Simultaneous determination of mycotoxins and pesticides by UHPLC-HRMS in aquaculture feed from intensive farms

Posted on 10 November, 2025 by fqm302

Background: Intensive aquaculture in southern Spain is growing, leading to increased use of plant-based in- gredients in fish feed. While this shift results in more nutritionally balanced feeds, it also introduces the risk of contaminants like mycotoxins and pesticides, which can negatively affect feed production, fish health, and pose potential risks to consumers. However, studies on these contaminants in aquaculture feed are limited. Results: In this study, a multiclass method was developed to determine 27 mycotoxins and 11 pesticides in aquaculture feed by ultrahigh performance liquid chromatography coupled with high resolution mass spec- trometry (UHPLC-HRMS). The method involved a previous ultrasound-assisted solid-liquid extraction using an acidified mixture of acetonitrile and water, followed by a dispersive solid-phase extraction with MgSO4 and C18. Characterization of the method was satisfactory for almost all compounds, with recovery values between 70 and 120 % and relative standard deviations (RSDs) below 20 %. The quantification limits were comparable to those reported in studies focusing on a single family of compounds. The method was applied to the analysis of aquaculture feeds with diverse compositions, revealing a high incidence of the mycotoxins enniatins, as well as the pesticides tebuconazole and pirimiphos methyl. Conclusion: The possible source of these contaminants and residues was investigated by analysing the main raw materials of the feeds, attempting to link their origin with the wheat and soy. Additionally, non-targeted analysis identified six additional pesticides, and one pesticide adjuvant not included in the targeted analysis, highlighting the importance of such studies.

Posted in Animal feed, Mass spectrometry, Mycotoxins, PID2021-127804OB-I00, PROYEXCEL_00195, Pesticides, RYC2023-044255-I, UHPLC, USLE, dSPE | Tagged , , , | Comments Off on Simultaneous determination of mycotoxins and pesticides by UHPLC-HRMS in aquaculture feed from intensive farms

A greener approach for determining cyanotoxins in vegetables: From NADES-air-assisted dispersive liquid-liquid microextraction to capillary electrophoresis–mass spectrometry analysis

Posted on 18 September, 2025 by fqm302

This study presents a sustainable analytical method for the determination of 12 cyanotoxins including 10 microcystins (−LR, -YR, -LF, -LW, −LA, -HilR, -HtyR, -LY, -WR, [D-Asp3]MC-LR), nodularin, and anabaenopeptin A in vegetables, based on air-assisted dispersive liquid-liquid microextraction (AA-DLLME) using a thymol:menthol natural deep eutectic solvent (NADES) in a 1:1 M ratio. Analyte separation and detection were performed using capillary zone electrophoresis coupled with tandem mass spectrometry (CZE-MS/MS), a technique that allows analysis with minimal solvent consumption and low waste generation, reducing the environmental impact as well as lowering costs. The injection solvent was carefully studied to ensure the compatibility of NADES-extraction with CZE-MS/MS analysis. The method demonstrated high linearity and extraction efficiency for apolar cyanotoxins, achieving satisfactory recoveries and detection limits suitable for their monitoring in vegetable samples. In pepper samples, low limits of quantification (1.5–12.4 ng/g, fresh weight), and satisfactory recoveries (63–105 %) were obtained. Matrix effects were generally below 16 %, and both intra- and inter-day precision remained under 15 %. Application to tomato and cucumber confirmed the method’s robustness in other high-water-content vegetable matrices. MC-HilR was detected at 0.9 ng/g in a field-grown pepper sample. The greenness of the method was evaluated using AGREE (0.52), AGREEprep (0.58), SPMS (7.68), and ComplexGAPI, confirming substantial improvements over conventional and regulatory protocols. Overall, the method represents a miniaturized, efficient, and environmentally friendly approach for monitoring cyanotoxins in vegetables, aligned with the principles of green analytical chemistry.

