Quantification of lactate from various metabolic pathways and quantification issues of lactate isotopologues and isotopmers

Abstract 13C-labeled glucose combined with chromatography and mass spectrometry enables us to decipher the percentage of lactate generated from various metabolic pathways. We showed that lactate derived from glycolysis, pentose phosphate pathway, Krebs cycle, and other sources accounted for 82–90%,...

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Autores principales:	Wei Zhang, Cheng Guo, Kezhi Jiang, Minfeng Ying, Xun Hu
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/f3c9ce25c6a34fad9c17cdf43f83d2f0
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spelling	oai:doaj.org-article:f3c9ce25c6a34fad9c17cdf43f83d2f02021-12-02T11:40:14ZQuantification of lactate from various metabolic pathways and quantification issues of lactate isotopologues and isotopmers10.1038/s41598-017-08277-32045-2322https://doaj.org/article/f3c9ce25c6a34fad9c17cdf43f83d2f02017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-08277-3https://doaj.org/toc/2045-2322Abstract 13C-labeled glucose combined with chromatography and mass spectrometry enables us to decipher the percentage of lactate generated from various metabolic pathways. We showed that lactate derived from glycolysis, pentose phosphate pathway, Krebs cycle, and other sources accounted for 82–90%, 6.0–11%, 0.67–1.8% and 1.5–7.9%, respectively, depending on different types of cells. When using glucose isotopomers ([1-13C]-, [3-13C]-, [4-13C]-, and [6-13C]glucose) or isotopologues ([1,2-13C2]- and [1,2,3-13C3]glucose) for tracing, the ratio of lactate derived from glucose carbon 1, 2, 3 over 4, 5, 6 via glycolysis varied significantly, ranging from 1.6 (traced with [1,2-13C2]glucose) to 0.85 (traced with [6-13C]glucose), but the theoretical ratio should be 1. The odd results might be caused by intramolecular 13C, which may significantly affect lactate fragmentation under tandem mass spectrometry condition, leading to erroneous quantification. Indeed, the fragmentation efficiency of [U-13C]lactate, [2,3-13C]lactate, and [3-13C]lactate were 1.4, 1.5 and 1.2 folds higher than lactate, respectively, but [1-13C]lactate was similar to lactate, suggesting that carbon-13 at different positions could differentially influence lactate fragmentation. This observed phenomenon was inconsistent with the data based on theoretical calculation, according to which activation energies for all lactate isotopomers and isotopologues are nearly identical. The inconsistency suggested a need for further investigation. Our study suggests that calibration is required for quantifying metabolite isotopolugues and isotopomers.Wei ZhangCheng GuoKezhi JiangMinfeng YingXun HuNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-12 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Wei Zhang Cheng Guo Kezhi Jiang Minfeng Ying Xun Hu Quantification of lactate from various metabolic pathways and quantification issues of lactate isotopologues and isotopmers
description	Abstract 13C-labeled glucose combined with chromatography and mass spectrometry enables us to decipher the percentage of lactate generated from various metabolic pathways. We showed that lactate derived from glycolysis, pentose phosphate pathway, Krebs cycle, and other sources accounted for 82–90%, 6.0–11%, 0.67–1.8% and 1.5–7.9%, respectively, depending on different types of cells. When using glucose isotopomers ([1-13C]-, [3-13C]-, [4-13C]-, and [6-13C]glucose) or isotopologues ([1,2-13C2]- and [1,2,3-13C3]glucose) for tracing, the ratio of lactate derived from glucose carbon 1, 2, 3 over 4, 5, 6 via glycolysis varied significantly, ranging from 1.6 (traced with [1,2-13C2]glucose) to 0.85 (traced with [6-13C]glucose), but the theoretical ratio should be 1. The odd results might be caused by intramolecular 13C, which may significantly affect lactate fragmentation under tandem mass spectrometry condition, leading to erroneous quantification. Indeed, the fragmentation efficiency of [U-13C]lactate, [2,3-13C]lactate, and [3-13C]lactate were 1.4, 1.5 and 1.2 folds higher than lactate, respectively, but [1-13C]lactate was similar to lactate, suggesting that carbon-13 at different positions could differentially influence lactate fragmentation. This observed phenomenon was inconsistent with the data based on theoretical calculation, according to which activation energies for all lactate isotopomers and isotopologues are nearly identical. The inconsistency suggested a need for further investigation. Our study suggests that calibration is required for quantifying metabolite isotopolugues and isotopomers.
format	article
author	Wei Zhang Cheng Guo Kezhi Jiang Minfeng Ying Xun Hu
author_facet	Wei Zhang Cheng Guo Kezhi Jiang Minfeng Ying Xun Hu
author_sort	Wei Zhang
title	Quantification of lactate from various metabolic pathways and quantification issues of lactate isotopologues and isotopmers
title_short	Quantification of lactate from various metabolic pathways and quantification issues of lactate isotopologues and isotopmers
title_full	Quantification of lactate from various metabolic pathways and quantification issues of lactate isotopologues and isotopmers
title_fullStr	Quantification of lactate from various metabolic pathways and quantification issues of lactate isotopologues and isotopmers
title_full_unstemmed	Quantification of lactate from various metabolic pathways and quantification issues of lactate isotopologues and isotopmers
title_sort	quantification of lactate from various metabolic pathways and quantification issues of lactate isotopologues and isotopmers
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/f3c9ce25c6a34fad9c17cdf43f83d2f0
work_keys_str_mv	AT weizhang quantificationoflactatefromvariousmetabolicpathwaysandquantificationissuesoflactateisotopologuesandisotopmers AT chengguo quantificationoflactatefromvariousmetabolicpathwaysandquantificationissuesoflactateisotopologuesandisotopmers AT kezhijiang quantificationoflactatefromvariousmetabolicpathwaysandquantificationissuesoflactateisotopologuesandisotopmers AT minfengying quantificationoflactatefromvariousmetabolicpathwaysandquantificationissuesoflactateisotopologuesandisotopmers AT xunhu quantificationoflactatefromvariousmetabolicpathwaysandquantificationissuesoflactateisotopologuesandisotopmers
_version_	1718395679501451264

Quantification of lactate from various metabolic pathways and quantification issues of lactate isotopologues and isotopmers

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