REAGENTS- INITIAL CONCENTRATION Pipette into labelled centrifuge tubes :
Via Acuto 30, – 00131 – Rome – ITALY Reagent 1 Good buffer, pH 10 100 mmol/L Standard - L
Tel: +39-(0)6-4190735 Polyethylenglycol-6000 (PEG6000) 30 % Sample 500 - L
Fax: +39-(0)6-4190735 - +39-(0)6-233223561 Sodium azide 0,05 - 0.1% Reagent 1 300 L
Ordini OnLIne:www.bio-gamma.com Reagent 2 Cholesterol HDL standard 50 mg/dL Mix thoroughly and after 10’ at +15-25°C centrifuge at 3000 rpm for 10’.
Measure the HDL cholesterol HDL on the clear supernatant with the reagent of the
HDL CHOLESTEROL PEG 6000 REAGENTS – STORAGE AND STABILITY cholesterol CHOD-PAP (COD.CCD016-CCD017).
Method with precipitanting reagent PEG 6000.
Kit for measurement of HDL cholesterol concentration in serum and in Kit: Store Reagent 1 at +15-25°C. ANALYTICAL PROCEDURE CHOLESTEROL CHOD-PAP (COD.CCD016-CCD017)
plasma. Store Reagent 2 at +2-8°C. Do not refrigerate.
Stable until the expiry date shown on the label. Wavelenght 520 (500 o 546)nm
Opened reagents: Reagents are stable after opening until the expiry date Lightpath 1 cm
CCD021 4x50 mL shown on the bottle if it's closed immediately after the Temperature 37°C
take of the sample and protected from direct light and Measurement Against reagent blank
stored at the reported temperature. Reaction End point (increase)
PRINCIPLE OF THE METHOD BLANK STANDARD SAMPLE
REAGENTS - PREPARATION
Lipoproteins of fractions VLDL (very low density lipoproteins) and LDL (low density Working Reagent 1000 1000 1000 L
The reagent s are stable liquid ready to use.
lipoproteins) and the chylomicrons are quantitative precipitated at buffered pH by Distilled water 50 - - L
addition of polyethylenglycol-6000 (PEG6000). After centrifuge, is determinated the Sample - - 50 L
REAGENTS - PRECAUTION AND WARNING Standard - 50 - L
HDL cholesterol concentration with the enzimatic colorimetric CHOD-PAP method.
Mix thoroughly and incubate for 5 minutes at 37°C.
- The present method descrives the manual use of this kit. Measure the absorbance of the Sample (E Sample) and the Standard (E Standard)
METROLOGICAL CHARACTERISTICS CHOLESTEROL CHOD-PAP - Quality control data sheet of the reagent are available upon request. Refer to the against the blank Reagent.
(COD.CCD016-CCD017) batch number on the label. The colour is stable at least 60 minutes at +15-25°C, protected from direct light.
INTERFERENCES ADDITIONAL EQUIPMENT CALCULATIONS
Doesn't interfere until the concentration of:
Bilirubin 20 mg/dL
Hemoglobin 600 mg/dL Pipettes Serum:
Cholestero l HDL(mg/dL) (mmol/L)
Control: Low High Cuvette (Lightpath = 1cm). Where:
n: 20 20 Thermostatic water at 37°C [STD] = concentration of standard cholesterol HDL (50mg/dL)
MEAN: 177,50 194,25 mg/dL
D.S.: 1,53 1,76
C.V.(%): 0,86 0,90 SAFETY PRECAUTIONS QUALITY CONTROL
Control: Low High Reagent 1 Refering to european law this product is not classified as a dangerous Each laboratory should estabilish its own internal Quality Control scheme and procedures
n: 20 20 substance. The reagents contain inactive components such as detergent for corrective action if controls do not recover within the acceptable tolerances.
MEAN: 176,77 196,68 mg/dL
and preservatives. The total concentration of these components is lower
D.S.: 3,51 4,39
C.V.(%): 1,99 2,22 than the limits reported by 67/548/CEE and 88/379/CEE directives and BIBLIOGRAPHY
CORRELATION next modifications about classification, packaging and labelling of
Results obtained on the same samples with an equivalent method have done the following data: dangerous substances.
Burstein, M., Scholnick, H.R., and Morgin, R.: Rapid method for the isolation of
y = 0,9462x + 2,8499 However, the reagents should be handled with caution, avoiding
lipoproteins from human serum by precipitation with polyanions, J. Lipid Res. 11:583-595,
2 = 0,9912 swallowing and contact with skin, eyes and mucous membranes.
n = 20 The use of laboratory reagents according to good laboratory pratice is
Wiebe, P.A., and Smith, S.J., Clin. Chem. 31:746-750, 1985.
LINEARITY: 800 mg/dL recommended (*).
Vilkari, J., Scand. J. Clin. Lab. Invest. 36:265-268, 1976.
SENSITIVITY: 2 mg/dL Castelli, W.P., Doyle, J.T., Gordon, T., et al., Circulation 55:767-772, 1977.
MEASURED RANGE: 2 - 800 mg/dL Reagent 2 Harmful
Tietz, NW., Clinical Guide To Laboratory Tests, W.B. Saunders Co., Philadelphia, 1986.
SAMPLE -Isooctylphenoxypolyethoxyethanol SYMBOLS
Serum, plasma. - Risk of serious damage to eyes (R 41)
Safety phrases Read use istruction Manufactured by
Note: - In case of contact with eyes, rinse immediately with plenty of water and
- Avoid use of hemolyzed samples. seek medical advise (S 26)
- After the collection of the sample , separate as soon as possible serum or plasma - Wear eye/face protection (S 39) In Vitro Diagnostic
Size/number of tests
from red cells. Medical Device
- Avoid use of anticoagulants such as heparin, citrate, fluoride and oxalate. * EU-Dir 1999/11 Commission Directive of March 1999 adapting to technical progress the
- Cholesterol HDL in the sample is stable for 14 hours at room temperature (+15- principles of good laboratory pratice as specified in Council Directive 87/18/EEC. CE mark declaring compliance with Catalogue number
30°C), 1 week at +2-8°C. the In Vitro Diagnostics Medical
Device Directive, 98/79/EC
WASTE MANAGEMENT Batch number
Store at +15-25°C
Please refer to local legal requirements.
MAN 41 – 58,7 mg/dL (1.06 – 1.52 mmol/L) ANALYTICAL PROCEDURE Vial content
WOMAN 45 - 65 mg/dL (1.26 – 1.94 mmol/L) Store at +2-8°C
Allow reagents to reach working temperature +15-25°C before using.
Every laboratory is advised to establish the refernce interval in relation to its own