Total Cholesterol

Total Cholesterol

Cromatest Reagent

PRINCIPLE

Enzymatic colorimetric method
ENDPOINT

This method for the measurement of total cholesterol in serum involves the use of three enzymes: cholesterol esterase (CE), cholesterol oxidase (CO) and peroxidase (POD). In the presence of the former the mixture of phenol and 4-aminoantipyrine (4-AA) are condensed by hydrogen peroxide to form a quinoneimine dye proportional to the concentration of cholesterol in the sample.

REAGENT COMPOSITION

4 x 100 ml
R1 Monoreagent:  PIPES 200 mmol/L pH 7.0, sodium cholate
1 mmol/L, cholesterol esterase > 250 U/L, cholesterol
oxidase > 250 U/L, peroxidase > 1 KU/L, 4-aminoantipyrine
0.33 mmol/L, phenol 4 mmol/L, non-ionic tensioactives 2
g/L (w/v). Biocides.

CAL Cholesterol standard: Cholesterol 200 mg/dL (5.18 mmol/L).
Organic matrix based primary standard. Concentration
value is traceable to Standard Reference Material 909b.  1 x 3 ml

STORAGE AND STABILITY

Store at 2-8ºC.

REAGENT PREPARATION
The Monoreagent and the Standard are ready-to-use.

SAMPLES
Serum, EDTA or heparinized plasma free of hemolysis.
Cholesterol in serum or plasma is stable up to 5 days at 2-8ºC and
for a few months at –20ºC.

MATERIALS REQUIRED

• Photometer or colorimeter capable of measuring absorbance
  at 500 ± 10 nm.
• Constant temperature incubator set at 37ºC.
• Pipettes to measure reagent and samples.

Total Protein Reagents per box