TBS-pyrrole as an "universal" reference to quantify artemisinin and structurally-diverse natural products in plants extracts by NMR.

The commercial production of artemisinin and other valuable bioactive natural products depends on their plant sources, which may provide variable amounts of the compound depending on plant variety, the period of the year, abiotic stress and other factors. Therefore, it requires a method for large-scale, low-cost natural product quantification. The standard HPLC and UHPLC methods are accurate but the analysis are costly and require different optimization for structurally-diverse products. An alternative method using NMR with TBS-pyrrole as a novel "universal" reference affords a simple, fast method to quantify many different products. The method is shown with antimalarial artemisinin, whose yield using conventional and novel extraction procedures was determined by standard UHPLC-MS procedures and by our NMR protocol, with similar quantification results. The novel reference compound does not interfere with artemisinin or extract signals, only needs a small amount of the extract, is accurate and operationally simple, and a large volume of samples can be processed in little time. Moreover, bioactive terpenes, steroids, alkaloids, aromatic compounds, and quinones, among others, were quantified in a model vegetal extract with this "universal" reference with excellent accuracy.

Assessment of endocrine disruptor pollutants and their metabolites in environmental water samples using a sustainable natural deep eutectic solvent-based analytical methodology.

In this work, an evaluation of the occurrence of fifteen phthalates, four metabolites and one adipate in different groundwater, seawater and wastewater samples has been carried out due to their relevance on human health as they act as endocrine disruptors. For this purpose, a sustainable, fast and easy-handling vortex-assisted liquid-liquid microextraction method using a natural hydrophobic deep eutectic solvent based on menthol and carvacrol as extraction agent, combined with ultra-high performance liquid chromatography-mass spectrometry technique, has been developed and applied for the first time. An optimization was performed to evaluate four important factors affecting the extraction performance, and an analytical validation was carried out in terms of matrix effect, linearity, extraction efficiency, and sensitivity. Recovery values were obtained in the range 72-119% for all analytes (except for monoethyl phthalate: 61.1-72.3%) with relative standard deviation values lower than 17%. Limits of quantification were found between 0.91 and 8.09 µg L-1. As a result of the assessment of 31 different environmental water samples, monoethyl phthalate, diethyl phthalate, dibutyl phthalate and bis (2-ethylhexyl) phthalate were detected and quantified at different concentrations in the range 2.59-21.17 µg L-1 in 6 samples, and diallyl phthalate, butyl benzyl phthalate, dipentyl phthalate, dicyclohexyl phthalate, dihexyl phthalate and bis (2-ethylhexyl) adipate were detected in 20 more, showing the exposition of the population to these hazardous substances.

Monitoring of the presence of plasticizers and effect of temperature and storage time in bottled water using a green liquid-liquid microextraction method.

In this work, a natural deep eutectic solvent was used for the liquid-liquid microextraction of fourteen phthalates and one adipate from bottled waters. The methodology was validated in terms of matrix effect, linearity, recovery, and limits of quantification (LOQs). Optimum extraction conditions (10 mL of water at pH 8.0 with 100 µL of thymol: menthol 2:1 (n/n) as solvent) provided satisfactory determination coefficients (≥ 0.9977), recovery values (82-127%), and LOQs (0.018-0.523 µg/L). The effects of temperature and storage time on plasticizer presence were studied for 36 different brands stored at 4 °C, room temperature, and 45 °C, and analyzed at 0, 24, 48, 72 h, and 1 week. Only diethyl-, dibutyl-, bis-(2-ethylhexyl) phthalates, and bis-(2-ethylhexyl) adipate were detected. The results showed that there is no relationship between the storage conditions, the bottle material or water carbonation, and the occurrence of these plasticizers, suggesting that residues are introduced during production or by the water supply. The estimated daily intake was lower than the total daily intake set by the European Food Safety Authority.

Assessment of pesticide residues contamination in cereals and pseudo-cereals marketed in the Canary Islands.

In this work, the development of an analytical method for the evaluation of a wide variety of pesticide residues in cereals and pseudo-cereals widely consumed has been carried out. A QuEChERS method was used as extraction and clean-up procedure prior to separation and quantification of the target analytes by ultra-high performance liquid chromatography coupled to mass spectrometry using a single quadrupole-time-of-flight analyser. The methodology was validated for oat, rye, spelt, barley and quinoa matrices, following the European Commission guidelines (SANTE/11312/2021), achieving good absolute recovery values in the range 60-124% with relative standard deviation values lower than 20% and providing limits of quantification of the method in the low mg/kg range, in accordance with the maximum residue limits established by European policies. Finally, considering the importance of cereals in general population diet, the analysis of 60 samples was carried out in order to assure their safe consumption.

