Abstract
Biologically important 2-hydroxy carboxylates such as lactate, malate, and 2-hydroxyglutarate exist in two enantiomeric forms that cannot be distinguished under achiral conditions. The D and L (or R, S) enantiomers have different biological origins and functions, and therefore, there is a need for a simple method for resolving, identifying, and quantifying these enantiomers. We have adapted and improved a chiral derivatization technique for nuclear magnetic resonance (NMR), which needs no chromatography for enantiomer resolution, with greater than 90% overall recovery. This method was developed for 2-hydroxyglutarate (2HG) to produce diastereomers resolvable by column chromatography. We have applied the method to lactate, malate, and 2HG. The limit of quantification was determined to be about 1 nmol for 2HG with coefficients of variation of less than 5%. We also demonstrated the method on an extract of a renal carcinoma bearing an isocitrate dehydrogenase-2 (IDH2) variant that produces copious quantities of 2HG and showed that it is the D enantiomer that was exclusively produced. We also demonstrated in the same experiment that the lactate produced in the same sample was the L enantiomer.
Original language | English |
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Pages (from-to) | 12286-12291 |
Number of pages | 6 |
Journal | Analytical Chemistry |
Volume | 94 |
Issue number | 36 |
DOIs | |
State | Published - Sep 13 2022 |
Bibliographical note
Funding Information:This work was supported in part by 5P20GM121327, Shared Resource(s) of the University of Kentucky Markey Cancer Center P30CA177558, and endowment funds to T.W.-M.F. and A.N.L. We thank Alyssa Clarke for assistance with some experiments.
Publisher Copyright:
© 2022 American Chemical Society.
ASJC Scopus subject areas
- Analytical Chemistry