Posted in CZE, Cucumber, Cyanotoxins, Electrophoresis, NADES, PID2021-127804OB-I00, PROYEXCEL_00195, Pepper, Tomato | Tagged , , , | Comments Off on A greener approach for determining cyanotoxins in vegetables: From NADES-air-assisted dispersive liquid-liquid microextraction to capillary electrophoresis–mass spectrometry analysis

Addressing chemical risks in beekeeping products: development of a combined ultrasound-assisted extraction and QuEChERS-based method for the simultaneous determination of mycotoxins and pesticides in bee pollen and honey.

Posted on 23 June, 2025 by fqm302

This work presents an ultrasound assisted extraction (UAE) combined with a QuEChERS approach, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis, for the determination of 20 mycotoxins and 10 pesticides in bee pollen and honey samples. The method was validated according to SANTE/11312/2021 guideline. Linearity was satisfactory (r > 0.99), with LODs ranging from 0.03 to 13.3 μg kg−1 and LOQs from 0.11 to 44.4 μg kg−1, except for five analytes in pollen. Trueness, expressed as apparent recoveries and evaluated at three concentration levels, was between 70 and 120 %, except for three compounds in honey and eight in pollen. RSDs for precisions were below 20 %. The method was applied to 28 honey and six pollen samples. Acetamiprid and azoxystrobin were detected in 29 % and 32 % of the honey samples, with concentration ranges of 1.26–16.2 μg kg−1 and 1.31–1.56 μg kg−1, respectively. No contaminants were found in any of the pollen samples.

Posted in Honey, Mass spectrometry, Mycotoxins, PID2021-127804OB-I00, PROYEXCEL_00195, Pollen, QuEChERS, UHPLC | Tagged , , | Comments Off on Addressing chemical risks in beekeeping products: development of a combined ultrasound-assisted extraction and QuEChERS-based method for the simultaneous determination of mycotoxins and pesticides in bee pollen and honey.

Toxic cyanopeptides monitoring in thermal spring water by capillary electrophoresis tandem mass spectrometry.

Posted on 4 February, 2025 by fqm302

Cyanobacteria are an ancient group of prokaryotes capable of oxygenic photosynthesis. Recently, thermal crises symptoms in hot springs have been associated with acute cyanopeptides poisoning. The aim of this work is to develop a fast, easy and reliable method to monitor the presence of toxic cyanopeptides in geothermal waters. The analytical method based on capillary zone electrophoresis coupled with tandem mass spectrometry (CZE-MS/MS) was developed for the simultaneous determination of 14 cyanopeptides in less than 7.5 min. A basic 50 mM ammonium acetate buffer at pH 10.2 was selected as the background electrolyte, positive electrospray ionization (ESI+) was employed for all compounds, and a salting-out assisted liquid–liquid extraction (SALLE) protocol with acetonitrile as an extraction solvent and MgSO4 as an auxiliary salting-out agent was optimized as sample treatment. Six natural hot springs in the province of Granada (Andalucía, Spain) were sampled at the beginning of the summer season (June) and at the end (September). Biomass collected at two sample points (Santa Fe and Zújar) contained cyanobacteria cells from the genera Phormidium, Leptolyngbya, and Spirulina. Nevertheless, cyanotoxins covered by this work were not found in any of the water samples analyzed. The greenness and transferability of the method was evaluated highlighting its sustainability and applicability.

Posted in CZE, Cyanotoxins, Electrophoresis, FASS, Mass spectrometry, PID2021-127804OB-I00, PROYEXCEL_00195, RED2022-134079-T, SALLE, Stacking, Thermal spring | Tagged , , , | Comments Off on Toxic cyanopeptides monitoring in thermal spring water by capillary electrophoresis tandem mass spectrometry.

Liquid chromatography-tandem mass spectrometry for the determination of multiple mycotoxins in serum through suspect screening and targeted approaches: Advancing human mycotoxin biomonitoring.

