Development of 2D and 3D front face fluorescence spectroscopy for monitoring ultrasound treatment in the removal of pesticides residues from fresh lettuces at the laboratory and pilot scales.

Pesticide residues in vegetables are potentially toxic components to humans and can cause serious health problems. To remove pesticide residues from fresh agricultural products and improve consumer food safety, various pesticide removal methods have been investigated over the past decades. In this study, the effectiveness of laboratory and pilot scale ultrasonic cleaning on the removal of boscalid and pyraclostrobin residues from lettuce was examined. 2D fluorescence spectroscopy, 3D fluorescence spectroscopy represented by excitation-emission matrix (EEM), and parallel factor analysis (PARAFAC) were used to characterize and discriminate the fluorescence signatures of these pesticides in the cleaning water to determine the effectiveness of the ultrasonic cleaning method as a function of the level of pesticide removal. The 2D fluorescence results showed that the rate of removal of boscalid by ultrasonics at the laboratory scale increased with the cleaning time. The ultrasonic treatment showed a higher cleaning efficiency compared to only soaking in distilled water for 10 min. The same trends were observed at the pilot scale. The EEM also showed differences in the concentration of pesticides removed by ultrasonication between the different parts of the lettuce, the concentration was higher in the upper part than the lower part. This study showed that ultrasonication is an effective technique for the removal of pesticide residues on lettuce, and it also showed the significant potential of fluorescence spectroscopy coupled with PARAFAC for the discrimination and characterization of pesticides.

Impact of storage period and lipid unsaturation on the kinetic of 5-hydroxymethylfurfural and furfural generation in pound cakes.

This work aimed at studying the influence of formulation and storage on the formation of Maillard compounds called 5-hydroxymethylfurfural (HMF) and furfural in pound cakes formulated with rapeseed oil (RO) and palm oil (PO). A progressive humidification of the crust and a dryness of the crumb were observed during storage. Lightness (L*) decreased for both PO and RO pound cakes in crumb as well as crust. The accumulation of HMF was found to be higher in the crust (2.21-38.5 mg kg-1) in comparison with the crumb (0.78 to 10.29 mg kg-1). Similar results were found for furfural where the concentration range was between 0.98 and 5.67 mg kg-1 in the crumb while between 2.1 and 38.39 mg kg-1 in the crust. Thus, the formation of HMF and furfural depended on formulation. The PLS-DA showed a positive correlation between the kinetic of HMF and furfural migration and the storage period.

Potentiality of front-face fluorescence and mid-infrared spectroscopies coupled with partial least square regression to predict lipid oxidation in pound cakes during storage.

The potentialities of front-face fluorescence (FFF) and mid-infrared (MIR) spectroscopies coupled with partial least square regression (PLSR) were compared to predict the lipid oxidation of pound cakes. The level of lipid oxidation in pound cakes determined using classical methods showed some changes. Similarly, the fluorescence emission (305-490 nm) and excitation (252-390 nm) spectra and MIR spectra scanned in the 4000-700 cm-1 region showed some changes in pound cakes as a function of both storage time and the type of oil used in the formulation. The application of PLSR to the MIR spectra, provided excellent predictive results for free fatty acid (R2 = 0.97) and peroxide values (R2 = 0.87). Similar results were obtained from both tryptophan and MIR spectra for the prediction of TOTOX (R2 > 0.86) demonstrating the efficiency of the MIR and FFF spectroscopies to qualify and quantify the level of lipid oxidation in pound cakes.

Identification of a discriminative metabolomic fingerprint of potential clinical relevance in saliva of patients with periodontitis using 1H nuclear magnetic resonance (NMR) spectroscopy.

Periodontitis is characterized by the loss of the supporting tissues of the teeth in an inflammatory-infectious context. The diagnosis relies on clinical and X-ray examination. Unfortunately, clinical signs of tissue destruction occur late in the disease progression. Therefore, it is mandatory to identify reliable biomarkers to facilitate a better and earlier management of this disease. To this end, saliva represents a promising fluid for identification of biomarkers as metabolomic fingerprints. The present study used high-resolution 1H-nuclear magnetic resonance (NMR) spectroscopy coupled with multivariate statistical analysis to identify the metabolic signature of active periodontitis. The metabolome of stimulated saliva of 26 patients with generalized periodontitis (18 chronic and 8 aggressive) was compared to that of 25 healthy controls. Principal Components Analysis (PCA), performed with clinical variables, indicated that the patient population was homogeneous, demonstrating a strong correlation between the clinical and the radiological variables used to assess the loss of periodontal tissues and criteria of active disease. Orthogonal Projection to Latent Structure (OPLS) analysis showed that patients with periodontitis can be discriminated from controls on the basis of metabolite concentrations in saliva with satisfactory explained variance (R2X = 0.81 and R2Y = 0.61) and predictability (Q2Y = 0.49, CV-AUROC = 0.94). Interestingly, this discrimination was irrespective of the type of generalized periodontitis, i.e. chronic or aggressive. Among the main discriminating metabolites were short chain fatty acids as butyrate, observed in higher concentrations, and lactate, γ-amino-butyrate, methanol, and threonine observed in lower concentrations in periodontitis. The association of lactate, GABA, and butyrate to generate an aggregated variable reached the best positive predictive value for diagnosis of periodontitis. In conclusion, this pilot study showed that 1H-NMR spectroscopy analysis of saliva could differentiate patients with periodontitis from controls. Therefore, this simple, robust, non-invasive method, may offer a significant help for early diagnosis and follow-up of periodontitis.

