High-performance liquid chromatographic assay for L-glyceric acid in body fluids. Application in primary hyperoxaluria type 2.
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.
Petrarulo
M
M
Renal Stone Laboratory, Ospedale Mauriziano Umberto I, Turin, Italy.
Marangella
M
M
Cosseddu
D
D
Linari
F
F
D001826:Body Fluids / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000592:standards / Q000706:statistics & numerical data; D005988:Glyceric Acids / Q000032:analysis* / Q000097:blood / Q000652:urine; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006960:Hyperoxaluria, Primary / Q000097:blood* / Q000652:urine; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D009243:NAD / Q000378:metabolism; D010070:Oxalates / Q000097:blood; D019815:Oxalic Acid; D010659:Phenylhydrazines / Q000378:metabolism; D011773:Pyruvates / Q000378:metabolism; D012016:Reference Values
D005988:Glyceric Acids; D010070:Oxalates; D010659:Phenylhydrazines; D011773:Pyruvates; C030299:phenylhydrazine; D009243:NAD; C042971:glyceric acid; C012375:hydroxypyruvic acid; D019815:Oxalic Acid; D007770:L-Lactate Dehydrogenase