Analysis of alkylphenols, bisphenols and alkylphenol ethoxylates in microbial-fermented functional beverages and bottled water: Optimization of a dispersive liquid-liquid microextraction protocol based on natural hydrophobic deep eutectic solvents.

In this work, the analysis of alkylphenols, bisphenols and alkylphenol ethoxylates in bottled waters, kombuchas and water kefir was performed through a vortex-assisted dispersive liquid-liquid microextraction method based on a natural hydrophobic eutectic solvent. For this purpose, mixtures of monoterpenes and fatty acids were employed. Different factors affecting extraction were optimized and the method was validated in terms of matrix effect, linearity, limits of detection and recovery. Recovery values varied between 70.0 and 124.3% (except for 4-tert-butylphenol: 67.0% and 4-n-nonylphenol: 60.8% in water kefir) and limits of detection were in the range 0.10 ng/L - 2.99 µg/L. Finally, 8 bottled waters, 8 kombuchas and 4 water kefirs were analyzed and 4-tert-octylphenol monoethoxylate was detected in water (20.28 ± 0.99 - 62.08 ± 3.63 µg/L). This is the first application analyzing xenobiotic contaminants in kombucha and water kefir and the first time in which the three types of compounds are simultaneously extracted by dispersive liquid-liquid microextraction.

Quality assessment of environmental water by a simple and fast non-ionic hydrophobic natural deep eutectic solvent-based extraction procedure combined with liquid chromatography tandem mass spectrometry for the determination of plastic migrants.

A non-ionic hydrophobic natural deep eutectic solvent (HNADES) based on thymol and menthol was proposed for the liquid-liquid microextraction of fourteen phthalates and one adipate from environmental water samples. Separation, identification, and quantification were achieved by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry. The main factors affecting the extraction efficiency were thoroughly studied. Sample pH of 8 and 100 µL of thymol:menthol at molar ratio 2:1 were selected as the best conditions, while ionic strength and type of dispersant solvent were not relevant for the extraction of the target compounds. The whole methodology was validated for treated wastewater, runoff, and pond water matrices, using di-n-butyl phthalate-3,4,5,6-d4 and dihexyl phthalate-3,4,5,6-d4 as surrogates. Recovery ranged from 70 to 127% with relative standard deviation values lower than 14%. Limits of quantification of the method were in the range 0.042-0.425 µg/L for treated wastewater, 0.015-0.386 µg/L for runoff, and 0.013-0.376 µg/L for pond water. The methodology was applied for the analysis of real treated wastewater, runoff, and pond water samples from different places of Tenerife and Gran Canaria (Canary Islands) finding the presence of diethyl phthalate, diallyl phthalate, dipropyl phthalate, benzylbutyl phthalate, di-n-butyl phthalate, bis-(2-n-butoxyethyl) phthalate, di-n-pentyl phthalate, dicyclohexyl phthalate, and bis-(2-ethylhexyl) phthalate at concentrations between 105.2 and 3414 ng/L.

Sustainable polypyrrole-based magnetic-microextraction of phthalates from jellies and apple-based beverages prior to tandem mass spectrometry analysis.

Synthesised polypyrrole-coated Fe3O4 magnetic nanoparticles have been successfully characterised and applied as sorbent for the magnetic-micro-dispersive solid-phase extraction of eleven phthalic acid esters from jelly and apple-based beverage matrices widely consumed by the population and, especially, by children. Sorbent was synthesised through chemical coprecipitation and subsequently characterised by different techniques. The influence of several parameters on the extraction efficiency was exhaustively evaluated using a step-by-step strategy. The separation and quantification of the selected phthalates were performed by ultra-high performance liquid chromatography coupled to tandem mass spectrometry. The validation of the methodology was carried out for jellies and apple-based beverages, employing dihexyl phthalate-3,4,5,6-d4 as the surrogate standard. Relative recovery values were in the range 70-114% for both matrices and relative standard deviations below 20% were obtained. The limits of quantification of the method were found in the range 0.147-0.416 µg/L. Feasibility of the developed methodology was proved by the analysis of commercialised jelly and apple-based beverage products.

Application of a Liquid-Liquid Microextraction Method Based on a Natural Hydrophobic Deep Eutectic Solvent for the Extraction of Plastic Migrants from Kombuchas.

A vortex-assisted liquid-liquid microextraction, based on a natural hydrophobic deep eutectic solvent made from the monoterpene thymol and octanoic fatty acid, was employed for the analysis of 11 phthalate esters and one adipate in kombucha (a tea-based fermented beverage). Separation and determination were performed using an ultra-high performance liquid chromatography (UHPLC) system coupled to a single quadrupole mass spectrometer. Confirmatory analyses were carried out through UHPLC tandem mass spectrometry. The full method was validated in terms of matrix effect, matrix-matched calibration, sensitivity, recovery, limits of detection and quantification and repeatability. Satisfactory determination coefficients for quadratic calibration curves (≥0.9938), recovery values (67-120%) and limits of detection (0.07-5.45 µg/L) were obtained. Analysis of 26 kombucha samples reported concentrations for dibutyl phthalate and dimethyl phthalate in the range between the limit of quantification (LOQ) and 16.18 ± 1.14 µg/L, although these phthalates were also detected under the LOQ in some of the analyzed samples. Only one of the samples bottled in plastic containers (7) did not present residues while only five of the 19 samples in glass bottles contained any plasticizer. However, the highest concentration was found in a kombucha bottled in food-grade glass. This work represents the first application in which phthalates and adipates are analyzed in kombuchas.

