HRISHABH B BOOND

DEMONSTRATOR
G.S.V.M. MEDICAL COLLGE KANPUR
Transamination
i. Transamination is the exchange of the alpha amino group
between one alpha amino acid and another alpha keto
acid, forming a new alpha amino acid.

amino acid 1 + keto acid 2 → amino acid 2 + keto acid 1

ii. As an example, amino group is interchanged between

alanine and glutamic acid . In almost all cases, the amino
group is accepted by alpha ketoglutaric acid so that
glutamic acid is formed.
What is SGPT & SGOT ?
 These are amino transferases.

 Amino transferases are the group of enzymes which

catalyses the interconversion of amino acids to 2 oxo
acids by transfer of amino groups.

 Also the production of new amino acid and a new

keto acid.
SGOT
 SGOT is known as Serum Glutamate Oxaloacetate
Transaminase
 Now a days know as Aspartate Amino Transaminase
(AST).
 Requires pyridoxal phosphate (Vitamin B6) for its activity
as coenzyme.
 Occurs in high concentration in heart, liver, skeletal
muscles, kidney and erythrocytes.
SGOT
 Normal range :- 5 to 40 IU/L

 In Myocardial infraction, rise in concentration may occur

within 24-36 hours after the onset of symptoms.

 Peak level is a high as 160 units within 36-48 hours

 Return to normal in within 4-6 days

 These days preference is given to Troponins as a very

good markers for MI thann SGOT
Estimation of SGOT
 Method :- UV kinetic method

 Requirements :-
 Test tube

 Reagent (SGOT)

 Serum sample

 Micropipette

 Biochemistry analyser
Principle
 It is based on the oxidation of NADH to NAD+. Enzyme
SGOT/AST converts L- aspartate to oxaloacetate. The
oxaloacetate so formed is then reduced to malate by
enzyme malate dehydrogenase (MDH) with simuntaneous
oxidation of NADH to NAD+ .
 Since NADH has a absorbance peak at 340 nm and NAD+
has little absorbance at this wavelength, AST activity is
measured as it is directly proportional to the decrease in the
wave length
Principle
 L-aspartate + 2-oxoglutarate SGOT oxaloacetate +
L-glutamate

 Oxaloacetate + NADH MDH Malate + NAD+

 Serum pyruvate + NADH LDH L-Lactate + NAD+

Here above reaction is only proceeds to completely

reduce the pyruvate so that it can not interfere in the
assay.
Reagents
 2-oxoglutarate

 L-aspartate

 Malate dehydrogenase

 Lactate dehydrogenase

 NADH

 Tris buffer (pH = 7.8 at 25°c)

Procedure
 Add 100µl of serum in 1000µl of working reagent.

 Mix well and aspirate the sample and result is

calculated
Calculation
 Convert the absorbance into IU

 AST = ∆ Abs/min * Total reaction volume * 1000

(IU/L) sample volume * Absorptivity

∆ Abs/min * 1.10 * 1000

0.10 * 6.22
 P = Cuvette Light path = 1 cm.

 AST = ∆ Abs/min * 1768.488

Clinical intrepretations
AST activities increased in-
 Myocardial Infraction

 Severe arrhytmias and severe angina

 Acute pancreatitis

 Progessive muscular dystrophy

 Recent brain trauma and its necrosis

 Metastatic tumor of liver

 Pulmonary infection

 Drugs
Clinical intrepretations
AST activities decreased in-

 Liver cirrhosis

 Malabsorption syndrome
SGPT
 SGPT is known as Serum Glutamate Pyruvate
Transaminase.
 Now a days known as Alanine amino transaminase
(ALT).
 It is a cytosolic enzyme synthesized in liver and it is
more liver specific enzyme.
 Normal serum level of ALT for male is 13–35 U/L and
for female is 10–30 U/L
SGPT
 SGPT is known as Serum Glutamate Pyruvate
Transaminase.

 Now a days known as Alanine transaminase (ALT).

 It is a cytosolic enzyme synthesized in liver and it isb

more liver specific enzyme.
Estimation of SGPT
 Method :- UV kinetic method

 Requirements :-
 Test tube

 Reagent (SGOT)

 Serum sample

 Micropipette

 Biochemistry analyser
Principle
 ALT acitivity is measured by the determining the rate of
oxidation of NADH to NAD+,which is indicated by
decrease in absorbance at 340 nm. GPT/ALT converts
L-alanine to pyruvate. Then pruvate is reduced to
lactate by lactate dehydrogenase with simultaneous
oxidation of NADH to NAD+ at 340 nm.
Principle
 ALT activity is directly proportional to the decrease in
absorbance at 340 nm due to oxidation of NADH to
NAD+.

