Determination of plastic polyester oligomers in real samples and their bioeffects.

Plastics are ubiquitously, becoming part of our everyday life. Recently, the issue of human exposure to micro- and nanoplastic particles and potentially resulting toxicological consequences has been broached, triggered by the discovery of microplastics in foodstuff and dietary exposure via contaminated food and beverages. Within this EU-FORA fellowship project, a determination and quantification of plastic polyester plastics oligomers in food samples was performed to assess exposure at these categories of 'nanoplastics', evaluating them as potential contaminants or as indicators and marker compounds of the exposure to specific nanoplastics/microplastics from polyesters, such as polyethylene terephthalate (PET) and polybutylene terephthalate (PBT). UHPLC-TOF-MS/MS analysis has been set-up for 10 PET and PBT oligomers and analysis has been performed in foods and drinks. Moreover, the project focused also on the effects of these oligomers in in vitro and ex vivo experiments. These data would be combined with EFSA Comprehensive Food Consumption Database, for the exposure and risk assessment of these 'Nanoplastics'.

Analysis of Migrant Cyclic PET Oligomers in Olive Oil and Food Simulants Using UHPLC-qTOF-MS.

Oligomers are a particular category of non-intentionally added substances (NIAS) that may be present in food contact materials (FCMs), such as polyethylene terephthalate (PET), and consequently migrate into foods. Here, an ultra-high-pressure liquid chromatography quadruple time-of-flight mass spectrometry (UHPLC-qTOF-MS) method was developed for the analysis of 1st series cyclic PET oligomers in virgin olive oil (VOO) following a QuEChERS clean-up protocol. Oligomer migration was evaluated with two different migration experiments using bottles from virgin and recycled PET: one with VOO samples stored in household conditions for a year and one using the food simulant D2 (95% v/v ethanol in water) at 60 °C for 10 days. Calibration curves were constructed with fortified VOO samples, with the LOQs ranging from 10 to 50 µg L-1 and the recoveries ranging from 86.6 to 113.0%. Results showed no migration of PET oligomers in VOO. However, in the simulated study, significant amounts of all oligomers were detected, with the migration of cyclic PET trimers from recycled bottles being the most abundant. Additional substances were tentatively identified as linear derivatives of PET oligomers. Again, open trimer structures in recycled bottles gave the most significant signals.

Untargeted screening and in silico toxicity assessment of semi- and non-volatile compounds migrating from polysaccharide-based food contact materials.

The chemical safety of representative polysaccharide films made with pea starch, organocatalytic acetylated pea starch and pectin was investigated at different migration conditions (20 °C/10 days, 70 °C/2 h) using two official simulants signifying hydrophilic (simulant A, 10% ethanol) or lipophilic (simulant D1, 50% ethanol) foods. Migrating semi-volatile and non-volatile compounds were identified and semi-quantified by ultra-high performance liquid chromatography-trap ion mobility time-of-flight mass spectrometry (UHPLC-TIMS-TOF-MS/MS), whereas their toxicity was evaluated by in silico models based on qualitative structure activity (QSAR). Physicochemical analysis revealed polymer wash-off into the simulants. Migration testing at 70 °C for 2 h using simulant D1 resulted in detectable concentrations of glycerol (≤72.1 mg/kg), monoacetylated maltose (≤6.5 mg/kg), and dibutyl phthalate (DBP) (≤0.5 mg/kg, compliant with the existing legislative migration limits) in samples containing acetylated starch. Migrating 3-ß-galactopyranosyl glucose (≤8.9 mg/kg) and 2,5-diketo-d-gluconic acid (≤4.9 mg/kg) were detected at 20 °C/10 days. In-silico toxicity emphasized no significant toxicity and categorized organocatalytic acetylated pea starch of no safety concern.

Liquid chromatography-mass spectrometry metabolite library for metabolomics: Evaluating column suitability using a scoring approach.

Untargeted metabolomic studies require an extensive set of analyte (metabolic) information to be obtained from each analyzed sample. Thus, highly selective, and efficient analytical methodologies together with reversed-phase (RP) or hydrophilic interaction liquid chromatography (HILIC) are usually applied in these approaches. Here, we present a performance comparison of five different chromatographic columns (C18, C8, RP Amide, zicHILIC, OH5 HILIC phases) to evaluate their sufficiency of analysis for a large analyte library, consisting of 817 authentic standards. By taking into account experimental chromatographic parameters (i.e. retention time, peak tailing and asymmetry, FWHM, signal-to-noise ratio and peak area and intensity), the proposed column scoring approach provides a simple criterion that may assist analysis in the select of a stationary phase for those metabolites of interest. RPLC methods offered better results regarding metabolic library coverage, while the zicHILIC stationary phase delivered a bigger number of properly eluted compounds. This study demonstrates the importance of choosing the most suitable configuration for the analysis of different metabolic classes.

Hepatic Metabolic Profiling of Lifelong Exercise Training Rats.

Regular physical exercise has been investigated as a primary preventive measure of several chronic diseases and premature death. Moreover, it has been shown to synchronize responses across multiple organs. In particular, hepatic tissue has proven to be a descriptive matrix to monitor the effect of physical activity. In this study, we performed an untargeted metabolomics-based analysis of hepatic tissue extracts from rats that have undergone either lifelong or chronic exercise training. For this purpose, 56 hepatic samples were collected and were analyzed by UHPLC-TOF-MS in negative ionization mode. This approach involved untargeted metabolite detection on hepatic tissue extracts accompanied by an in-house retention time/accurate mass library enabling confident metabolite identification. Unsupervised (PCA) and supervised (OPLS-DA) multivariate analysis showed significant metabolic perturbation on a panel of 28 metabolites, including amino acids, vitamins, nucleotides, and sugars. The training regime employed in this study resulted in a probable acceleration of the bioenergetic processes (glycolysis, glycogen metabolism), promoted catabolism of purines, and supplied biosynthetic precursors via the pentose phosphate pathway and pentose and glucuronate interconversions. Overall, the applied methodology was able to discriminate the different training schedules based on the rat liver metabolome.