Comparison of Pesticide Residue Levels in Red Wines from Canary Islands, Iberian Peninsula, and Cape Verde.

In this work, the QuEChERS method coupled to liquid chromatography-time-of flight-mass spectrometry and gas chromatography-triple quadrupole-mass spectrometry were applied for the evaluation of pesticide residues and risk assessment in red wines. The methodologies were successfully validated for 173 pesticides. Recovery values were in the range 75-100% for almost all pesticides and limits of quantification were between 2.60 and 21.39 µg/kg, which are in good agreement with the maximum residue limits (MRLs) established by the European Commission for pesticides in wine grapes. Finally, the analysis of 84 red wine samples from the Canary Islands, the Iberian Peninsula, and Cape Verde was carried out, which found the presence of 31 pesticide residues. However, the risk assessment disclosed that despite the large number of pesticides and the concentrations found, which in some cases exceeded the MRLs, the consumption of these wines, without considering a possible cumulative effect, does not entail a risk to the consumers.

Nano-liquid chromatography combined with a sustainable microextraction based on natural deep eutectic solvents for analysis of phthalate esters.

The separation of 11 phthalic acid ester (PAEs) was carried out by nano-liquid chromatography coupled to ultraviolet and MS detection. Preliminary experiments were achieved in order to select suitable stationary phases and chromatographic conditions. The baseline separation was obtained, for all compounds, with an XBridgeTM C18 column in less than 15 min, working in step gradient mode. The sensitivity of the method was improved by on-column focusing. PAEs were extracted from alcoholic and nonalcoholic beverages using vortex-assisted emulsification dispersive liquid-liquid microextration and natural deep eutectic solvents. The whole method was validated in terms of linearity, sensitivity, precision, recovery, and repeatability. Combination of both off-line sample preparation preconcentration and large injection volume led to obtain LOQs in the range 5-47 ng/mL. The developed nano-LC-UV method was extended to MS detection to confirm the presence of PAEs in some beverages commercialized in different types of packaging.

A simple, fast and easy methodology for the monitoring of plastic migrants in alcoholic and non-alcoholic beverages using the QuEChERS method prior to gas chromatography tandem mass spectrometry.

In this work, the development of a simple, fast and reliable method for the evaluation of a group of twelve plastic migrants in alcoholic and non-alcoholic beverages widely consumed by the population has been carried out. For that, a modified QuEChERS method for the extraction and preconcentration of the target compounds has been used prior to their separation and quantification by gas chromatography coupled to triple quadrupole tandem mass spectrometry. The whole methodology was validated for beer, cider and grape juice matrices, using dibutyl phthalate-3,4,5,6-d4 as surrogate. Recovery ranged from 75 to 120% for all matrices with relative standard deviation values lower than 20%, and the limits of quantification of the method were achieved in the range 0.034-1.415 µg/L. Finally, the analysis of different beer, cider and grape juice samples commercialised in different supermarkets of Tenerife was carried out, finding the presence of four of the evaluated phthalates in the range 0.14-1.1 µg/L in some of the evaluated beers, six of them in several cider samples, in the range 0.3-2.1 µg/L, and one in the range 1.2-1.5 µg/L in three of the analysed grape juices.

A green and simple procedure based on deep eutectic solvents for the extraction of phthalates from beverages.

In this work, a green, inexpensive, simple and fast deep eutectic solvent (DES)-based dispersive liquid-liquid microextraction was evaluated, for the first time, for the extraction of phthalates (i.e. benzylbutyl phthalate, diisobutyl phthalate, diisopentyl phthalate, di-n-pentyl phthalate, di-(2-ethylhexyl) phthalate, di-n-octyl phthalate, diisononyl phthalate, diisodecyl phthalate) from different beverages. Separation and determination were achieved by high performance liquid chromatography-diode-array detection while confirmation was carried out by tandem mass spectrometry. The main factors affecting the extraction such as type and volume of DES and emulsifier, pH and ionic strength, were optimised. Choline chloride:phenol-based DES showed the best results. The methodology was validated for tea, apple-based beverage and pineapple juice. Recovery values ranged from 84 to 120% with relative standard deviation values lower than 11%. Limits of detection of the method were in the range 5.1-14.2 µg L-1 for tea, 5.3-17.8 µg L-1 for apple beverages and 5.9-15.6 µg L-1 for pineapple juices.

