Sweeping-micellar electrokinetic chromatography with tandem mass spectrometry as an alternative methodology to determine neonicotinoid and boscalid residues in pollen and honeybee samples

En este trabajo se propone por primera vez un método electroforético basado en cromatografía capilar electrocinética micelar acoplada con espectrometría de masas en tándem (MEKC-MS/MS) para la determinación simultánea de 9 neonicotinoides (NNI) junto con el pesticida boscalid en muestras de polen y abejas. La separación se llevó a cabo empleando perfluorooctanoato amónico (50 mM, pH 9'0), que sirvió tanto de surfactante volátil como de tampón electroforético compatible con la detección por espectrometría de masas. Se llevó a cabo la preconcentración en línea de los analitos mediante un procedimiento conocido como "barrido" para mejorar la eficacia de la separación y la sensibilidad. Además, se desarrolló un Quechers miniaturizado como tratamiento de muestra. Ello implicó un menor consumo de disolvente orgánico y el empleo de Z-Sep+ como sorbente dispersante en la etapa de limpieza. La detección se realizó con un triple cuadrupolo en electroespray positivo y mediante monitorización de reacción multiple (MRM). Para conseguir una sensibilidad y precisión satisfactorias se optimizaron los principales parámetros que afectan a la detección como la composición del líquido adicional (etanol, agua, ácido fórmico: 50, 49'5, 0'5 v/v/v) y otras variables relacionadas con el electroespray. Se establecieron curvas de calibrado procedimentales en muestras de polen y abejas. Se consiguieron límites de cuantificación por debajo de 11'6 µg/kg y 12'5 µg/kg, respectivamente. La precisión expresada como desviación estándar relativa (RSD) fue inferior al 15'2 % y las recuperaciones fueron mayores al 70 % en ambos tipos de muestras. Se encontraron dos muestras positivas de polen que contenían imidacloprid y tiametoxam. Imidacloprid se encontró también en una muestra de abejas. Los resultados obtenidos resaltan la viabilidad del método propuesto que supone una alternativa útil, sensible, eficiente y respetuosa con el medioambiente para la determinación de NNI y boscalid en muestras de polen y abejas. In this work, it is proposed for the first time an electrophoretic approach based on micellar electrokinetic chromatography coupled with tandem mass spectrometry (MEKC-MS/MS) for the simultaneous determination of nine neonicotinoids (NNIs) together with the fungicide boscalid in pollen and honeybee samples. The separation was performed using ammonium perfluorooctanoate (50 mM, pH 9) as both volatile surfactant and electrophoretic buffer compatible with MS detection. A stacking strategy for accomplishing the on-line pre-concentration of the target compounds, known as sweeping, was carried out in order to improve separation efficiency and sensitivity. Furthermore, a scaled-down QuEChERS was developed as sample treatment, involving a lower organic solvent consumption and using Z-Sep+ as dispersive sorbent in the clean-up step. Regarding the detection mode, a triple quadrupole mass spectrometer was operating in positive ion electrospray mode (ESI+) under multiple reaction monitoring (MRM). The main parameters affecting MS/MS detection as well as the composition of the sheath-liquid (ethanol/ultrapure water/formic acid, 50:49.5:0.5 v/v/v) and other electrospray variables were optimized in order to achieve satisfactory sensitivity and repeatability. Procedural calibration curves were established in pollen and honeybee samples with LOQs below 11.6 µg/kg and 12.5 µg/kg, respectively. Precision, expressed as RSD, lower than 15.2% and recoveries higher than 70% were obtained in both samples. Two positive samples of pollen were found, containing imidacloprid and thiamethoxam. Imidacloprid was also found in a sample of honeybees. The obtained results highlight the applicability of the proposed method, being an environmentally friendly, efficient, sensitive and useful alternative for the determination of NNIs and boscalid in pollen and honeybee samples.Ministerio de Ciencia e Innovación y Agencia Estatal de Investigación (/10.13039/501100011033/ FEDER “Una manera de hacer Europa”): EQC2018-004453-P y RTI2018-097043-B-I00.Junta de Andalucía-Programa Operativo FEDER (B-AGR-202-UGR20)Red española de excelencia en tratamiento de muestra (RED2018-102522-T) financiada por MCIN/AEI /10.13039/501100011033.B.H. agradece el apoyo del proyecto EFSA-CDN (No. CZ.02.1.01/0.0/0.0/16_019/0000841) confinanciado por ERDF y una ayuda Erasmus+.LCR le da las gracias a Francisco Gerardo C.M y Pasión R.S por su apoyo técnico durante la etapa de muestreo.Financiación para publicación en acceso abierto a cargo de Universidad de Granada/CBUA