Use of front face fluorescence for monitoring lipid oxidation during ageing of cakes.

Front face fluorescence spectroscopy coupled with chemometric tools was used as a useful tool for the monitoring of sponge cakes freshness, produced at the pilot scale, during ageing (i.e. 1, 3, 6, 9, 16, and 20days). The fluorescence emission spectra were acquired in the 340-490nm and 390-680nm after excitation at 325 and 380nm, respectively, while excitation spectra (250-390nm) were scanned after emission at 410nm. The primary and secondary products of lipid oxidation were also determined on the same cakes. The principal component analysis (PCA) applied to the each spectral collection obtained after excitation at 325 and 380nm and emission at 410nm allowed a clear discrimination of cakes according to their ageing. A high correlation between the intensity of fluorescence at 521nm and the p-anisidine values was observed since squared correlation coefficient of 0.73 was obtained. The results showed that fluorescence spectroscopy could be used as a powerful tool for the evaluation of cake freshness throughout storage.

Monitoring changes in sponge cakes during aging by front face fluorescence spectroscopy and instrumental techniques.

In the present study, sponge cakes, produced at the pilot scale, were monitored during aging (i.e., 1, 3, 6, 9, 16, and 20 days) by three different analytical techniques. For the texture analyzer, the hardness and elasticity of crumb cakes were found to significantly increase and decrease, respectively, throughout aging. Color parameters (L*, a*, and b*) showed only slight change throughout aging, and a high correlation (R(2) = 0.88) was observed between the whiteness and the yellowness. Tryptophan fluorescence spectra (excitation, 290 nm; emission, 305-490 nm) recorded on cakes exhibited three maxima located at 382, 435, and 467 nm that were attributed to maximum emission of tryptophan (382 nm) and fluorescent Maillard reaction products (435 and 467 nm). The principal component analysis (PCA) applied to the tryptophan spectra allowed a clear discrimination of cakes aged for 1, 3, and 6 days from those aged for 9, 16, and 20 days. Finally, canonical correlation analysis (CCA) performed on the textural and tryptophan fluorescence spectral data sets showed that the two groups of variables were highly correlated because the squared canonical coefficients for canonical variates were 0.99, indicating that cake texture determined at the macroscopic level by texture analyzer is a reflection of its structure at the molecular level determined by fluorescence spectroscopy.

Analytical model for site-specific isotope fractionation in 13C during sorption: determination by isotopic 13C NMR spectrometry with vanillin as model compound.

The aim of this study was to conceive a reactive transport model capable of providing quantitative site-specific enrichment factors for fractionation in (13)C isotopic content during sorption. As test compound the model treats vanillin, for which the (13)C isotopic content at natural abundance at each of the 8 carbon positions can be measured by quantitative (13)C nuclear magnetic resonance spectrometry. This technique determines the isotope ratios with a resolution better than ±1‰ (0.1%) at each carbon position. Site-specific isotope fractionations were recorded in chromatography column experiments with silica RP-18 as stationary phase. The one dimensional reactive transport model accounted for the sorption/desorption behavior of 8 individual (13)C-isotopomers and one (12)C-isotopomer of vanillin and reproduced satisfactorily the bulk (average over the whole compound) fractionation observed during elution. After model calibration, the enrichment factors were fitted for each carbon site where a significant fractionation was recorded. To show the interest of such a transport model for environmental studies, the model, extended to three dimensions, was exploited to simulate reactive transport in an aquifer. These results show that significant (13)C isotope fractionation is expected for 4 out of 8 (13)C-isotopomers in vanillin, and illustrate that bulk isotope ratios measured by conventional compound specific isotope analysis and mass spectrometry would hardly document significant isotope fractionations in vanillin. It is concluded that modeling of site-specific isotope ratios in molecules is a priori feasible and may help to quantify unknown processes in the environment.

