HDL- C Direct
Direct Method
METHOD / PRINCIPLE
ANALYTICAL PROCEDURE
LDL, VLDL and Chylomicrons in the sample are masked
by specific complexing agents. The HDL fraction is released by detergent and is determined by enzymaticcolorimetric end point method.
This is the standard procedure, Instrument
Applications are available upon request.
REAGENTS
Wavelength
Temperature
Cuvette
R1:
Good’s buffer, pH 7.0
H-DAOS
Complexing agents / detergents
R2:
Good’s buffer, pH 7.0
CE*
COD**
POD
4-AA***
20 mmol/L
0.8 mmol/L
600 nm (sub 700)
37°C
1 cm light path
Read against reagent blank
20 mmol/L
> 250 U/L
> 5000 U/L
> 11000 U/L
2.6 mmol/L
(RBL)
STABILITY
RBL
CALIBRATOR SAMPLE
Reagent R1
240 µL
240 µL
240 µL
Calibrator
3 µL
Sample
3 µL
Mix well, incubate for 5 min, read A1 and then add
Reagent R2
80 µL
80 µL
80 µL
Mix well, incubate for 5 min,then read A2
The sealed reagents are stable up to the indicated expiry
date if stored at 2° to 8°C.
A = (A2 – A1) Sample or Calibrator - (A2 – A1) RBLl
* Cholesterolesterase
**Cholesteroloxidase
*** 4-Aminoantipyrin
PREPARATION
R1 and R2 are ready for use. Store at 2-8°C.
Do not freeze !
CALCULATION
with calibrator
Stability after opening :
At least 3 months
at 2° to 8 °C, if contamination is avoided
HDL - C [mg / dl] 
On board stability is at least 4 weeks at 2 – 8°C
A sample
 Conc. Cal. [mg / dl]
A calibrator
WARNINGS AND PRECAUTIONS
1.When discarding the reagents, dispose them
according to local or federal regulations.
2.Avoid contact with eyes and skin. If contacted,
flush eyes or rinse skin with a large amount of
water. If irritation persists, consult a physician.
CALIBRATION & QUALITY CONTROL
For the calibration of automated analyzers Greiner special
calibrator or the Multicalibrator is recommended, for
quality control use Greiner Unitrol-I and -II .
NOTES
HDL-C Calibrator is not provided with the kit. Refer
to the instruction sheet in the Calibrator kit.
Page 1/2
Greiner Diagnostic GmbH - Unter Gereuth 10 – D-79353 Bahlingen – Germany
www.greiner-diagnostic.com
PERFORMANCE
BIBLIOGRAPHY
- Analytical range
The measurable range is 1-200 mg/dL HDL-cholesterol.
If the obtained value exceeds 200 mg/dL, repeat the test
using serum diluted 1/10 with 0.9% sodium chloride.
Multiply result by 10.
- Sensitivity
The lowest detectable value is 1 mg/dL
1.
2.
- Precision
Within series
n = 30
Mean value
[mg/dL]
CV
[%]
32.6
47.3
68.5
0,65
0,78
0,65
Sample 1
Sample 2
Sample 3
3.
4.
- Correlation
A comparative study has been performed between the
Greiner method and another commercial reagent on 100
human serum samples. The parameters of linear
regression are as follows:
5.
Rifai N, Bachorik PS, Albers JJ. Lipids, lipoproteins
and apolipoproteins. In: Burtis CA, Ashwood ER,
editors. Tietz Textbook of Clinical Chemistry. 3rd ed.
Philadelphia: W.B Saunders Company; 1999. p.
809-61.
Recommendation of the Second Joint Task Force of
European and other Societies on Coronary
Prevention. Prevention of coronary heart disease in
clinical practice. Eur Heart J 1998;19: 1434-503.
Wiebe DA, Warnick GR. Measurement of highdensity lipoprotein cholesterol. In: Rifai N, Warnick
GR, Dominiczak MH, eds. Handbook of lipoprotein
testing. Washington: AACC Press, 1997:p.127-44.
Nauck M, Maerz W, Wieland H. New
immunoseparation-based homogenous assay for
HDL-cholesterol compared with three homogenous
and two heterogeneous methods for HDLcholesterol. Clin Chem 1998;44:1443-51.
Schaefer EJ, McNamara J. Overview of the
diagnosis and treatment of lipid disorders. In: Rifai
N, Warnick GR, Dominiczak MH, eds. Handbook of
lipoprotein
testing.
Washington:
AACC
Press;1997.p.25-48.
Correlation coefficient
r = 0.996
Linear regression
y = 1.000 x + 2.0 mg/dL
INTERFERENCES
SYMBOLS USED
- Ascorbic Acid: no interference up to 100 mg/dL
- Bilirubin: no interference up tol 40 mg/dL
- Triglycerides: no interference up to 1000 mg/dL
- Hemoglobin: no interference up to 500 mg/dL
For in vitro diagnostic medical use
Batch Code
EXPECTED VALUES
Normal values are > 35 mg/dl (0.9 mmol/l)
Clinical Interpretation:
Epidemiological studies have shown that for men values
< 35 mg/dl (< 0.9 mmol/l), and for women
values < 43 mg/dl (< 1.0 mmol/l) stand for increased
coronary risk factors, especially if triglyrerides
are > 180 mg/dl (> 2 mmol/l).
Use by
Temperature limitation
Greiner Diagnostic GmbH - Unter Gereuth 10 – D-79353 Bahlingen – Germany
www.greiner-diagnostic.com