Biomonitoring of mycotoxins and exposomics-informed profiling in extreme preterm infants from the NeoVIP cohort

Extremely premature infants are a highly vulnerable patient group that might be exposed to a vast number of dietary and environmental contaminants, which may impact their health trajectories. This study assessed xenobiotic exposures in 26 extremely premature infants, with a special focus on dietary mycotoxins, using a dual analytical approach combining four sensitive, targeted LC-MS/MS methods and one non-targeted high-resolution mass spectrometry (HR-MS) assay. A total of 187 matched biological samples comprising breast milk (n = 35), plasma (n = 48), urine (n = 52), and stool (n = 52) collected at 7 and 28 days post-partum were subjected to mycotoxin analysis by quantitative LC-MS/MS. In addition, the LC-HRMS workflow using data-dependent MS2 acquisition was applied to a small subset of 20 selected plasma samples to provide first insights into a broader chemical exposure characterization through suspect screening and non-targeted analysis (NTA). The targeted analyses revealed that most mycotoxin exposure biomarkers were absent in the samples. However, ochratoxin A was quantified in all breast milk and plasma samples (median values of 43 ng/L and 135 ng/L) and in 27% of urine samples (median: 20 ng/L). Citrinin was detected in 34% of breast milk and 6% of stool samples (median values of 10 ng/L and 2.5 ng/g). Alternariol monomethyl ether was detected in 94% of breast milk, 12% of stool, and 31% of tested urine specimens. In the non-targeted analysis, 356 features were annotated with confidence Levels 2a, 2b, and 3, while 20 compounds were identified at Level 1 with authentic reference standards. This included key toxicants like per- and polyfluorinated substances (PFAS), bisphenol A, cotinine, and phthalates. Albeit a small cohort, this study demonstrated that extremely premature infants were exposed to a mixture of dietary, environmental, and lifestyle-related exposures. While many of the analyzed mycotoxins, including the carcinogenic aflatoxins, were not detected and indicated low overall mycotoxin exposure, plastics- and industry-related chemicals were detected at comparatively high levels, warranting further investigation into their potential health implications.