TY - JOUR
T1 - Resolving Enantiomers of 2-Hydroxy Acids by Nuclear Magnetic Resonance
AU - Lane, Andrew N.
AU - Lin, Penghui
AU - Crooks, Daniel R.
AU - Linehan, W. Marston
AU - Fan, Teresa W.M.
N1 - Publisher Copyright:
© 2022 American Chemical Society.
PY - 2022/9/13
Y1 - 2022/9/13
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85137913643&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85137913643&partnerID=8YFLogxK
U2 - 10.1021/acs.analchem.2c00490
DO - 10.1021/acs.analchem.2c00490
M3 - Article
AN - SCOPUS:85137913643
SN - 0003-2700
VL - 94
SP - 12286
EP - 12291
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 36
ER -