LC/MS (TOF) Analysis of DL-α-Hydroxyglutaric Acid Enantiomers on Astec® CHIROBIOTIC® R, Fast Separation
CONDITIONS
column
Astec CHIROBIOTIC® R, 10 cm x 4.6 mm I.D., 5μm particles (13022AST)
mobile phase
5:95, (water:methanol) with 0.3% acetic acid and 0.1% ammonium hydroxide
flow rate
1 mL/min
pressure
1700 psi (117 bar)
column temp.
25 °C
detector
MS, ESI(-) TOF, SIM, m/z 147.03, split 100:1
injection
10 μL
sample
DL-a-Hydroxyglutaric acid, 1 mg/mL in 50:50, water:methanol
Description
Analysis Note
Chiral 2-hydroxyglutarates (2-OHG) are important molecular signatures of both healthy and diseased biological cells, their specific biochemical pathways and inborn errors of metabolism. The chiral differentiation and quantification of D-2-OHG and L-2-OHG is key for characterizing neurometabolic disorders like the 2-hydroxyglutaric acidurias that cause neurological impairment early in life. In this application, an Astec® CHIROBIOTIC® R column run under polar ionic mobile phase conditions achieved a rapid separation of the D and L enantiomers of 2-OHG under conditions amenable for sensitive LC-MS/MS detection.
Legal Information
Astec is a registered trademark of Merck KGaA, Darmstadt, Germany
CHIROBIOTIC is a registered trademark of Sigma-Aldrich Co. LLC