6, Rue Ibn Ennafis - Z.I.

Lac 3 Tunisie
HDL-CHOLESTEROL DIRECT
Tél. : 71 182 500 - Fax : 71 182 250
www.biomaghreb.com

IN VITRO DIAGNOSTIC USE

IVD

CLINICAL SIGNIFICANCE LIMITS

The principal role of high density lipoprotein (HDL) in lipid metabolism is the uptake and transport Do not use the reagents if they are cloudy or after the expiry date.
of cholesterol from peripherical tissue to the liver through a process known as reverse cholesterol
transport. Low HDL cholesterol levels are strongly associated with an increased risk of coronary ADDITIONAL EQUIPMENT
heart disease and coronary artery disease. Hence, the determination of serum HDL-Cholesterol is • Basic equipment of the medical analysis laboratory ;
a useful tool in identifying high-risk patients. Increased Total Cholesterol/HDL-Cholesterol ratio is
• Spectrophotometer or Clinical Biochemistry Analyzer.
significant of an increased risk of atheroslerosis.
LINEARITY
PRINCIPLE The reaction is linear from 25mg/dl up to 200mg/dl.
“Selective detergent and accelerator» methodology Above this concentration, dilute the sample 1+1 with a
Direct method, without pre-treatment of the specimen. 9 g/l NaCl solution and repeat the determination.
During the first phase, LDL, VLDL, and chylomicron particles release free cholesterol which under- Multiply the result by 2.
goes an enzymatic reaction, producing hydrogen peroxide, which is degraded by the reaction with
POD and DSSmT. No coloured derivatives are formed. PROCEDURE
During the second phase, a specific detergent solubilises the HDL cholesterol. Under the combined
action of CO and CE, the POD + 4- AAP couple develops a coloured reaction proportional to the HDL Wavelength…………………………..............600-700 nm
cholesterol concentration. The reading is taken at 600 nm. Tank: ……………………………………………1 cm thick
LDL= Low Density Lipoproteins Temperature...............................................…………37°C
HDL = High Density Lipoprotein Adjusting the spectrophotometer zero with distilled water
VLDL= Very Low Density Lipoproteins - POD = Peroxidase
Blank Calibrator Dosage
CO = Cholesterol Oxidase - CE = Cholesterol Esterase – 4
AAP = 4-Aminoantipyrine - AAO = Ascorbate Oxidase Reagent R1 300 µl 300 µl 300 µl
DSSmT = N, N-bis(sulphobutyl)-m-toluldine-disodium Calibrator -- 3 µl --

REAGENT COMPOSITION Sample -- -- 3 µl

Stir well, leave to stand for 5 minutes at 37°C.
GOOD pH = 7 Record absorbances A1 at 600 nm against the reagent blank.
Cholesterol oxidase < 1000 U/I Add Blank Calibrator Dosage
Reagent 1
Peroxidase < 1300 U/ l
DSBmT < 1mM Reagen R2 100 µl 100 µl 100 µl
GOOD pH = 7 Mix well, let stand 5 minutes at 37°C.
Cholestérol oxydase < 1500 U/l Record absorbances A2 against the reagent blank
Reagent 2 4 Amino Antipyrine < 1mM
Detergent < 2% CALCULATION
Ascorbate oxidase < 3000 U/ l
Calculating the increase in absorbance A = A2-A1
Reagent 3 HDLc/LDLc calibrator: freeze-dried human serum
A sample
SAFETY CAUTIONS A calibrator
x Calibrator Concentration
Biomaghreb reagents are intended for use by qualified personnel for in vitro use (do not pipette by
mouth). = mg/dl of HDL direct
- Refer to the current SDS available on request or at www.biomaghreb.com;
- Verify the integrity of the reagents before use; and with mg/dl x 0,0259 = mmol/l
- Disposal of waste: comply with the legislation in force.
For safety reasons, treat any specimen or reagent of biological origin as potentially infectious. REFERENCE VALUES
Respect the legislation in force.
Men Women
REAGENT PREPARATION
Reagents R1 and R2 are ready for use. Low risk > 50mg/dl > 60mg/dl
- Reconstitute the vial of the HDLc/LDLc calibrator with 1 ml of distilled water, then homogenize the Moderate risk 35-50mg/dl 45-60mg/dl
contents of the bottle gently. High risk <35mg/dl <45mg/dl
- Wait 30 minutes before use.
For safety reasons, treat the calibrator as potentially infectious. -Concentrations tested (mg/dl) without significant interference (+10%):
Conjugated bilirubin: 60 mg/dl
SAMPLE PREPARATION
Total Bilirubin: 60 mg/dl
The patient must be taken after at least 12h -14h fasting
-Plasma : collected on EDTA or sodium heparinate or lithium ; citrate must not be used. Hemoglobin : 1000 mg/dl
Separate plasma from blood cells by centrifugation within 3 hours after collection. Ascorbic Acid: 100 mg/dl
-Serum: Centrifugally separate the Serum from the blood cells within 3 hours after collection.
Sera and plasma should not be left at room temperature for more than 14 hours. Fat (intralipid) 1800 mg/dl
HDL-cholesterol is stable in the specimen: -The reagent may interfere with the magnesium determination.
- 7 days at 2-8°C ;
- 1 month at -20°C. REFERENCES
PRESERVATION AND STABILITY - bNaito N K HDL.Cholesterol, Kaplon A et al.Clin Chem the C.V.Mosby Co.St Louis. Toronto. Prince-
ton 1984 : 1207 - 1213 and 437.
Store at 2-8°C, in the original bottle, tightly stoppered and protected from light.
- Us Naffonal Cholesterol Education Program of the National instites of Health.
•B efore opening : if stored under the recommended conditions, the reagents are stable until
the expiry date indicated on the label. - Young DS. Effects of drugs on Clinical Lab. tests, 4th ed AACC Press.1995.
•A fter opening and in the absence of contamination: Reagents R1 and R2 are stable for 8 - Young DS. Effects ofdisease on Clinical Lab. Tests, 4th ed AACC 2001.
weeks at 2-8°C. - Burits A et al. Tietz Texibook of Clinical Chemistry, 3rd ed AACC 1999.
•A fter reconstitution: The calibrator is stable 2 weeks at 2-8°C and 3 months at -20°C. - Tietz N W et al. Clinical Guide to laboratory tests, 3rd ed AACC 1995. PCT/JP97/04442.

Manufacturer Use by In Vitro Diagnostic Temperature Catalogue number See insert Store away from light Sufficient Batch number
Limitation for < n > essais

FT En 47 Date of updating : 10/ 2020 Version B