Journal
ANALYTICAL CHEMISTRY
Volume 93, Issue 11, Pages 4809-4817Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.0c03769
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Funding
- Gordon and Betty Moore Foundation [GBMF5503]
- Simons Foundation International [409923]
- National Science Foundation [1656311]
- Direct For Biological Sciences
- Division Of Integrative Organismal Systems [1656311] Funding Source: National Science Foundation
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Utilizing pre-extraction derivatization and ultrahigh performance liquid chromatography electrospray ionization tandem mass spectrometry, targeted dissolved metabolites in seawater and saline culture media were successfully detected and quantified. This quantitative method has been optimized for 73 common metabolites, with 50 requiring derivatization for quantification due to low extraction efficiencies.
Dissolved metabolites serve as nutrition, energy, and chemical signals for microbial systems. However, the full scope and magnitude of these processes in marine systems are unknown, largely due to insufficient methods, including poor extraction of small, polar compounds using common solid-phase extraction resins. Here, we utilized pre-extraction derivatization and ultrahigh performance liquid chromatography electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS) to detect and quantify targeted dissolved metabolites in seawater and saline culture media. Metabolites were derivatized with benzoyl chloride by their primary and secondary amine and alcohol functionalities and quantified using stable isotope-labeled internal standards (SIL-ISs) produced from C-13(6)-labeled benzoyl chloride. We optimized derivatization, extraction, and sample preparation for field and culture samples and evaluated matrix-derived biases. We have optimized this quantitative method for 73 common metabolites, of which 50 cannot be quantified without derivatization due to low extraction efficiencies. Of the 73 metabolites, 66 were identified in either culture media or seawater and 45 of those were quantified. This derivatization method is sensitive (detection limits = pM to nM), rapid (similar to 5 min per sample), and high throughput.
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