ALANINE TRANSAMINASE (ALT) SGPT

ALT is a blood test that determines the level of the enzyme alanine
transaminase (ALT) in the blood often part of an initial screening for liver
disease.

The liver plays several important roles: It stores fuel from food, makes
proteins, and helps remove toxins from the body. The liver also makes bile, a
fluid that helps in digestion. Proteins called enzymes help the liver build and
break down proteins. ALT (or SGPT, which stands for serum glutamic-pyruvic
transaminase) is one of these enzymes. It's found in particularly large
amounts in the liver and plays an important role in metabolism, the process
that converts food into energy.

Normally, ALT is found inside liver cells. But if the liver is inflamed or injured,
ALT is released into the bloodstream. Measuring blood levels of ALT can give
doctors important information about how well the liver is functioning and
whether a disease, drug, or other problem is affecting it.

Why It's Done

The ALT test may be ordered if you are experiencing symptoms of liver
disease, including jaundice (yellowish skin or eyes), dark urine, nausea,
vomiting, or abdominal pain. It may also be ordered to help diagnose
infections of the liver such as viral hepatitis (ALT levels are high with acute
hepatitis) or to monitor patients taking medications that cause liver-related
side effects.

The ALT test is often performed as part of a full hepatic function panel that
includes other liver enzyme tests. Looking at ALT levels along with those of
other liver enzymes — such as aspartate aminotransferase (AST) and alkaline
phosphatase (ALP) — can give doctors more specific information about liver
problems

Preparation & Expectations

What is involved in preparation for the test?

Generally, no preparation is needed for an ALT blood test. Test is performed
in blood serum the only information required is the drugs you are taking
because there are certain drugs that alter the ALT levels.

Results and Values

What do the test results mean?

Normally, ALT levels range from 6 to 59 units per liter (U/L). Elevated levels of
ALT may indicate one of the following conditions:

hepatitis, or inflammation of the liver disease

 alcoholic liver
 cholestasis, or congestion of the bile ducts
 cirrhosis, or scarring of the liver with loss of function
 death of liver tissue
 cancer of the liver
 noncancerous tumor of the liver
 use of medicines or drugs toxic to the liver

Risks

The ALT test is considered a safe procedure. However, as with many medical
tests, some problems can occur with having blood drawn:
 fainting or feeling lightheaded
 hematoma (blood accumulating under the skin causing a lump or
bruise)
 pain associated with multiple punctures to locate a vein
 SERUM ALANINE AMINOTRANSFERASE ESTIMATION

PRINCIPLE:

The reaction system is as follows:

GPT
L-alanine + 2-oxoglutarate---------------- pyruvate + L-glutamate

LDH
Pyruvate + NADH + H ------------------ l-lactate + NAD+
+

PROVIDED REAGENTS:

Buffer: TRIS buffer solution, pH 7.5 with L-alanine

Substrate: Vials containing 2-oxoglutarate, reduced NADH and LDH

INSTRUCTIONS FOR USE:

Provided reagents are ready to use. They may be used separately or as Monoreagent,
mixing 4 parts Buffer and 1 part Substrate.

SAMPLE:

• Collection: Collect anticoagulated blood.

• Additives: Use concentrated EDTA anticoagulants or dried balance
anticoagulants.
• Stability and storage instructions: ALT in serum is stable in refrigerator up to 3
days without preservatives. Do not freeze.

REQUIRED MATERIAL (non- provided):

• Spectrophotometer.
• Micropipettes and pipettes for measuring the stated volumes.
• Spectrophotometer cuvettes or Photo colorimeter tubes.
• Watch or timer.
• Water bath

ASSAY CONDITIONS:

• Wave-length: 340nm
 • Reaction temperature: 25, 30 or 370C
• Reaction time: 4 min.
• Sample volume: 100 µl.
Sample and reagent volumes may be proportionally changed without altering the
corresponding calculation factors.

PROCEDURE:

MONOREAGENT TECHNIQUE:
In a cuvette at 30-370C, place:

Monoreagent 1.0ml

Preincubate for a few minutes, and then add:

Sample 100 µl

Mix immediately and simultaneously start stop watch. After 90 seconds record initial
absorbance and then at 1, 2 and 3 minutes after the first reading. Determine average
change in absorbance/min (∆A/min), subtracting each reading from the previous one and
averaging these values. Use this mean for calculations.

CALCULATIONS:

A1 =

A2 =

A3 =

A4 =

∆A = (A2 – A1) + (A4 –A3)

----------------------------
2

ALT(U/l) = ∆A/min × factor (factor = 1746)