Screening gut microbial trimethylamine production by fast and cost-effective capillary electrophoresis
The present work is aimed to develop a simple, rapid, and cost-effective CE method for the determination of trimethylamine (TMA) from bacterial origin. Optimum separation of TMA from the other components of the bacterial culture was achieved using a fused silica capillary (27 cm × 75 μm ID) and a background electrolyte solution that consisted of 0.75 M formic acid at pH 2.05. Analytical characteristics of the proposed method were evaluated through the study of its specificity, linearity, precision, accuracy, robustness, and detection/quantitation limit values. The method was linear over the range 25–2000 μM (R2 = 0.9998). The LOD and LOQ were 9 μM and 27 μM, respectively. Intra-day and inter-day RSD were ≤ 0.24% and ≤ 1.3% for migration time, respectively. Intra-day and inter-day RSD for peak area were ≤ 2.44% and ≤ 3.51%, respectively. The method showed a good accuracy with recovery percentages ranging from 95.45 to 102.21%. The method was successfully applied for the determination of microbial conversion of l-carnitine to TMA. The method shows great potential in high-throughput screening applications to assess the functionality of the gut microbiota to produce TMA.
KeywordsTrimethylamine Capillary electrophoresis TMA-producing microorganism Carnitine monooxygenase Gut microbial metabolite
The authors wish to thank Prof. Jonas Bergquist from Uppsala University for the donation of the P/ACE 5010 capillary electrophoresis system and Dr. Alberto Valdés for his support during the donation of the equipment. The authors also thank Prof. Rosario Muñoz (ICTAN-CSIC) and Dr. Héctor Rodríguez (CIC bioGUNE) for their kind donation of bacteria strains.
This work was funded by the Spanish Ministry of Science, Innovation and Universities (project AGL2017-89055-R).
Compliance with ethical standards
The ethical standards of the respective institutions as directed by the institutional biosafety and ethical committees have been adhered to.
Conflict of interest
The authors declare that they have no conflict of interest.
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