Chromatographic separation and continuously referenced, on-line monitoring of creatine kinase isoenzymes by use of an immobilized-enzyme microreactor
We describe a new concept in continuously referenced monitoring of the isoenzyme activities of creatine kinase (EC 2.7.3.2) after liquid-chromatographic separation. After separation on a diethylaminoethyl-Sephacel column, the three isoenzymes of creatine kinase undergo a series of upled enzyme reactions, ultimately resulting in the formation of ultraviolet-detectable NADPH. A major advantage of this detection system is the immobilized-enzyme microreactor (2 x 17 mm), which may be removed and stored refrigerated when not in use. A split-stream configuration allows self-blanking of endogenous ultraviolet-absorbing constituents in authentic sera samples, which would otherwise make definitive diagnosis and quantitation difficult or impossible. This detection system is applicable to the automated analysis of creatine kinase isoenzymes in the clinical laboratory. 5 figures; 42 references.
- Research Organization:
- Oak Ridge National Lab., TN
- OSTI ID:
- 6781907
- Journal Information:
- Clin. Chem.; (United States), Vol. 24:8; Conference: Annual symposium on advanced analytical concepts for the clinical laboratory, Oak Ridge, TN, USA, 16 Mar 1978
- Country of Publication:
- United States
- Language:
- English
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ORGANIC
PHYSICAL AND ANALYTICAL CHEMISTRY
ADP
CHEMICAL REACTIONS
CHEMICAL REACTORS
MONITORING
COENZYMES
CHEMICAL REACTION YIELD
GLUCOSE
ISOENZYMES
CATALYTIC EFFECTS
QUANTITATIVE CHEMICAL ANALYSIS
PHOSPHOCREATINE
BLOOD SERUM
CATALYSIS
CREATINE
DIAGNOSTIC TECHNIQUES
ENZYMES
HEART
LIQUID COLUMN CHROMATOGRAPHY
METABOLISM
PHOSPHOTRANSFERASES
SEPARATION PROCESSES
ULTRAVIOLET SPECTRA
ALDEHYDES
AMINO ACIDS
BODY
CARBOHYDRATES
CARBOXYLIC ACIDS
CARDIOVASCULAR SYSTEM
CHEMICAL ANALYSIS
CHROMATOGRAPHY
HEXOSES
MONOSACCHARIDES
NUCLEOTIDES
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
ORGANS
SACCHARIDES
SPECTRA
TRANSFERASES
YIELDS
400105* - Separation Procedures