Muhammad Asif Zeb

Lecturer MLT
IPMS,KMU
Measurements of the activity of
enzymes in plasma are of value in
the diagnosis and management of a
wide variety of diseases.
 Small amounts of intracellular
enzymes are present in the blood as a
result of normal cell turnover.

When damage to cells occurs,

increased amounts of enzymes will be
released and their concentrations in
the blood will rise.
 However, such increases are not
always due to tissue damage.
Other possible causes include:

increased cell turnover

cellular proliferation (E.g. neoplasia)

increased enzyme synthesis (enzymes

induction)

obstruction to secretion
Enzyme assays usually depend on the
measurement the catalytic activity of the
enzyme, rather than the concentration of the
enzyme protein itself.

One international unit is the amount of

enzyme that will convert one micromole
of substrate per minute per litre of
sample and is abbreviated as U/L.
A major disadvantage in the use of enzymes
for the diagnosis of tissue damage is their
lack of specificity to a particular tissue or
cell type.
Many enzymes are common to more than
one tissue.
 Alanine transaminase or ALT is a transaminase
enzyme (EC 2.6.1.2).

It is also called serum glutamic pyruvic

transaminase (SGPT) or alanine
aminotransferase (ALT).

ALT is found in serum and in various body

tissues, but is most commonly associated with
the liver.
 It is the reaction between an amino acid and an alpha-
keto acid.

The amino group is transferred from the former to the

latter; this results in the amino acid being converted to
the corresponding -keto acid, while the reactant -
keto acid is converted to the corresponding amino
acid.

Transamination in biochemistry is accomplished by

enzymes called transaminases or aminotransferases.

This process is an important step in the synthesis of

some non-essential amino acids (amino acids that are
not supplied from the diet).
More specifically, AST, ALT, and alkaline
phosphatase are called the liver enzymes
and they typically are used to detect
damage or injury to the liver (not its
function).
 ALT is exclusively in cytoplasm of many
tissue like liver, heart, kidney, lungs.

But highest concentration is present in liver.

 It is commonly measured clinically as a part of a
diagnostic liver function test, to determine liver
health.

Diagnostically, it is almost always measured in

units/liter (U/L).

Normal level in serum:

ALT < 45 units/ml
Elevated levels:
Viral or autoimmune hepatitis.

Cirrhosis.

Lack of blood flow to the liver (liver ischemia).

Death of liver tissue (liver necrosis).
Liver tumor.
Use of drugs that are poisonous to the liver.

Biliary duct problems.

For this reason, ALT is commonly used as a way

of screening for liver problems.
 When elevated ALT levels are found in the
blood, the possible underlying causes can be
further narrowed down by measuring other
enzymes.

For example, elevated ALT levels due to

liver-cell damage can be distinguished from
biliary duct problems by measuring
alkaline phosphatase.
 The Alanine Transaminase (ALT) spectrophotometric
assay uses enzymatic reaction:
GPT
L-Alanine + -ketoglutaric acid L-Glutamic acid +
Pyruvic

LDH
Pyruvic acid + NADH + H+ L-Lactic acid +
NAD+

The absorbance is measured at 505 nm, is

proportional to the level of ALT enzyme in the
sample.
Why Get Tested?
To screen for liver damage and/or to help diagnose
liver disease.

Sample Required?
Serum.

When is it ordered?
To evaluate a person who has symptoms of a liver
disorder:
Weakness, fatigue, Loss of appetite
Nausea, vomiting
Abdominal swelling and/or pain
Jaundice
Dark urine, light colored stool
Itching (pruritus)
What does the test result mean?

Very high levels of ALT (more than 10

times the highest normal level) are usually
due to acute hepatitis, often due to a virus
infection.

Inacute hepatitis, ALT levels usually stay

high for about 12 months but can take as
long as 36 months to return to normal.

ALT levels are usually not as high in

chronic hepatitis, often less than 4 times
the highest normal level.
 Other causes of moderate increases in ALT
include obstruction of bile ducts, cirrhosis
and with tumors in the liver.

Inmost types of liver diseases, the ALT level

is higher than AST.
 Aspartate transaminase (AST) also
called serum glutamic oxaloacetic
transaminase (SGOT) (EC 2.6.1.1) is
similar to alanine transaminase (ALT) in that
it is another enzyme associated with liver
parenchymal cells.
Two isoenzymes are present in humans.

GOT1, the cytosolic isoenzyme derives

mainly from red blood cells and heart.

GOT2, the mitochondrial isoenzyme is

predominantly present in liver.

Normal Level:
AST <40 units/ml
 It is raised in acute liver damage. It is also
present in red blood cells and cardiac
muscle, skeletal muscle and may be
elevated due to damage to those sources
as well.
AST was defined as a biochemical marker
for the diagnosis of acute myocardial
infarction in 1954. However the use of
AST for such a diagnosis is now redundant
and has been superseded by the cardiac
troponins.
AST (SGOT) is commonly measured
clinically as a part of diagnostic liver
function tests, to determine liver health.
 Glutamic-Pyruvic Transaminase (GPT) is found
in significant quantities in liver, kidney, and
skeletal muscle, in decreasing order.

When liver cells are damaged, GOT and GPT

levels rise especially early in the disease.

In hepatitis, transaminase levels rise several

days before jaundice begins.

The enzyme levels are especially useful in

assessing subtle and early changes in biliary
obstruction and active cirrhosis.
Principle:
GOT
L-Aspartate + -ketoglutarate L-Glutamate +
Oxaloacetate

The oxaloacetate obtained is measured in its

derative from 2,4 dinitropheny hydrazone.
The absorbance is measured at 505 nm, is
proportional to the level of ALT enzyme in the
sample.

Sample:
Serum.
 Glutamic-Oxaloacetic Transaminase (GOT)
occurs in large concentrations in the heart
and liver with moderate amounts in skeletal
muscle, kidneys, and pancreas.
GOT levels can be used to diagnose
myocardial infarction within 10-48 hours.
Other conditions with elevated GOT include
arrhythmias and severe angina of the heart,
and liver damage.