LC-MS/MS methodology for simultaneous determination of patulin and citrinin in urine and plasma applied to a pilot study in colorectal cancer patients

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