High-performance liquid chromatographic assay for L-glyceric acid in body fluids. Application in primary hyperoxaluria type 2.

Abstract

We describe a liquid chromatographic technique to determine L-glycerate in body fluids. The method is based on the derivatisation of the L-glycerate by incubation with lactate dehydrogenase and nicotinamide-adenine dinucleotide in the presence of phenylhydrazine. Oxidation of L-glycerate forms beta-hydroxypyruvate which is converted in turn into the related phenylhydrazone. The UV-absorbing derivative is determined using reversed-phase high performance liquid chromatography. The sensitivity was 5 mumol/l and 50 microliters of sample were required. The imprecision relative standard deviation was 4.5% and the recovery was 96.5 +/- 6.8% for L-glycerate in plasma. L-Glycerate concentrations in urine and plasma were less than 5 mumol/l in both normal individuals and patients with glycolic aciduria. In a patient with systemic oxalosis and normal plasma glycolate, plasma L-glyceric acid was 887 mumol/l.

• Authors: M Petrarulo, M Marangella, D Cosseddu, F Linari
• Journal: Clinica chimica acta; international journal of clinical chemistry (Clin Chim Acta) Vol. 211 Issue 3 Pg. 143-53 (Oct 30 1992) ISSN: 0009-8981 [Print] Netherlands
• PMID: 1458609 (Publication Type: Journal Article)

Chemical References

• Glyceric Acids
• Oxalates
• Phenylhydrazines
• Pyruvates
• phenylhydrazine
• NAD
• glyceric acid
• hydroxypyruvic acid
• Oxalic Acid
• L-Lactate Dehydrogenase

Topics

• Body Fluids (chemistry)
• Chromatography, High Pressure Liquid (methods, standards, statistics & numerical data)
• Glyceric Acids (analysis, blood, urine)
• Humans
• Hydrogen-Ion Concentration
• Hyperoxaluria, Primary (blood, urine)
• L-Lactate Dehydrogenase (metabolism)
• NAD (metabolism)
• Oxalates (blood)
• Oxalic Acid
• Phenylhydrazines (metabolism)
• Pyruvates (metabolism)
• Reference Values