Antibody-Hapten Recognition at the Surface of Functionalized Liposomes Studied by SPR: Steric Hindrance of Pegylated Phospholipids in Stealth Liposomes Prepared for Targeted Radionuclide Delivery.

Targeted PEGylated liposomes could increase the amount of drugs or radionuclides delivered to tumor cells. They show favorable stability and pharmacokinetics, but steric hindrance of the PEG chains can block the binding of the targeting moiety. Here, specific interactions between an antihapten antibody (clone 734, specific for the DTPA-indium complex) and DTPA-indium-tagged liposomes were characterized by surface plasmon resonance (SPR). Non-PEGylated liposomes fused on CM5 chips whereas PEGylated liposomes did not. By contrast, both PEGylated and non-PEGylated liposomes attached to L1 chips without fusion. SPR binding kinetics showed that, in the absence of PEG, the antibody binds the hapten at the surface of lipid bilayers with the affinity of the soluble hapten. The incorporation of PEGylated lipids hinders antibody binding to extents depending on PEGylated lipid fraction and PEG molecular weight. SPR on immobilized liposomes thus appears as a useful technique to optimize formulations of liposomes for targeted therapy.

Evidence of 13C non-covalent isotope effects obtained by quantitative 13C nuclear magnetic resonance spectroscopy at natural abundance during normal phase liquid chromatography.

Quantitative isotopic (13)C NMR at natural abundance has been used to determine the site-by-site (13)C/(12)C ratios in vanillin and a number of related compounds eluted from silica gel chromatography columns under similar conditions. Head-to-tail isotope fractionation is observed in all compounds at the majority of carbon positions. Furthermore, the site-specific isotope deviations show signatures characteristic of the position and functionality of the substituents present. The observed effects are more complex than would be obtained by simply summing the individual effects. Such detail is hidden when only the global (13)C content is measured by mass spectrometry. In particular, carbon positions within the aromatic ring are found to show site-specific isotope fractionation between the solute and the stationary phase. These interactions, defined as non-covalent isotope effects, can be normal or inverse and vary with the substitution pattern present.

Quantitative isotopic 13C nuclear magnetic resonance at natural abundance to probe enzyme reaction mechanisms via site-specific isotope fractionation: the case of the chain-shortening reaction for the bioconversion of ferulic acid to vanillin.

Isotope fractionation is a powerful technique by which to probe the reaction mechanism of enzymes. The effect of a heavy isotope on the reaction energetics can be used to predict transition state architecture and reaction mechanism. In order to examine simultaneously the isotope fractionation in (13)C at multiple sites within the substrate and product molecules without any need for site-selective isotope enrichment, a technique exploiting quantitative isotopic nuclear magnetic resonance (NMR) spectrometry at natural abundance (NAQ-NMR) has been developed. Here we report the first application of this technique to the study of an enzyme-catalyzed reaction, the bioconversion of ferulic acid to vanillin in cultures of Streptomyces setonii. We were able to show that the NAQ-NMR methodology is sufficiently precise and robust to measure the isotope shifts in the (13)C/(12)C ratios in both substrate and product of this biotransformation, thereby permitting meaningful data to be obtained even at carbon positions that take part only indirectly in the reaction and show only secondary isotope fractionation. The results obtained provide direct evidence in support of the current hypothesis for the reaction mechanism of the enzyme hydroxycinnamoyl-CoA hydratase/lyase, notably the proposed involvement of the quinone methide enolate of feruloyl-CoA as intermediate in the catalytic pathway.

Accurate quantitative 13C NMR spectroscopy: repeatability over time of site-specific 13C isotope ratio determination.

The stability over time (repeatability) for the determination of site-specific 13C/12C ratios at natural abundance by quantitative 13C NMR spectroscopy has been tested on three probes: enriched bilabeled [1,2-13C2]ethanol; ethanol at natural abundance; and vanillin at natural abundance. It is shown in all three cases that the standard deviation for a series of measurements taken every 2-3 months over periods between 9 and 13 months is equal to or smaller than the standard deviation calculated from 5-10 replicate measurements made on a single sample. The precision which can be achieved using the present analytical 13C NMR protocol is higher than the prerequisite value of 1-2 per thousand for the determination of site-specific 13C/12C ratios at natural abundance (13C-SNIF-NMR). Hence, this technique permits the discrimination of very small variations in 13C/12C ratios between carbon positions, as found in biogenic natural products. This observed stability over time in 13C NMR spectroscopy indicates that further improvements in precision will depend primarily on improved signal-to-noise ratio.