Clinical Chemistry - Amylase Determination

Question 1

What enzyme reaction class does Amylase belong?

Answer 1

Hydrolases

Question 2

What is the importance of Amylase?

Answer 2

Digestion of starch, it converts to its end product which are simple sugars.

Question 3

Amylase is the breakdown of?

Answer 3

Starch and glycogen with glucose, maltose and dextrin

Question 4

Where does amylase filtered?

Answer 4

In glomerulus (kidney)

Question 5

True/False: Does enzyme amylase found in the urine?

Answer 5

True

Question 6

Where does it is mainly found?

Answer 6

Acinar cells of the pancreas and salivary glands.

Question 7

What minor source organs that can we found amylase?

Answer 7

Small intestine, skeletal muscle, liver and fallopian tube

Question 8

What is the significant diagnosis for amylase?

Answer 8

Acute pancreatitis and salivary gland lesions.

Others: Intra abdominal diseases, renal insufficiency, diabetic ketoacidosis, and macroamylasemia

Question 9

What is the substrate and product of amylase determination?

Answer 9

SUBSTRATE: 2-chloro-4-nitrophenyl-maltotrioside

it reacts directly with alpha-amylase in the sample.

PRODUCT: 2-chloro-4-nitrophenol

Question 10

Does amylase determination method requires ancillary enzymes?

Answer 10

False

Question 11

What are the activators of Amylase determination?

Answer 11

Calcium and Chloride

Question 12

The amylase concentration in Amylase pancreatitis rises, peak and returns within?

Answer 12

Arise: 2 to 12 hours
Peak: 24 hours
Back to normal: 3 to 5 days

Question 13

What are the isoenzymes of Amylase?

Answer 13

P-type: (P1, P2, P3) - predominate in urine

S-type: (S1, S2, S3) - predominate in serum

Question 14

What amylase isoenzyme markedly increases in acute pancreatitis?

Answer 14

P3

Question 15

Which amylase isoenzyme migrates faster?

Answer 15

S-type

Question 16

What is the inhibitor of S-type amylase isoenzyme?

Answer 16

Wheat germ lectin

Question 17

True/False: Can the T-AMS be measure with and without the presence of lectin?

Answer 17

True, it is the combination of S-AMS and P-AMS

Question 18

What are the Amylase Methods? Define each method.

Answer 18

Amyloclastic - measures the disappearance of starch substrate
Saccharogenic - measures the appearance of the product
Chromogenic - it measures the increasing color from the production of product coupled with chromogenic dye
Continuous monitoring - coupling of several enzyme systems

Question 19

Amyloclastic is required to reach it time at?

Answer 19

Chromic point: fast disappearance (high amylase)

Question 20

Another term for amyloclastic?

Answer 20

Idiometric

Question 21

What indicator is used in amyloclastic?

Answer 21

Iodine (forming dark blue color)

Question 22

What is the end-product amyloclastic method?

Answer 22

When amylase catalyzes the hydrolysis of starch-iodine complex it will form glucose, maltose, dextrin and released iodine.

Question 23

The disappearance of blue color result into?

Answer 23

The reduction of blue color and reduction of amylase concentration. As the decrease in color is directly proportional to the amylase concentration.

Question 24

In Saccharogenic method, it is reported in what units?

Answer 24

Somogyi units: wherein the mg of glucose in 30 minutes at 37 degree Celsius at specific assay conditions

Question 25

How Saccharogenic method processed?

Answer 25

Starch substrate is hydrolyzed by AMS to its constituent carbohydrate molecule (simple sugars) containing simple sugars with reducing property.

Question 26

What method of AMS that employs starch substrate with chromogenic dye forming insoluble dye-substrate complex (H2O soluble)?

Answer 26

Chromolytic/ Chromogen Labeled Substrate Method/Colorimetric

Question 27

What AMS method measures colorimetrically?

Answer 27

Chromolytic/Chromogen labelled substrate method/Chromogenic

Question 28

An AMS Method that involves kinetic measurement, which measures the change in absorbance of NAD+.

Answer 28

Coupled enzyme reaction/Continuous Monitoring Method

Question 29

What absorbance and pH does Coupled Enzyme reaction measures it absorbance?

Answer 29

340 nm; 6.9 pH

Question 30

What is the substrate and indicator reaction of Continuous monitoring/ Couple enzyme reaction method?

Answer 30

Substrate: maltotriose/ maltopentose/ maltopentaose | Indicator reaction: conversion of NAD to NADH

Question 31

In the Amylase determination reaction, what substrate is used?

Answer 31

2-chloro-4-nitrophenyl-maltotrioside (CNPG3) reacts with amylase

Question 32

In the Amylase determination reaction, what product is released?

Answer 32

2-chloro-4-nitrophenol (CNP)

Question 33

In the Amylase determination reaction, what absorbance and temperature is AMS measured?

Answer 33

405 nm at 37 Celsius

Question 34

In the Amylase determination reaction, what reagents is used?

Answer 34

CNPG3, Calcium acetate, sodium chloride, potassium thiocyanate, sodium azide

Question 35

In the Amylase determination reaction, what buffer is used?

Answer 35

MES: Morpholino Ethane Sulfonic Acid (6 pH)

Question 36

When can we add the working reagent?

Answer 36

It can be added once the spectrophotometer is ready.

Question 37

In AMS Determination it is allowed to used hemolyzed samples.

Answer 37

No.

Question 38

What inhibits amylase activity?

Answer 38

Triglycerides, especially in Acute pancreatitis patient with hyperlipidemia.

Question 39

What factors that causes false decrease in AMS activity?

Answer 39

Triglycerides, Oxalates, Citrates

Question 40

If plasma is used as specimen for AMS Determination, what anticoagulant is used?

Answer 40

Heparinized plasma

Question 40

What factors that causes false increase in AMS activity? Why?

Answer 40

Morphine and Opiates, it causes constriction in sphincter of Oddi and pancreatic ducts This causes regurgitation of AMS in serum

Question 41

What is the reference range of AMS concentration in serum?

Answer 41

28-100 unit per liter

Question 42

What is the reference range of AMS concentration in urine?

Answer 42

1 to 15 U/hour