Posted on 8 January, 2025 by fqm302

Human biomonitoring (HBM) is accepted as an effective way to assess human exposure to mycotoxins and to investigate their impact on human health. Ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) has emerged as a powerful tool for the quantitative and qualitative analysis of multiple mycotoxins in various matrices. However, there is a need for selective, sensitive and high-throughput methods using limited sample volumes. Here, the potential of suspect screening and targeted analysis by UHPLC-MS/MS for HBM of multiple mycotoxins in serum samples using an optimized sample pretreatment to ensure the highest coverage and the lowest limit of detection was investigated. A suspect screening method for 144 mycotoxins and metabolites using UHPLC-high-resolution MS/MS followed by data analysis using an in-house library was developed. This library was incorporated into the UNIFI software and contained MS/MS information from the literature as well as retention times determined experimentally (when commercial standards were available) or predicted using the retention time prediction Retip R package (when no commercial standards were available). The suspect screening and targeted methods were validated using pooled human serum spiked with 41 mycotoxins and metabolites standards and applied for HBM of multiple mycotoxins in serum samples from ten Ethiopian adults. In those ten samples, five different mycotoxins were detected using the suspect screening method, including ochratoxin B, for which no standard was available, while seven different mycotoxins were detected using the targeted method. The targeted method excelled in quantitative determination due to the lower limit of quantification for most mycotoxins. In contrast, the screening method, which does not require standards for qualitative analysis, may be particularly suitable for qualitative and semiquantitative determination of a large number of mycotoxins, including metabolites, in a large number of serum samples. In view of the complementarity of the two approaches, the developed suspect screening and targeted methods for mycotoxin analysis can be regarded as promising tools to better understand mycotoxin exposure and their metabolism in humans and address the associated health risks.

Posted in Human biomonitoring, Human serum, Mass spectrometry, Mycotoxins, PROYEXCEL_00195, Protein precipitation, UHPLC | Tagged , | Comments Off on Liquid chromatography-tandem mass spectrometry for the determination of multiple mycotoxins in serum through suspect screening and targeted approaches: Advancing human mycotoxin biomonitoring.

Emerging mycotoxin occurrence in chicken feed and eggs from Algeria.

Posted on 4 December, 2024 by fqm302

Poultry farming has developed into one of Algeria’s most productive industrial farming because of the growing demand for sources of protein among Algerian society. Laying hen feed consists mainly of cereals, which can be contaminated with molds and subsequently with their secondary metabolites known as mycotoxins. These later can pose a serious danger to the production and quality of eggs in the commercial layer industry. This work focuses on the detection of emerging mycotoxins, mainly enniatins (ENNs) and beauvericin (BEA), in poultry feed and eggs from different locations in Algeria. Two different QuEChERS-based extractions were established to extract ENNs and BEA from chicken feed and eggs. The determination of mycotoxin occurrence was achieved by a UHPLC-MS/MS method using 0.1% (v/v) formic acid in water and MeOH as mobile phase, an ESI interface operating in positive mode, and a triple quadrupole mass spectrometer operating in MRM for the detection. Matrix-matched calibration curves were carried out for both matrices, obtaining good linearity (R2 > 0.99). The method performance was assessed in terms of extraction recovery (from 87 to 107%), matrix effect (from − 47 to − 86%), precision (RSD < 15%), and limits of quantitation (≤ 1.1 µg/kg for feed and ≤ 0.8 µg/kg for eggs). The analysis of 10 chicken feed samples and 35 egg samples composed of a 10-egg pool each showed that ENN B1 was the most common mycotoxin (i.e., found in 9 feed samples) with contamination levels ranging from 3.6 to 41.5 µg/kg, while BEA was detected only in one feed sample (12 µg/kg). However, eggs were not found to be contaminated with any mycotoxin at the detection limit levels. Our findings indicate that the searched mycotoxins are present in traces in feed and absent in eggs. This can be explained by the application of a mycotoxin binder. However, this does not put a stop on the conduction of additional research and ultimately setting regulations to prevent the occurrence of emerging mycotoxins.

