amylase INTRODUCTION Amylase found in pancreas and salivary gland It is break down the polysaccharide into monosaccharide as this equation General principle Starch amylase maltose Glucose ID: 918870
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Slide1
Lab 2Amylase
Quantitative determination of
amylase
Slide2INTRODUCTION
*Amylase found in pancreas and salivary gland.
It is break down the polysaccharide into monosaccharide as this equation( General principle):
Starch amylase maltose + Glucose
Slide3Salivary and Pancreatic Amylase
In the body, the highest concentrations of amylase are found in the saliva and pancreas.
Salivary amylase
(ptyalin, S-type): hydrolysis of starches while the food is in the mouth and esophagus.
Pancreatic amylase (P-type): synthesized by the acinar cells and then secreted into the intestinal tract by way of pancreatic duct system. Small amounts of amylase are normally present in the blood. However, increased amounts may be released into the blood
when the pancreas is injured, inflamed, or blocked.
Slide4Introduction:
The determination of amylase activity in serum and urine is the most commonly performed for the diagnosis of acute pancreatitis.
In acute pancreatitis , amylase levels are elevated for longer periods of time in urine than in serum.
Determining the ratio of the amylase and creatinine clearances is important in following the course of the pancreatitis.
Slide5Acute and chronic pancreatitis
Acute pancreatitis occurs when the pancreas suddenly becomes inflamed but then gets better.
Chronic pancreatitis is persistent inflammation of the pancreas or a combination of persistent inflammation with repeated attacks of acute pancreatitis.
Causes: alcohol abuse or gallstones.
Symptoms of a pancreatic disorder: abdominal
pain, fever, loss of appetite, or nausea.
Slide6Clinical diagnosis
Elevation of enzyme in:
Acute or chronic pancreatitis.
It reflects biliary or gastrointestinal disease .It is increase in salivary gland lesions such as mumps.Amylase is the only enzyme seen normally in urine
Slide7PRINCIPLE of Kinetic Method
Amylase hydrolyzed p-
Nitrophenyl
D-Maltoheptaoside (PNPG7) top-Nitrophenylmaltotriose (PNPG3) and Maltotetraose
. Glucoamylase hydrolyzes PNPG3 to p-Nitrophenylglycoside (PNPG1) and glucose.Then PNPG1 is hydrolyzed by glucosidase to glucose and p-Nitrophenol
, which produces a yellow color. The rate of increase in absorbance is measured at 405 nm and is proportional to the amylase activity in the sample.
Slide8Procedure
WR
-ml
1
Incubate at 37 C for 5 min.
Sample-µl
25
Incubate at 37 C for 90
sec then read Abs. at 405 nm.
Continue read Abs. every 30 sec for 2 min.
Calculate
A/min.
WR
-ml
1
Incubate at 37 C for 5 min.
Sample-µl
25
Slide9Calculation and normal range∆A/min X 4824 = activity of amylase U/L
Normal range:
16-108 U/L