CORMAY HDL DIRECT_a

CORMAY HDL DIRECT
DIAGNOSTIC KIT
FOR DETERMINATION OF
HDL-CHOLESTEROL CONCENTRATION
(DIRECT METHOD)
Kit name
CORMAY HDL DIRECT mini
CORMAY HDL DIRECT 30
CORMAY HDL DIRECT 60
CORMAY HDL DIRECT 120
CORMAY HDL DIRECT
Cat. No
2-217
2-181
2-182
2-183
2-179
INTRODUCTION
Plasma lipoproteins are spherical particles containing varying
amounts of cholesterol, triglycerides, phospholipids and proteins.
The relative protein and lipid determine the density of these
lipoproteins and provide the basis on which to begin their
classification. The classes are: chylomicron, very-low-density
lipoprotein (VLDL), low-density-lipoprotein (LDL) and high-density
lipoprotein (HDL). The principle role of HDL in lipid metabolism is
the uptake and transport of cholesterol from peripheral tissues to the
liver. Low HDL cholesterol (HDL-C) levels are strongly associated
with an increased risk of coronary artery disease.
METHOD PRINCIPLE
The assay consists of 2 distinct reaction steps:
1. Elimination of chylomicron, VLDL and LDL by cholesterol
esterase, cholesterol oxidase and subsequently catalase.
cholesterol esterase
cholesterol ester
cholesterol + fatty acid
cholesterol oxidase
cholesterol + O2
cholestenone + H2O2
catalase
H2O + O2
2 H2O2
2. Specific measurement of HDL-Cholesterol after release of HDLCholesterol by detergents in 2-Reagent. In the second reaction
catalase is inhibited by sodium azide in 2-Reagent.
Concentrations in the test
1-Reagent
Good’s buffer (pH 6.6)
cholesterol esterase
cholesterol oxidase
catalase
N-(2-hydroxy-3-sulfopropyl)-3,5dimethoxyaniline (HDAOS)
2-Reagent
Good’s buffer (pH 7.0)
peroxidase
4–aminoantipyrine (4-AA)
100 mmol/l
1400 U/l
800 U/l
600 kU/l
0.6 mmol/l
100 mmol/l
3 kU/l
4 mmol/l
Warnings and notes
Product for in vitro diagnostic use only.
The reagent contains < 0.1% sodium azide as a preservative.
Avoid contact with skin and mucous membranes.
ADDITIONAL EQUIPMENT
automated analyser;
general laboratory equipment;
SPECIMEN
Serum, heparinized or EDTA plasma.
Blood should be collected only if the patient has been fasting for 12 14 hours.
Serum and plasma can be stored up to 6 days at 2 - 8°C. Sample are
stable for 1 year when stored at for -70°C. Samples may be frozen
once. If any samples show precipitates, centrifuge before using.
Nevertheless it is recommended to perform the assay with freshly
collected samples.
PROCEDURE
wavelength
temperature
600 nm
37°C
CORMAY HDL DIRECT is intended for automated analysers.
Applications for particular analysers are available on request.
cholesterol esterase
cholesterol ester
cholesterol + fatty acid
REFERENCE VALUES4
cholesterol oxidase
cholesterol + O2
cholestenone + H2O2
peroxidase
2 H2O2 + 4-AA + HDAOS
quinone pigment + 4 H2O
The colour intensity is proportional to the HDL-cholesterol
concentration when measured at 600 nm.
REAGENTS
Package
CORMAY
HDL
DIRECT
mini
CORMAY
HDL
DIRECT
30
CORMAY
HDL
DIRECT
60
CORMAY
HDL
DIRECT
120
CORMAY
HDL
DIRECT
1-Reagent
2 x 30 ml
3 x 30 ml
3 x 50 ml
3 x 100 ml
4 x 30 ml
2-Reagent
2 x 10 ml
1 x 30 ml
1 x 50 ml
1 x 100 ml
4 x 10 ml
Working reagent preparation and stability
The reagents are ready to use.
The reagents are stable up to the kit expiry date printed on the
package when stored at 2-8°C. The reagents are stable for
8 weeks on board the analyser at 2-10°C. Protect from light and
contamination!
