A radioenzymatic ultramicro method applicable to the measurement of a wide range of metabolites
Journal Article
·
· Anal. Biochem.; (United States)
A radioenzymatic method suitable for the estimation of nanomole amounts of a wide range of metabolites is described. The method involves the conversion of (1-/sup 14/C)..cap alpha..-ketoglutarate to (1-/sup 14/C)glutamate by NADH or NADPH generated stoichiometrically in preliminary reactions specific for the metabolite under assay. The labeled glutamate is then decarboxylated with glutamate decarboxylase and the liberated /sup 14/CO/sub 2/ trapped and measured by scintillation counting. Examples of the variety of substrates that can be estimated by this method include procedures for the determination of lactate, glucose, ammonia, pyruvate, malate, sorbitol, and NAD(P)H. However, most enzymatic spectrophotometric or fluorimetric metabolite assays should be readily adaptable for radioenzymatic assay. In validation of the method, glucose and lactate were measured in whole blood or plasma, and the values obtained by the radioenzymatic method were found to correlate well with established clinical procedures. The method lends itself to both clinical and basic biochemical applications, can be used over a very wide range of metabolite concentrations, and is sufficiently sensitive to allow the measurement of an amount of metabolite as low as 0.25 nmol.
- Research Organization:
- Flinders Univ. of South Australia, Bedford Park
- OSTI ID:
- 5580560
- Journal Information:
- Anal. Biochem.; (United States), Journal Name: Anal. Biochem.; (United States) Vol. 118:2; ISSN ANBCA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBON 14
CARBON ISOTOPES
CARBON-CARBON LYASES
CARBOXY-LYASES
CARBOXYLIC ACIDS
CHEMICAL REACTIONS
COENZYMES
COUNTING TECHNIQUES
DECARBOXYLASES
DECARBOXYLATION
DICARBOXYLIC ACIDS
ENZYMES
EVEN-EVEN NUCLEI
GLUTAMIC ACID
GLUTARIC ACID
ISOTOPES
LIGHT NUCLEI
LYASES
METABOLITES
NADH2
NADP
NUCLEI
NUCLEOTIDES
ORGANIC ACIDS
ORGANIC COMPOUNDS
RADIOASSAY
RADIOISOTOPES
REDUCTION
SCINTILLATION COUNTING
USES
YEARS LIVING RADIOISOTOPES
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBON 14
CARBON ISOTOPES
CARBON-CARBON LYASES
CARBOXY-LYASES
CARBOXYLIC ACIDS
CHEMICAL REACTIONS
COENZYMES
COUNTING TECHNIQUES
DECARBOXYLASES
DECARBOXYLATION
DICARBOXYLIC ACIDS
ENZYMES
EVEN-EVEN NUCLEI
GLUTAMIC ACID
GLUTARIC ACID
ISOTOPES
LIGHT NUCLEI
LYASES
METABOLITES
NADH2
NADP
NUCLEI
NUCLEOTIDES
ORGANIC ACIDS
ORGANIC COMPOUNDS
RADIOASSAY
RADIOISOTOPES
REDUCTION
SCINTILLATION COUNTING
USES
YEARS LIVING RADIOISOTOPES