Occurrence of Ergot Alkaloids in Barley and Wheat from Algeria

The natural occurrence of six major ergot alkaloids, ergometrine, ergosine, ergotamine, ergocornine, ergokryptine and ergocristine, as well as their corresponding epimers, were investigated in 60 cereal samples (barley and wheat) from Algeria. Ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) and a QuEChERS extraction method were used for sample analysis. The results revealed that 12 out of 60 samples (20%) were contaminated with ergot alkaloids. Wheat was the most contaminated matrix, with an incidence of 26.7% (8 out of 30 samples). The concentration of total ergot alkaloids ranged from 17.8 to 53.9 µg/kg for barley and from 3.66 to 76.0 µg/kg for wheat samples. Ergosine, ergokryptine and ergocristine showed the highest incidences in wheat, while ergometrine was the most common ergot in barley.Spanish Ministry of Science, Innovation and Universities (Project Ref.: RTI2018-097043-B-I00

Ion mobility-mass spectrometry to extend analytical performance in the determination of ergot alkaloids in cereal samples

This work evaluates the potential of ion mobility spectrometry (IMS) to improve the analytical perfor- mance of current liquid chromatography-mass spectrometry (LC-MS) workflows applied to the determi- nation of ergot alkaloids (EAs) in cereal samples. Collision cross section (CCS) values for EA epimers are reported for the first time to contribute to their unambiguous identification. Additionally, CCS values have been inter-laboratory cross-validated and compared with CCS values predicted by machine-learning mod- els. Slight differences were observed in terms of CCS values for ergotamine, ergosine and ergocristine and their corresponding epimers (from 3.3 to 4%), being sufficient to achieve a satisfactory peak-to-peak res- olution for their unequivocal identification. A LC-travelling wave ion mobility (TWIM)-MS method has been developed for the analysis of EAs in barley and wheat samples. Signal-to-noise ratio (S/N) was im- proved between 2.5 and 4-fold compared to the analog LC-TOF-MS method. The quality of the extracted ion chromatograms was also improved by using IMS.European Commission 795946MCIN/AEI (European Regional Development Fund-ERDF, a way to build Europe) PID2021-127804OB-I0

Nanofibrous Online Solid-Phase Extraction Coupled with Liquid Chromatography for the Determination of Neonicotinoid Pesticides in River Waters

Polymeric nano- and microfibers were tested as potential sorbents for the extraction of five neonicotinoids from natural waters. Nanofibrous mats were prepared from polycaprolactone, polyvinylidene fluoride, polystyrene, polyamide 6, polyacrylonitrile, and polyimide, as well as microfibers of polyethylene, a polycaprolactone nano- and microfiber conjugate, and polycaprolactone microfibers combined with polyvinylidene fluoride nanofibers. Polyimide nanofibers were selected as the most suitable sorbent for these analytes and the matrix. A Lab-In-Syringe system enabled automated preconcentration via online SPE of large sample volumes at low pressure with analyte separation by HPLC. Several mat layers were housed in a solvent filter holder integrated into the injection loop of an HPLC system. After loading 2 mL sample on the sorbent, the mobile phase eluted the retained analytes onto the chromatographic column. Extraction efficiencies of 68.8-83.4% were achieved. Large preconcentration factors ranging from 70 to 82 allowed reaching LOD and LOQ values of 0.4 to 1.7 and 1.2 to 5.5 mu g center dot L-1, respectively. Analyte recoveries from spiked river waters ranged from 53.8% to 113.3% at the 5 mu g center dot L-1 level and from 62.8% to 119.8% at the 20 mu g center dot L-1 level. The developed methodology proved suitable for the determination of thiamethoxam, clothianidin, imidacloprid, and thiacloprid, whereas matrix peak overlapping inhibited quantification of acetamiprid.STARSS project - ERDF CZ.02.1.01/0.0/0.0/15_003/0000465Grant Agency of the Czech Republic 20-19297SSpanish Government RED2018-102522-TDivision of Analytical Chemistry of the European Chemical SocietyProject Junta de Andalucia-Programa Operativo FEDER B-AGR-202-UGR2

New analytical strategies for the determination of insecticide residues and ergot alkaloids in environmental and food samples

