Pharmaceuticals released into the aquatic and soil environments can be absorbed by plants and soil organisms, potentially leading to the formation of unknown metabolites that may negatively affect these organisms or contaminate the food chain. The aim of this study was to identify pharmaceutical metabolites through a triplet approach for metabolite structure prediction (software-based predictions, literature review, and known common metabolic pathways), followed by generating in silico mass spectral libraries and applying various mass spectrometry modes for untargeted LC-qTOF analysis. Therefore, Eisenia fetida and Lactuca sativa were exposed to a pharmaceutical mixture (atenolol, enrofloxacin, erythromycin, ketoprofen, sulfametoxazole, tetracycline) under hydroponic and soil conditions at environmentally relevant concentrations. Samples collected at different time points were extracted using QuEChERS and analyzed with LC-qTOF in data-dependent (DDA) and data-independent (DIA) acquisition modes, applying both positive and negative electrospray ionization. The triplet approach for metabolite structure prediction yielded a total of 3762 pharmaceutical metabolites, and an in silico mass spectral library was created based on these predicted metabolites. This approach resulted in the identification of 26 statistically significant metabolites (p < 0.05), with DDA + and DDA - outperforming DIA modes by successfully detecting 56/67 sample type:metabolite combinations. Lettuce roots had the highest metabolite count (26), followed by leaves (6) and earthworms (2). Despite the lower metabolite count, earthworms showed the highest peak intensities, closely followed by roots, with leaves displaying the lowest intensities. Common metabolic reactions observed included hydroxylation, decarboxylation, acetylation, and glucosidation, with ketoprofen-related metabolites being the most prevalent, totaling 12 distinct metabolites. In conclusion, we developed a high-throughput workflow combining open-source software with LC-HRMS for identifying unknown metabolites across various sample types.

• Keywords
• High-resolution mass spectrometry, In silico spectral library, Liquid chromatography, Metabolite identification in Eisenia fetida and Lactuca sativa, Pharmaceuticals, Software prediction,
• MeSH
• Chromatography, Liquid methods   MeSH
• Mass Spectrometry methods   MeSH
• Soil Pollutants analysis   metabolism   MeSH
• Pharmaceutical Preparations metabolism   chemistry   analysis   MeSH
• Oligochaeta * metabolism   chemistry   MeSH
• Computer Simulation MeSH
• Lactuca * metabolism   chemistry   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Soil Pollutants MeSH
• Pharmaceutical Preparations MeSH

The presence of human and veterinary pharmaceuticals (PhACs) in the environment poses potential risks. To comprehensively assess these risks, robust multiresidual analytical methods are essential for determining a broad spectrum of PhAC classes in various environmental compartments (soil, plants, and soil organisms). This study optimized extraction methods for analyzing over 40 PhACs from various matrices, including soil, lettuce, and earthworms. A four-step ultrasonic extraction method with varying extraction conditions and subsequent solid phase extraction was developed for soil samples. QuEChERS methods were optimized for extracting PhACs from lettuce and earthworm samples, addressing a literature gap in these less-studied matrices. The quantification of PhACs in soil, lettuce, and earthworm extracts was performed using a single LC-MS/MS method. Following thorough method validation, earthworms and lettuce were exposed to a mixture of 27 pharmaceuticals in a soil environment. The method validation results demonstrated the robustness of these methods for a broad spectrum of PhACs. Specifically, 29 out of 42 PhACs were extracted with an average efficiency > 50% and RSD < 30% from the soil; 40 out of 42 PhACs exhibited average efficiency > 50% and %RSD < 30% from the earthworms, while 39 out of 42 PhACs showed average efficiency > 50% and RSD < 30% from the lettuce. Exposure experiments confirmed the viability of these methods for quantifying a diverse range of PhACs in different environmental compartments. This study presents three thoroughly validated methods for determining more than 40 PhACs in diverse matrices, enabling a comprehensive assessment of PhAC dissemination in the environment.