CORMAY HDL DIRECT page 1
40 – 60 mg/dl
1.04 – 1.55mmol/l
As HDL cholesterol is affected by a number of factors such as
smoking, exercise, hormones, age and sex, each laboratory should
establish its own reference ranges for local population.
serum / plasma
QUALITY CONTROL
For internal quality control it is recommended to use CORMAY
LIPID CONTROL 1 (Cat. No 5-179) and CORMAY LIPID
CONTROL 2 (Cat. No 5-180) or CORMAY SERUM HN
(Cat. No 5-172) and CORMAY SERUM HP (Cat. No 5-173) with
each batch of samples.
For the calibration of automatic analysers the CORMAY HDL/LDL
CALIBRATOR (Cat. No 5-178) is recommended.
The calibration curve should be prepared every 8 weeks or with
change of reagent lot number.
PERFORMANCE CHARACTERISTICS
These metrological characteristics have been obtained using
automatic analyser Biolis 24i Premium. Results may vary if a
different instrument is used.
Sensitivity : 3.94 mg/dl (0.102 mmol/l).
Linearity: up to 400 mg/dl (10.36 mmol/l).
For higher concentration of HDL cholesterol dilute the sample
with physiological saline before assaying. Multiply the result
obtained from the manual dilution by the appropriate dilution
factor.
Specificity / Interferences
Bilirubin up to 20 mg/dl, ascorbate up to 62 mg/l, haemoglobin
up
to
5 g/dl and triglycerides up to 500 mg/dl do not interfere with the
test.
Precision (Biolis 24i Premium)
Repeatability (run to run)
Mean
n = 20
[mg/dl]
level 1
55.18
level 2
21.70
SD
[mg/dl]
0.92
0.45
CV
[%]
1.66
2.09
Reproducibility (day to day)
n = 80
level 1
level 2
SD
[mg/dl]
2.96
0.85
CV
[%]
4.91
3.75
Mean
[mg/dl]
60.32
22.67
Method comparison
A comparison between HDL cholesterol values determined at Biolis
24i Premium (y) and at ADVIA 1650 (x) using 48 samples gave
following results:
y = 0.9848 x + 2.7175 mg/dl;
R= 0.9855
(R - correlation coefficient)
LITERATURE
1. National Institutes of Health Consensus Development
Conference Statement: Triglyceride, High Density Lipoprotein
and Coronary Heart Disease. Washington D.C. Feb 26-28, 1992.
2. Izawa S., Okada M., Matusui H., and Horita Y. J. Medicine and
Pharmaceutical Sci., 1385-1388, 37 (1997).
3. Shih WJ, Bachorik PS, Haga JA, Myers GL, Stein EA; Clinical
Chemistry, 2000; 46:3:351 – 364.
4. Alan H.B. Wu: Tietz Clinical Guide to Laboratory Tests, 4th ed.
WB Saunders, 564 (2006).
5. Jacobs D., et al. In Laboratory and Test Handbook; Jacobs D.S;
Kasten B.L., De Mott W.R., Wolfson W.L., Eds; Lexi-Comp Inc:
Hudson (Cleveland), 1990; P.219.
6. Gordon T. et al.: Am. J. Med., 62; 707 (1977).
7. Wiliams P., Robinson D., Baily A.: Lancet, 1/72 (1979).
8. Goto A.M.: Hospital Practice, 23; Suppl., 1, 4 (1988).
9. Crouse J.R. et al.: J. Lipid Res., 26; 566 (1985).
10. Badmion J.J., Badmion L., Fuester V.: Journal of Clinical
Investigation, 85:1234-41 (1990).
11. Castelli W.P. et all.: Circulation, 55; 767 (1977).
12. Barr D.P., Russ E.M., Eder H.A.: Am. J. Med., 11; 480 (1951).
13. Kannel W.B., Castelli W.P., Gordon T.: Ann. Intern. Med., 90:85
(1979).
Date of issue: 06. 2013.
WASTE MANAGEMENT
Please refer to local legal requirements.
MANUFACTURER
PZ CORMAY S.A.
22 Wiosenna Street,
05-092 Łomianki, POLAND
tel.: +48 (0) 22 751 79 10
fax: +48 (0) 22 751 79 14
http://www.cormay.pl
06/13/06/13
CORMAY HDL DIRECT page 2

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