A green approach for the automatic quantitative analysis of additives in plastic samples using in-tube extraction dynamic headspace sampling technique coupled to GC-MS/MS.

BACKGROUND: Many of the chemicals frequently used as additives have been recognised as hazardous substances, and therefore their analysis is necessary to evaluate plastic contamination risk. Additives analysis in plastic samples is usually performed by methods involving high volumes of toxic solvents or having high detection limits. In this work, a novel, fast, solventless and reliable green method was developed for the automated analysis of plastic additives from plastic samples. The proposed method consists of in-tube extraction dynamic headspace sampling (ITEX-DHS) combined with gas chromatography (GC) and mass spectrometry (MS/MS) determination. RESULTS: Several parameters affecting the ITEX-DHS extraction of 47 additives in plastic samples (including phthalates, bisphenols, adipates, citrates, benzophenones, organophosphorus compounds, among others) were optimised. The use of matrix-matched calibration, together with labelled surrogate standards, minimises matrix effects, resulting in recoveries between 70 and 128%, with good quantitation limits (below 0.1 µg g-1 for most compounds) and precision (<20%). The method proposed can be applied to any type of polymer, but due to the existence of the matrix effect, calibrates with the adequate matrix should be performed for each polymer. SIGNIFICANCE: This method represents an effective improvement compared to previous methods because it is fast, solvent-free, fully automated, and provides reliable quantification of additives in plastic samples.

Bioaccumulation of trace metals in the plastisphere: Awareness of environmental risk from a European perspective.

The term "Plastisphere" refers to the biofilm layer naturally formed by microorganisms attaching to plastic surfaces. This layer possesses the capability to adsorb persistent organic and inorganic pollutants, particularly trace metals, which are the focus of this research study. Immersion experiments were concurrently conducted in five locations spanning four European countries (France, Ireland, Spain, and Italy) utilising eight distinct polymers. These immersions, repeated every three months over a one-year period, aimed to evaluate the baseline bioaccumulation of 12 trace metals. The study underscores the intricate nature of metal bioaccumulation, influenced by both micro-scale factors (such as polymer composition) and macro-scale factors (including geographical site and seasonal variations). Villefranche Bay in France exhibited the lowest metals bioaccumulation, whereas Naples in Italy emerged as the site where bioaccumulation was often the highest for the considered metals. Environmental risk assessment was also conducted in the study. The lightweight nature of certain plastics allows them to be transported across significant distances in the ocean. Consequently, evaluating trace metal concentrations in the plastisphere is imperative for assessing potential environmental repercussions that plastics, along with their associated biota, may exert even in locations distant from their point of emission.

Organotin compounds in seafood by ultrasonic assisted extraction and gas chromatography-triple quadrupole tandem mass spectrometry.

Although restricting environmental quality values for organotin compounds (OTs) are set by Directive 2013/39/EU of the European Parliament, marine environment remains being affected due to maritime circulation at global scale. Fish and seafood accumulate OTs, making fish and seafood consumption the main source of OTs in humans. Because of the fish and seafood matrices complexity and the required low limits of detection, a robust and fast procedure for the quantification of OTs in fish and seafood, using ultrasound-assisted extraction and gas chromatography-tandem mass spectrometry, was validated and applied. Detection (2.7 µg Sn kg-1) and quantification (8.0 µg Sn kg-1) limits, repeatability and intermediate precision (<10%), accuracy by analysing ERM®-CE477 Mussel Tissue and analytical recoveries (65-122%) were assessed. Multivariate analysis shown that the matrix effect for some OTs displayed good negative correlation with the fat and protein content. Health risk assessment of OTs intake revealed no serious risk for human consumption.

Improved methodology for microplastic extraction from gastrointestinal tracts of fat fish species.

