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Amylase Test Procedure

Amylase is an enzyme, or special protein, produced by your pancreas and salivary glands. The pancreas is an organ located behind your stomach. It creates various enzymes that help break down food in your intestines.

Urine Amylase Test

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Summary:

A Amylase is secreted by the pancreas into the duodenum where it aids the catabolism of carbohydrates to simple sugars. Damage to the pancreas or obstruction to the pancreatic duct causes the enzyme to enter the blood stream. Elevated levels are found in acute pancreatitis, perforated / penetrating peptic ulcers, paraotitis (mumps). Patients with chronic pancreatic disorders having pancreatic cell destruction do not have high levels as less amylase is produced by the pancreas.

Principle:

A Amylase catalyses the hydrolysis of a 2 – chloro – 4 nitro phenol salt to chloro nitrophenol (CNP). The rate of hydrolysis is measured as an increase in absorbance due to the formation of chloro nitrophenol which is proportional to the a Amylase activity in the sample.

Contents:

L1. Reagent2x10ml , 2x30ml

Storage / stability:

Contents are stable at 2-8°C till the expiry mentioned on
the label.

Reagent Preparation:

Reagents are ready to use. Do not pipette with mouth.

Sample material:

Serum, Heparinised Plasma, urine.
A Amylase is reported to be stable in the sample for 5 days at 2-8°C. Separate serum from clot as soon as possible.

System Parameters:

ReactionKinetic
Wavelength405 / (Hg405) / violet
Zero SettingDistilled water
lncub. Temp37°c
lncub. Time
Delay Time60 sec.
Read Time180 sec.
No. of read.4
Interval60 sec.
Sample Vol. 0.02 ml / 0.01 ml
Reagent Vol. 1.00ml
Standard Factor 3954 / 7830
React. Slope Increasing
Linearity 1000 U/L
UnitsU/L

Procedure:

Serum / Plasma Sample:

Pipette into a clean dry test tube labelled as Test (T)

Addition SequenceT (37°C)
Amylase Reagent ( L1 )1.0 ml
Sample0.02 ml (20μl)
Mix well and read the initial absorbance A,after 1 minute & repeat the absorbance reading after every 1, 2, & 3 minutes. Calculate the mean absorbance change per minute (M/min.)

Urine Sample:

Pipette into a clean dry test tube labelled as Test (T)

Addition SequenceT (37°C)
Amylase Reagent ( L1 )1.0 ml
Sample0.01 ml (10μl)
Mix well and read the initial absorbance A,after 1 minute & repeat the absorbance reading after every 1, 2, & 3 minutes. Calculate the mean absorbance change per minute (M/min.)

Calculations:

a Amylase Activity in U /L (Serum) = M/min.x 3954
a Amylase Activity in U /L (Urine) = M/min.x 7830

Normal value:

The value may vary from lab to lab.

Serum:

  • Adult = 19 to 86 U/L
  • Newborn = 6 to 65 U /L

Urine:

  • Spot Urine : Upto 48 U/l
  • 24 hrs Urine: Upto 17 U/l

Amylase Increased Level Is Seen In:

  • Acute Pancreatitis and Acute exacerbation of chronic Pancreatitis.
  • Pancreatic cyst and pseudocyst.
  • Acute alcoholic poisoning.
  • Salivary glands diseases like mumps, or duct obstruction or parotitis.
  • Maybe in acute cholecystitis, intestinal obstructions.
  • Increased in obstruction of the common bile duct, pancreatic duct, or ampulla of Vater.
  • Ruptured Ectopic pregnancy.
  • Renal failure due to reduced excretion by the kidneys.
  • Diabetic ketoacidosis.
  • In peritonitis.
  • In some of the lung and o to 65 ovarian tumors.
  • Alcohol intake.
  • May be seen in abdominal trauma, head injury, viral infections, and postoperative patient.

Amylase Level Decreased In:

  1. Extensive marked destruction of pancreas e.g. Acute Fulminant Pancreatitis.
  2. Chronic Pancreatitis and pancreatic cancer.
  3. In pancreatectomy.
  4. Cirrhosis and severe liver diseases like hepatitis.
  5. Perforating or penetrating peptic ulcers.
  6. Toxemia of pregnancy.
  7. Advanced cystic fibrosis.

Related Articles:

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