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Screening gut microbial trimethylamine production by fast and cost-effective capillary electrophoresis

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Abstract

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.

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Abbreviations

BGE:

Background electrolyte

FMO:

Flavin-containing monooxygenase

TMA:

Trimethylamine

TMAO:

Trimethylamine N-oxide

References

  1. Wang Z, Klipfell E, Bennett BJ, Koeth R, Levison BS, Dugar B, et al. Gut flora metabolism of phosphatidylcholine promotes cardiovascular disease. Nature. 2011;472:57–63.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Koeth RA, Wang Z, Levison BS, Buffa JA, Org E, Sheehy BT, et al. Intestinal microbiota metabolism of L-carnitine, a nutrient in red meat, promotes atherosclerosis. Nat Med. 2013;19:576–85.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Zhu Y, Jameson E, Crosatti M, Schäfer H, Rajakumar K, Bugg TD, et al. Carnitine metabolism to trimethylamine by an unusual Rieske-type oxygenase from human microbiota. Proc Natl Acad Sci U S A. 2014;111:4268–73.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Martínez-del Campo A, Bodea S, Hamer HA, Marks JA, Haiser HJ, Turnbaugh PJ, et al. Characterization and detection of a widely distributed gene cluster that predicts anaerobic choline utilization by human gut bacteria. mBio. 2015;6:e00042–15.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Rath S, Heidrich B, Pieper DH, Vital M. Uncovering the trimethylamine-producing bacteria of the human gut microbiota. Microbiome. 2017;5:1–14.

    Article  Google Scholar 

  6. Beck W, Engelhardt H. Capillary electrophoresis of organic and inorganic cations with indirect UV detection. Chromatographia. 1992;33:313–6.

    Article  CAS  Google Scholar 

  7. Timm M, Jørgensen BM. Simultaneous determination of ammonia, dimethylamine, trimethylamine and trimethylamine-n-oxide in fish extracts by capillary electrophoresis with indirect UV-detection. Food Chem. 2002;76:509–18.

    Article  CAS  Google Scholar 

  8. Johnson DW. A flow injection electrospray ionization tandem mass spectrometric method for the simultaneousmeasurement of trimethylamine and trimethylamine N-oxide in urine. J Mass Spectrom. 2008;43:495–9.

    Article  CAS  PubMed  Google Scholar 

  9. Veeravalli S, Karu K, Phillips IR, Shephard EA. A highly sensitive liquid chromatography electrospray ionization mass spectrometry method for quantification of TMA, TMAO and creatinine in mouse urine. MethodsX 2017;4:310–319.

  10. Awwad HM, Geisel J, Obeid R. Determination of trimethylamine, trimethylamine N-oxide, and taurine in human plasma and urine by UHPLC-MS/MS technique. J Chromatogr B. 2016;1038:12–8.

    Article  CAS  Google Scholar 

  11. Hou W, Zhong D, Zhang P, Li Y, Lin M, Liu G, Yao M, Liao Q, Xie Z. A strategy for the targeted metabolomics analysis of 11 gut microbiota-host co-metabolites in rat serum, urine and feces by ultra high performance liquid chromatography-tandem mass spectrometry. J Chromatogr A 2016;1429:207–217.

  12. Vaz FAS, Neves LNO, Marques R, Sato RT, Oliveira MAL. Chromophoreasy, an Excel-based program for detection and integration of peaks from chromatographic and electromigration techniques. J Braz Chem Soc. 2016;27:1899–911.

    CAS  Google Scholar 

  13. Ahmed IAM, Maimaiti A, Mori N, Yamanaka N, Taniguchi T. Simultaneous determination of β-alanine betaine and trimethylamine in bacterial culture and plant samples by capillary electrophoresis. JAST. 2014;5:38–45.

    Article  Google Scholar 

  14. Romano KA, Vivas EI, Amador-Noguez D, Rey FE. Intestinal microbiota composition modulates choline bioavailability from diet and accumulation of the proatherogenic metabolite Trimethylamine-N-Oxide. mBio. 2015;6:e02481–14.

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Acknowledgments

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.

Funding

This work was funded by the Spanish Ministry of Science, Innovation and Universities (project AGL2017-89055-R).

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Authors and Affiliations

  1. Molecular Nutrition and Metabolism, Institute of Food Science Research (CIAL, CSIC), c/ Nicolás Cabrera 9, 28049, Madrid, Spain

    Virginia García-Cañas & Carolina Simó

  2. Área de Microbiología, Laboratorio Clínico Central, P° de Europa 34, 28702, San Sebastián de los Reyes, Madrid, Spain

    Esteban Aznar

Authors
  1. Virginia García-Cañas
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  2. Esteban Aznar
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  3. Carolina Simó
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Correspondence to Carolina Simó.

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García-Cañas, V., Aznar, E. & Simó, C. Screening gut microbial trimethylamine production by fast and cost-effective capillary electrophoresis. Anal Bioanal Chem 411, 2697–2705 (2019). https://doi.org/10.1007/s00216-019-01716-2

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  • DOI: https://doi.org/10.1007/s00216-019-01716-2

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