Resolving Enantiomers of 2-Hydroxy Acids by Nuclear Magnetic Resonance

Andrew N. Lane, Penghui Lin, Daniel R. Crooks, W. Marston Linehan, Teresa W.M. Fan

Research output: Contribution to journalArticlepeer-review

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 languageEnglish
Pages (from-to)12286-12291
Number of pages6
JournalAnalytical Chemistry
Volume94
Issue number36
DOIs
StatePublished - 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

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