Simultaneous Measurement of Urinary Trimethylamine (TMA) and Trimethylamine <i>N</i>-Oxide (TMAO) by Liquid Chromatography–Mass Spectrometry
Trimethylamine (TMA) is a gut microbial metabolite—rendered by the enzymatic cleavage of nutrients containing a TMA moiety in their chemical structure. TMA can be oxidized as trimethylamine <i>N</i>-oxide (TMAO) catalyzed by hepatic flavin monooxygenases. Circulating TMAO has been demons...
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| Format: | Article |
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MDPI AG
2020-04-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/25/8/1862 |
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| author | Xun Jia Lucas J. Osborn Zeneng Wang |
| author_facet | Xun Jia Lucas J. Osborn Zeneng Wang |
| author_sort | Xun Jia |
| collection | DOAJ |
| description | Trimethylamine (TMA) is a gut microbial metabolite—rendered by the enzymatic cleavage of nutrients containing a TMA moiety in their chemical structure. TMA can be oxidized as trimethylamine <i>N</i>-oxide (TMAO) catalyzed by hepatic flavin monooxygenases. Circulating TMAO has been demonstrated to portend a pro-inflammatory state, contributing to chronic diseases such as cardiovascular disease and chronic kidney disease. Consequently, TMAO serves as an excellent candidate biomarker for a variety of chronic inflammatory disorders. The highly positive correlation between plasma TMAO and urine TMAO suggests that urine TMAO has the potential to serve as a less invasive biomarker for chronic disease compared to plasma TMAO. In this study, we validated a method to simultaneously measure urine TMA and TMAO concentrations by liquid chromatography–mass spectrometry (LC/MS). Urine TMA and TMAO can be extracted by hexane/butanol under alkaline pH and transferred to the aqueous phase following acidification for LC/MS quantitation. Importantly, during sample processing, none of the nutrients with a chemical structure containing a TMA moiety were spontaneously cleaved to yield TMA. Moreover, we demonstrated that the acidification of urine prevents an increase of TMA after prolonged storage as was observed in non-acidified urine. Finally, here we demonstrated that TMAO can spontaneously degrade to TMA at a very slow rate. |
| first_indexed | 2024-03-10T20:22:56Z |
| format | Article |
| id | doaj.art-2656871f6e86433e80a56eb71f4e739a |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T20:22:56Z |
| publishDate | 2020-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-2656871f6e86433e80a56eb71f4e739a2023-11-19T21:58:31ZengMDPI AGMolecules1420-30492020-04-01258186210.3390/molecules25081862Simultaneous Measurement of Urinary Trimethylamine (TMA) and Trimethylamine <i>N</i>-Oxide (TMAO) by Liquid Chromatography–Mass SpectrometryXun Jia0Lucas J. Osborn1Zeneng Wang2Department of Cardiovascular & Metabolic Sciences, Lerner Research Institute, Cleveland Clinic, 9500 Euclid Ave, Cleveland, OH 44195, USADepartment of Cardiovascular & Metabolic Sciences, Lerner Research Institute, Cleveland Clinic, 9500 Euclid Ave, Cleveland, OH 44195, USADepartment of Cardiovascular & Metabolic Sciences, Lerner Research Institute, Cleveland Clinic, 9500 Euclid Ave, Cleveland, OH 44195, USATrimethylamine (TMA) is a gut microbial metabolite—rendered by the enzymatic cleavage of nutrients containing a TMA moiety in their chemical structure. TMA can be oxidized as trimethylamine <i>N</i>-oxide (TMAO) catalyzed by hepatic flavin monooxygenases. Circulating TMAO has been demonstrated to portend a pro-inflammatory state, contributing to chronic diseases such as cardiovascular disease and chronic kidney disease. Consequently, TMAO serves as an excellent candidate biomarker for a variety of chronic inflammatory disorders. The highly positive correlation between plasma TMAO and urine TMAO suggests that urine TMAO has the potential to serve as a less invasive biomarker for chronic disease compared to plasma TMAO. In this study, we validated a method to simultaneously measure urine TMA and TMAO concentrations by liquid chromatography–mass spectrometry (LC/MS). Urine TMA and TMAO can be extracted by hexane/butanol under alkaline pH and transferred to the aqueous phase following acidification for LC/MS quantitation. Importantly, during sample processing, none of the nutrients with a chemical structure containing a TMA moiety were spontaneously cleaved to yield TMA. Moreover, we demonstrated that the acidification of urine prevents an increase of TMA after prolonged storage as was observed in non-acidified urine. Finally, here we demonstrated that TMAO can spontaneously degrade to TMA at a very slow rate.https://www.mdpi.com/1420-3049/25/8/1862trimethylamine (TMA)trimethylamine <i>N</i>-oxide (TMAO)biomarkerurineliquid chromatography–mass spectrometry (LC/MS) |
| spellingShingle | Xun Jia Lucas J. Osborn Zeneng Wang Simultaneous Measurement of Urinary Trimethylamine (TMA) and Trimethylamine <i>N</i>-Oxide (TMAO) by Liquid Chromatography–Mass Spectrometry Molecules trimethylamine (TMA) trimethylamine <i>N</i>-oxide (TMAO) biomarker urine liquid chromatography–mass spectrometry (LC/MS) |
| title | Simultaneous Measurement of Urinary Trimethylamine (TMA) and Trimethylamine <i>N</i>-Oxide (TMAO) by Liquid Chromatography–Mass Spectrometry |
| title_full | Simultaneous Measurement of Urinary Trimethylamine (TMA) and Trimethylamine <i>N</i>-Oxide (TMAO) by Liquid Chromatography–Mass Spectrometry |
| title_fullStr | Simultaneous Measurement of Urinary Trimethylamine (TMA) and Trimethylamine <i>N</i>-Oxide (TMAO) by Liquid Chromatography–Mass Spectrometry |
| title_full_unstemmed | Simultaneous Measurement of Urinary Trimethylamine (TMA) and Trimethylamine <i>N</i>-Oxide (TMAO) by Liquid Chromatography–Mass Spectrometry |
| title_short | Simultaneous Measurement of Urinary Trimethylamine (TMA) and Trimethylamine <i>N</i>-Oxide (TMAO) by Liquid Chromatography–Mass Spectrometry |
| title_sort | simultaneous measurement of urinary trimethylamine tma and trimethylamine i n i oxide tmao by liquid chromatography mass spectrometry |
| topic | trimethylamine (TMA) trimethylamine <i>N</i>-oxide (TMAO) biomarker urine liquid chromatography–mass spectrometry (LC/MS) |
| url | https://www.mdpi.com/1420-3049/25/8/1862 |
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