RESUMEN
Liquid chromatography combined with high-resolution mass spectrometry (LC-HRMS) is a frequently applied technique for suspect screening (SS) and non-target screening (NTS) in metabolomics and environmental toxicology. However, correctly identifying compounds based on SS or NTS approaches remains challenging, especially when using data-independent acquisition (DIA). This study assessed the performance of four HRMS-spectra identification tools to annotate in-house generated data-dependent acquisition (DDA) and DIA HRMS spectra of 32 pesticides, veterinary drugs, and their metabolites. The identification tools were challenged with a diversity of compounds, including isomeric compounds. The identification power was evaluated in solvent standards and spiked feed extract. In DDA spectra, the mass spectral library mzCloud provided the highest success rate, with 84% and 88% of the compounds correctly identified in the top three in solvent standard and spiked feed extract, respectively. The in silico tools MSfinder, CFM-ID, and Chemdistiller also performed well in DDA data, with identification success rates above 75% for both solvent standard and spiked feed extract. MSfinder provided the highest identification success rates using DIA spectra with 72% and 75% (solvent standard and spiked feed extract, respectively), and CFM-ID performed almost similarly in solvent standard and slightly less in spiked feed extract (72% and 63%). The identification success rates for Chemdistiller (66% and 38%) and mzCloud (66% and 31%) were lower, especially in spiked feed extract. The difference in success rates between DDA and DIA is most likely caused by the higher complexity of the DIA spectra, making direct spectral matching more complex. However, this study demonstrates that DIA spectra can be used for compound annotation in certain software tools, although the success rate is lower than for DDA spectra.
RESUMEN
Data on the fate of chemical substances in the environment after e.g. manure application is mandatory input for risk assessment in perspective of a more circular biobased economy. Such fate studies include a persistence study to determine a half-life value and a mobility study. It is recognized that not only the native substance should be considered, but that also degradation products should be included that might exert a similar effect as the native substance. We report a tiered fate study strategy that starts with a persistence study. For non-persistent substances a study is performed to determine if degradation products have a similar effect as the native compound. If so, a procedure using high resolution mass spectrometry is suggested to identify the potentially active degradation products. Based on the outcomes, substances are divided into three categories: (I) persistent, (II) degradable to inactive products or (III) degradable to active products. Even though the priority is with category I and III, for all substances and possible degradation products a mobility study is proposed. The fate strategy is successfully applied to ten antimicrobially active substances originating from the tetracyclines, sulfonamides, diaminopyrimidines, fluoroquinolones, macrolides and lincosamides. The fluoroquinolones, tetracyclines and trimethoprim were relatively persistent. The sulfonamides, macrolides and lincomycin (the latter also depending on soil type) degraded relatively quickly. Tylosin A proved to degrade to antimicrobially active degradation products which were tentitatively identified as tylosin C, tylosin A acid, tylosin B acid and tylosin C acid.
Asunto(s)
Contaminantes del Suelo , Suelo , Antibacterianos , Estiércol , Contaminantes del Suelo/análisis , Tetraciclinas/análisis , TilosinaRESUMEN
RATIONALE: The separation of isomeric compounds with major differences in their physiochemical and pharmacokinetic properties is of particular importance in pharmaceutical R&D. However, the structural assessment and separation of these compounds with current analytical techniques and methods are still a challenge. In this study, we describe strategies to separate the various structural and stereo-isomers. METHODS: The separation of ten structural and stereo-isomers was investigated using Trapped and Travelling Wave ion mobility spectrometry (TIMS and TWIMS). Different strategies including adduct ion formation with Na, Li, Ag and Cs as well as fragmentation before and after the ion mobility cell were applied to separate the isomeric compounds. RESULTS: All the counter ions (in particular Na) strongly coordinated with the test analytes in all the IMS systems. The highest resolving power was achieved for the sodium and lithium adducts using TIMS-time-of-flight (TOF). However, some separation was attained on a Synapt HDMS system with its unique potential to monitor the ion mobility of the product ions. The elution order of the adduct ions was the same in all instruments, in which, unexpectedly, the para-substituted isomer of the [M + Na]+ species had the lowest collision cross section followed by the meta- and ortho-isomers. CONCLUSIONS: The formation of adduct ions could facilitate the separation of structural and even stereo-isomers by generating different molecular conformations. In addition, fragmenting isomers before or after the ion mobility cell is a valuable strategy to separate and also to assess the structures of adducts and different conformers.