CHOLESTEROL LDL

COD 11585 1 x 80 mL
DIRECT
STORE AT 2-8ºC
Reagents for measurement of LDL cholesterol concentration CHOLESTEROL LDL DIRECT
Only for in vitro use in the clinical laboratory
DETERGENT

PRINCIPLE OF THE METHOD QUALITY CONTROL

A specific detergent solubilizes the cholesterol from high density lipoproteins (HDL), very low It is recommended to use the Lipid Control Serum level I (cod. 18040) and II (cod. 18041) to
density lipoproteins (VLDL) and chylomicrons. The cholesterol esters are broken down by verify the performance of the measurement procedure.
cholesterol esterase and cholesterol oxidase in a non-color forming reaction. The second Each laboratory should establish its own internal Quality Control scheme and procedures for
detergent, present in the reagent B, solubilizes cholesterol from low density lipoproteins (LDL) in corrective action if controls do not recover within the acceptable tolerances.
the sample. The LDL cholesterol is then spectrophotometrically measured by means of the
coupled reactions described below1. METROLOGICAL CHARACTERISTICS
chol. esterase
Cholesterol esters + H2O Cholesterol + Fatty acid − Detection limit: 0.28 mg/dL = 0.007 mmol/L
chol. oxidase − Linearity limit: 990 mg/dL = 25.6 mmol/L.
Cholesterol + ½ O2 + H2O Cholestenone + H2O2 − Repeatibility (within run):
peroxidase
Mean Concentration CV n
2 H2O2 + 4 – Aminoantipyrine + DSBmT Quinoneimine + 4 H2O
146 mg/dL = 3.78 mmol/L 0.7 % 20
CONTENTS AND COMPOSITION 210 mg/dL = 5.43 mmol/L 0.6 % 20

A. Reagent. 1 x 60 mL. MES buffer > 30 mmol/L, cholesterol esterase < 1.5 U/mL, cholesterol − Reproducibility (run to run):
oxidase < 1.5 U/mL, 4-aminoantipyrine 0.5 mmol/L, ascorbate oxidase < 3.0 U/L, peroxidase
Mean Concentration CV n
> 1 U/mL, detergent, pH 6.3.
143 mg/dL = 3.70 mmol/L 2.0 % 40
B. Reagent. 1 x 20 mL. MES buffer > 30 mmol/L, N,N-bis(4-sulfobutyl)-m-toluidine (DSBmT)
207 mg/dL = 5.35 mmol/L 1.7 % 40
1 mmol/L, detergent, pH 6.3.
S. HDL/LDL calibrator. Human serum. Concentration is given on the label. − Trueness: Results obtained with this reagent did not show systematic differences when
Components from human origin have been tested and found to be negative for the presence of compared with reference reagents . Details of the comparison experiments are available on
antibodies anti-HIV and anti-HCV, as well as for Hbs antigen. However, they should be handled request.
cautiously as potentially infectious. − Interferences: Lipemia (triglycerides 12.9 g/L), hemoglobin (5 g/L) and bilirubin (20 mg/dL) do
not interfere. Other drugs and substances may interfere3.
STORAGE
These metrological characteristics have been obtained using an analyzer. Results may vary if a
Store at 2-8ºC. different instrument or a manual procedure are used.
Reagents are stable until the expiry date shown on the label when stored tightly closed and if
contaminations are prevented during their use. DIAGNOSTIC CHARACTERISTICS
Indications of deterioration: Presence of particulate material, turbidity. LDL is the main lipoprotein transporting cholesterol from liver to tissues.
REAGENT PREPARATION Increased plasma LDL-cholesterol concentrations are positively correlated with the incidence of
atherosclerosic diseases, basis of myocardial infarction and cerebrovascular accidents4,5.
Reagents are provided ready to use.
HDL/LDL calibrator: reconstitute with 1.0 mL of distilled water . Stable for 1 week at 2-8ºC or for There are several disease states or environmental influences associated with increased levels of
2 months at –18ºC when frozen in aliquots. LDL-cholesterol: nephrosis, diabetes, obesity, some drugs and smoking4,5.
Clinical diagnosis should not be made on the findings of a single test result, but should integrate
ADDITIONAL EQUIPMENT both clinical and laboratory data.
− Thermostatic water bath at 37ºC
NOTES
− Analyzer, spectrophotometer or photometer with cell holder thermostatable at 37ºC and able
to read at (main wavelength) 546 ± 20 nm and (sub-wavelength) 700 nm ± 50 nm. 1. Sample and Reagent volumes may be varied as long as the same ratio is maintained.
2. These reagents may be used in several automatic analysers. Instructions for many of them
SAMPLES are available on request.
Serum, EDTA-treated plasma or sodium heparinized plasma collected by standard procedures.
BIBLIOGRAPHY
LDL cholesterol in serum is stable for 5 days at 2-8ºC.
1. Nauck M, Warnick GR, Rifai N. Methods for measurement of LDL-cholesterol: a critical
PROCEDURE assessment of direct measurement by homogeneous assays versus calculation. Clin Chem
2002; 48: 236-54.
1. Bring the Reagents and the photometer to 37ºC.
2. National Cholesterol Educarion Program Expert Panel. Third report of the National
2. Pipette into a cuvette: (Note 1) Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and
Reagent A 750 µL
Treatment of High Blood Cholesterol in Adults (ATP III). NIH Publication. Bethesda: National
Serum/Calibrator 7 µL
Heart, Lung, and Blood Institute; 2001.
3. Young DS. Effects of drugs on clinical laboratory tests, 4th ed. AACC Press, 1995.
3. Mix and insert the cuvette into the photometer. Start the stopwatch. After 3-5 minutes, read
the absorbance (A1) at 546/700 nm against distilled water. 4. Tietz NW. Clinical guide to laboratory tests, 3rd ed. Saunders Co, 1999.
4. Pipette into a cuvette: 5. Friedman and Young. Effects of disease on clinical laboratory tests, 3th ed. AACC Press,
1997.
Reagent B 250 µL

Mix.
5. After 5 minutes, read the absorbance (A2) at 546/700 nm.

CALCULATIONS
The cholesterol LDL concentration is calculated using the following general formula:
(A2 – A1) Sample
x C Calibrator = C Sample
(A2 – A1) Calibrator

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 risk2.

Up to 100 mg/dL = 2.59 mmol/L Optimal

100-129 mg/dL = 2.59-3.34 mmol/L Near optimal/above optimal
130-159 mg/dL = 3.37-4.12 mmol/L Borderline High
160-189 mg/dL = 4.14 –4.90-mmol/L High
> 190 mg/dL = 4.92 mmol/L Very High

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