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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Nature Portfolio
2017-08-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-017-08277-3 |
| _version_ | 1818841813751955456 |
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| 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 |
| collection | DOAJ |
| 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. |
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| format | Article |
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| last_indexed | 2024-12-19T04:32:03Z |
| publishDate | 2017-08-01 |
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| series | Scientific Reports |
| spelling | doaj.art-f3c9ce25c6a34fad9c17cdf43f83d2f02022-12-21T20:35:51ZengNature PortfolioScientific Reports2045-23222017-08-017111210.1038/s41598-017-08277-3Quantification of lactate from various metabolic pathways and quantification issues of lactate isotopologues and isotopmersWei Zhang0Cheng Guo1Kezhi Jiang2Minfeng Ying3Xun Hu4Cancer Institute (a Key Laboratory For Cancer Prevention & Intervention, China National Ministry of Education), The Second Affiliated Hospital, Zhejiang University School of MedicineCancer Institute (a Key Laboratory For Cancer Prevention & Intervention, China National Ministry of Education), The Second Affiliated Hospital, Zhejiang University School of MedicineKey Laboratory of Organosilicon Chemistry and Material Technology, Hangzhou Normal UniversityCancer Institute (a Key Laboratory For Cancer Prevention & Intervention, China National Ministry of Education), The Second Affiliated Hospital, Zhejiang University School of MedicineCancer Institute (a Key Laboratory For Cancer Prevention & Intervention, China National Ministry of Education), The Second Affiliated Hospital, Zhejiang University School of MedicineAbstract 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.https://doi.org/10.1038/s41598-017-08277-3 |
| spellingShingle | 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 Scientific Reports |
| title | 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_short | 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 |
| url | https://doi.org/10.1038/s41598-017-08277-3 |
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