Sara Palmieria, Fabiola Eugelioa, Francesco Della Vallea, Federico Fantia, Manuel Sergib, Marcello Mascinia, Michele Del Carloa, Dario Compagnonea

aDepartment of Biosciences and Agri-Food and Environmental Technologies, University of Teramo, 64100 Teramo, TE
bDepartment of Chemistry, La Sapienza University of Rome, 00185 Roma, RM

Phytoprostanes (PhytoPs) are non-enzymatic products of lipid peroxidation derived from α-linolenic acid. While PhytoPs are not essential for the metabolic activity of living cells, they are considered components of the oxidative damage detection systems and act as excellent biomarkers for oxidative degradation in plant-based foods. Among these PhytoPs, in the literature is reported a limited number of analytical methods resulting from homemade chemical synthesis few of them are available in commerce. Various methodologies are employed to qualitatively and quantitatively detect PPs in foods. Recent advancements in the identification of PhytoPs primarily employing UHPLC-MS/MS, allowing for rapid and precise measurement [2]. 

Additionally, methods integrating biosensors and high-resolution mass spectrometry are emerging, promising to further refine quantification in complex matrices like food and biological samples. The extraction of PhytoPs can be limited by factors such as the complexity of the matrix, the stability of the compounds, and the efficiency of the extraction method. In recent years, researchers introduced molecularly imprinted polymers (MIPs) extraction for target compounds or for class selective extraction [3]. In this work, an alternative strategy for fast MIPs synthesis was proposed and apply to the PhytoPs compounds, in commercial food samples analysed by UHPLC-MS/MS, as targeted method. A Orbitrap IQ-X tribrid mass spectrometry was employed to confirm the results using the high resolution and mass accuracy, allowing for precise identification of PhytoPs. MIPs were synthesized via a low-cost and rapid (5 min) sonochemical free-radical polymerization, using 4-cyclopentene-1,3-dione as a dummy template. 

To this aim, we tested methacrylic acid (MAA) and methacrylamide (MMA) as monomers, using ethylene glycol dimethacrylate as the cross-linker and 2,2 azobisisobutyronitrile as the initiator. MIP MAA- based in dispersion solid phase extraction (dSPE) yielded the best result than the other. The evaluation of the performance of MAA-MIP-dSPE were performed with 5 of isoprostanes standard (8R-Isoprostane (8-ISOR), 8S isoprostane (8-ISOS), 8-keto-isoprostane (8 KETO), 11β- isoprostane (11β) and 5IPF2A-VI), obtaining satisfactory recovery and good reproducibility. Proposed MIP was tested on food samples proving the selective extraction of PhytoP forms by their putative identification.

Acknowledgements: this research/publication/volume was funded the European Union – Next Generation EU. Project Code: ECS00000041; Project CUP: C43C22000380007; Project Title: Innovation, digitalization and sustainability for the diffused economy in Central Italy - VITALITY;

Keywords: phytoprostanes, molecularly imprinted polymers, UHPLC-MS/MS, foods

References

1. Lipan L, Collado-González J, et al., JAFC, 68 (2020): 7214-7225. DOI: 10.1021/acs.jafc.0c02268 

2. Mersni M, Zhou B, et al., Fitoterapia 172 (2024): 105717. DOI: 10.1016/j.fitote.2023.105717 

3. Chapuis F, Pichon V, et al, J. Chromatogr. A 999(2003), 23-33. DOI:10.1016/S0021-9673(03)00552-1