Delos Santos , Jesspeed Marvel

Dimapilis, Almira

Aspartate Transaminase (AST) is also

called Serum Glutamic Oxaloacetic
Transaminase (SGOT).

Alanine Transaminase (ALT) is also

called Serum Glutamic Pyruvic
Transaminase (SGPT).

SGOT and SGPT are one of the blood tests

that are most commonly performed to
assess the function of the liver.

ALT

AST Determination using Malate

Dehydrogenase (MDH)

ALT Determination using Lactate

Dehydrogenase (LDH)

Simultaneous oxidation of NADH to NAD is

measured by a change in absorbance at
340nm,

Absorbance Transamination Activity.

Pipette into 25C, 30 C

Cuvettes
Sample
Working
Reagent

200 ul
1000 ul

37 C

100 ul
1000 ul

Mix and read the absorbance after 1 minute.

Read the absorbance again exactly after 1, 2
and 3 minutes.

ALT
Enzyme reagent

Tris buffer (pH=7.5) -100 mmol/L

L-alanine - 500 mmol/L
LDH - 1200 U/L
NADH - 0.18 mmol/K

Starting reagent
-ketoglutarate - 15 mmol/L

Lactate Dehydrogenase
An enzyme that catalyzes the conversion of lactate
to pyruvate
Some of the organs relatively rich in LDH are the
heart, kidney, liver, and muscle.

Malate Dehydrogenase
Enzyme in the citric acid cycle that catalyzes the
conversion of malate into oxaloacetate.

Tris Buffer
Commonly known as
trishydroxymethylaminomethane
2-amino-2-hydroxymethyl-1,3-propanediol
Effective pH range between 6.5 and 9.7.
An effective buffer for slightly basic solutions.

L-aspartate
Classified as an acidic amino acid.
Non-essential- produced from oxaloacetate by
transamination.
Carries reducing equivalents in the malateaspartate shuttle, which utilizes the ready
interconversion of aspartate and oxaloacetate,
which is the oxidized (dehydrogenated)
derivative of malic acid.

NADH
in metabolism, it is used as a reducing agent
that donate electrons
The main role of NAD(P)+ in metabolism is
the transfer of electrons from one redox
reaction to another.
The energy is transferred to NAD+ by
reducing this to NADH, as part of glycolysis
and the citric acid cycle.

-Ketoglutarate
Keto acid produced by deamination of glutamate
Key intermediate in the Krebs cycle, coming
after isocitrate and before succinyl CoA
Anaplerotic reactions can replenish the cycle at
this juncture by synthesizing Alpha-ketoglutarate

Results and Discussion

SGOT absorbance at 1st sample

Time elapsed (min)

Absorbance

0.017

0.011

0.005

SGOT absorbance at 2nd sample

Time elapsed (min)

Absorbance

0.017

0.007

0.005

SGPT absorbance of 1st sample

Time elapsed (min)

Absorbance

0.031

0.041

0.047

SGPT absorbance of 2nd sample

Time elapsed (min)

Absorbance

0.228

0.213

0.194

Formula:
Hg 334nm

340 nm

Hg 365

U/l (25C, 30C) = A/min x

971

952

1765

U/l (37C) = A/min x

1780

1745

3235

Where, A/min is the average of difference of two

consecutive absorbances per time elapsed
e.g. A/min=[(a1-a2)+(a2-a3)]/2

SGOT:

1st sample
sample1 = A/min x 952
sample1 = [(a1-a2)+(a2-a3)]/2 x 952
sample1 = [(0.017-0.011)+(0.011-0.005)]/2 x 952
sample1 = 5.71 U/l

NV: Male = up to 18 U/l

Female = up to 15 U/l

Interpretation: Within normal values

SGOT:

2nd sample
sample2 = A/min x 952
sample2 = [(a1-a2)+(a2-a3)]/2 x 952
sample2 = [(0.017-0.007)+(0.007-0.005)]/2 x 952
sample2 = 5.71 U/l

NV: Male = up to 18 U/l

Female = up to 15 U/l

Interpretation: Within normal values

Increased SGOT:

Acute pancreatitis

Muscle dystrophy

Myocardial infarction

Hepatocellular disorder

SGPT:
1st sample
sample1 = A/min x 952
sample1 = [(a1-a2)+(a2-a3)]/2 x 952
sample1 = [(0.031-0.041)+(0.041-0.047)]/2 x 952
sample1 = 7.62 U/l
NV: Male = up to 18 U/l
Female = up to 15 U/l
Interpretation: Within normal values

SGPT:
2nd sample
sample2 = A/min x 952
sample2 = [(a1-a2)+(a2-a3)]/2 x 952
sample2 = [(0.228-0.213)+(0.213-0.194)]/2 x 952
sample2 = 16.18 U/l
NV: Male = up to 18 U/l
Female = up to 15 U/l
Interpretation: Slightly elevated due to
hemolysis

Increased ALT:

Hepatic parenchymal disease

Viral hepatitis

Obstructive jaundice

Heart failure

Acute myocardial infarct

Muscular dystrophy

Skeletal muscle disorders

Normally present in serum and are in high

concentrations in some conditions
function in the synthesis and degradation of
amino acids.
diagnosing liver diseases, myocardial
infarction and musculoskeletal diseases.

They are often determined simultaneously

because in diagnosing a certain condition,
there maybe a single transaminase or both are
elevated.
close proximity in the organs which they can
be found. ALT is primarily found in the liver
while AST is found in many body tissues
including the heart, muscle, kidney, brain and
lungs.