- Acknowledgements: This research was funded Project PID2021-1278040BI00 funded by Spanish MCIN/ AEI /10.13039/501100011033 and by “ERDF A way of making Europe” and PROYEXCEL_00195 funded by Consejería de Universidad, Investigación e Innovación, Junta de Andalucía. RCM thanks to the predoctoral contract from Project PID2021-1278040B-I00.
- Authors: R. Carmona-Molero, L. Carbonell-Rozas, F.J. Lara, A.M. García-Campaña, M. del Olmo-Iruela.
- Reference: Microchemical Journal 218 (2025) 115244.
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.
