CHOLESTEROL

(CHOD / POD METHOD)

INTENDED USE:
The reagent kit is intended for the “in vitro” quantitative
determination of Cholesterol in serum/plasma.
PROCEDURE:
Pipette into clean dry test tubes labeled as Blank (B), Standard (S)
CLINICAL SIGNIFICANCE: and Test (T) :
Cholesterol is the main lipid found in blood, bile and brain tissue. It
is the main lipid associated with arteriosclerotic vascular Addition Sequence B. S. T.
diseases. It is required for the formation of steroids and cellular
membranes. The liver metabolizes the cholesterol and it is Enzyme reagent 1 ml 1 ml 1 ml
transported in the blood stream by lipoproteins. Increased levels Standard - 10 µl -
are found in hypercholesterolaemia , hyperlipidaemia Sample - - 10 µl
hypothyroidism, uncontrolled diabetes, nephrotic syndrom and
cirrhosis. Decreased levels are found in malabsorption,
malnutrition, hyperthyroidism anemias and liver diseases. Mix well and incubate at 37° C for 5 mins. Measure
absorbance of the Standard (Abs.S) and Test (Abs.T) against
Reagent Blank at 505 nm.
PRINCIPLE:
Cholesterol esterase hydrolyses esterified cholesterols to free
cholesterol. The free cholesterol is oxidised to form hydrogen CALCULATION : Abs. T
peroxide which further reacts with phenol and 4-aminoantipyrine Cholesterol mg/dl = X 200
by the catalytic action of peroxidase to form a red coloured Abs.S
quinoneimine dye complex. Intensity of the colour formed is
directly propotional to the amount of cholesterol present in the NORMAL VALUE :
sample. Serum : 130 - 250 mg/dl
Each Laboratory should establish it's own normal range
REACTION: representing its patient population.
Cholesterol Esterase
Cholesterol esters + H2O Cholesterol + Fatty LINEARITY :
Acids The procedure is linear upto 1000 mg/dl. If the value exceeds this
Cholesterol Oxidase limit, dilute the serum with normal saline (NaCl 0.9%) and repeat
Cholesterol + O2 Cholestenone + H2O2 the assay. Multiply result by dilution factor.

Peroxidase QUALITY CONTROL :

H2O2 + Phenol + 4 amino antipyrine Red quinonimine For accuracy it is necessary to run known controls with every
dye + H2O assay.
CONTENTS:
Reagent 1 : Cholesterol Enzyme Reagent LIMITATION & PRECAUTIONS :
Reagent 2 : Cholesterol Standard 200 mg/dl. 1. Storage condition as mentioned on the kit to be adhered.
Reagent 3 : Cholesterol Precipitating Reagent (Free) 2.Do not freeze or expose reagents to high temperature.
Protect from direct sun light as it may affect the performance of
MATERIALS REQUIRED BUT NOT PROVIDED: the Kit.
- Clean & Dry Glassware. 3. The Enzyme reagent contains Sodium Azide.
- Laboratory Glass Pipettes or Micropipettes & Tips. 4. Before the assay bring all the reagents to room temperature .
- Colorimeter or Bio-Chemistry Analyzer. 5. Avoid contamination of the reagents during the assay process.
6. Use clean glassware free from dust or debris.
SAMPLES:
Serum, heparinized/ EDTA plasma. Cholesterol is reported to be BIBLIOGRAPHY :
stable for 7 days when stored at 2 - 8°C. 1. Richmond, N.Clin. Chem. 1973: 19 : 1350 - 1356
2. Overview and Recommendations : Current Status of Blood
PREPARATION OF REAGENT & STABILITY : Cholesterol Measurement clin.chem. 34/1, 193-201 (1988)
All Reagents are ready to use and are Stable till the expiry date 3. Allain C.C and al. Clin. Chem., 20 (1974), 470.
mentioned on the label, when stored at 2 - 8 °C.
CODE NO. PACK SIZE Reagent 1 Reagent 2 Reagent 3
GENERAL SYSTEM PARAMETERS: S06 2 x 30 ml 2 x 30 ml 3.0 ml 3.0 ml
Reaction type : End point S06D 2 x 50 ml 2 x 50 ml 3.0 ml 3.0 ml
Wave Iength : 505 nm. (490-530nm) S06E 6 x 50 ml 6 x 50 ml 3.0 ml 3.0 ml
Temperature : 37°C
Incubation : 5 minutes IVD
Reagent volume : 1.0 ml. UNIONPROJET S.R.L.S.
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Sample volume : 10µl. ISO 9001:2015 ISO 13485:2003

Standard concentration : 200 mg/dl

Zero setting : Reagent blank
Light path : 1 cm
HDL CHOLESTEROL
INTENDED USE:
The reagent kit is intended for the in-vitro quantitative
determination of HDL Cholesterol in serum/plasma.

MATERIALS REQUIRED BUT NOT PROVIDED:-

- Clean & Dry Glassware.
- Laboratory Glass Pipettes or Micropipettes & Tips.
- Colorimeter or Bio-Chemistry Analyzer.

SAMPLES:
Unhemolysed Serum, heparinized plasma or EDTA plasma.

GENERAL SYSTEM PARAMETERS:

Reaction type : End point
Wave Iength : 510 nm. (490-530nm)
Temperature : 37°C
Incubation : 5 minutes
Reagent volume : 1.0 ml
Sample volume : 10µl
Standard concentration : 200 mg/dl
Zero setting : Reagent blank
Light path : 1 cm

PREPARATION:
Take 0.5 ml of serum / plasma in to glass tube. Add 50 µl
precipitating reagent. Mix well, leave it at R.T. for 10 minutes.
Centrifuge at 3000 r.p.m. for 10 minutes.Take the clear
supernatant for HDL cholesterol estimation.
PROCEDURE:
1. Bring all the reagents of assay to room temperature.
2. Working assay table
B. S. T.

Enyzyme reagent 1 ml 1 ml 1 ml

Standard - 10µ l -

Supernatant sample - - 10 µl
0
Mix well and incubate for 5 minutes at 37 c
Measure the absorbance of test & Std at 510 nm.

CALCULATION:
Absorbance of HDL Test
HDL Cholesterol mg/dl = x 200
Absorbance of std

NORMAL VALUE:
HDL cholesterol : Men :30-60 mg% , Women :40-70 mg%

NOTE:
If Triglycerides level is more then 300 mg/dl in sample please
dilute the same test sample tube 1:1 with normal saline, add
50 µl precipitating reagent & use the diluted sample for HDL
cholesterol estimation. Final results should be multiplied by the
dilution factor (Two).

IVD www.jas-anz.com.au/register

ISO 9001:2015 ISO 13485:2003