COD 11805 COD 11505 COD 11506 COD 11539 CHOLESTEROL

1 x 50 mL 1 x 200 mL 1 x 500 mL 1x1L

STORE AT 2-8ºC
Reagents for measurement of cholesterol concentration CHOLESTEROL
Only for in vitro use in the clinical laboratory CHOLESTEROL OXIDASE/PEROXIDASE

PRINCIPLE OF THE METHOD QUALITY CONTROL

Free and esterified cholesterol in the sample originates, by means of the coupled reactions It is recommended to use the Biochemistry Control Serum level I (cod. 18005, 18009 and 18042)
described below, a coloured complex that can be measured by spectrophotometry1,2. and II (cod. 18007, 18010 and 18043) to verify the performance of the measurement procedure.
chol. esterase Each laboratory should establish its own internal Quality Control scheme and procedures for
Cholesterol ester + H2O Cholesterol + Fatty acid corrective action if controls do not recover within the acceptable tolerances.
chol. oxidase
Cholesterol + ½ O2 + H2O Cholestenone + H2O2 METROLOGICAL CHARACTERISTICS
peroxidase
2 H2O2 + 4 – Aminoantipyrine + Phenol Quinoneimine + 4 H2O − Detection limit: 0.3 mg/dL = 0.008 mmol/L
− Linearity limit: 1000 mg/dL = 26 mmol/L. For higher values dilute sample 1/2 with distilled
CONTENTS water and repeat measurement.
COD 11805 COD 11505 COD 11506 COD 11539 − Repeatibility (within run):
A. Reagent 1 x 50 mL 1 x 200 mL 1 x 500 mL 1x1L Mean Concentration CV n
S. Standard 1 x 5 mL 1 x 5 mL 1 x 5 mL 1 x 5 mL
121 mg/dL = 3.13 mmol/L 1.1 % 20
COMPOSITION 257 mg/dL = 6.66 mmol/L 0.9 % 20

A. Reagent. Pipes 35 mmol/L, sodium cholate 0.5 mmol/L, phenol 28 mmol/L, cholesterol − Reproducibility (run to run):
esterase > 0.2 U/mL, cholesterol oxidase > 0.1 U/mL, peroxidase > 0.8 U/mL,
4-aminoantipyrine 0.5 mmol/L, pH 7.0. Mean Concentration CV n

S. Cholesterol Standard. Cholesterol 200 mg/dL (5,18 mmol/L). Aqueous primary standard. 121 mg/dL = 3.13 mmol/L 1.9 % 25
257 mg/dL= 6.66 mmol/L 1.0 % 25
STORAGE
− Trueness: Results obtained with this reagent did not show systematic differences when
Store at 2-8ºC. compared with reference reagents (Note 2). Details of the comparison experiments are
Reagent and Standard are stable until the expiry date shown on the label when stored tightly available on request.
closed and if contaminations are prevented during their use. − Interferences: Lipemia (triglycerides 10 g/L) does not interfere. Bilirubin (>10 mg/dL) and
Indications of deterioration: hemoglobin (>5 g/L) may affect the results. Other drugs and substances may interfere4.
− Reagent: Presence of particulate material, turbidity, absorbance of the blank over 0.200 at These metrological characteristics have been obtained using an analyzer. Results may vary if a
500 nm (1 cm cuvette). different instrument or a manual procedure are used.
− Standard: Presence of particulate material, turbidity.
DIAGNOSTIC CHARACTERISTICS
REAGENT PREPARATION Cholesterol is a steroid of high molecular weight and possesses the cyclopentanophenanthrene
Reagent and Standard are provided ready to use. skeleton. Dietary cholesterol is partially absorbed and it is also synthesized by the liver and
other tissues. Cholesterol is transported in plasma by lipoproteins. It is excreted unchanged into
ADDITIONAL EQUIPMENT bile or after transformation to bile acids.
− Thermostatic water bath at 37ºC Increased total cholesterol values are associated with a progressively escalating risk of
atherosclerosis and coronary artery disease5,6.
− Analyzer, spectrophotometer or photometer able to read at 500 ± 20 nm
Clinical diagnosis should not be made on the findings of a single test result, but should integrate
SAMPLES both clinical and laboratory data.

Serum or plasma collected by standard procedures. NOTES

Cholesterol is stable for 7 days at 2-8ºC. Heparin, EDTA, oxalate and fluoride may be used as 1. This reagent may be used in several automatic analysers. Instructions for many of them are
anticoagulants. available on request.
PROCEDURE 2. Calibration with the provided aqueous standard may cause a matrix related bias, specially in
some analyzers. In these cases, it is recommended to calibrate using a serum based
1. Bring the Reagent to room temperature. standard (Biochemistry Calibrator, cod. 18011 and 18044).
2. Pipette into labelled test tubes: (Note 1)
BIBLIOGRAPHY
Blank Standard Sample
1. Allain CC, Poon LS, Chan CSG, Richmond W and Fu PC. Enzymatic determination of total
Cholesterol Standard (S)  10 µL  serum cholesterol. Clin Chem 1974; 20: 470-475.
Sample   10 µL
Reagent (A) 1.0 mL 1.0 mL 1.0 mL
2. Meiattini F, Prencipe L, Bardelli F, Giannini G and Tarli P. The 4-hydroxybenzoate/4-
aminophenazone chromogenic system used in the enzymic determination of serum
3. Mix thoroughly and incubate the tubes for 10 minutes at room temperature (16-25ºC) or for 5 cholesterol. Clin Chem 1978; 24: 2161-2165.
minutes at 37ºC. 3. National Cholesterol Education Program Expert Panel. Third report of the National
4. Measure the absorbance (A) of the Standard and Sample at 500 nm against the Blank. The Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and
colour is stable for at least 2 hours. Treatment of High Blood Cholesterol in Adults (ATP III). NIH Publication. Bethesda: National
Heart, Lung, and Blood Institute; 2001.
CALCULATIONS 4. Young DS. Effects of drugs on clinical laboratory tests, 5th ed. AACC Press, 2000.
The cholesterol concentration in the sample is calculated using the following general formula: 5. Tietz Textbook of Clinical Chemistry and Molecular Diagnostics, 4th ed. Burtis CA, Ashwood
A Sample ER, Bruns DE. WB Saunders Co, 2005.
x C Standard = C Sample 6. Friedman and Young. Effects of disease on clinical laboratory tests, 4th ed. AACC Press,
A Standard
2001.
If the Cholesterol Standard provided has been used to calibrate (Note 2):
A Sample x 200 = mg/dL cholesterol
A Standard x 5.18 = mmol/L cholesterol

REFERENCE VALUES
The following uniform cut-off points have been established by the US National Cholesterol
Education Program and have also been adopted in many other countries for the evaluation of
coronary artery disease risk3.
Up to 200 mg/dL = 5.2 mmol/L Desirable
200-239 mg/dL = 5.2-6.21 mmol/L Borderline High
> 240 mg/dL = > 6.24 mmol/L High

M11505i-20 BioSystems S.A. Costa Brava, 30. 08030 Barcelona (Spain) 04/2011
Quality System certified according to
EN ISO 13485 and EN ISO 9001 standards