Potassium hydroxide (KOH) digestion protocols are currently applied to separate microplastics from biological samples, allowing efficient digestion with minor degradation of polymers in a time- and cost-effective way. For biota samples with high-fat content, KOH reacts with triglycerides generating an overlying soap layer, making difficult the digestion and solubilization and subsequent microplastics extraction. Here we studied the addition of Tween-20 in different concentrations to evaluate the effect on the soap layer of post-digested samples. Addition of 10 % of Tween-20 presented higher flow rate during filtration, being set as optimal value. Incorporation of Tween-20 in the extraction procedure increased recovery rates of LDPE, PC and PET and appears to have a protective effect on PC and PET degradation. Tween-20 did not interfere in FTIR spectrum of polymers available in the marine environment. Being low-toxic, makes addition of Tween-20 a simple and economical way to optimize KOH digestion protocols for microplastics extraction.

Weathering-independent differentiation of microplastic polymers by reflectance IR spectrometry and pattern recognition.

The presence and effects of microplastics in the environment is being continuously studied, so the need for a reliable approach to ascertain the polymer/s constituting them has increased. To characterize them, infrared (IR) spectrometry is commonly applied, either reflectance or attenuated total reflectance (ATR). A common problem when considering field samples is their weathering and biofouling, which modify their spectra. Hence, relying on spectral matching between the unknown spectrum and spectral databases is largely defective. In this paper, the use of IR spectra combined with pattern recognition techniques (principal components analysis, classification and regression trees and support vector classification) is explored first time to identify a collection of typical polymers regardless of their ageing. Results show that it is possible to identify them using a reduced suite of spectral wavenumbers with coherent chemical meaning. The models were validated using two datasets containing artificially weathered polymers and field samples.

A reliable method for the isolation and characterization of microplastics in fish gastrointestinal tracts using an infrared tunable quantum cascade laser system.

Societal and environmental concern due to frequent reports of microplastics in fish stomachs raised as they may accumulate along the trophic chain. The request for analysing microplastics in fish stresses two major analytical issues: sample treatment and final characterization. The, so far, workhorse for chemical characterization is infrared spectroscopy which is time-consuming. Here, a quantum cascade laser-based device is used to accelerate the characterization stage. Its novelty poses new challenges for sample processing and particle handling because the unknown particles must be transferred to a reflective slide. In this study, three sample digestion protocols (alkaline-oxidative with H2O2, and alkaline-oxidative with NaClO and enzymatic-oxidative) and three different procedures to transfer the filter cake to reflective slides are compared. A simplified enzymatic-oxidative digestion (validated through an interlaboratory exercise) combined with a Syncore® automatic evaporation system and a Laser Direct Infrared Imaging (LDIR) device is proposed first time as a reliable and relatively fast method to treat gastrointestinal tracts of fish. Analytical recoveries were studied using samples of Scomber scombrus and they were ca. 100% for big -i.e., >500 µm- and ca. 90% for medium -i.e., 200-300 µm- particles and ca. 75% for 10 µm thick fibres.

Monitorization of polyamide microplastics weathering using attenuated total reflectance and microreflectance infrared spectrometry.

The EU goal to reduce marine plastic litter by ca. 30% by 2020 stressed the need to deploy analytical methods to ascertain the polymeric nature of a residue. Furthermore, as plastics age under natural conditions and usual databases do not include their weathered spectra, (micro)plastics in environmental samples may be unidentified. In this paper, polyamide (nylon) microplastics weathering was monitored because of its ubiquity in household commodities, clothes, fishery items and industry, whose residues end up frequently in the environment. Infrared spectra (ATR and microreflectance) and Scanning Electron Microscopy (SEM) images were collected periodically while exposing nylon to controlled weathering. It was seen that ATR was more sensitive than microreflectance to monitor the structural evolution of polyamide and that the spectra and the surface of weathered microplastics showed remarkable differences with the pristine material, which stresses the need for considering its evolution when identifying microplastics in environmental studies. The evolution of six band ratios related to the chemical evolution of this polymer are presented. SEM images revealed the formation of secondary microplastics at the most advanced weathering stages of polyamide.

