Determination of Urinary Mycotoxin Biomarkers Using a Sensitive Online Solid Phase Extraction-UHPLC-MS/MS Method

Abstract

In the course of assessing the human exposure to mycotoxins, biomarker-based approaches have proven to be important tools. Low concentration levels, complex matrix compositions, structurally diverse analytes, and the large size of sample cohorts are the main challenges of analytical procedures. For that reason, an online solid phase extraction-ultra high-performance liquid chromatography-tandem mass spectrometry (online SPE-UHPLC-MS/MS) method was developed, allowing for the sensitive, robust, and rapid analysis of 11 relevant mycotoxins and mycotoxin metabolites in human urine. The included spectrum of analytes comprises aflatoxin M1 (AFM1), altenuene (ALT), alternariol monomethyl ether (AME), alternariol (AOH), citrinin (CIT) and its metabolite dihydrocitrinone (DH-CIT), fumonisin B1 (FB1), ochratoxin A (OTA), and zearalenone (ZEN) as well as α- and β-zearalenol (α- and β-ZEL). Reliable quantitation was achieved by means of stable isotope dilution, except for ALT, AME and AOH using matrix calibrations. The evaluation of method performance displayed low limits of detection in the range of pg/mL urine, satisfactory apparent recovery rates as well as high accuracy and precision during intra- and interday repeatability. Within the analysis of Zimbabwean urine samples (n = 50), the applicability of the newly developed method was shown. In addition to FB1 being quantifiable in all analyzed samples, six other mycotoxin biomarkers were detected. Compared to the occurrence rates obtained after analyzing the same sample set using an established dilute and shoot (DaS) approach, a considerably higher number of positive samples was observed when applying the online SPE method. Owing to the increased sensitivity, less need of sample handling, and low time effort, the herein presented online SPE approach provides a valuable contribution to human biomonitoring of mycotoxin exposure.

Assessment of multiple mycotoxin exposure and its association with food consumption: a human biomonitoring study in a pregnant cohort in rural Bangladesh

“…The applied method is based on a dilute and shoot approach, covering a broad spectrum of compounds but focusing on the previously detected highly relevant mycotoxins OTA and CIT / HO-CIT. For other analytes, a moderate sensitivity was accepted as no more sensitive methods with comparable efficiency were available at the time of sample analysis (Schmidt et al 2021a ). As a consequence, the LODs of some mycotoxins were comparably high, allowing only the detection of very high exposure scenarios but not the baseline level of mycotoxin uptake present in Bangladesh.…”

“…Current state-of-the-art biomarker analysis methods enable relatively easy and objective detection and quantification of major mycotoxins from bio-samples (Escrivá et al 2017 ; Ezekiel et al 2022 ; Šarkanj et al 2018 ). Examples of major mycotoxins investigated include aflatoxins (AFs), fumonisin B1 (FB1), ochratoxin A (OTA), citrinin (CIT), zearalenone (ZEN), and deoxynivalenol (DON) (Al-Jaal et al 2019 ; Gerding et al 2015 , 2014 ; Schmidt et al 2021a , 2021b ; Solfrizzo et al 2014 ; Warth et al 2014 ). Biomonitoring of exposure biomarkers is also considered advantageous, as it provides information on the internal dose of exposure via all routes, including dietary intake and exposures via the respiratory tract and the skin (Degen 2011 ; Fromme et al 2016 ).…”

Citrinin-Induced Hepatotoxicity in Mice Is Regulated by the Ca2+/Endoplasmic Reticulum Stress Signaling Pathway

“…Mycotoxins are toxic secondary metabolites produced by molds, contaminating more than 25% of the world's agricultural products (e.g., grains, fruits, and nuts) and causing serious threat to human and animal health. Thus, mycotoxins draws the wide attention of the international society as a global problem [1,2].…”

Assessment of Human Exposure to Five Alternaria Mycotoxins in China by Biomonitoring Approach

“…The signal suppression/enhancements (SSEs) for AOH, AME and TeA were 54.4%, 70.0% and 58.7%, respectively, verifying significant matrix effects. Thus, matrix-matched calibration curves were established as an alternative accurate quantitation strategy for the lack of the isotope-labelling standards [28]. As urine composition may vary among individuals, it should bear in mind that the uncertainties related to the calculations of the concentrations of Alternaria mycotoxins might be relatively higher.…”