ACCUCARETM
CHOLESTEROL HDL
Precipitant Phosphotungstic Acid Reagent
ORDER INFORMATION
Catalog No: Product Description: Pack Size:
HDL 50 CHOLESTEROL HDL 2 X 25 ML


METHOD
Colorimetric endpoint test


PRINCIPLE
Low density lipoproteins (LDL and VLDL) and chylomicron fractions are precipitated by buffered phosphotungstic acid reagent. After centrifugation, the cholesterol concentration in the HDL (high density lipoprotein) fraction, remains in the supernatant. Cholesterol included in this phase, is determined by an enzymatic method.


SAMPLE:
Serum or EDTA plasma.


REFERENCE VALUES
MEN 41.0 - 58.7 mg/dl
WOMEN 48.8 - 75.0 mg/dl


QUALITY CONTROL
Accutestrol N - H


REAGENTS
Reagent I : Buffered phosphotungstic acid reagent
Cholesterol standard : 50 mg/dl


REAGENT PREPARATION
Ready to use.

STABILITY :
the contents are ready to use and is stable upto the expiry date when stored at 2-8°C.

PRECIPITATION METHOD

Pipette into centrifugate tubes

SAMPLE REAGENT
500 µl 500 µl
Mix and allow to stand for 5 min. centrifugate for 10 min at 3000 rpm and determine the cholesterol content by the CHOD-PAP method. Only clear supernatant must be used.

AUTOMATED PARAMETERS
Wave length 520 nm
Reaction Type End Point
Cuvette 1 cm light path
Reaction Temperature 37°C
Reaction Type Increasing
Measurement Against Reagent Blank
Sample/Reagent Ratio 1: 20
Incubation 5 minutes
Maximum Reagent Blank 0.50
Low Normal at 37°C 41 mg/dl
High Normal at 37°C 75 mg/dl
Linearity at 37°C 200 mg/dl

MANUAL PROCEDURE

 PIPETTE INTO TEST TUBE
  BLANK STD SAMPLE
Sample - - 50 µl
Standard - 50 µl -
Reagent 1000 µl 1000 µl 1000 µl

Mix incubate for 5 min at 37°C (10 min at 20 - 25°C).
Measure absorbance of sample (Ac) and standard (As) against
the reagent blank.
The colour is stable for 90 min at 20 - 25°C.

CALCULATION AND LINEARITY

Ac/As x Conc. Standard x 2 = mg% HDL in serum
2 = dilution factor of the Precipitation step

The method is linear to a concentration of 200 mg/dl.

REFERENCES
Denacherp N.M., Hijans A.G.M., Vos-janseen H.E., Vant Laar A.P., Clin. Chem., 26 1775 (1980)
Casteli W.P., : Current Prescribing, 3, 39 (1977).
Lab Care Diagnostics (India) Pvt. Ltd.
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Phone: 00 91 22 2554 2109/ 00 91 22 2554 1558 Fax: 00 91 22 2554 3541
E-mail : accucare@labcarediagnostics.com