Isabel Durán-Merás

Usefulness of fluorescence excitation-emission matrices in combination with PARAFAC, as fingerprints of red wines.

The possibility of using front-face fluorescence spectroscopy to characterize red wines was investigated, and a tentative identification of their main fluorescent components was attempted. Fifty-seven red wine samples from different origins were included in the present study. Their fluorescence excitation-emission matrices (EEMs) were registered directly on 3-mL aliquots of untreated samples. The assayed excitation and emission ranges were 245-340 and 300-500 nm, respectively. The set of 57 EEMs was analyzed by means of parallel factor analysis (PARAFAC). Thus, the spectral excitation and emission profiles of possible pure fluorescence components (PARAFAC loadings) and the relative contribution of each component to the individual EEMs (PARAFAC scores) were obtained. The red wine system contained four main fluorescence components, and the excitation and emission loadings had maxima at the wavelength pairs 260/380, 275/323, 330/410, and 280/364 nm, respectively. A tentative identification of fluorophores was done by matching PARAFAC score values with HPLC measurements on the same 57 samples, as well as fluorescence measurements on pure compounds typically present in red wine. It was found that the third component was highly correlated with concentrations of catechin and epicatechin. When the PARAFAC score values were plotted against each other, they did to some extent discriminate the wines according to origin (country) and grape variety.

Multicomponent determination of flavour enhancers in food preparations by partial least squares and principal component regression modelling of spectrophotometric data.

Three multivariate calibration methods, partial least squares (PLS-1 and PLS-2) and principal component regression (PCR), were applied to the simultaneous determination of three flavour enhancers (inosine 5-monophosphate, guanosine 5-monophosphate and monosodium glutamate), in mixtures by ultraviolet/visible absorption spectrophotometry. The absorption and first- and second-derivative absorption spectra of the ternary mixtures were used to perform the optimization of the calibration matrices by the PLS and PCR methods. The results obtained by the application of the different chemometric approaches are discussed and compared. No significant advantages were found for the prior differentiation step. The proposed method was applied satisfactorily to the determination of inosine 5-monophosphate and guanosine 5-monophosphate in several food preparations, in the presence of monosodium glutamate.

Cyclodextrin-induced fluid solution room-temperature phosphorescence from acenaphthene in the presence of 2-bromoethanol

Abstract A 1:1 stoichiometric ratio and a formation constant of 130 1 mol −1 were obtained for the binary inclusion complex between acenaphthene and β-cyclodextrin (CD). On addition of 2-bromoethanol, effective quenching of the fluorescence emission and enhancement of the room-temperature phosphorescence (RTP) of acenaphthene is observed. The use of 2-bromoethanol as an external “heavy atom” to obtain room-temperature phosphorescence in fluid solutions is discussed. The RTP emission from acenaphthene included in β-CD was optimized and characterized. The apparent formation constant of the ternary associate was determined to be 880 1 mol −1 by the use of RTP enhancement in the presence of 1% 2-bromoethanol. The data indicate a ternary association between acenaphthene, β-CD and the bromo alcohol. The use of sodium sulfite, for chemical deoxygenation, to obtain RTP emission from a non-heavy atom-containing luminophor is shown to be feasible. The kinetics of the deoxygenation reaction were established and a photochemical catalytic effect on the reaction was demonstrated.

Determination of salicylic acid and its metabolites in urine by derivative synchronous spectrofluorimetry

The simultaneous determination of salicylic acid in binary and/or ternary mixtures and its two main urinary metabolites is proposed. Mixtures of salicylic, salicyluric and gentisic acids are resolved by synchronous spectrofluorimetry, in combination with first-derivative measurements. The urine is extracted with diethyl ether in acid medium. Salicylic and salicyluric acids are re-extracted into glycine-sodium hydroxide buffer solution of pH 11.6 and determined at that pH, and salicylic and gentisic acids are re-extracted into boric acid-sodium hydroxide buffer solution of pH 8.5 and determined at pH 6.

Spectrofluorimetric determination of nalidixic acid based on host–guest complexation with γ-cyclodextrin

The characteristics of host–guest complexation between γ-cyclodextrin and nalidixic acid were investigated by fluorescence spectrometry. A 1 : 1 stoichiometry of the complex was established and an association constant of 292 l mol–1 was calculated by using the changes in the fluorescence of nalidixic acid that occurred when it was included in the hydrophobic cyclodextrin cavity. A spectrofluorimetric method for the determination of nalidixic acid was developed, with a range of application between 0.1 and 2 µg ml–1. The method was applied satisfactorily to the determination of nalidixic acid in a pharmaceutical preparation and in urine.

