Unraveling biomarkers of exposure for tenuazonic acid through urinary metabolomics.

Mycotoxins are secondary metabolites produced by fungi such as Aspergillus, Alternaria, and Penicillium, affecting nearly 80% of global food crops. Tenuazonic acid (TeA) is the major mycotoxin produced by Alternaria alternata, a prevalent pathogen affecting plants, fruits, and vegetables. TeA is notably prevalent in European diets, however, TeA biomarkers of exposure and metabolites remain unknown. This research aims to bridge this knowledge-gap by gaining insights about human TeA exposure and metabolization. Nine subjects were divided into two groups. The first group received a single bolus of TeA at the Threshold of Toxicological Concern (TTC) to investigate the presence of TeA urinary biomarkers, while the second group served as a control. Sixty-nine urinary samples were prepared and analyzed using UPLC-Xevo TQ-XS for TeA quantification and UPLC-Orbitrap Exploris for polar metabolome acquisition. TeA was rapidly excreted during the first 13 h and the fraction extracted was 0.39 ± 0.22. The polar metabolome compounds effectively discriminating the two groups were filtered using Orthogonal Partial Least Squares-Discriminant Analysis and subsequently annotated (n = 122) at confidence level 4. Finally, the urinary metabolome was compared to in silico predicted TeA metabolites. Nine metabolites, including oxidized, N-alkylated, desaturated, glucuronidated, and sulfonated forms of TeA were detected.

earlyMYCO: A Pilot Mother-Child Cohort Study to Assess Early-Life Exposure to Mycotoxins-Challenges and Lessons Learned.

Early-life exposure occurs during gestation through transfer to the fetus and later, during lactation. Recent monitoring data revealed that the Portuguese population is exposed to mycotoxins, including young children. This study aimed to develop a pilot study to assess the early-life exposure to mycotoxins through a mother-child cohort, and to identify the associated challenges. Participants were recruited during pregnancy (1st trimester) and followed-up in three moments of observation: 2nd trimester of pregnancy (mother), and 1st and 6th month of the child's life (mother and child), with the collection of biological samples and sociodemographic and food consumption data. The earlyMYCO pilot study enrolled 19 mother-child pairs. The analysis of biological samples from participants revealed the presence of 4 out of 15 and 5 out of 18 mycotoxins' biomarkers of exposure in urine and breast milk samples, respectively. The main aspects identified as contributors for the successful development of the cohort were the multidisciplinary and dedicated team members in healthcare units, reduced burden of participation, and the availability of healthcare units for the implementation of the fieldwork. Challenges faced, lessons learned, and suggestions were discussed as a contribution for the development of further studies in this area.

Untargeted Metabolomics in Forensic Toxicology: A New Approach for the Detection of Fentanyl Intake in Urine Samples.

The misuse of fentanyl, and novel synthetic opioids (NSO) in general, has become a public health emergency, especially in the United States. The detection of NSO is often challenged by the limited diagnostic time frame allowed by urine sampling and the wide range of chemically modified analogues, continuously introduced to the recreational drug market. In this study, an untargeted metabolomics approach was developed to obtain a comprehensive "fingerprint" of any anomalous and specific metabolic pattern potentially related to fentanyl exposure. In recent years, in vitro models of drug metabolism have emerged as important tools to overcome the limited access to positive urine samples and uncertainties related to the substances actually taken, the possible combined drug intake, and the ingested dose. In this study, an in vivo experiment was designed by incubating HepG2 cell lines with either fentanyl or common drugs of abuse, creating a cohort of 96 samples. These samples, together with 81 urine samples including negative controls and positive samples obtained from recent users of either fentanyl or "traditional" drugs, were subjected to untargeted analysis using both UHPLC reverse phase and HILIC chromatography combined with QTOF mass spectrometry. Data independent acquisition was performed by SWATH in order to obtain a comprehensive profile of the urinary metabolome. After extensive processing, the resulting datasets were initially subjected to unsupervised exploration by principal component analysis (PCA), yielding clear separation of the fentanyl positive samples with respect to both controls and samples positive to other drugs. The urine datasets were then systematically investigated by supervised classification models based on soft independent modeling by class analogy (SIMCA) algorithms, with the end goal of identifying fentanyl users. A final single-class SIMCA model based on an RP dataset and five PCs yielded 96% sensitivity and 74% specificity. The distinguishable metabolic patterns produced by fentanyl in comparison to other opioids opens up new perspectives in the interpretation of the biological activity of fentanyl.

The Impact of COVID-19 Pandemic and Lockdown on Alcohol Consumption: A Perspective From Hair Analysis.

Introduction and Aims: The increase in stress levels, social confinement, and addiction's physical consequences play an essential role in the proliferation of drug abuse. In this context, the Covid-19 pandemic produced remarkable effects on those individuals prone to addictions, especially to alcohol. Alcohol is linked to multiple dangerous conditions such as social issues, severe medical conditions, and road accidents. The determination of ethylglucuronide (EtG) in hair is frequently performed to test and monitor chronic excessive alcohol intake conditions, as it allows differentiation among low-risk/moderate drinkers, and excessive/chronic drinkers. Our study aimed to explore hair EtG levels in a controlled population to assess the impact of Covid-19 lockdown on alcohol intake along March-May 2020. Materials and Methods: EtG levels were measured in all hair samples collected in the months following April 2020 to evaluate the behaviors related to alcohol intake along with the time frame from March to May 2020. The measured concentration distributions for each month were compared with those reported in the same month during the previous 4 years (2016-2019). The dataset was built to highlight possible differences between genders, and the different categories of alcohol consumption, separately. Results: The samples collected from April to August 2020 (500 < N <1,100 per month) showed an increase in the percentage of subjects classified as abstinent/low-risk drinkers (from 60 up to 79%) and a decrease of subjects classified as moderate and chronic drinkers (-12 and -7%, respectively) when compared to the previous 4 years. A decrease in the overall mean value of EtG in the period April-June 2020 was observed, while the EtG levels of both June and July 2020 provided an increasing trend for chronic/excessive consumers (+27 and +19% for June and July 2020, respectively). A peculiar rise in the EtG levels of moderate and chronic/excessive female consumers was observed along April-June 2020, too. Discussion and Conclusions: Behavioral and social studies generally report a decrease in alcohol consumption during the Covid-19 lockdown. However, people already suffering from drug or alcohol addictions before Covid-19 pandemic seemingly enhance their harmful behavior. Our data from April to August 2020 are consistent with both suppositions. Our observations confirm once again the utility of EtG to investigate the patterns of alcohol consumption in the population.