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. 2019 Dec 20;9:19568. doi: 10.1038/s41598-019-55951-9

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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LC-MS measurement of pyruvate and acetate concentrations. Reactions were carried out with 300 µM each of pyruvate and H₂O₂ in DPBS at 37 °C. At the indicated time, aliquots of the reaction mixture were removed for LC-MS analysis. (A,B) LC-MS chromatograms showing the detection of pyruvate and acetate, respectively, in the samples at the indicated times. (C,D) Concentrations of pyruvate and acetate, respectively, were calculated according to standard curves, graphed vs. time, and fit to second-degree polynomial equations. (E) Data are graphed as the sum of pyruvate and acetate concentrations vs. time, showing an unchanged total concentration over time. (F) Data are graphed as the inverse concentration of pyruvate vs. time and fit to a linear equation. The slope of the line indicates the k of the reaction. Data were obtained from 3 separate experiments and presented as mean ± SD.