Phthalates, organotin compounds and per-polyfluoroalkyl substances in semiconfined areas of the Spanish coast: Occurrence, sources and risk assessment.

In this work two sensitive areas of the Spanish coast located in the Atlantic (Ria de Vigo) and Mediterranean (Mar Menor lagoon) have been studied regarding their contamination by phthalates, organotin compounds and per-polyfluoroalkyl substances (seawater and sediments) in two different campaigns (spring and autumn in 2015). PFAS and OTCs were detected in seawater and sediments at low concentrations (few ng L-1 or ng g-1), whereas PAEs were detected at levels two orders of magnitude higher, particularly in Mar Menor lagoon due to its semi-confined characteristics. However, PAEs and OTCs concentration in sediments were higher in Ría de Vigo than in Mar Menor lagoon as a consequence of the influence of the important urban nuclei and port in that area. The ecological risk assessment revealed that in both areas tributyltin, dibutyltin and diethylphthalate pose a significant risk in sediments, whereas in seawater tributyltin in both areas resulted in a high risk.

Development of a fast and efficient method to analyze microplastics in planktonic samples.

Microplastics (MPs) affect plankton (a basis of the trophic chain) and planktivorous fish can ingest them through food confusion or by trophic transmission. Consensus to determine MPs in plankton is lacking and, so, three digestion treatments were evaluated: Alkaline (potassium hydroxide) and enzymatic (protease plus lipase) digestions, both combined with a hydrogen peroxide stage; and an oxidative method using a surfactant (sodium dodecyl sulfate) plus hydrogen peroxide. The alkaline method using potassium hydroxide was found to damage polystyrene. MPs were identified with a stereomicroscope and characterized by reflectance infrared microscopy in semi-automatic mode (using dedicated multi-well aluminium plates). Analytical recoveries for polypropylene, polystyrene, polyethylene, polyamide, polyvinyl chloride and polyethylene terephthalate were higher than 75%, 82% and 83% for the alkaline, enzymatic and oxidative treatments, respectively. The enzymatic method was successfully validated in a European interlaboratory exercise and the oxidative method was demonstrated to be a reliable, fast and cheaper alternative.

A matrix solid-phase dispersion-gel permeation chromatography-programmed temperature vaporisation-GC-MS procedure for the analysis of polycyclic aromatic hydrocarbons in mussel samples.

A simultaneous matrix solid-phase dispersion extraction-gel permeation chromatography cleanup with programmed temperature vaporisation GC-MS determination is proposed for the analysis of parent and alkylated polycyclic aromatic hydrocarbons (PAHs) in mussel samples. The parameters affecting the extraction and elution efficiency were studied. Final conditions were: 0.5 g of mussel sample, 0.5 g diatomaceous earth as dispersant; and 4 g of Bio-Beads S-X3 for the gel permeation chromatography. Ethyl acetate/hexane (1:1, v/v) was selected for the elimination of the lipid fraction and the elution of PAHs. The detection and quantification limits achieved with this procedure were between 0.01 µg/kg and 0.67 µg/kg and 0.02 µg/kg and 1.93 µg/kg, respectively. The linearity of the method ranged between 5 µg/kg and 1000 µg/kg for most of the studied PAHs. The method was validated by the analysis of mussel tissue reference material (SRM 2977). The repeatability, intermediate precision and accuracy of the method obtained are in excellent agreement with the certified values. The proposed method is simple, precise and robust; no special instruments or costly equipment are required, and a reduction in the total time of analysis, sample handling and solvent consumption is achieved in comparison with classic procedures.

Identification of contaminants of emerging concern with potential environmental risk in Spanish continental shelf sediments.

