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Patulin (PAT) is a common mycotoxin in fruit products, especially in apples and apple-based products. The European Commission has set maximum levels for PAT in food. Nevertheless, worrying PAT levels were recently recorded in diverse foods across the world. Therefore, a worldwide follow-up of PAT-levels in foods should be considered. Because of PAT's high probability in food products, the toxicological implications for humans need to be addressed as well. Recent studies proved adverse health effects of PAT, such as hepatotoxicity, gastrointestinal alterations and inmunotoxicity. In comparison to the toxicity of other mycotoxins such as ochratoxin A, PAT's immunotoxicity can be even more outspoken destructive. In addition, PAT is a low-molecular-weight and highly polar molecule, resulting in many analytical challenges for its detection. As the analytical techniques are continuously improving, PAT determination in multi-mycotoxin analysis has advanced using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) during the last year. Finally, the presence and toxicity of PAT requires a biomarker method to assess its exposure among the population. To date, however, there is no information regarding PAT biomarkers in biological samples. This short review highlights the PAT-occurrence profile, toxicological discoveries and analytical challenges of 2014 until to date.

Biomarker-driven research has been proposed as a successful method to assess the exposure of individuals to xenobiotics, including mycotoxins, through estimation of their metabolites in biological fluids. A methodology to determine patulin (PAT) and citrinin (CIT) in human urine and plasma using liquid chromatography coupled to tandem mass spectrometry was developed and validated in the present study. Selectivity/specificity, linearity, limit of detection and quantification, apparent recovery, intraday- and interday-precision and measurement uncertainty were investigated for validation purposes. Finally, the method was used to analyze human urine (n = 100) and plasma (n = 100) case-control samples, where 50 samples originated from colorectal cancer patients and 50 from age/sex-matched controls. This case-control study revealed that PAT was not detected in urine samples, however occurred in 25% of the analysed plasma samples with an average concentration of 11.62 ± 6.67 ng/mL in the positive samples. CIT was found in urine samples (74%) and plasma samples (36%) with average concentrations in the positive samples of 0.45 ± 0.24 ng/mL and 0.49 ± 0.2 ng/mL respectively. No statistically significant difference of PAT and CIT concentration among colorectal cancer and control patients (p > 0.05) was observed. Highlights: Development and validation of an LC-MS/MS method to determine patulin and citrinin in biological fluids; Patulin was detected in plasma (25%) for the first time; Citrinin was present in 74% of Tunisian urine samples; Citrinin was present in 36% of Tunisian plasma samples. The authors acknowledge the support of the Tunisian Ministry of Education and Scientific Research, the Toxicology and Environment Research Laboratory LR12SP07 (CAMUR, Tunis, Tunisia) and MYTOXSOUTH (https://mytoxsouth.org) (Ghent University and Global Minds International Thematic Network). Dr. Arnau Vidal expresses thanks to The Research Foundation Flanders(Wetenschappelijk Oderzoek- Vlaanderen, FWO) for the postdoctoral grant (12Y1719N). info:eu-repo/semantics/publishedVersion