samples by front-surface long-wavelength fluoroimmunoassay using nile blue-doped silica nanoparticles as labels.

A heterogeneous immunoassay for monensin determination in milk samples using a tracer formed by anti-monensin antibodies bound to nile blue (NB)-doped silica nanoparticles (NPs), 96-well microplates as solid supports and long-wavelength fluorescence measurements is described for the first time. The assay relies on the competition of the monensin present in the samples with a monensin-bovine serum albumin conjugate, which was immobilized onto the well surface, for the active sites of anti-monensin antibodies. After subsequent incubation and washing steps, the fluorescence of the bound tracer fraction is measured onto the dry surface of the well. An antigen capture format was also assayed by immobilizing anti-sheep IgG previously to the incubation of sheep anti-monensin antibodies and using a tracer formed by monensin bound to nile blue-doped silica NPs, which competes with the analyte for binding the immobilized antibody. Although the fluorescence signal obtained in both formats can be correlated to the analyte concentration, better results were obtained using the antibody capture format. After the optimization of the system using this format, the method features a detection limit of 0.015 µg L(-1) and a dynamic range from 0.05 to 5 µg L(-1). The precision, assayed at two different analyte concentrations, 0.2 and 1 µg L(-1), and expressed as relative standard deviation, gave values of 5.9% and 4.0%, respectively. The method was satisfactorily applied to the analysis of milk samples, which only required a simple extraction step in order to remove the proteins from samples, giving recoveries in the range 83.3-107.5%.

Synthesis and characterization of oxazine-doped silica nanoparticles for their potential use as stable fluorescent reagents.

The synthesis process to obtain silica nanoparticles (NPs) doped with two oxazine dyes, nile blue and cresyl violet, has been investigated using a modification of the reverse micelle microemulsion method and a procedure based on the Stöber method. A micellar medium provided by the non-ionic surfactant Triton X-100 in a hexanol:water mixture and an ethanol:water mixture, have been used to provide the synthesis medium in each case. Tetraethoxysilane has been used as the initiator of the polymerization and condensation reactions after its hydrolysis in basic medium using ammonium hydroxide. Dye-silane precursor NPs have been also synthesized in order to compare their potential advantages against the NPs obtained by the direct encapsulation of the oxazine dyes. Size distribution and fluorescence of the synthesized NPs, which were monitored using Transmision Electron Microscopy (TEM) and a microplate reader, respectively, depend on the molar ratio and total concentration of the reagents involved in the synthesis. NPs obtained using the developed synthesis procedures had sizes below 400 nm in most instances and the best luminescent properties were observed for NPs with sizes ranging from 100 to 300 nm. Lower sizes result in a decrease in the fluorescence intensities of these nanomaterials. Parameters related with the luminescence features of these NPs were calculated in order to compare the feasibility of both synthesis approaches. The repeatability of the reverse-micelle microemulsion procedure performed in different days gave a relative standard deviation of 10% for the fluorescence intensity values.

Analytical innovations in the detection of phenolics in wines.

A liquid chromatographic method with online photometric and luminescent detection for the determination of 18 phenolic compounds in wines is reported. Photometric detection is performed at four wavelengths, namely, 256, 280, 320, and 365 nm, using a diode array detection system. The luminescent detection is achieved by means of a postcolumn derivatization reaction of 10 of these compounds with terbium(III) in the presence of synergistic agents, such as ethylenediaminetetraacetic acid (EDTA) and n-octyltriphosphine oxide (TOPO). A micellar medium provided by the surfactants sodium dodecyl sulfate and Triton X-100 was used for the determination of the luminescent chelates at lambdaex 317 and lambdaem 545 nm. The long wavelength emission of lanthanide chelates can minimize interferences from background sample matrix, which usually emit at shorter wavelengths. The analytical features of the photometric and fluorometric methods, such as dynamic ranges of the calibration graphs, detection limits, and precision data, have been obtained. The practical usefulness of the developed methods is demonstrated by the analysis of Spanish and Italian wine samples (red, rosé, oloroso, and white), which were diluted and directly injected into the chromatographic system. The accuracy of both methods was checked by assaying a recovery study, which was performed at three different analyte levels for each type of sample.

Long-wavelength fluorescence polarization immunoassay: determination of amikacin on solid surface and gliadins in solution.