Liquid chromatography-mass spectrometry method for the determination of polyethylene terephthalate and polybutylene terephthalate cyclic oligomers in blood samples.

Food contact materials (FCM) polyethylene terephthalate (PET) and polybutylene terephthalate (PBT) used extensively in food packaging may contain cyclic oligomers which may migrate into food and thus cause toxic effects on human health. A simple, fast, and sensitive ultra-high-performance liquid chromatography method quadrupole time-of-flight mass spectrometer was developed for the analysis of 7 cyclic oligomers in post-mortem blood samples. The targeted analytes were separated on a Waters BEH C18 (150 × 2.1 mm, 1.7 µm) analytical column by gradient elution. Calibration curves were constructed based on standard solutions and blood samples and Student's t-test was applied to evaluate the matrix effect. The LODs ranged from 1.7 to 16.7 µg mL-1, while the method accuracy was assessed by recovery experiments and resulting within the range 84.2-114.6%. Such an analytical method for the determination of PET and PBT cyclic oligomers in biological samples is reported for the first time. The developed methodology allows the determination of these oligomers in blood providing a useful analytical tool to assess the exposure and thus the potential hazard and health risks associated with these non-intentionally added substances (NIAS) from PET and PBT FCM through food consumption. The method was validated and successfully applied to the analysis of 34 post-mortem whole blood samples. Polyethylene terephthalate trimer was detected in four of them, for the first time in literature.

Selective Voltammetric Detection of Ascorbic Acid from Rosa Canina on a Modified Graphene Oxide Paste Electrode by a Manganese(II) Complex.

Voltammetric techniques have been considered as an important analytical tool applied to the determination of trace concentrations of many biological molecules including ascorbic acid. In this paper, ascorbic acid was detected by square wave voltammetry, using graphene oxide paste as a working electrode, modified by a film of a manganese(II) complex compound. Various factors, such as the effect of pH, affecting the response characteristics of the modified electrode were investigated. The relationship between the peak height and ascorbic acid concentration within the modified working electrode was investigated, using the calibration graph. The equation of the calibration graph was found to be: I = 0.0550γac + 0.155 with R2 = 0.9998, where I is the SWV current and γac is the mass concentration of ascorbic acid. The LOD and LOQ of the proposed method were determined to be 1.288 µg/L and 3.903 µg/L, respectively. Several compounds, such as riboflavin, biotin, and ions, such as Fe and Cu, were tested and it seemed that they did not interfere with the analytic signal. The proposed procedure was successfully applied in the determination of ascorbic acid in Rosa canina hips.

Development and validation of an ultra high performance liquid chromatography-tandem mass spectrometry method for the determination of phthalate esters in Greek grape marc spirits.

An Ultra High Performance Liquid Chromatography - Tandem Mass Spectrometry method has been developed for the analysis of 12 phthalate esters in Greek grape marc spirits. The phthalates were separated on a U-VDSpher PUR 100 C18-E (100 mm x 2.0 mm, 1.8 µm) column by gradient elution. The analytes were ionized by positive electrospray ionization using the multiple reaction monitoring mode. The standard addition method was used for quantification and the Student's t-test was carried out to evaluate the matrix effect. The accuracy of the method was assessed by recovery experiments resulting in values from 81.6 to 109.6%. The detection limits ranged from 0.3 to 33.3 µg L-1.The proposed method was validated and successfully applied to the analysis of 45 samples collected from Greece and Cyprus. All phthalate esters proved to be present at least once in the analysed grape marc spirits samples, except only in cases of diphenyl phthalate and diisodecyl phthalate, while for the regulated phthalates only bis (2-ethylhexyl) phthalate was quantified above the legislative concentration limits.

Wine and grape marc spirits metabolomics.

INTRODUCTION: Mass spectrometry (MS)-based and nuclear magnetic resonance (NMR) spectroscopic analyses play a key role in the field of metabolomics due to their important advantages. The use of metabolomics in wine and grape marc spirits allows a more holistic perspective in monitoring and gaining information on the making processes and thus it can assist on the improvement of their quality. OBJECTIVES: This review surveys the latest metabolomics approaches for wine and grape marc spirits with a focus on the description of MS-based and NMR spectroscopic analytical techniques. METHODS: We reviewed the literature to identify metabolomic studies of wine and grape marc spirits that were published until the end of 2017, with the key term combinations of 'metabolomics', 'wine' and 'grape marc spirits'. Through the reference lists from these studies, additional articles were identified. RESULTS: The results of this review showed that the application of different metabolomics approaches has significantly increased the knowledge of wine metabolome and grape marc spirits; however there is not yet a single analytical platform that can completely separate, detect and identify all metabolites in one analysis. CONCLUSIONS: The authentication and quality control of wines and grape marc spirits has to be taken with caution, since the product's chemical composition could be affected by many factors. Despite intrinsic limitations, NMR spectroscopy and MS based strategies remain the key analytical methods in metabolomics studies. Authenticity, traceability and health issues related to their consumption are the major research initiatives in wine and grape marc spirits metabolomics analysis.