Innovative Integration of LC-HRMS and In-Silico Libraries for Pharmacetical Metabolite Annonation and MALDI-HRMS Spatial Residue Analysis in Environmental Samples

Innovative Integration of LC-HRMS and In-Silico Libraries for Pharmacetical Metabolite Annonation and MALDI-HRMS Spatial Residue Analysis in Environmental Samples

Abstrakt

This study presents an integrated analytical approach to investigate pharmaceutical uptake and metabolite formation in environmental organisms exposed to contaminated soil and water. Earthworms (Eisenia fetida) and lettuce (Lactuca sativa L.) were used as model organisms to examine how veterinary and human pharmaceuticals accumulate, distribute, and transform within terrestrial system. Spatial distribution of pharmaceutical residues in earthworm tissues was visualized using optimized MALDI-HRMS imaging, revealing rapid uptake within the first day and uniform distribution throughout body tissues, suggesting combined dermal and ingestion-based uptake mechanisms. For metabolite annotation in organisms, samples were extracted using the QuEChERS method optimized for parent compounds. Subsequent analyses were performed using LC-ESI-qTOF with both DDA and DIA in positive and negative ionization modes. A high-throughput workflow was introduced, incorporating open-source tools for predicting pharmaceutical metabolite structures, generating in silico MS/MS spectral libraries, processing LC-HRMS data, and statistical evaluation. Across six parent pharmaceuticals, 3,762 potential metabolites were predicted, and 26 statistically significant metabolites were annotated in earthworms and lettuce samples. Together, these methods provide a comprehensive framework for characterizing pharmaceutical residues and their metabolites in environmental matrices, improving risk assessment and advancing understanding of pharmaceutical pollution in terrestrial ecosystems.

This study presents an integrated analytical approach to investigate pharmaceutical uptake and metabolite formation in environmental organisms exposed to contaminated soil and water. Earthworms (Eisenia fetida) and lettuce (Lactuca sativa L.) were used as model organisms to examine how veterinary and human pharmaceuticals accumulate, distribute, and transform within terrestrial system. Spatial distribution of pharmaceutical residues in earthworm tissues was visualized using optimized MALDI-HRMS imaging, revealing rapid uptake within the first day and uniform distribution throughout body tissues, suggesting combined dermal and ingestion-based uptake mechanisms. For metabolite annotation in organisms, samples were extracted using the QuEChERS method optimized for parent compounds. Subsequent analyses were performed using LC-ESI-qTOF with both DDA and DIA in positive and negative ionization modes. A high-throughput workflow was introduced, incorporating open-source tools for predicting pharmaceutical metabolite structures, generating in silico MS/MS spectral libraries, processing LC-HRMS data, and statistical evaluation. Across six parent pharmaceuticals, 3,762 potential metabolites were predicted, and 26 statistically significant metabolites were annotated in earthworms and lettuce samples. Together, these methods provide a comprehensive framework for characterizing pharmaceutical residues and their metabolites in environmental matrices, improving risk assessment and advancing understanding of pharmaceutical pollution in terrestrial ecosystems.

Pharmaceutical uptake, Metabolite formation, Earthworm, Lettuce, MALDI-HRMS imaging, LC-ESI-qTOF, Terrestrial ecosystem, mass spectrometry

Pharmaceutical uptake, Metabolite formation, Earthworm, Lettuce, MALDI-HRMS imaging, LC-ESI-qTOF, Terrestrial ecosystem, mass spectrometry

FUČÍK, J.; FUČÍK, S.; JAROŠOVÁ, R.; REXROTH, S.; BAUMEISTER, A.; NIEHOFF, A.; HAMPLOVÁ, M.; NAVRKALOVÁ, J.; ZLÁMALOVÁ GARGOŠOVÁ, H.; MRAVCOVÁ, L.

05.06.2025

73rd Conference on Mass Spectrometry and Allied Topics (ASMS 2025)

Baltimore (USA)

4

https://www.asms.org/conferences/annual-conference/annual-conference-homepage