Posted in Animal feed, Eggs, Mass spectrometry, Mycotoxins, QuEChERS, UHPLC | Tagged , , | Comments Off on Emerging mycotoxin occurrence in chicken feed and eggs from Algeria.

HILIC-MS and CE-MS as complementary analytical approaches to assess the impact of exposure to polychlorinated biphenyls on the polar serum metabolome of pigs.

Posted on 7 October, 2024 by fqm302

Hydrophilic interaction liquid chromatography (HILIC) and capillary electrophoresis (CE) coupled to mass spectrometry (MS) are two of the most widely used techniques to explore and characterize the polar metabolome. Both platforms have some peculiar characteristics that make them more suitable for studying different kind of metabolites and different parts of the metabolome. Consequently, their combined use is recommended to extend the coverage of the polar metabolome, particularly when multiple metabolic pathways are expected to be perturbed, as in toxicological studies related to exposure to environmental pollutants such as polychlorinated biphenyls (PCBs). It is unclear whether chronic exposure to low doses leads to effects on the most polar metabolites.

In this work, both HILIC-MS and CE-MS platforms have been used to characterize the polar metabolome of blood serum from pigs exposed to low-doses (20 ng/kg body weight (b.w.)/day) of Aroclor 1260 (a technical mixture of PCBs). The use of both platforms aims to cover large parts of the polar metabolome to better characterize the effects of PCB exposure. The results showed that the combined use of HILIC-MS and CE-MS effectively extended the coverage of the metabolome and the interpretability of the data, highlighting their complementarity in metabolic pathways analysis. The analysis of the polar metabolome evidenced a strong effect of exposure on amino acid metabolism, affecting 10 of the 20 proteic amino acids, and a consistent overflow in the urea cycle.

This work highlights the need to implement analytical multi-platforms to better address metabolomics studies, showing the complementarity of two widely used analytical techniques (i.e. HILIC-MS and CE-MS) to study the polar metabolome in a risk assessment framework.

Posted in Exposomics, HILIC, Metabolomics, PID2020–120020RA-I00, PROYEXCEL_00195, Pig serum, Polychlorinated biphenyls | Tagged , , , | Comments Off on HILIC-MS and CE-MS as complementary analytical approaches to assess the impact of exposure to polychlorinated biphenyls on the polar serum metabolome of pigs.

Analytical challenges and opportunities in the study of endocrine disrupting chemicals within an exposomics framework.

Posted on 29 July, 2024 by fqm302

Exposomics aims to measure human exposures throughout the lifespan and the changes they produce in the human body. Exposome-scale studies have significant potential to understand the interplay of environmental factors with complex multifactorial diseases widespread in our society and whose origin remain unclear. In this framework, the study of the chemical exposome aims to cover all chemical exposures and their effects in human health but, today, this goal still seems unfeasible or at least very challenging, which makes the exposome for now only a concept. Furthermore, the study of the chemical exposome faces several methodological challenges such as moving from specific targeted methodologies towards high-throughput multitargeted and non-targeted approaches, guaranteeing the availability and quality of biological samples to obtain quality analytical data, standardization of applied analytical methodologies, as well as the statistical assignment of increasingly complex datasets, or the identification of (un)known analytes.

This review discusses the various steps involved in applying the exposome concept from an analytical perspective. It provides an overview of the wide variety of existing analytical methods and instruments, highlighting their complementarity to develop combined analytical strategies to advance towards the chemical exposome characterization. In addition, this review focuses on endocrine disrupting chemicals (EDCs) to show how studying even a minor part of the chemical exposome represents a great challenge. Analytical strategies applied in an exposomics context have shown great potential to elucidate the role of EDCs in health outcomes. However, translating innovative methods into etiological research and chemical risk assessment will require a multidisciplinary effort. Unlike other review articles focused on exposomics, this review offers a holistic view from the perspective of analytical chemistry and discuss the entire analytical workflow to finally obtain valuable results.

Posted in Endocrine disrupting chemicals, Exposomics, PROYEXCEL_00195, Review | Tagged | Comments Off on Analytical challenges and opportunities in the study of endocrine disrupting chemicals within an exposomics framework.

