PREPARATION With Factor:

 The reagent is ready to use. A505 nm Sample x 320 = mg/Dl HDLc in the sample.
A546 nm Sample x 475 = mg/Dl HDLc in the sample
STORAGE AND STABILITY
 All the components of the kit are stable until the
expiration date on the label when stored tightly Calculation of LDL-cholesterol
HDL cholesterol According to the Friedewald Formula:
closed at 2-8°C, protected from light and
Precipitating reagent contaminations prevented during their use. Triglycerides
 Do not use reagents over the expiration date. LDL cholesterol = Total cholesterol - -HDL cholesterol

For in -vitro diagnostic use only  Signs of reagent deterioration: 5

 Presence of particles and turbidity. QUALITY CONTROL
EQUIPMENTS NEEDED BUT NOT PROVIDED Control sera are recommended to monitor the
Store at 2° to 8°C  Spectrophotometer or colorimeter measuring at performance of assay procedures. If control values are
505 nm (500-550). found outside the defined range, check the instrument,
INTENDED USE  Matched cuvettes 1.0 cm light path. reagents and calibrator for problems.
For the measurement of cholesterol concentration in  General laboratory equipment. Each laboratory should establish its own Quality Control
human serum or plasma. scheme and corrective actions if controls do not meet
SAMPLES the acceptable tolerances.
INTRODUCTION
Serum or plasma: REFERENCE VALUES
HDL particles carry cholesterol from the cells back to the
Free of hemolysis. Removed from the blood clot as soon HDL- Cholesterol Men Women
liver.
as possible.
HDL is known as “good cholesterol” because high levels
Stability: HDL Cholesterol is stable for 7 days at 2-8°C.
are thought to lower the risk of heart disease. Lower risk > 55 mg/dL > 65 mg/dL
A low HDL cholesterol levels, is considered a greater heart
PROCEDURE
disease risk. Standard risk 35-55 mg/dL 45-65 mg/dL
Precipitation
Clinical diagnosis should not be made on a single test
1. Pipette into a centrifuge tube:
result; it should integrate clinical and other laboratory
R (Precipitating Reagent) 100 µL Increased risk < 35 mg/dL < 45 mg/dL
data.
PRINCIPLE OF THE METHOD Sample 1.0 mL
LDL- Cholesterol
The very low density (VLDL) and low density (LDL)
lipoproteins from serum or plasma are precipitated by
2. Mix well; allow to stand for 10 min at room Suspected 150 mg/dL
phosphotungstate in the presence of magnesium ions.
After removed by centrifugation the clear supernatant temperature.
containing high density lipoproteins (HDL) is used for the 3. Centrifuge at 4000 r.p.m. for 20 min or 2 min at Increased 190 mg/dL
determination of HDL cholesterol. 12000 r.p.m..
4. Collect the supernatant and test HDLc.
REAGENTS
These values are for orientation purpose; each laboratory
R Phosphotungstic acid 14 mmol/L TEST should establish its own reference range.
Precipitating Reagent Magnesium chloride 2 mmol/L Following the Cholesterol reagent instructions.
50mg/dl CALCULATIONS PERFORMANCE CHARACTERISTICS
HDL CHOLESTEROL Cholesterol
With Calibrator: Measuring range:
From detection limit of 1.57 mg/dL to linearity limit of
Calibrator (A) Sample x (Calibrator conc.) = mg/dL HDLc in
275 mg/dL.
(A) Standard the sample If the results obtained were greater than linearity limit,
dilute the sample 1/2 with NaCl 9 g/L and multiply the
result by 2. 4th ed AACC Press, 1995.
5. Young DS. Effects of disease on Clinical Lab. Tests,
Precision: 4th ed. AACC 2001.
Intra-assay Inter-assay 6. Burtis A. et al. Tietz Textbook of Clinical
(n=20) (n=20) Chemistry, 3rd ed. AACC 1999.
7. Tietz N W et al. Clinical Guide to Laboratory Tests,
Mean 75.8 33.9 95.2 182
3rd ed. AACC 1995.
(mg/dL)

SD 0.89 0.85 2.59 3.04

CV (%) 1.18 2.51 2.72 1.68 ATLAS Medical

William James House,
Sensitivity: Cowley Road, Cambridge, CB4 4WX, UK
Tel: ++44 (0) 1223 858 910
1 mg/dL = 0.0015 A. Fax: ++44 (0) 1223 858 524
Accuracy:
Results obtained using ATLAS reagents (y) did not show PPI413A01
systematic differences when compared with other Rev C (19.10.2015)
commercial reagents (x). Catalogue Number Store at
The results obtained using 50 samples were the
For In-Vitro Diagnostic
following: use
Caution
Correlation coefficient (r): 0.99. Number of tests in the Read product insert
Regression equation: y= 0,9944x – 1, 2346. pack before use
The results of the performance characteristics depend on Lot (batch) number Manufacturer
the analyzer used.
Fragile, handle with
Expiry date
care
INTERFERENCES
No interferences were observed with triglycerides up to Manufacturer fax Do not use if
number package is damaged
4 g/L.
A list of drugs and other interfering substances with HDL Manufacturer
telephone number
cholesterol determination has been reported by Young et.
al.

REFERENCES
1. Naito H K. High-density lipoprotein (HDL)
cholesterol. Kaplan A et al. Clin Chem The C.V.
Mosby Co. St Louis. Toronto. Princeton 1984;
1207-1213 and 437.
2. Grove T H. Effect of reagent pH on Determination
of HDL Cholesterol by precipitation with Sodium
Phosphotungstate-magnesium Clin Chem 25:560,
1979.
3. US National Cholesterol Education Program of the
National Institutesof Health.
4. Young DS. Effects of drugs on Clinical Lab. Tests,