 L-alanine + 2-oxoglutarate ALT Pyruvate +

L-Glutarate

 Pyruvate + NADH LDH L-Lactate + NAD+

Reagents
 2-oxoglutarate

 L-alanine

 Lactate dehydrogenase (LDH)

 NADH

 Tris buffer (pH = 7.8 at 25°c)

Procedure
 Add 100µl of serum in 1000µl of working reagent.

 Mix well and aspirate the sample and result is

calculated
Calculation
 Convert the absorbance into IU

 ALT = ∆ Abs/min * Total reaction volume * 103

(IU/L) sample volume * Absorptivity

 P = Cuvette Light path = 1 cm.

 ALT = ∆ Abs/min * 1768

Clinical intrepretations
ALT activities increased in-
 Acute viral hepatitis

 Toxic hepatitis (due to overdose of drug)

 Tumor of liver

 Highly increased in hepatic jaundice

 Obstructive jaundice

 Ischemia of liver

 Extensive trauma, muscle disease, shock

Clinical intrepretations
ALT activities decreased in-

 Liver cirrhosis

 Malabsorption syndrome during abnormal metabolism

of pyrimidine.
 Elevation of ALT is more in cases of hepatic disease
compared to AST. But AST may be more than ALT in
alcoholic liver disease. In alcoholic liver disease, the
actual values show only mild elevation; but a ratio of
AST/ALT more than two is quite suggestive.
 Moderate elevation of amino transferases often
between 100–300 U/L is seen in alcoholic hepatitis,
autoimmune hepatitis and nonalcoholic chronic
hepatitis .
 Minor elevation less than 100U/L is seen in chronic
viral hepatitis (hepatitis C), fatty liver and in
nonalcoholic steatohepatitis (NASH). In chronic
hepatitis and cirrhosis of liver, serum ALT poorly
correlates with the degree of liver cell damage.
Clinical Case Study
 Alanine aminotransferase (ALT):
 Formerly called glutamate-pyruvate
transaminase, ALT is present in many tissues.
 The enzyme catalyzes the transfer of the amino
group of alanine to α-ketoglutarate, resulting in
the formation of pyruvate and glutamate.
 The reaction is readily reversible. However,
during amino acid catabolism, this enzyme (like
most aminotransferases) functions in the
direction of glutamate synthesis.
 Thus, glutamate, in effect, acts as a “collector” of
nitrogen from alanine.
 Aspartate aminotransferase (AST):
 AST formerly called glutamate-oxaloacetate
transaminase, AST is an exception to the rule that
aminotransferases funnel amino groups to form
glutamate.
 During amino acid catabolism, AST transfers amino
groups from glutamate to oxaloacetate, forming
aspartate, which is used as a source of nitrogen in
the urea cycle
 [Note: The AST reaction is also reversible.]
 Liver disease:
 Plasma AST and ALT are elevated in nearly all liver
diseases, but are particularly high in conditions that cause
extensive cell necrosis, such as severe viral hepatitis, toxic
injury, and prolonged circulatory collapse.
 ALT is more specific than AST for liver disease, but the
latter is more sensitive because the liver contains larger
amounts of AST.
 Serial enzyme measurements are often useful in
determining the course of liver damage.
Clinical Case Study
 A middle aged man complained of fatigue. There was
vague abdominal pain. On examination, there was
enlargement of the liver. Laboratory results obtained
were as follows – Serum AST – 120 U/L, ALT – 80 U/L,
ALP – 68 U/L, GGT – 170 U/L, Bilirubin – 1.0 mg/dL,
glucose – 60 mg/dL, Uric acid – 8.0 mg/dL. CBC and
urinalysis were normal. What is the diagnosis? Interpret
the biochemical data on the basis of your diagnosis.
Clinical Case Study
 A patient presented with acute chest pain of half
hour duration. The biochemical analysis reports are
as follows Blood Glucose – 350 mg%, Serum
Cholesterol – 288 mg%, SGOT – 55 U/L, SGPT – 15
U/L. CPK and LDH were elevated. Give your
provisional diagnosis. What are the other markers
which can be estimated in this case?