TM

5 x 25 mL , 5 x 100 mL , 4 x 250 mL
CHOLESTEROL (S.L) 11403002, 11403003, 11403007

Intended Use Precaution

This reagent is intended for in vitro quantitative determination of Cholesterol in To avoid contamination, use clean laboratory wares. Avoid direct
serum or plasma. exposure of working reagent to light.
- CHOD-PAP methodology Waste Management
- Linear upto 600 mg/dL Reagent must be disposed off in accordance with local regulation.
- Contains LCF (Lipid clearing factor) which minimizes rerun Sample
Clinical Significance Fresh serum / plasma (free of haemolysis)
It is the main lipid found in the blood, bile & brain tissues. It is also one of the Interferences
most important steroids of the body & is a precursor of many steroid hormones. No interference for
Two thirds of cholesterol present in the blood is esterified. The liver metabolizes Bilirubin up to 10 mg/dL
the cholesterol & it is transported in the blood stream by lipoproteins.
Ascorbic acid up to 50 mg/dL
Increased levels are found in hypercholesterolaemia, hyperl ipidaemia, Haemoglobin up to 1000 mg/dL
hypothyroidism, uncontrolled diabetes, nephritic syndrome & cirrhosis.
Materials Provided
Decreased levels are found in malabsorption, malnutrition, hyperthyroidism, Cholsterol Reagent & Standarad.
anaemia & liver diseases.
Materials required but not provided
Principle
l Pipettes& Tips
Enzymatic colorimetric determination of total cholesterol according to the l Test Tubes & racks
following reactions.
l Timer
Cholesterol esterase l Incubator
Cholesterol ester +H2O > Cholesterol + fatty acids
l Analyzer
Cholesterol Oxidase
Cholesterol + O2—————————————> 4-Cholesten-3- one + H2O2 Test Parameter
Peroxidase
2H2O2 +Phenol+4-Aminoantipyrine ———————> - Red quinone + 4H2O Mode of Reaction End point
Slope of reaction Increasing
Kit Components
Wavelength I 505 (492 -550) nm
Reagent/ Product Product Product Description Wavelength II 630 nm
Component code: 11403002 code: 11403003 code: 11403007 0
Temperature 37 C
Cholesterol 5 x 25 mL 5 x 100 mL 4 x 250 mL Pipes Buffer
Reagent (pH 6.70) 50 mmol/L Standard Concentration 200 mg/dL
Phenol- 24 mmol/L Blank Reagent
Sodium Cholate Linearity 600 mg/dL
0.5 mmol/L
4 -Aminoantipyrine Incubation time 5 min
0.5 mmol/L Sample volume 10 µL
Cholesterol Esterase Reagent volume 1000 µL
>180 U/L
Cholesterol Oxidase Cuvette 1 cm light path
>200 U/L
Peroxidase >1000 U/L Application parameters for various instrument are available. Please contact
customer support department for specific information.

Cholesterol 1 x 4 mL 1 x 4 mL 1 x 4 mL Cholesterol std.

Unit Conversion
Standard Conc. 200 mg/dL
Risk & safety Unit (Conventional) Unit (SI) Conversion to SI unit
Material safety data sheets (MSDS) will be provided on request. mg/dL mmol/L x0.026
Reagent Preparation
Cholesterol Reagent & standard are ready to use. Calibration
Agappe multicalibrator is recommended for calibration of this assay on fully auto
Reagent Storage and Stability analyzers.
The sealed reagents are stable upto the expiry date stated on the label when stored Provided Standard is recommended for calibration of this assay on semi auto
at 2- 80C and protected from light. analyzers.
Open Vial Stability Procedure notes
o
Once opened the reagent is stable up to 4 weeks at 2- 8 C if contamination is avoided
On-board Calibration Stability Laboratory procedure For Semi Auto
Blank Standard Sample
On board calibration stability is 20 days.
Working Reagent 1000 µL 1000 µL 1000 µL
Reagent Deterioration
Standard - 10 µL -
Turbidity or precipitation in any kit component indicates deterioration and the
component must be discarded. Values outside the recommended acceptable range Sample - - 10 µL
for the Agappe Qualicheck Norm & Path control may also be an indication of 0
Mix, and incubate for 5 min.at 37 C. Measure the absorbance of sample and
reagent instability and associated results are invalid. Sample should be retested standard against reagent blank
using fresh vial of reagent.

SYMBOLS USED ON THE LABELS

IN VITRO DIAGNOSTIC USE SEE PACKAGE INSERT FOR PROCEDURE LOT NUMBER MANUFACTURER’S ADDRESS MANUFACTURING DATE EXPIRY DATE TEMPERATURE LIMIT

AGAPPE DIAGNOSTICS LTD.

‘Agappe Hills’, Dist. Ernakulam, Kerala, India-683 562.
Tel. +91 484 2867 000 | Customer Support No.: 1800 425 7151(Toll Free) ISO 9001 : 2015
customersupport@agappe.in | www.agappe.com REV. NO.: ADL/IFU/CHO/LIQ/R02 EN ISO 13485: 2016
 TM

5 x 25 mL , 5 x 100 mL , 4 x 250 mL
CHOLESTEROL (S.L) 11403002, 11403003, 11403007

3. Precision
Calculation Absorbance of sample Intra Run
Cholesterol Conc. (mg/dL) = x 200
Absorbance of standard Control Level 1 Control Level 2
Quality control n 20 20
It is recommended to use Agappe Qualicheck Norm and Path(11601001) to verify Mean ( mg/dL) 239.3 95.7
the performance of the assay.Each laboratory has to establish its own internal
quality control scheme and procedure for corrective action,if control do not recover SD 7.11 3.23
within the acceptable range. CV(%) 2.97 3.37

Reference Range
It is recommended that each laboratory has to establish its own reference values. Inter Run
The following value may be used as guide line. Control Level 1 Control Level 2
Serum / Plasma : 150 – 220 mg/dL n 20 20
Results obtained for patient samples are to be correlated with clinical finding of Mean (mg/dL) 236.60 94.58
patient for interpretation and diagnosis. SD 4.96 2.89
Performance CV % 2.10 3.06
1. Linearity
This reagent is linear upto 600 mg/dL. Accuracy (mg/dL)
If the concentration is greater than linearity (600 mg/dL), dilute the sample Control Expected Value Measured Value
with normal saline and repeat the assay. Multiply the result with dilution factor.
Control Level 1 263 ± 54.0 268
2. Comparison Control Level 2 105 ± 24.0 109
A comparison study has been performed between Agappe reagent and another Qualichek Norm 97 ± 18.0 94.4
internationally available reagent yielded a correlation coefficient of r2= 0.998
and a regression equation of y =1.0019x. Qualichek Path 195 ± 15.0 195.7

4.Sensitivity
Lower detection Limit is 3 mg/dL.
Bibliography
Allain, C.C., et al.; Clin.Chem 20 (1974), 470

SYMBOLS USED ON THE LABELS

IN VITRO DIAGNOSTIC USE SEE PACKAGE INSERT FOR PROCEDURE LOT NUMBER MANUFACTURER’S ADDRESS MANUFACTURING DATE EXPIRY DATE TEMPERATURE LIMIT

AGAPPE DIAGNOSTICS LTD.

‘Agappe Hills’, Dist. Ernakulam, Kerala, India-683 562.
Tel. +91 484 2867 000 | Customer Support No.: 1800 425 7151(Toll Free) ISO 9001 : 2015
customersupport@agappe.in | www.agappe.com REV. NO.: ADL/IFU/CHO/LIQ/R02 EN ISO 13485: 2016