AL- Bayan University

College of Pathological Analysis

Technologies
BioChemistry
2st Year 2020 / 2021

Lab 4

Total Cholesterol Test

Assistant Lecturer
Mustafa Saleem
 Dr. Mustafa Saleem

Cholesterol is : a (a steroid compound) that is essential for life. It

forms the membranes for cells in all organs and tissues in the body.
It is used to make hormones that are essential for development,
growth, and reproduction. It forms bile acids that are needed to
absorb nutrients from food. The test for cholesterol measures total
cholesterol that is carried in the blood by lipoproteins
Cholesterol has three principal functions within the body:
1. It is a structural component of all cell membranes.
2. used to manufacture steroid hormones and Vitamin D; and is
used
to produce bile acids, which facilitate the digestion and
absorption of fats in the diet.
3. The body produces its own supply of cholesterol, mainly in
the liver. It is also present in foods of animal origin, such as
eggs, meat and dairy products.

Total cholesterol:
A small amount of cholesterol circulates in the blood in complex
particles called lipoproteins. Each particle contains a
combination of protein, cholesterol, triglyceride, and
phospholipid molecules and the particles are classified by their
density into:
1.Chylomicrons:Transport dietary Triglyceride(TG) and cholesterol
from the intestines to the liver.
2.high-density lipoproteins (HDL), HDL-C particles, sometimes
called "good" cholesterol, carry excess cholesterol away for
disposal and LDL-C particles, or "bad" cholesterol, deposit
cholesterol in tissues and organs.
3. low-density lipoproteins (LDL), A group of related particles that
transport endogenous TG and cholesterol from the liver to the
tissues.
4.very low-density lipoproteins (VLDL).
Principle of total Cholesterol
Cholesterol is measured enzymatically in serum or plasma in a
series of coupled reactions that hydrolyze cholesteryl esters
and oxidize the 3-OH group of cholesterol. One of the reaction
by products, H2O2 is measured quantitatively in a peroxidase
catalyzed reaction that produces a color. Absorbance is
measured at 500 nm. The color intensity is proportional to
cholesterol concentration. The reaction sequence is as follows:

Cholesteryl ester +H2O-------cholesterylester hydrolase----------

>cholesterol + fatty acid
Cholesterol + O2 ---- cholesterol oxidase -----------> cholest-4-en-3-
one + H2O2
2H2O2+4-aminophenazone+phenol-----peroxidase…------->4-(p-
benzoquinone- monoimino)- phenazone + 4 H2O.
Elevated levels of cholesterol increase the risk for coronary heart
disease (CHD). Cholesterol is measured to help assess the patient's
risk status and to follow the progress of patient's treatment to lower
serum cholesterol concentrations. Desirable cholesterol levels are
considered to be those below 200 mg/dL in adults and below 170
mg/dL in children

How can you calculate total cholesterol in the Body ?

You should know total cholesterol mean
1.cylomicron .(equal zero in fasting)
2. HDL-C.
3.LDL-C.
4.VLDL-C.
 Dr. Mustafa Saleem

The patient fasting two hour at least before analysis total cholesterol.
But lipid profile mean total cholesterol + TG
The patient fasting from 14 to 16 hours before analysis lipid profile.

Total cholesterol= HDL-C +LDL-C+ VLDL-C

LDL-C= Total Cholesterol - [VLDL-C + HLDL-
C]
VLDL= TG mg/dl
5
VLDL= TG
2.2 mmol/L
If the result of TG > 340mg/dl
You must be use kit to find the result of
VLDL-C. and LDL-C
Risk Factor = LDL-C < 2.6
HDL-C
Normal values