ACCUCARETM
CHOLESTEROL
Enzymatic colorimetric test
ORDER INFORMATION
Catalog No: Product Description: Pack Size:
CHOL 50 CHOLESTEROL 5 X 10 ML
CHOL 100 CHOLESTEROL 5 X 20 ML
CHOL 200 CHOLESTEROL 4 X 50 ML

METHOD
 Enzymatic colorimetric test

PRINCIPLE Cholesterol esters are hydrolyzed by Cholesterol esterase to produce cholesterol. Hydrogen Peroxide is then produced from oxidation of cholesterol by cholesterol oxidase. The indicator quinoneimine is formed from hydrogen peroxide and 4 - aminoantipyrine in the presence of phenol and peroxide. The absorption of the red quinoneimine dye is proportional to the concentration of cholesterol in the sample.

SAMPLE: Serum heparinised plasma or EDTA plasma. Stability : 6 days at 4 - 25°C ; 4 months at -20°C

REFERENCE VALUES
< 200 mg/dl Normal
220 - 239 mg/dl Borderline - High
> 240 mg/dl High

QUALITY CONTROL :
Accutestrol N - H

REAGENTS
Reagent I : Buffer reagent
Reagent II : Enzyme reagent
Reagent III : Cholesterol standard : 200 mg/dl

REFERENCES Teitz N.W fundamentals of Clin. Chem,
Young D.S, Naito, HK.et.al. (1973), 10.79.

REAGENT PREPARATION
Dissolve one vial of enzyme reagent with amount of buffer reagent as specified on the enzyme.
 STABILITY : 120 days at 2-8°C. Store protected from light.

AUTOMATED PARAMETERS
Wave length 520 nm
Reaction Type End Point
Cuvette 1 cm light path
Reaction Temperature 37°C
Measurement Against Reagent blank
Sample/Reagent Ratio 1: 100
Incubation 5 minutes
Maximum Blank Absorbance 0.50
Low Normal at 37°C < 200 mg/dl
High Normal at 37°C > 240 mg/dl
Linearity at 37°C 500 mg/dl

MANUAL PROCEDURE

PIPETTE INTO TEST TUBE
  BLANK STD SAMPLE
SAMPLE - - 10 µl
STANDARD - 10 µl -
REAGENT 1000 µl 1000 µl 1000 µl

Mix , Incubate 5 mins at 37°C (or 10 mins at 20 - 25°C) Measure absorbance of the sample (AT) and standard (AS) against reagent blank. The colour is stable for 90 mins at 20 - 25°C

CALCULATION AND LINEARITY
AT/AS x conc. Standard = mg/dl Total Cholesterol

The method is linear to a concentration of 500 mg/dl. Dilute samples above this concentration 1:1 with 0.9 % saline solution and repeat assay. Multiply the result by 2.
Lab Care Diagnostics (India) Pvt. Ltd.
16-A, 'A' Wing , 1st Floor Sita Estate Mahul RoadAziz Baug, Chembur Mumbai 400 074. India
Phone: 00 91 22 2554 2109/ 00 91 22 2554 1558 Fax: 00 91 22 2554 3541
E-mail : accucare@labcarediagnostics.com