Reagent for measuring alanine aminotransferase activity
View Patent ↗A reagent for measuring an alanine aminotransferase activity comprising L-alanine, 2-oxoglutaric acid, lactate dehydrogenase, and reduced nicotinamide adenine dinucleotide, characterized by further comprising a substance having an activity of inhibiting lactate dehydrogenase activity is disclosed. Further, a method for measuring an alanine aminotransferase activity, characterized by bringing a sample to be analyzed, which may contain alanine aminotransferase, into contact with L-alanine, 2-oxoglutaric acid, lactate dehydrogenase, reduced nicotinamide adenine dinucleotide, and a substance having an activity of inhibiting a lactate dehydrogenase activity is disclosed. According to the reagent and method, an increase in the reagent blank reaction, i.e., an increase in the initial absorbance, can be suppressed.
1. In a method for measuring an alanine aminotransferase activity in a sample, said method comprising the steps of: bringing the sample into contact with L-alanine, 2-oxoglutaric acid, lactate dehydrogenase, and reduced nicotinamide adenine dinucleotide,
generating pyruvic acid and L-glutamic acid from L-alanine and 2-oxoglutaric acid as substrates by alanine aminotransferase,
converting pyruvic acid and reduced nicotinamide adenine dinucleotide to lactic acid and oxidized nicotinamide adenine dinucleotide by lactate dehydrogenase, and
measuring a decreased amount of reduced nicotinamide adenine dinucleotide or an increased amount of oxidized nicotinamide adenine dinucleotide,
the improvement comprising bringing the sample into contact with L-alanine, 2-oxoglutaric acid, lactate dehydrogenase, and reduced nicotinamide adenine dinucleotide in the presence of a substance having an activity of inhibiting a lactate dehydrogenase activity, said substance being selected from the group consisting of oxamic acid, oxalic acid, oxalacetic acid, pyruvic acid, phosphoenolpyruvic acid, sodium dodecyl sulfate, lactic acid, and hydroxyglutaric acid, and salts thereof, and the concentration of said substance in the measuring system being a concentration exhibiting the lactate dehydrogenase activity which does not affect the measurement of the alanine aminotransferase activity.
2. The method according to claim 1 , wherein the substance having an activity of inhibiting a lactate dehydrogenase activity is oxamic acid or a salt thereof.
3. The method according to claim 2 , wherein a concentration of oxamic acid or a salt thereof is 0.005 to 5 mmol/L as a final concentration in the measuring system.
4. The method for measuring an alanine aminotransferase activity according to claim 1 , wherein a concentration of lactate dehydrogenase is 100 U/L or more as a final concentration in the measuring system.