Systematic identification of trimethoprim metabolites in lettuce

Abstract

Antibiotics are some of the most widely used drugs. Their release in the environment is of great concern since their consumption is a major factor for antibiotic resistance, one of the most important threats to human health. Their occurrence and fate in agricultural systems have been extensively investigated in recent years. Yet whilst their biotic and abiotic degradation pathways have been thoroughly researched, their biotransformation pathways in plants are less understood, such as in case of trimethoprim. Although trimethoprim has been reported in the environment, its fate in higher plants still remains unknown. A bench-scale experiment was performed and 30 trimethoprim metabolites were identified in lettuce (Lactuca sativa L.), of which 5 belong to phase I and 25 to phase II. Data mining yielded a list of 1018 ions as possible metabolite candidates, which was filtered to a final list of 87 candidates. Molecular structures were assigned for 19 compounds, including 14 TMP metabolites reported for the first time. Alongside well-known biotransformation pathways in plants, additional novel pathways were suggested, namely, conjugation with sesquiterpene lactones, and abscisic acid as a part of phase II of plant metabolism. The results obtained offer insight into the variety of phase II conjugates and may serve as a guideline for studying the metabolization of other chemicals that share a similar molecular structure or functional groups with trimethoprim. Finally, the toxicity and potential contribution of the identified metabolites to the selective pressure on antibiotic resistance genes and bacterial communities via residual antimicrobial activity were evaluated.

Keywords: Antibiotics; Conjugates; High-resolution mass spectrometry; Non-target screening; Plant metabolites.

Fig. 1

Characteristic fragmentation pathways of TMP with proposed structures for the neutral form. The main reactions included the fragmentation of methoxy groups, such as loss of CH₃ (− 15.0235), C₂H₆ (− 30.0469), CH₅O (− 33.0340), C₂H₅O (− 44.0469), and C₂H₅O₂ (− 61.0288)

Fig. 2

Proposed biotransformation pathways of TMP in lettuce. Metabolites labelled with red colour belong to the phase I, whilst the rest to the phase II of plant metabolism