The distribution of per- and polyfluoroalkyl substances (PFAS), alkylphenols, organotin compounds, phthalates, alkylated polycyclic aromatic hydrocarbons, current-use pesticides (CUPs) and personal care products (PCPs) was characterized in 29 surface sediments from two Spanish Iberian continental shelf areas (14 on the Atlantic and 15 on the Mediterranean coasts). Concretely, 115 organic contaminants were determined and a specific methodology was used for each contaminant group, including contaminants of emerging concern (CECs) and traditional ones, such as polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and organochlorinated pesticides (OCPs). PAHs, alkylated PAHs, alkylphenols and phthalates were found in all samples, showing mean concentrations per group higher than 20 ng/g (16-4974 ng/g d.w.) in the subregions under consideration (Galician, Cantabrian, Levantine-Balearic and Strait-Alboran). CUPs and PCPs were found in the majority of samples at very low concentrations of ng/g (1.4-46.8 ng/g d.w.), whereas organotins and PFAS were found principally in sediments from the Mediterranean subregions (2.5-3.9 ng/g d.w.). Different distribution patterns were observed for the contaminant groups and subregions under consideration as a consequence of the diverse predominant sources (industrial, urban, transport and agricultural activities) and environmental behavior (mainly hydrophobicity and persistence). Risk assessment confirmed the impact of phthalates, alkylphenols, PAHs and PCBs on Atlantic ecosystems and of alkylphenols, chlorpyrifos, phthalates, TBT, PAHs, OCPs and PCBs on the Mediterranean ones. Furthermore, the presence of CUPs, PCPs and PFAS in sediments from the Spanish continental shelf located between 2 and 31 km from the coast suggested that those contaminants may also provoke adverse effects on coastal marine ecosystems between their sources and their depositional areas. CAPSULE: Alkylphenols, phthalates and organotins may provoke adverse effects on Spanish coastal marine ecosystems from their sources to the sediment depositional areas.

PAHs, pesticides, personal care products and plastic additives in plastic debris from Spanish Mediterranean beaches.

In this study the role of plastic debris as a pollution vector has been evaluated by determining the concentrations of hydrophobic organic contaminants in polymers from three Western Mediterranean coastal areas as well as their potential transfer to seawater. Plastic debris was sampled at three Iberian Peninsula Southeastern beaches, each affected by different predominant anthropogenic activities (tourism, agriculture, urban activities, transport and industry). Plastic debris was characterized by attenuated total reflection Fourier-transform infrared spectrometry. The organic contaminants were extracted from plastics by ultrasonic extraction with methanol and quantified by stir bar sorptive extraction coupled to gas chromatography-mass spectrometry (GC-MS). In two areas, the desorption of these contaminants from plastic debris to seawater during 24 h was also evaluated. The contaminant groups considered in this study (polycyclic aromatic hydrocarbons (PAHs), personal care products (PCPs), current use pesticides (CUPs), organochlorinated compounds (OCPs, including polychlorinated biphenyls and organochlorinated pesticides) and plastic additives were found in polymers from the three areas. The most abundant contaminants were plastic additives and PCPs, underlining the relevance of the leaching of plastic components, and urban and tourism activities as typical pollution sources in the coastal areas. In general, large piece-to-piece variability was found for all polymers and areas mainly as a consequence of their different origin, exposition time, use and surface-to-volume ratio. This fact difficulted the visualization of significant differences between polymers or areas, but for CUPs, whose concentrations were significantly higher in Cape Cope than in the other areas due to the influence of close agricultural activities. PCPs and CUPs were desorbed partially in seawater for 24 h, particularly the most hydrophilic compounds such as triazines and other CUPs. However, a significant fraction of other contaminants (mainly PAHs) was retained, which suggests they can be transported far away from their origin.

Potential transfer of organic pollutants from littoral plastics debris to the marine environment.