The versatility of the fluorescence polarization immunoassay (FPIA) is increased by using two long-wavelength labels, Nile Blue and a ruthenium(II) chelate. The first label has been used to study the potential of FPIA on a solid surface using dry reagent technology. The aminoglycoside antibiotic amikacin has been used as an analyte model, and the method has been applied to the analysis of serum samples. The second label has been used to show the practical application of FPIA to the determination of macromolecules, using gliadins as an analyte model, which have been determined in gluten-free food. Very low amounts of anti-amikacin antibodies and amikacin-Nile Blue tracer were immobilized onto nitrocellulose membranes, for the development of the amikacin method, and the consumption of reagents is lower than in conventional FPIA. Only the addition of the standard or sample extract at an adequate pH is required at the analysis time. The analyte displaces the tracer from the tracer-antibody immunocomplex, obtaining a decrease in the fluorescence polarization proportional to the analyte concentration. The gliadin-Ru(II) chelate tracer shows a relatively long lifetime, which allows the observation of differences in fluorescence polarization values between the tracer-antibody complex and the tracer alone. The dynamic range of the calibration graphs for both analytes is 0.5-10 microg mL-1 and the detection limits are 0.1 and 0.09 microg mL-1 for amikacin and gliadins, respectively. The study of the precision gave values of relative standard deviations lower than 5 and 1.5% for the amikacin and gliadin methods, respectively. Amikacin was determined in human serum samples using a previous deproteinization step with acetonitrile, obtaining recovery values in the range 83.4-122.8%. The gliadin method was applied to the analysis of gluten-free food samples by using a previous extraction step. The recovery study gave values between 94.3 and 105.0%.

Determination of fluoroquinolones in milk samples by postcolumn derivatization liquid chromatography with luminescence detection.

A liquid chromatography (LC) method with luminescence detection for the determination of eight quinolone antibiotics is reported. The system encompasses three consecutive steps: (a) chromatographic separation using reverse-phase mode (RP-LC), (b) postcolumn derivatization reaction, and (c) luminescence detection by monitoring fluorescence (FL) and time-resolved (TR) signals. The derivatization step is based on the reaction between quinolones and terbium(III) to form luminescent chelates, which were determined at lambda(ex) 340 and lambda(em) 545 nm (FL mode) or at lambda(ex) 281 and lambda(em) 545 nm (TR mode). Dynamic ranges of the calibration graphs, obtained with standard solutions of analytes and FL and TR modes, respectively, were 190-3500 and 316-2000 ng mL-1 for marbofloxacin, 8-3500 and 8.1-1500 ng mL-1 for ciprofloxacin, 6.2-3500 and 13-1500 ng mL-1 for danofloxacin, 7.4-3500 and 8.4-1500 ng mL-1 for enrofloxacin, 14-3500 and 20-2000 ng mL-1 for sarafloxacin, 12.5-3500 and 13.9-1200 ng mL-1 for difloxacin, 7.6-3500 and 13-3000 ng mL-1 for oxolinic acid, and 9-2000 and 130-3000 ng mL-1 for flumequine. Limit of detection values obtained using FL and TR modes, respectively, were 60 and 95 ng mL-1 for marbofloxacin, 2 and 2.4 ng mL-1 for ciprofloxacin, 1.9 and 3.9 ng mL-1 for danofloxacin, 2.2 and 2.5 ng mL-1 for enrofloxacin, 3.8 and 7 ng mL-1 for sarafloxacin, 4 and 4.2 ng mL-1 for difloxacin, 2.3 and 4 ng mL-1 for oxolinic acid, and 2.7 and 40 ng mL-1 for flumequine. The precision was established at two concentration levels of each analyte and expressed as the percentage of relative standard deviation with values ranging between 1.9 and 7.8%. The validation procedure for the analysis of samples was carried out using European Community recommendations, and the decision limit and detection capability were calculated for bovine whole milk. The method was applied to whole, semiskimmed, and skimmed milk samples spiked with the target analytes, and the recoveries ranged between 93.3 and 106.0%.

Usefulness of ytterbium(III) as analytical reagent for total sulfite determination in white wine samples.

Ytterbium(III) is used as reagent for the determination of sulfite by measuring the formation of the Yb(III)-sulfite complex through the variation of the light scattering intensity with time. The low solubility of this complex causes an efficient dispersion of the radiation at 490 nm, which is measured at 980 nm. Each kinetic datum is automatically obtained in only 0.5 s by stopped-flow mixing technique. The application of the initial rate method using a long emission wavelength minimizes the potential interference of fluorescent background signals from the sample matrix. The dynamic range of the calibration graph is 1-250 microg/mL, and the calculated detection limit is 0.35 microg/mL. The precision, expressed as relative standard deviation, is <6%. The method has been applied to the determination of total sulfites in white wine samples, which requires only the sample dilution and the use of two aliquots to improve selectivity. However, the matrix effect found for red wines precludes the application of the method to the direct analysis of these samples. Analytical recoveries ranged from 96.0 to 106.7%. The results obtained with the proposed method agreed with those provided by the p-rosaniline method. Unlike this method, in which toxic reagents are required, the use of ytterbium(III) as analytical reagent shows the advantage of its low acute toxic rating.