Storage:Store at -20℃,6 months
Determination of Significance:
Acetoacetic acid (AcAc) is one of the important components of ketone body. It is about 20% of the total ketone body in normal people. It is a strong organic acid produced by oxidation of fatty acids. Normal content of acetoacetic acid is harmless to human body. In diabetic patients, the amount of acetoacetic acid is accumulated due to the decrease of carbohydrate use or starvation due to the metabolic disorder of sugar. Acetoacetic acid can be converted into acetone as well as acetone β- Hydroxybutyric acid.
Measurement Principle:
At pH 7.0 and 37℃, β-Hydroxybutyrate dehydrogenase (HBDH) reduced AcAc to 3-hydroxybutyrate or decarboxylated to acetone, and NADH was oxidized to NAD+. In the presence of 1-mPMS, WST-1 can react with NADH to produce water-soluble formazan with a characteristic absorption peak at 450nm. The content of AcAc can be calculated by detecting the wavelength change at 450 nm.
Self Provided:
Spectrophotometer/microplate reader, desk centrifuge, pipette, micro glass cuvette/96 well flat-bottom plate, mortar/homogenizer/cell ultrasonic crusher, ice and distilled water.
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