Plastic polymers act as passive samplers in air system and concentrate hydrophobic organic contaminants by sorption or specific interactions, which can be transported to other systems such as the marine environment. In this study plastic debris was sampled in the surrounding area of a Mediterranean lagoon in order to determine the concentration of persistent and emerging organic contaminants. More specifically, desorption of 91 regulated and emerging organic contaminants (polycyclic aromatic hydrocarbons, polychlorinated biphenyls, organochlorinated pesticides, current-use pesticides, personal care products, other pesticides and plastic additives) was characterized for the first 24 h from different polymers to seawater and the remaining content of these contaminants was also extracted by ultrasonic extraction with methanol. All samples were analyzed by Stir Bar Sorptive Extraction coupled to GC/MS. A significant fraction of sorbed contaminants in polymers was desorbed in the first 24 h, particularly for triazines and organophosphorus pesticides due to their lower hydrophobicity than other considered analytes. The remaining contaminants contained in plastics can be also transferred to seawater, sediments or biota. Considering 24 h desorbed fraction plus the remaining methanol extracted fraction, the highest transfer levels corresponded to personal care products, plastic additives, current-use pesticides and PAHs. This is the first study to show the relevance of the transport of organic contaminants on plastic debris from littoral areas to the marine environment.

Cirugía ortognática y rehabilitación implantosoportada simultáneas mediante implantes subperiósticos o supraóseos personalizados / Simultaneous orthognathic surgery and implant-supported rehabilitation using customized subperiosteal or supraosseous implants

Presentamos un caso clínico donde, utilizando implantes subperiósticos o supraóseos, es decir, estructuras metálicas tridimensionales personalizadas de titanio fabricadas de forma digital, hemos conseguido realizar dos objetivos a la vez: 1) la corrección de una severa deformidad facial, y 2) dotar a la paciente de fijaciones o implantes para la rehabilitación implantosoportada de su edentulismo maxilar completo. No tenemos constancia de ningún caso similar publicado en la literatura a día de hoy.Creemos que esta nueva técnica, relativamente sencilla, puede ser una alternativa a las cirugías complejas que involucran el empleo de complicados injertos óseos, con el consiguiente ahorro en tiempo y morbilidad para los pacientes que asocian hipoplasia maxilar y edentulismo con grave atrofia ósea. (AU)

We present a clinical case in which, using subperiosteal or supraosseous implants, that is to say, three-dimensional personalized metallic titanium structures manufactured digitally, we have managed to achieve two objectives at the same time: 1) correction of a severe facial deformity, and 2) furnish the patient with fixations or implants for the implant-supported rehabilitation of her complete maxillary edentulism. We are not aware of any similar case published in the literature to date.We believe that this new, relatively simple technique can be an alternative to complex surgeries that involve the use of complicated bone grafts, with the consequent savings in time and morbidity for patients who associate maxillary hypoplasia and edentulism with severe bone atrophy. (AU)

Triphenyltin induces imposex in Nucella lapillus through an aphallic route.

Triphenyltin (TPT) was used until recently as a biocide in antifouling systems and nowadays is still applied as an agriculture pesticide in some countries. This compound is known to cause imposex (the imposition of male characters in females of gastropod molluscs) in a very limited number of species, when compared with tributyltin (TBT), the universally recognized imposex-causing agent. In this study, we tested if TPT could induce imposex in females of the dog-whelk Nucella lapillus. Experimental groups of 40 females were injected with a volume of 2µL/g of soft tissue wet weight (ww) of one of the following treatments, using DMSO as a solvent carrier: DMSO (solvent control); 1µg/g ww of TBT (positive control); 0.2, 1 and 5µg/g ww of TPT and a non-injected group (negative control). Concentrations were confirmed in the organism tissues by means of chemical analyses of a pool of 10 specimens at T0 and then after the imposex analysis at T56days. After 8-week trial, results pointed out statistically significant differences between treatments, with both TPT and TBT positively inducing imposex. However, imposex development in TPT-injected females differed from that of TBT, since females that developed imposex presented an aphallic condition (no penis development) while the TBT-treated females developed standard imposex (with penis formation). These results suggest that TPT and TBT act differently in the sequential process of female masculinization, casting new insights about the hypothetical pathways underlying imposex development.