Quantification of lactate enantiomers in human sweat samples using two-dimensional liquid chromatography
Lactate (LA) is primarily produced by the reduction of pyruvate in the human body and is crucial for energy production via anaerobic glycolysis. Although the D-LA concentration is considerably lower than that of L-LA, a significant increase in D-LA concentration alone has been reported in some disea...
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Elsevier
2025-11-01
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| Series: | Journal of Chromatography Open |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2772391725000416 |
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| author | Kazushi Mori Makoto Tsunoda |
| author_facet | Kazushi Mori Makoto Tsunoda |
| author_sort | Kazushi Mori |
| collection | DOAJ |
| description | Lactate (LA) is primarily produced by the reduction of pyruvate in the human body and is crucial for energy production via anaerobic glycolysis. Although the D-LA concentration is considerably lower than that of L-LA, a significant increase in D-LA concentration alone has been reported in some diseases. Quantifying LA enantiomers in human biofluids has the potential for disease diagnosis. Sweat has recently been recognized as a novel biological alternative to blood because it can be sampled non-invasively. Therefore, in this study, heart-cutting two-dimensional liquid chromatography (2D-LC) using a highly sensitive fluorescence detection method was developed for the analysis of LA enantiomers in small amounts of human sweat. LA was derivatized with 4-nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ) and separated from endogenous compounds using an octadecyl silica column in the first dimension. Subsequently, the NBD-PZ-LA peak was fractionated and enantiomerically separated in the second dimension on a chiral column. Sufficient linearities (R² > 0.999) were observed in the ranges of 1–100 and 10–1000 µM for NBD-PZ-D-LA and NBD-PZ-L-LA, respectively. The corresponding limits of quantification were 0.97 and 1.12 µM. The precision values were 1.04 %–12.03 %, and the accuracies were 85.6 %–100.4 %. The developed method was successfully applied to ∼5 µL of human sweat collected from five healthy subjects. The concentrations of D-LA and L-LA in sweat were 30.29 ± 20.18 µM and 23.69 ± 12.15 mM, respectively. The developed 2D-LC system should be clinically applicable to LA enantiomer analysis in human sweat as a non-invasive biomarker. |
| format | Article |
| id | doaj-art-db7cd5488ea84433b26a6a844a57723b |
| institution | Kabale University |
| issn | 2772-3917 |
| language | English |
| publishDate | 2025-11-01 |
| publisher | Elsevier |
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| series | Journal of Chromatography Open |
| spelling | doaj-art-db7cd5488ea84433b26a6a844a57723b2025-08-20T03:58:14ZengElsevierJournal of Chromatography Open2772-39172025-11-01810024310.1016/j.jcoa.2025.100243Quantification of lactate enantiomers in human sweat samples using two-dimensional liquid chromatographyKazushi Mori0Makoto Tsunoda1Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo 113-0033, JapanCorresponding author.; Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo 113-0033, JapanLactate (LA) is primarily produced by the reduction of pyruvate in the human body and is crucial for energy production via anaerobic glycolysis. Although the D-LA concentration is considerably lower than that of L-LA, a significant increase in D-LA concentration alone has been reported in some diseases. Quantifying LA enantiomers in human biofluids has the potential for disease diagnosis. Sweat has recently been recognized as a novel biological alternative to blood because it can be sampled non-invasively. Therefore, in this study, heart-cutting two-dimensional liquid chromatography (2D-LC) using a highly sensitive fluorescence detection method was developed for the analysis of LA enantiomers in small amounts of human sweat. LA was derivatized with 4-nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ) and separated from endogenous compounds using an octadecyl silica column in the first dimension. Subsequently, the NBD-PZ-LA peak was fractionated and enantiomerically separated in the second dimension on a chiral column. Sufficient linearities (R² > 0.999) were observed in the ranges of 1–100 and 10–1000 µM for NBD-PZ-D-LA and NBD-PZ-L-LA, respectively. The corresponding limits of quantification were 0.97 and 1.12 µM. The precision values were 1.04 %–12.03 %, and the accuracies were 85.6 %–100.4 %. The developed method was successfully applied to ∼5 µL of human sweat collected from five healthy subjects. The concentrations of D-LA and L-LA in sweat were 30.29 ± 20.18 µM and 23.69 ± 12.15 mM, respectively. The developed 2D-LC system should be clinically applicable to LA enantiomer analysis in human sweat as a non-invasive biomarker.http://www.sciencedirect.com/science/article/pii/S2772391725000416BiofluidsEnantiomer separationFluorescenceNon-invasive sampling |
| spellingShingle | Kazushi Mori Makoto Tsunoda Quantification of lactate enantiomers in human sweat samples using two-dimensional liquid chromatography Journal of Chromatography Open Biofluids Enantiomer separation Fluorescence Non-invasive sampling |
| title | Quantification of lactate enantiomers in human sweat samples using two-dimensional liquid chromatography |
| title_full | Quantification of lactate enantiomers in human sweat samples using two-dimensional liquid chromatography |
| title_fullStr | Quantification of lactate enantiomers in human sweat samples using two-dimensional liquid chromatography |
| title_full_unstemmed | Quantification of lactate enantiomers in human sweat samples using two-dimensional liquid chromatography |
| title_short | Quantification of lactate enantiomers in human sweat samples using two-dimensional liquid chromatography |
| title_sort | quantification of lactate enantiomers in human sweat samples using two dimensional liquid chromatography |
| topic | Biofluids Enantiomer separation Fluorescence Non-invasive sampling |
| url | http://www.sciencedirect.com/science/article/pii/S2772391725000416 |
| work_keys_str_mv | AT kazushimori quantificationoflactateenantiomersinhumansweatsamplesusingtwodimensionalliquidchromatography AT makototsunoda quantificationoflactateenantiomersinhumansweatsamplesusingtwodimensionalliquidchromatography |