estimation of total lipid in serum,

8/8/2019 Estimation of Total Lipid in Serum,

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Estimation of Total Lipid in Serum,

Estimation of Triglycerides in Serum&

Estimation of Cholesterol in Serum ,

Estimation of HDL Cholesterol in Serum


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Introduction:

A lipid panel or lipid profile is a

blood test that measures lipids-fats and

fatty substances used as a source of

energy by the body.


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This Panel Measures:

1. Total cholesterol level.

2. Triglyceride level.

3. HDL cholesterol level. This is the "good"

cholesterol.

4. LDL cholesterol level. This is the "bad"

cholesterol.


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Other measurements that may be done for a

lipid panel include:

5.Very-low-density lipoprotein (VLDL) cholesterol level.

6. The ratio of total cholesterol to HDL.

7. The ratio ofLDL to HDL.


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Lipids: are insoluble (does not dissolve) in water but

are soluble (dissolves) in alcohol and other solvents. They

are an important part of cells, and they help keep your

body working normally. Lipid disorders, such as high

cholesterol, may lead to life-threatening illnesses, such as

coronary artery disease (CAD), heart attack or stroke.


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WhyIt Is Done?

1.To help determine the risk of heart disease.

2. To screen for a lipid disorder.

3. To evaluate the success of lipid-lowering lifestyle changes

such as diet and exercise.

4. To determine the effectiveness of drug therapy.


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Note:

1. Very Low Density Lipoprotein (VLDL).

2. High density lipoprotein (HDL)

3. Low density lipoprotein (LDL).

Each one of these particles contains a mixture of

cholesterol, protein, and triglyceride, but in varying

amounts unique to each type of particle.


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LDL (bad cholesterol) contains the highest amount of

cholesterol and it causes heart disease. HDL (good

cholesterol) contains the highest amount of protein and

it protects against heart disease.The HDL transport

cholesterol from the tissues of the body to the liver.


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Very Low Density Lipoprotein (VLDL) is one of

three major lipoprotein particles. VLDL contains the

highest amount of triglyceride. It is possible to

estimate the amount of VLDL cholesterol by

dividing the triglyceride value (in mg/dL) by 5.


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This calculation is not valid when the triglyceride is

greater than 400 mg/dl.A normal VLDL cholesterol

level is between 5 and 40 milligrams per deciliter.


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Lipid Profile Value:

High RiskBorderlinedesirableAdult Value

240 mg/dl200-240 mg/dl200 mg/dlCholesterol

400 -1000

mg/dl

200-400 mg/dl200 mg/dlTriglycerides

35 mg/dl35-45 mg/dl60 mg/dlHDL-

Cholesterol

160 mg/dl130-160 mg/dl 130 mg/dlLDL-Cholesterol


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Estimation ofTriglycerides in Serum

Principle:

It is Enzymatic Colorimetric, Determined of triglycerides

after enzymatic hydrolysis with lipases. The indictor

quinonemine is formed from hydrogen peroxide and 4-

aminoantipyrine and 4-chlorophenol under the catalytic

influence of peroxidaes.


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1.Triglycerides Lipase Glycerol + Fatty acid

2. Glycerol + ATP Glycerol Kinase Glycerol-3-Phosphate +ADP

3. Glycerol-3-Phosphate + O2 Glycerol-3-Phosphate oxidase

dihydroxyacetonephosphate + H2O2

4. H2O2 +4-aminoantipyrine + 4-chlorophenol Peroxidaes

quinonemine + 2H2O + HCL


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Reagents:

The reagents and standard provided in the kit are

ready for use.

Specimen:Serum of fasting10 -12h or plasma.


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Assay Parameters:

Wavelength: Hg505 nm.

Optical path: 1 cm.

Temperature: 20-25C

Measurement: Against reagent blank


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Protocol:

1. Take three cuvettes label the first one as reagent

blank, the second as standard and the last one as sample.

2. To the reagent cuvette, add1000 l of the reagent only.

3. To the standard cuvette, add10 l standard and1000

l reagent.4. To the sample cuvette, add10 l sample and1000 l

reagent.


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5. Mix the three cuvettes and incubate for10 min.

at 20-25C.

6. Measure the absorbance of the sample and thestandard against the reagent blank within 30 min at

505 nm.

7. The measured value will yield the value of change

in absorbance orA.


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Calculation of theTriglycerides Concentration:

C = 200 xA sample /ASTD [mg/dl ]

Interpretation:

High RiskBorderlineDesirableAdult Value

400-1000

mg/dl

200-400

mg/dl200 mg/dlTriglycerides


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Estimation of Cholesterol in Serum

Principle:

It is Enzymatic Colorimetric, Determined of cholesterol

after enzymatic hydrolysis and oxidation. The indictor

quinonemine is formed from hydrogen peroxide and 4-

aminophenazone in the presence of phenol and

peroxidase.


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1. Cholesterolester + H2O Cholesterolesterase

Cholesterol + Fatty acid

2. Cholesterol + O2 Cholesteroloxidase

Cholesterol-3-one + H2O2

3. 2H2O2 +4-aminophenazone + phenol Peroxidaes

quinonemine + 4H2O


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Reagents:

The reagents and standard provided in the kit are

ready for use.

Specimen:

Serum of fasting10 -12h or EDTA plasma.


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Assay Parameters:

Wavelength: Hg505 nm.

Optical path: 1 cm.

Temperature: 20-25C

Measurement: Against reagent blank


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Protocol:

1. Take three cuvettes label the first one as reagent

blank, the second as standard and the last one as sample.

2. To the reagent cuvette, add1000 l of the reagent only.

3. To the standard cuvette, add10 l standard and1000

l reagent.4. To the sample cuvette, add10 l sample and1000 l

reagent.


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5. Mix the three cuvettes and incubate for10 min.

at 20-25C.

6. Measure the absorbance of the sample and thestandard against the reagent blank within 30 min at

505 nm.

7. The measured value will yield the value of change

in absorbance orA.


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Calculation of the Cholesterol Concentration:

C = 200 xA sample /ASTD [mg/dl ]

Interpretation:


240 mg/dl200-240

mg/dl

200 mg/dlCholesterol


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Estimation of HDL Cholesterol in Serum

Phosphotungestic Precipitation Method,CHOD PAP Method.

Principle:

The chylomicrons, VLDL (very low density lipoproteins),

IDL (Intermediate density lipoproteins), and LDL (low

density lipoproteins) (parts of lipoprotein) are precipitated

by addition of phosphotungstic acid and magnesium

chloride. After centrifugation the supernant fluid contains

the HDL fraction, which is assayed for HDL cholesterol.


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Calculation of HDL cholesterol Concentration:

With Factor:

C = 180 xA sample [mg/dl]

With Standard:

C = 150 xA sample /ASTD [mg/dl]

LDL cholesterol = total cholesterol HDL cholesterol (0.20

triglycerides) (mg / dl)


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ormal Ranges in Serum or Plasma:


35mg/dl

35

-45

mg/dl60 mg/dlHDL-Cholesterol

160 mg/dl130-160

mg/dl

130 mg/dlLDL-

Cholesterol

estimation of total lipid in serum,

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