RTI2018-097043-B-I00 (Spanish Ministry of Science, Innovation, and Universities). Towards a safe consumption of nutraceuticals: analytical evaluation of the presence of emerging mycotoxins and cyanotoxins by advanced techniques (NUTRATOX). Main researchers: A.M. García Campaña, M. del Olmo Iruela. 1/01/2019 - 31/12/2021; 72.600,00 €. AGL2015-70708-R (Spanish Ministry of Science, Innovation, and Universities). Chemical hazards in nutraceuticals: advanced analytical proposals for the determination of mycotoxins and pesticides. Main researchers: A.M. García Campaña, L. Gámiz Gracia. 1/01/2016 - 31/09/2019; 84.700 €. RED2018-10522-T (Spanish Ministry of Science, Innovation, and Universities). Spanish Network of Excellence in Sample preparation - Sample Preparation Task Force and Network, supported by the Division of Analytical Chemistry of the European Chemical Society. Coordinator: M.S. Cárdenas (U. Córdoba)/Main researcher UGR: A.M. García Campaña. AGL2017-90787-REDT (Spanish Ministry of Science, Innovation, and Universities). National Network on mycotoxins and toxigenic moulds and their decontamination processes (MICOFOOD). Coordinator: J. Mañés (U. Valencia)/Main researcher UGR: A.M. García Campaña. Youth Employment Operational Program co-funded by the Ministry of Science, Innovation and Universities of the Junta de Andalucía and the European Social Fund (ESF). Mobility Program between Andalusian and Iberoamerican universities from the Iberoamerican Association of Postgraduate Universities (AUIP, 2019). International Mobility Program 2020 for PhD students from the University of Granada.In this Thesis, different analytical strategies have been proposed as efficient alternatives for the monitoring of several insecticides and ergot alkaloids, considered as relevant chemical hazards in food and environmental samples.En esta tesis se han propuesto diferentes estrategias analíticas como alternativas eficientes para la determinación de diferentes insecticidas y alcaloides ergóticos, considerados como peligros químicos de gran importancia en muestras alimentarias y medioambientales.Tesis Univ. Granada.RTI2018-097043-B-I00 (Spanish Ministry of Science, Innovation, and Universities)AGL2015-70708-R (Spanish Ministry of Science, Innovation, and Universities)RED2018-10522-T (Spanish Ministry of Science, Innovation, and Universities)AGL2017-90787-REDT (Spanish Ministry of Science, Innovation, and Universities)Youth Employment Operational Program co-funded by the Ministry of Science, Innovation and Universities of the Junta de Andalucía and the European Social Fund (ESF)Mobility Program between Andalusian and Iberoamerican universities from the Iberoamerican Association of Postgraduate Universities (AUIP, 2019)International Mobility Program 2020 for PhD students from the University of Granad

Occurrence of Ergot Alkaloids in Barley and Wheat from Algeria

The natural occurrence of six major ergot alkaloids, ergometrine, ergosine, ergotamine, ergocornine, ergokryptine and ergocristine, as well as their corresponding epimers, were investigated in 60 cereal samples (barley and wheat) from Algeria. Ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) and a QuEChERS extraction method were used for sample analysis. The results revealed that 12 out of 60 samples (20%) were contaminated with ergot alkaloids. Wheat was the most contaminated matrix, with an incidence of 26.7% (8 out of 30 samples). The concentration of total ergot alkaloids ranged from 17.8 to 53.9 µg/kg for barley and from 3.66 to 76.0 μg/kg for wheat samples. Ergosine, ergokryptine and ergocristine showed the highest incidences in wheat, while ergometrine was the most common ergot in barley

Determination of the Main Ergot Alkaloids and Their Epimers in Oat-Based Functional Foods by Ultra-High Performance Liquid Chromatography Tandem Mass Spectrometry

An ultra-high performance liquid chromatography coupled to tandem mass spectrometry method is proposed for the determination of the major ergot alkaloids (ergometrine, ergosine, ergotamine, ergocornine, ergokryptine, ergocristine) and their epimers (ergometrinine, ergosinine, ergotaminine, ergocorninine, ergokryptinine, and ergocristinine) in oat-based foods and food supplements. A modified QuEChERS (quick, easy, cheap, effective, rugged, and safe) procedure was applied as sample treatment, reducing the consumption of organic solvent and increasing sensitivity. This method involved an extraction with acetonitrile and ammonium carbonate (85:15, v/v) and a clean-up step based on dispersive solid-phase extraction, employing a mixture of C18/Z-Sep+ as sorbents. Procedural calibration curves were established and limits of quantification were below 3.2 µg/kg for the studied compounds. Repeatability and intermediate precision (expressed as RSD%) were lower than 6.3% and 15%, respectively, with recoveries ranging between 89.7% and 109%. The method was applied to oat-based products (bran, flakes, flour, grass, hydroalcoholic extracts, juices, and tablets), finding a positive sample of oat bran contaminated with ergometrine, ergosine, ergometrinine, and ergosinine (total content of 10.7 µg/kg).Spanish Ministry of Science, Innovation, and Universities (project reference: RTI2018-097043-B-I00

Multi-mycotoxin determination in plant-based meat alternatives and exposure assessment