Assessing human exposure to pesticides and mycotoxins: optimization and validation of a method for multianalyte determination in urine samples.

Posted on 9 April, 2024 by fqm302

Humans are exposed to an increasing number of contaminants, with diet being one of the most important exposure routes. In this framework, human biomonitoring is considered the gold standard for evaluating human exposure to chemicals. Pesticides and mycotoxins are chemicals of special concern due to their health implications. They constitute the predominant border rejection notifications for food and feed in Europe and the USA. However, current biomonitoring studies are focused on a limited number of compounds and do not evaluate mycotoxins and pesticides together. In this study, an analytical method has been developed for the determination of 30 pesticides and 23 mycotoxins of concern in urine samples. A salting-out liquid–liquid extraction (SALLE) procedure was optimized achieving recoveries between 70 and 120% for almost all the compounds and limits as lower as when QuEChERS was applied. The compounds were then determined by liquid chromatography coupled to triple quadrupole mass spectrometry. Different chromatographic conditions and analytical columns were tested, selecting a Hypersild gold aQ column as the best option. Finally, the method was applied to the analysis of 45 urine samples, in which organophosphate and pyrethroid pesticides (detection rates (DR) of 82% and 42%, respectively) and ochratoxin A and deoxynivalenol (DR of 51% and 33%, respectively) were the most detected compounds. The proposed analytical method involves the simultaneous determination of a diverse set of pesticides and mycotoxins, including their most relevant metabolites, in human urine. It serves as an essential tool for biomonitoring the presence of highly prevalent contaminants in modern society.

Posted in Biomonitoring, Mass spectrometry, Mycotoxins, PID2021-127804OB-I00, PROYEXCEL_00195, Pesticides, SALLE, UHPLC, Urine | Tagged , , | Comments Off on Assessing human exposure to pesticides and mycotoxins: optimization and validation of a method for multianalyte determination in urine samples.

Capillary electrophoresis tandem mass spectrometry to determine multiclass cyanotoxins in reservoir water and spinach samples.

Posted on 24 January, 2024 by fqm302

Cyanotoxins constitute a group of toxic secondary metabolites, the presence of which in any water body poses a major health risk. Moreover, advanced organisms such as edible plants exposed to these toxins, are a possible pathway for human exposure. Green analytical chemistry is demanding environmentally friendly analytical techniques. In this sense, we propose the use of capillary electrophoresis coupled to tandem mass spectrometry (CE-MS/MS) to determine a mixture of eight cyanotoxins belonging to three different classes: cyclic peptides (microcystin-LR, microcystin-RR and nodularin), alkaloids (cylindrospermopsin and anatoxin-a) and three isomeric non-protein amino acids (β-methylamino-l-alanine, 2,4-diaminobutyric acid and N-(2-aminoethyl)glycine). Separation was achieved by using an acidic background electrolyte consisting of 2 M formic acid and 20% acetonitrile in water. Parameters affecting MS/MS detection and the sheath-liquid interface were also studied. Finally, a combination of pH-junction, field-amplified sample stacking (FASS) and acid barrage as online preconcentration strategies, was employed to improve sensitivity and efficiency. The online preconcentration applied, in combination with a dual cartridge solid-phase extraction (SPE) system, allows to obtain limits of detection in the very low range of µg·L−1 for these multiclass cyanotoxins in reservoir water samples (from 0.005 to 0.10 µg·L−1). Furthermore, for the first time cyanotoxins are analysed in spinach samples through CE-MS/MS using the same SPE procedure, following lyophilisation and solid-liquid extraction with 6 mL 80 % aqueous MeOH.

Posted in CZE, Cyanotoxins, FASS, Mass spectrometry, PID2021-127804OB-I00, SPE, Spinach, Water, acid barrage, pH-junction | Tagged , , , | Comments Off on Capillary electrophoresis tandem mass spectrometry to determine multiclass cyanotoxins in reservoir water and spinach samples.