• Keywords
• Liquid chromatography, Mass spectrometry, Multiresidual analytical methods, Pharmaceutical pollution, QuEChERS, Solid-phase extraction,
• MeSH
• Chromatography, Liquid MeSH
• Solid Phase Extraction MeSH
• Soil Pollutants * analysis   MeSH
• Pharmaceutical Preparations analysis   MeSH
• Oligochaeta * MeSH
• Soil * chemistry   MeSH
• Lactuca * chemistry   MeSH
• Tandem Mass Spectrometry MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Soil Pollutants * MeSH
• Pharmaceutical Preparations MeSH
• Soil * MeSH

In recent years, there has been increasing concern over the environmental risks of the so called "Emerging pollutants (EPs)" that are defined as synthetic or naturally occurring chemicals that are not commonly monitored in the environment but which have the potential to enter the environment and cause adverse ecological and (or) human health effects [...].

• Publication type
• Editorial MeSH

Albendazole (ABZ), a widely used anthelmintic drug, enters the environment mainly via livestock excrements. To evaluate the environmental impact of ABZ, the knowledge of its uptake, effects and metabolism in all non-target organisms, including plants, is essential. The present study was designed to identify the metabolic pathway of ABZ and to test potential ABZ phytotoxicity in fodder plant alfalfa, with seeds and in vitro regenerants used for these purposes. Alfalfa was chosen, as it may meet manure from ABZ-treated animals in pastures and fields. Alfalfa is often used as a feed of livestock, which might already be infected with helminths. The obtained results showed that ABZ did not inhibit alfalfa seed germination and germ growth, but evoked stress and a toxic effect in alfalfa regenerants. Alfalfa regenerants were able to uptake ABZ and transform it into 21 metabolites. UHPLC-MS/MS analysis revealed three new ABZ metabolites that have not been described yet. The discovery of the parent compound ABZ together with the anthelmintically active and instable metabolites in alfalfa leaves shows that the contact of fodder plants with ABZ-containing manure might represent not only a danger for herbivorous invertebrates, but also may cause the development of ABZ resistance in helminths.

• Keywords
• UHPLC-MS/MS, anthelmintics, drug metabolism, drug phytotoxicity, drugs in the environment,
• MeSH
• Albendazole pharmacology   MeSH
• Anthelmintics pharmacology   MeSH
• Germination MeSH
• Animal Feed MeSH
• Medicago sativa drug effects   growth & development   metabolism   MeSH
• Metabolome * MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Albendazole MeSH
• Anthelmintics MeSH

In recent years interest has grown in the occurrence and the effects of pharmaceuticals in the environment. The aim of this work is to evaluate the risk of fertilizing crops with manure from livestock treated with anthelmintics. The present study was designed to follow the fate of the commonly used anthelmintic drug, ivermectin (IVM) and its metabolites in soybeans (Glycine max (L.) Merr.), a plant that is grown and consumed world-wide for its high content of nutritional and health-beneficial substances. In vitro plantlets and soybean plants, cultivated in a greenhouse, were used for this purpose. Our results showed the uptake of IVM and its translocation to the leaves, but not in the pods and the beans. Four IVM metabolites were detected in the roots, and one in the leaves. IVM exposure decreased slightly the number and weight of the beans and induced changes in the activities of antioxidant enzymes. In addition, the presence of IVM affected the proportion of individual isoflavones and reduced the content of isoflavones aglycones, which might decrease the therapeutic value of soybeans. Fertilization of soybean fields with manure from IVM-treated animals appears to be safe for humans, due to the absence of IVM in beans, the food part of plants. On the other hand, it could negatively affect soybean plants and herbivorous invertebrates.

• Keywords
• anthelmintics, antioxidant enzymes, biotransformation, drug metabolites, isoflavonoids,
• MeSH
• Antioxidants metabolism   MeSH
• Antiparasitic Agents pharmacology   MeSH
• Biological Transport MeSH
• Glycine max drug effects   growth & development   metabolism   MeSH
• Isoflavones metabolism   MeSH
• Ivermectin pharmacology   MeSH
• Plant Roots drug effects   growth & development   metabolism   MeSH
• Plant Leaves drug effects   growth & development   metabolism   MeSH
• Seeds drug effects   growth & development   metabolism   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Antioxidants MeSH
• Antiparasitic Agents MeSH
• Isoflavones MeSH
• Ivermectin MeSH