VLD 421: Veterinary Clinical Biochemistry and Laboratory Diagnosis-II (0+1)

ESTIMATION OF ALPHA AMYLASE

Amylase:  (CNPG₃ Method- Agappe Diagnostics)

• Amylase occurs in the salivary glands, pancrease. Alpha amylase is secreted by the pancrease from where it enters the duodenum, through the panceatic duct.

Sample

Serum, Plasma and Urine

Principle:

       5 CNPG₃    __Amylase___   3 CNP + 2CNPG2 + 3 Maltotriose +  Glucose .

               CNP _ 2- Chloro- 4 - nitrophenol

               CNP-G₂ - 2- Chloro- 4- nitrophenyl-a-maltoside.

Reagent composition

• MES buffer (pH 6.0) - 50 mmol/L
• CNPG₃                        - 2.27 mmol/L
• Calcium chloride        -  60 mmol/L
• Sodium chloride         - 70 mmol/L
• Activator                     - 900 mmol /L

Laboratory procedure

Working  Reagent                                               1000 μL

Sample                                                                      25 μL

Mix and incubate for 1 minute, at 37 C . Measue the change in absorbance per minute during 3 minutes.

Calculation 

 Alpha Amylase activity (IU/dl) = (Change in OD/ min ) x 3178

Clinical Significance

• Elevated in acute pancreatitis, peptic ulcers, biliary disease, intestinal obstrction.
• Decrease in chronic pancreatic disorders, pancreatic cell destruction.