Introduction Recently we have proposed the use of catalase coupled with aldehyde dehydrogenase to measure the action of H2O2 producing oxidoreductases (1,2). Meanwhile this principle has been extensively evaluated for the determination of uric acid in human serum and urine samples (3). The main advantages over comparable methods are: the reaction is completed in a few minutes and the use of NAD"1" or NADP* (3) permits the direct calculation of the substrate concentration from the absorbance value without reference to a standard solution. This is especially relevant if the preparation of primary standard solutions is problematical. In the following study the cholesterol concentration was determined with cholesterol oxidase (4) using the aldehyde dehydrogenase principle: cholesterol ester + H2O cholesterol + Q2 H2O2 Methanol acetaldehyde + NAD(P)* J. Clin. Chem. Clin. Biochem. / VoL 14,1976 / No. 8 cholesterol esterase cholesterol oxidäse catalase cholesterol + fatty acid cholest-4-en-3-one + H202 acetaldehyde + 2 H2O aldehyde dehydrogenase acetate+ NAD(P)H + H+ 29A Haeckel and Perlick: New enzymatic determination of cholesterol Mix; record absorbance after reaction has come to the end and extrapolate to absorbance value before the addition of cholesterol oxidase () Cholesterol oxidase (20 M!) Materials Most reagents required
Clinical Chemistry and Laboratory Medicine – de Gruyter
Published: Jan 1, 1976
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