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Finding unknown endocrine disrupting compounds through target pull-down assay filtration, effect direct analysis and ultra-high resolution mass spectrometry for a comprehensive efficient workflow.

Project description

Novel assays will efficiently identify toxic chemicals in environmental and human samples

The endocrine system creates hormones and releases them into the bloodstream, where they travel to target cells to modulate important processes related to growth, metabolism and reproduction, among other things. The pituitary gland, pancreas and ovaries are just a few of the endocrine system's organs. Endocrine-disrupting compounds can have very serious effects, and their increasing presence in the environment is a growing public health concern. The EU-funded PullEd-MS project is working on streamlining the assays used to determine which of the many compounds in a sample are responsible for the toxic response generated in humans. Various methods will also be employed to help determine the level of exposure of individuals.

Objective

There are thousands of anthropogenic compounds in general use across the globe and every year more chemicals are created. For every compound, that finds its way into the environment multiple degradation compounds can result due to environmental processes. These changes alter the toxicological properties and chemical mobility creating potentially unpredicted effects. Environmental monitoring using effect direct analysis (EDA) to identify toxicological endpoints combined with mass spectrometry (MS) to detect the most abundant active compounds is an established methodology. The latest MS instruments the ultra-high resolution MS (UHRMS) have allowed full scan acquisition to become the cutting edge in the detection and identification of unknown compounds. However, knowing which EDA to apply is still a challenge and when UHRMS scans detect thousands of compounds identifying which compounds caused the toxicological response is a challenge. This MSC proposal aims to develop the use of pull-down assays linked to specific nuclear receptors (NR) such as estrogen receptor α as a filter to bind only those chemicals with an affinity for that specific protein. By then applying only the target specific EDA and fractionation to identify the most bioactive fraction before UHRMS a more focused workflow for compound detection based on effect can be developed. This workflow will be tested using samples of varying complexity including wastewater, sediment, and human biological samples. These workflows will then be tested using water collected from the Global Water Network to identify novel compounds of toxicological effect in the environment and using human urine from pregnant mothers collected from the CELSPAC-TNG cohort for identification of human exposure. In developing this comprehensive methodology, the sample will have specific toxicological effects identified as well as chemical investigation providing a thorough NR specific determination of both known and unknown compounds.

Fields of science

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Coordinator

Masarykova univerzita
Net EU contribution
€ 242 521,68
Address
Zerotinovo namesti 9
601 77 Brno
Czechia

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Region
Česko Jihovýchod Jihomoravský kraj
Activity type
Higher or Secondary Education Establishments
Links
Total cost
€ 242 521,68

Partners (1)