Technical Summary: Acetoacetate Analysis (Bulletin ref. TB078716C)
 

A fast enzymatic analysis for plasma Acetoacetate.
 

Name:
Code:
Size:
 

Acetoacetate
GMRD-180
20 tests; 140 analyser cycles. The number of tests can be doubled by using half-volumes (micro-method).
 

Instruments:
 

All GM7 Series analysers
 

Principle:

i) In the presence of excess NADH and the enzyme 3-hydroxybutyrate dehydrogenase (3-HBDH), acetoacetate is reduced to 3-hydroxybutyrate in a brief pre-reaction:
 

ii) Under the conditions of the assay, the rate of oxidation of excess NADH by peroxidase (POD) is inversely proportional to the plasma acetoacetate concentration:
 

A sample blank allows for interference by pyruvate and other endogenous NADH consuming reactions.
 

Samples:
Sample volume:
Linearity:
Precision:

Accuracy:
 

Plasma, serum
50 µl
1000 µmol/L (100 µg/ml)
Typical within-run CV = 2 - 3 % @ 500 µmol/L

(Recovery data) 97 - 106 %
 

Reagent Stability:

Shelf life 6 months unopened when stored at 0 - 5°C. Reconstituted NADH reagent and standard, 10 - 14 days if stored at 0 - 5°C.