Nanomaterials as alternative dispersants for the multiresidue analysis of phthalates in soil samples using matrix solid phase dispersion prior to ultra-high performance liquid chromatography tandem mass spectrometry.

In this study, the application of different nanomaterials as dispersants in matrix solid phase dispersion has been evaluated for the extraction of fifteen phthalates from different environmental samples prior to their separation and quantification by ultra-high performance liquid chromatography coupled to triple quadrupole mass spectrometry. Within the evaluated nanomaterials, including graphene oxide, multi-walled carbon nanotubes and iron 1,3,5-benzenetricarboxylate metal-organic framework, the last one showed the best results in terms of extraction capacity and sample clean-up. The effects of the different parameters affecting the sample pretreatment efficiency were exhaustively evaluated. The whole methodology was validated for agricultural soil and sand, using dibutyl phthalate-3,4,5,6-d4 as surrogate. Recovery values ranged from 70 to 120% for both matrices with RSD values lower than 20% and the limits of quantification of the method achieved were in the range 0.14-2.7 µg/kg dry weight. Finally, the analysis of soil samples from different locations of Tenerife (Canary Islands, Spain) was carried out finding the presence of BBP, DIBP and DBP in the range 5-52 µg/kg dry weight in agricultural soils, and DIPP, DNOP and DINP in the range 2-101 µg/kg dry weight in sand samples.

Reduced graphene oxide-coated magnetic-nanoparticles as sorbent for the determination of phthalates in environmental samples by micro-dispersive solid-phase extraction followed by ultra-high-performance liquid chromatography tandem mass spectrometry.

In this work, the suitability of Fe3O4 nanoparticles coated with reduced-graphene oxide as sorbent was evaluated for the extraction of a group of fourteen phthalic acid esters (i.e. benzylbutyl phthalate (BBP), bis-2-n-butoxyethyl phthalate (DBEP), dibutyl phthalate (DBP), diisobutyl phthalate (DIBP), dicyclohexyl phthalate (DCHP), bis-2-ethoxyethyl phthalate (DEEP), diisodecyl phthalate (DIDP), diisononyl phthalate (DINP), bis-isopentyl phthalate (DIPP), bis-(2-methoxyethyl) phthalate (DMEP), dimethyl phthalate (DMP), di-n-octyl phthalate (DNOP), bis-n-pentyl phthalate (DNPP), dipropyl phthalate (DPP)) from environmental samples. Extraction was carried out using magnetic-micro dispersive solid-phase extraction while separation, identification and quantification of the target analytes were achieved by ultra-high-performance liquid chromatography coupled to triple quadrupole tandem mass spectrometry. The methodology was validated for three different types of water samples using dibutyl phthalate-3,4,5,6-d4 as internal standard for all of them. Recovery values ranged from 70 to 120% for the three matrices with relative standard deviation values lower than 20%. Limits of quantification of the method achieved were in the range 6-178 ng/L for all samples and analytes. The methodology was applied for the evaluation of real samples finding the presence of DMP, DPP, BBP, DIBP and DBP in some of the analysed matrices.

Determination of phthalic acid esters in different baby food samples by gas chromatography tandem mass spectrometry.

In this work, a new method has been developed for the determination of 14 phthalic acid esters (i.e., benzylbutyl phthalate (BBP), bis-2-n-butoxyethyl phthalate (DBEP), dibutyl phthalate (DBP), dicyclohexyl phthalate (DCHP), bis-2-ethoxyethyl phthalate (DEEP), diethyl phthalate (DEP), diisodecyl phthalate (DIDP), diisononyl phthalate (DINP), bis-isopentyl phthalate (DIPP), bis (2-methoxyethyl) phthalate (DMEP), dimethyl phthalate (DMP), di-n-octyl phthalate (DNOP), bis-n-pentyl phthalate (DNPP), dipropyl phthalate (DPP)) and one adipate (bis (2-ethylhexyl) adipate (DEHA)) in different baby foods. Separation was carried out by gas chromatography triple quadrupole tandem mass spectrometry while the previous extraction of the samples was carried out using the QuEChERS method. The methodology was validated for four baby food samples (two fruit compotes of different compositions and two meat and fish purees with vegetables) using dibutyl phthalate-3,4,5,6-d4 (DBP-d4) as internal standard. Determination coefficients (R2) of matrix-matched calibration curves were above 0.9922 in all cases while relative recovery values ranged between 70 and 120%, with relative standard deviation values below 19%. The limits of quantification of the method ranged between 0.03 and 1.11 µg/kg. Finally, the analysis of commercially available samples was carried out finding the presence of BBP, DEHA, DEP, DIDP, and DPP in some of the studied samples.