Rapid and sensitive on-line solid phase extraction-ultra high performance liquid chromatography–electrospray-tandem mass spectrometry analysis of pesticides in surface waters

A simple, fast and sensitive method has been developed for the determination of 37 LC-amenable pesticides in surface water samples. On-line solid phase extraction (SPE) coupled to ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS. was employed for pre-concentration and analysis of all compounds in 16min. SPE parameters were evaluated in order to increase sample throughput and detectability. Thus, injected sample volume, sample loading flow, carryover effects and reusability of the cartridges employed were studied, observing that 70 extractions can be performed with the same cartridge. Validation parameters were performed and good linearity (R(2)>0.99 in all cases) and precision (interday relative standard deviation values were lower than 14%) were obtained. Limits of detection (LOD) and limits of quantification (LOQ) were lower than 6.0 and 18.0ngL(-1) applying an injection sample volume of 1.5mL, respectively, with exception of thifensulfuron methyl, whose limits were 10.0 and 33.0ngL(-1), respectively. On-line SPE recoveries were evaluated for three concentration levels (0.01, 0.03, 0.10 and 0.20μgL(-1)) and acceptable values were found. The on-line SPE method was also compared with off-line SPE. Matrix effects were observed for majority of compounds and standard addition method was selected for analysis of real water samples. Finally, surface water samples were analyzed and, in all cases, the pesticide concentrations were below than the allowable limit in water for human consumption.

Synchronous fluorimetric determination of salicylic acid and diflunisal in human serum using partial least-squares calibration

The simultaneous determination of salicylic acid and diflunisal in human serum has been accomplished by synchronous fluorimetry, in combination with partial least-squares multivariate calibration. The total luminescence information of the analytes has been used to optimize the spectral data set for the calibration, by analysis of the three-dimensional excitation-emission matrices. The synchronous spectrum, maintaining a constant difference of Deltalambda = 128 nm between the emission and excitation wavelengths, has been selected as optimum to perform the determination. The method is based on the fluorescence of these compounds in chloroform containing 1% (v/v) acetic acid. Serum samples are treated with trichloroacetic acid to remove the proteins, and both analytes are extracted into chloroform-1% (v/v) acetic acid prior to the determination. For concentrations ranging from 60-240 mug ml(-1) of each drug, analytical recoveries range from 96% to 103% for salicylic acid and from 97% to 105% for diflunisal.

Development of a method for the determination of advanced glycation end products precursors by liquid chromatography and its application in human urine samples

A liquid chromatographic method with fluorimetric detection has been developed to determine the most abundant α-dicarbonyl compounds, generated as intermediates in the Maillards reaction, previous derivatization to high fluorescent pteridinic derivatives. Hence, the biomarkers D-glucosone, 3-deoxyglucosone, glyoxal, methylglyoxal, diacetyl, 2,3-pentanedione, and phenylglyoxal were quantified using a gradient elution mode. The experimental conditions of the derivatization reaction and mobile phase composition were optimized. Linearity ranges (peak area versus α-dicarbonyl compound concentration) from 1.0 to 100.0 ng mL(-1) were obtained. Detection limits were comprised between 0.3 and 11.0 ng mL(-1). The high sensitivity of the method allows the determination of α-dicarbonyl compounds present in human urine, such as D-glucosone, 3-deoxyglucosone, glyoxal, and methylglyoxal, that are used as biomarkers, in order to investigate their roles in several diseases, with special emphasis in diabetes mellitus. With the aim of avoiding the interferences due to pteridinic compounds present in urine, a cleanup step with an ISOLUTE ENV+ cartridge was carried out. The concentrations of these urinary biomarkers have been reported as a normalized ratio to urinary creatinine, and determined in healthy and in diabetic volunteers, of different ages and sex. In all urine samples, standard addition and external calibration procedures were applied and compared.

On line photochemically induced excitation-emission-kinetic four-way data Analytical application for the determination of folic acid and its two main metabolites in serum by U-PLS and N-PLS/residual trilinearization (RTL) calibration

The determination of folic acid and its two main serum metabolites, 5-methyltetrahydrofolic acid and tetrahydrofolic acid, has been accomplished using four-way data modelled by the third-order multivariate calibration methods unfolded and N-dimensional partial least-squares (U-PLS and N-PLS), in combination with the separate procedure known as residual trilinearization (RTL). The four-way data were acquired by following the photochemical reaction of these compounds by on line irradiation with a UV lamp. The excitation-emission matrices (EEMs) were recorded as a function of the irradiation time, using a fast scanning spectrofluorimeter. The method achieves selectivity from the different rates at which the corresponding photoproducts of the folic acid derivatives are formed and degraded. Several N-dimensional chemometric algorithms were used and the method was applied to the determination of these compounds in serum samples. The best algorithms to perform the multivariate calibration were U-PLS and N-PLS in combination with the separate residual trilinearization procedure, achieving the second-order advantage. The approach allows minimizing or eliminating traditionally time-consuming sample pre-treatments and can facilitate quantifying an analyte in its native environment.

Resolution of ternary mixtures of salicylic, salicyluric and gentisic acids by partial least squares and principal component regression: Optimization of the scanning path in the excitation-emission matrices

The resolution of ternary mixtures of salicylic, salicyluric and gentisic acids has been accomplished by partial least squares (PLS) and principal component regression (PCR) multivariate calibration. The total luminescence information of the compounds has been used to optimize the spectral data set to perform the calibration. A comparison between the predictive ability of the three multivariate calibration methods, PLS-1, PLS-2 and PCR, on three spectral data sets, excitation, emission and synchronous spectra, has been performed. The excitation spectrum has been the best scanning path for salicylic and salicyluric acid determinations, while the emission spectrum has been the best for the gentisic acid determination. The convenience of analysing the total luminescence spectrum information when using multivariate calibration methods on fluorescence data is demonstrated.