The aim of this study was to fill in the gap regarding the occurrence of mycotoxins in plant-based meat alter-natives. Hence, a multi-mycotoxin method (aflatoxins, ochratoxin A, fumonisins, zearalenone, and mycotoxins from the Alternaria alternata genera) was developed followed by an exposure assessment for the Italian con-sumers' exposure to mycotoxins. A total of 13 meat alternatives samples based on soy, pea, chickpea, lupin, and seitan were analysed. With the exception of seitan, all of the remaining samples were contaminated with one mycotoxin or mixtures of up to seven mycotoxins. The level of contamination was as low as 0.2 mu g/kg alternariol methyl ether and as high as 66.9 mu g/kg fumonisin B1. To analyse the exposure to mycotoxins due to plant-based meat alternatives consumption we used the consumption meat data from the Food and Agriculture Organization for Italian adult consumers and simulated a full replacement of meat with plant-based meat alternatives. Based on our model, consumption of plant-based meat alternatives led to a non-tolerable exposure to alternariol (hazard index (HI) > 1) in pea-based burger and soy + wheat-based steak, while samples contaminated with aflatoxins, respectively ochratoxin A, indicated a health concern related to liver and renal cancer (margin of exposure (MOE) < 10,000). This is the first study that presents the co-occurrence of mycotoxins in multiple plant -based meat alternatives. Moreover, these results indicate that there is a need for policymakers to consider the regulation of mycotoxins in plant-based meat alternatives in order to ensure consumers' safety

A natural deep eutectic solvent as a novel dispersive solvent in dispersive liquid-liquid microextraction based on solidification of floating organic droplet for the determination of pesticide residues

Current trends in analytical chemistry encourage the use of innocuous solvents to develop modern methods aligned with green chemistry. In this sense, natural deep eutectic solvents (NADESs) have emerged as a novel generation of green solvents which can be employed in sample treatments as an alternative to the toxic organic solvents commonly used so far. In this work, a new extraction method employs dispersive liquid-liquid microextraction based on a solid floating organic droplet (DLLME-SFO), by using a mixture composed of a less dense than water extraction solvent, 1-dodecanol, and a novel dispersive solvent, NADES. The methodology was proposed to extract and preconcentrate some pesticide residues (fipronil, fipronil-sulfide, fipronil-sulfone, and boscalid) from environmental water and white wine samples before analysis by liquid-chromatography coupled to ultraviolet detection (HPLC-UV). Limits of quantification (LOQs) lower than 4.5 μg L−1, recoveries above 80%, and precision, expressed as RSD, below 15% were achieved in both samples showing that the proposed method is a powerful, efficient, and green alternative for the determination of these compounds and, therefore, demonstrating a new application for NADES in sample preparation. In addition, the DLLME-SFOD-HPLC-UV method was evaluated and compared with other reported approaches using the Analytical GREEnness metric approach, which highlighted the greenness of the proposed method.Fil: Carbonell-Rozas, Laura. Universidad de Granada; EspañaFil: Canales, Maria Romina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Biología Agrícola de Mendoza. Universidad Nacional de Cuyo. Facultad de Ciencias Agrarias. Instituto de Biología Agrícola de Mendoza; ArgentinaFil: Lara, Francisco J.. Universidad de Granada; EspañaFil: García Campaña, Ana María. Universidad de Granada; EspañaFil: Silva, María Fernanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Biología Agrícola de Mendoza. Universidad Nacional de Cuyo. Facultad de Ciencias Agrarias. Instituto de Biología Agrícola de Mendoza; Argentin

Nanofibrous Online Solid-Phase Extraction Coupled with Liquid Chromatography for the Determination of Neonicotinoid Pesticides in River Waters

Polymeric nano- and microfibers were tested as potential sorbents for the extraction of five neonicotinoids from natural waters. Nanofibrous mats were prepared from polycaprolactone, polyvinylidene fluoride, polystyrene, polyamide 6, polyacrylonitrile, and polyimide, as well as microfibers of polyethylene, a polycaprolactone nano- and microfiber conjugate, and polycaprolactone microfibers combined with polyvinylidene fluoride nanofibers. Polyimide nanofibers were selected as the most suitable sorbent for these analytes and the matrix. A Lab-In-Syringe system enabled automated preconcentration via online SPE of large sample volumes at low pressure with analyte separation by HPLC. Several mat layers were housed in a solvent filter holder integrated into the injection loop of an HPLC system. After loading 2 mL sample on the sorbent, the mobile phase eluted the retained analytes onto the chromatographic column. Extraction efficiencies of 68.8–83.4% were achieved. Large preconcentration factors ranging from 70 to 82 allowed reaching LOD and LOQ values of 0.4 to 1.7 and 1.2 to 5.5 µg·L−1, respectively. Analyte recoveries from spiked river waters ranged from 53.8% to 113.3% at the 5 µg·L−1 level and from 62.8% to 119.8% at the 20 µg·L−1 level. The developed methodology proved suitable for the determination of thiamethoxam, clothianidin, imidacloprid, and thiacloprid, whereas matrix peak overlapping inhibited quantification of acetamiprid