Laboratory Exp. 8 AMYLASE Flashcards
What class does amylase belong?
Hydrolases
Starch is composed of?
Amylose & Amylopectin
It is long, unbranched, polypeptide chain with alpa-1,4-glycosidic bonds.
Amylose
What bond is found in amylose?
Alpha-1,4-glycosidic bonds
a branch of polypeptide chain with alpha-1,6-glycosidic bonds
Amylopectin
What bond is found in Amylopectin?
alpha-1,6-glycosidic bonds
It is the smallest enzyme that is readily filtered by glomerulus(renal)
Amylase
Amylase can appear in?
Urine
It is important enzyme in the physiologic digestion or starches
Amylase
What are the major source/tissue source of Amylase?
Pancreas and Salivary glands
What are the isoenzyme in amylase?
P-type isoamylase
S-type isoamylase
Amylase catalyze the breakdown of _____ and _____
Starch & Glycogen
Difference between glycogen and amylopectin?
Glycogen is more HIGHLY BRANCHED
What is the only bond attacked by alpha-AMY to produce degradation products consisting of glucose, maltose, and intermediate chains, called dextrin?
alpha-1,4-glycosidic bonds
What are the degradation products?
Glucose, Maltose, and Dextin
Tissue source with lesser concentration of serum AMY
Skeletal muscle, small intestine, fallopian tubes
Molecular weight of Amylase
50,000 to 55,000
Stability of Specimen in Amylase?
1 week at Room Temperature
What is the temperature in Several months:
2-8 Deg C
Does the anticoagulant Citrate, oxalate, and EDTA can used in Amylase Specimen collection? (T/F)
False
Status of serum AMY levels in axite pancreatitis: Time
Begin to rise:______
Peak:_____
Return to normal:______
:5-8 hrs
:24 hrs
:3-5 days
Other disorders causing elevated serum AMY level
•Salivary gland lesions(mumps and parotitis)
•Intra-abdominal diseases(intestinal obstruction, cholecystitis, appendicitis)
•Renal insufficiency
•Diabetic ketoacidosis
Cause of false increase:
-Macroamylasemia
-Lipemia
-Bilirubin Concentration >20mg/dL
-Insulin
-Morphine and Opiates
-Contamination with saliva and sweat
It measures the disappearance of starch substrate
AMYLOCLASTIC
Measures the appearance of the product
Saccharogenic
Measures the increasing color from production of product coupled with a chromogenic dye
Chromogenic
Coupling of several enzyme systems to monitor amylase activity
Continuous Monitoring
Methods to separate isoenzyme:
Physical Characteristics
Chromatography
Electrophoresis
Electric forms
Immune inhibition
Immune precipitation
selective inhibition
Where is P isoamylase derived from?
Pancreatic tissue
Where is S isoamylase derived from?
Salivary glands, Fallopian tube, & Lung
Method of Amylase
Kinetic Method by wallenfels et al
He introduced iodometric method in 1908?
Wohlegemuth
He introduced a procedure that standardized the amount of starch and iodine? What year?
Somogyi,1938
Somogyi’s work became the basis for the widely used _______ & __________ methods in 1956 & 1960
Amyloclastic and Saccharogenic
What did Wallenfels et al introduced?
p-nitrophenylglycosides
a substrate that terminal glucose blocked to reduce spontaneous degradation
p-nitrophenyl-D-maltohepataoside(PNPG7)
Reduce spontaneous degradation of the substrate by:
glucosidase & glucoamylase
Convert Somogyi units to IU
x 1.85
Linearity of Amylase Determination
1500IU/L
Factor in Amylase
4824
Expected Value
up to 96 IU/L
