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Clinical Biochemistry: Lipid Profile

The document discusses lipid profiles and cholesterol testing. A lipid profile measures total cholesterol, triglycerides, HDL, LDL, and VLDL in serum. These lipoproteins are tested to screen for hyperlipidemia, predict heart disease risk, and monitor treatment. The document then describes the manual enzymatic method used to quantify total cholesterol levels in serum or plasma samples.

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356 views

Clinical Biochemistry: Lipid Profile

The document discusses lipid profiles and cholesterol testing. A lipid profile measures total cholesterol, triglycerides, HDL, LDL, and VLDL in serum. These lipoproteins are tested to screen for hyperlipidemia, predict heart disease risk, and monitor treatment. The document then describes the manual enzymatic method used to quantify total cholesterol levels in serum or plasma samples.

Uploaded by

Qasm
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
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Clinical Biochemistry

Lipid Profile
Serum lipid profile include:
1. Total cholesterol
2. Triglyceride
3. High-density lipoprotein cholesterol (HDL-cholesterol)
4. Low-density lipoprotein cholesterol (LDL-cholesterol)
5. very low-density lipoprotein cholesterol (VLDL-cholesterol)

There are five major classes of lipoproteins :

1- Chylomicrons.
2- Very-low-density lipoproteins (VLDL).
3- Intermediate-density lipoproteins (IDL).
4- Low-density lipoproteins (LDL)
5- High-density lipoproteins (HDL).

Cholesterol levels are fairly constant, but triglyceride levels


fluctuate considerably from day to day and are highest 1- 4 hours
after meals.
Plasma lipoprotein particles contain variable proportions of four
major elements: cholesterol, triglycerides, phospholipids and
specific proteins called apoproteins.

Clinical Significance:

1- Screening and diagnosis of hyperlipidemia:

Types of hyperlipidemia

a- Primary hyperlipidemia : genetic or familial.


1- primary hypercholesterolemia
2- primary hypertriglyceridemia
3- primary combined hyperlipidemia

b- Secondary hyperlipidemia:
Certain conditions, medications or food stuffs may lead to
hypercholesterolemia or hypertriglyceridemia or both.
Most common causes of secondary hyperlipidemia are:

1) Uncontrolled diabetes mellitus.

2) Nephrotic syndrome (nephrosis).

3) Chronic obstructive liver disease.

4) Hypothyroidism.

5) Exogenous factors such as excessive intake of dietary fat,


chronic alcohol consumption, or corticosteroid drugs.

2- Prediction of coronary heart disease

increased levels of Serum total cholesterol and low-density


lipoprotein cholesterol (LDL- cholesterol) is directly related to a
greater incidence of coronary heart disease (CHD).

Elevated plasma triglycerides and very-low-density lipoproteins


(VLDL) is also directly associated with increased risk of CHD.

In contrast, high levels of high-density lipoprotein cholesterol


(HDL- cholesterol) have been found to be a protective factor for
the development of CHD, so that decreased levels constitute a
risk factor.

3- Diagnosis and monitoring treatment of atherosclerotic diseases.


Total Cholesterol:

Almost all of the endogenous cholesterol is synthesized by the liver and


gut.

Cholesterol exists in two forms:

- Esterified (75%)
- Non-esterified (Free): (25%).

• Cholesterol is an unsaturated alcohol of the steroid family of


compounds.
• It is essential for the normal function of all human and animal cells
and is a fundamental element of their cell membranes.
• It is also a precursor of various essential substances such as adrenal
and gonadal steroid hormones, vitamin D and bile acids.

Laboratory measurement of total serum cholesterol

Specimen Collection:

• Use serum or plasma.

• For plasma, use heparin or EDTA as anticoagulant.

• Collect blood from a fasting patient (at least 12 hours), non fasting blood
sample is acceptable if the purpose of the test is screening for
hyperlipidemia.

• Separate serum from blood cells within 2 hours.

• Cholesterol is stable in serum or plasma specimen for 5-7 days at 2-8 °C


Reagents:

Reagent 1: Buffer: phosphate Buffer, chloro-4-phenol ,Triton,


Sodium cholate, preservative .

Reagent 2: enzymes (Cholesterol oxidase , Cholesterol esterase,


peroxidase) and 4-amino-antipyrine .

Reagent 3: Standard Cholesterol 200 mg/ dl

Principle:
Enzymatic colourimetric method

Cholesterol esters cholesterol esterase Cholesterol + Free fatty acids

Cholesterol + O2 Cholesterol oxidase Cholesten 4 one 3 + H2O2

2H2O2 + Phenol + 4-amino-antipyrine Peroxidase quinoneimine (pink) + 4H₂O

The intensity of pink color of the complex (quinoneimine) is directly


proportional to total cholesterol concentration in the specimen. The
absorbance is measured at 500 nm.
Manual procedure:

Pipette into well identified test tubes Blank Standard Test

Working reagent (R1+R2) 1ml 1ml 1ml

Standard 10µl

Specimen 10µl

• Mix well, let stand for 5 minutes at 37 °C or 10 minutes at room


temperature.
• Record absorbance (A) of the blank (B), standard (S) and test (T) at 500
nm.
• Color is stable for 1 hour.

Calculation:

𝐀𝐛𝐬 𝐓𝐞𝐬𝐭 − 𝐀𝐛𝐬 𝐁𝐥𝐚𝐧𝐤


𝐂𝐨𝐧𝐜. 𝐨𝐟 𝐓𝐨𝐭𝐚𝐥 𝐂𝐡𝐨𝐥𝐞𝐬𝐭𝐞𝐫𝐨𝐥 = 𝐗 𝐒𝐭d 𝐂𝐨𝐧𝐜.
𝐀𝐛𝐬 𝐒𝐭𝐝 − 𝐀𝐛𝐬 𝐁𝐥𝐚𝐧𝐤

Abs = absorbance, it is measured by spectrophotometer.

Std = standard.

Cholesterol standard concentration = 200 mg/dl


Expected values:

Values for adults, estimated in relation to the risk of atherosclerotic


diseases:

• Recommended (normal) level < 200 mg/dl .

• Borderline high (low risk for CHD) 200 - 239 mg/dl

• High level (high risk for CHD) ≥ 240mg/dl

CHD = Coronary Heart Disease

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