Simultaneous Determination of Lactate and 3-Hydroxybutyrate Enantiomers in Mouse Urine Using a Three-Dimensional Hplc System

Project: A - Government Institutionb - National Science and Technology Council

Project Details


The structures of lactate and 3-hydroxybutyrate (3HB) are similar, and both of them have a hydroxyl group, a carboxyl group, and a chiral carbon. Thus, there are two enantisomers each of lactate and 3HB. D-Lactate is the metabolite of methylglyoxal, and 3HB, the product of fatty acid metabolism, could be metabolized into methylglyoxal. D-Lactate is related diabetes mellitus, acidosis, encephalopathy, and renal damage, and the level of D-lactate in serum increased in the condition of disease or damaged tissue. D-3HB is the most of ketone bodies in human body, so it has been studied with diabetes mellitus. Recently, L-3HB was found in the serum of mice with aristolochic acid-induced nephropathy. According to the similarity of chemical structure, methylglyoxal-related metabolism, elevation in pathological state, it is important to develop a more efficient system to determine the enantiomers of lactate and 3HB. Furthermore, we could study the association among the change in enantiomers of lactate and 3HB and disease/damaged tissue. We have developed a two-dimensional high performance liquid chromatograph system (2D-HPLC) for the determination of lactate and 3HB enantiomers, however, the recovery in the urine was not enough. In this project, we are going to establish an off-line three-dimensional HPLC system, which is composed of 2D-HPLC and ion-exchange HPLC, to analyze the enantiomers of lactate and 3HB in the first year; we will set up an on-line three dimensional-high performance liquid chromatography (3D-HPLC) to separate them in the second year. Comparing to 2D-HPLC, 3D-HPLC might purify the samples and improve the efficiency of separation. Moreover, this method might be applied to study other samples which is trace or difficult to be detected.
Effective start/end date8/1/187/1/19


  • lactate
  • 3-hydroxybutyrate
  • enantiomers
  • enantiomeric separation
  • 3D-HPLC


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