HDL CHOLESTEROL

Precipitation

Cat.No Package Size REFERENCE VALUES

HD450S15 4 x 50 mL incl. Standard
HD4100S75 4 x 100 mL incl. Standard Good Standard Increased
prognosis risk risk
HD2125S132 2 x 125 mL incl. Standard Male > 55 mg/dL 35 – 55 mg/dL < 35 mg/dL
> 0.55 g/L 0.35 – 0.55 g/L < 0.35 g/L
METHOD > 1.4 mmol/L 0.9 – 1.4 mmol/L < 0.9 mol/L
Precipitation, Phosphotungstic acid. Female > 65 mg/dL 45 – 65 mg/dL < 45 mg/dL
> 0.65 g/L 0.45 – 0.65 g/L < 0.45 g/L
> 1.7 mmol/L 1.2 – 1.7 mmol/L < 1.2 mmol/L
PRINCIPLE
Chylomicrons, Very Low Density Lipoproteins (VLDL)
and Low Density Lipoproteins (LDL) of serum are Note: It is recommended for each laboratory to establish and
maintain its own reference values. The given data are only an
precipitated by phosphotungstic acid and magnesium indication.
ions.
After centrifugation the High Density Lipoproteins
(HDL) are in the supernatant, and the HDL- ANALYTICAL PROCEDURES
cholesterol is determined by a cholesterol - Precipitation
determination.
Pipet into centrifuge tubes Macro Semi-Micro
REAGENT COMPOSITION Sample 500 µl 200 µl
Precipitant R1 1000 µl -
R1: Phosphotungstic acid 0,55 mmol/L Diluted Precipitant - 500 µl
Magnesium chloride 25 mol/L Mix well, incubate for 10 min at room temperature. Centrifuge
Standard: 50 mg/dl 10 min at 4000 r.p.m. or for 2 min at 10000 r.p.m.
Separate clear supernatant from residue in between 1 hour and
determine cholesterol concentration as follows
Additional reagent (required but not supplied):
Kit for Cholesterol determination.
- HDL-Cholesterol Determination
PRECAUTIONS
- For in vitro diagnostic use only Wavelength 546 nm Hg (500 nm)
- Avoid contamination by using clean laboratory Cuvette 1 cm
Temperature 20 - 37 °C
material (pipettes, plastic vials for analyzers,...). Measure against Reagent Blank (RBL)

STABILITY OF REAGENTS Pipet into cuvettes RBL Standard Sample

When stored at 2-8°C and protected from light, the Dist. Water 100 µl - -
Standard - 100 µl -
reagents are stable until the expiry date stated on the
Supernatant - - 100 µl
label. Cholesterol Rgt 1000 µl 1000 µl 1000 µl
Mix well ,incubate for 5 min at 37°C or for 10 min at roomtem-
PREPARATION AND STABILITY perature. Measure sample and standard against RBL within
OF WORKING REAGENT 1 hour => ∆A
Macro Assay : Reagent R1 is ready for use
Semi-Micro Assay : Dilute 4 parts of R1 with CALCULATION
1 part of dist. water
Stability : Until the expiration date printed on - with factor
the labels HDL-Cholesterol (mg/dl) = ∆ A x F1
Standard is ready for use : Use directly in the HDL-Cholesterol (mmol/l) = ∆ A x F2
testprocedure .
Do not precipitate – calculation formula comprises the Factors Macro Micro
dilution factor ! Wavelength F1 F2 F1 F2
Stability : Until the expiration date printed on 500 nm 180 4,65 210 5,43
Hg 546 nm 275 7,11 320 8,29
the labels

SAMPLES - with standard

Serum, plasma (EDTA or heparinized)

L-S 01/2010 page 1/2

Inmesco GmbH – Wiedtalstrasse 11&18 – D-53577 Neustadt/Wied – Germany

www.inmesco.de
 INTERFERENCES
Macromethod: Interferences are according to literature
∆A sample
HDL - C = 150 x [mg /dl]
∆A standard

BIBLIOGRAPHY
∆A sample
HDL - C = 3.87 x [mmol/l]
∆A standard
1. Rifai N, Bachorik PS, Albers JJ. Lipids, lipoproteins and
apolipoproteins. In: Burtis CA, Ashwood ER, editors. Tietz
Semi-Micromethod: Textbook of Clinical Chemistry. 3rd ed. Philadelphia: W.B
Saunders Company; 1999. p. 809-61.
2. Recommendation of the Second Joint Task Force of
∆A sample
HDL - C = 175 x [mg /dl] European and other Societies on Coronary Prevention.
∆A standard
Prevention of coronary heart disease in clinical practice. Eur
∆A sample
HDL - C = 4.52 x [m mol /l] Heart J 1998;19: 1434-503.
∆A standard
3. Wiebe DA, Warnick GR. Measurement of high-density
lipoprotein cholesterol. In: Rifai N, Warnick GR, Dominiczak
MH, eds. Handbook of lipoprotein testing. Washington: AACC
Press, 1997:p.127-44.
4. Schaefer EJ, McNamara J. Overview of the diagnosis and
CALIBRATION & QUALITY CONTROL treatment of lipid disorders. In: Rifai N, Warnick GR,
For the calibration of automated analyzers Inmesco Dominiczak MH, eds. Handbook of lipoprotein testing.
Multicalibrator Introcal is recommended, for quality Washington: AACC Press;1997.p.25-48.
control use Inmesco normal and abnormal control,
Intronorm I and Intronorm II or the special lipid
control . SYMBOLS USED

PERFORMANCE DATA (37°C)

For in vitro diagnostic medical use
- Analytical range
Refer to the performance data of the applied
cholesterol reagent. Batch Code

- Detection limit
Refer to the performance data of the applied
Use by
cholesterol reagent.

- Precision (n = 20) Temperature limitation

Within run Mean Standard- CV
[mg/dl] deviation [%]
[mg/dl]
Sample 1 15,3 0,22 1,44
Sample 2 27,8 0,19 0,67
Sample 3 22,6 0,33 1,44

Sample 1 15,0 0,08 0,54

- Correlation
A comparative study has been performed between
the Inmesco method and another commercial
reagent on 31 human serum samples. The
parameters of linear regression are as follows:

y = 1,060 x – 0,328 mg/dl r= 0,998.

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