Delos Santos , Jesspeed MarvelDimapilis, 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 Cuvettes

25ᵒC, 30 ᵒC 37 ᵒC

Sample 200 ul 100 ul

Working Reagent

1000 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 α-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 malate-aspartate 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.

α-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

SGOT absorbance at 2nd sample

Time elapsed (min) Absorbance

1 0.017

2 0.011

3 0.005

Time elapsed (min) Absorbance

1 0.017

2 0.007

3 0.005

SGPT absorbance of 1st sample

SGPT absorbance of 2nd sample

Time elapsed (min) Absorbance

1 0.031

2 0.041

3 0.047

Time elapsed (min) Absorbance

1 0.228

2 0.213

3 0.194

Formula:Hg 334nm 340 nm Hg 365

U/l (25°C, 30°C) = A/min x 971 952 1765

U/l (37°C) = 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 952sample1 = [(a1-a2)+(a2-a3)]/2 x 952sample1 = [(0.017-0.011)+(0.011-0.005)]/2 x 952sample1 = 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 952sample2 = [(a1-a2)+(a2-a3)]/2 x 952sample2 = [(0.017-0.007)+(0.007-0.005)]/2 x 952sample2 = 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 952sample1 = [(a1-a2)+(a2-a3)]/2 x 952sample1 = [(0.031-0.041)+(0.041-0.047)]/2 x 952sample1 = 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 952sample2 = [(a1-a2)+(a2-a3)]/2 x 952sample2 = [(0.228-0.213)+(0.213-0.194)]/2 x 952sample2 = 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.