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Clinical Chemistry 2 > Clinical Chemistry - Amylase Determination > Flashcards

Clinical Chemistry - Amylase Determination Flashcards

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1
Q

What enzyme reaction class does Amylase belong?

A

Hydrolases

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2
Q

What is the importance of Amylase?

A

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

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3
Q

Amylase is the breakdown of?

A

Starch and glycogen with glucose, maltose and dextrin

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4
Q

Where does amylase filtered?

A

In glomerulus (kidney)

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5
Q

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

A

True

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6
Q

Where does it is mainly found?

A

Acinar cells of the pancreas and salivary glands.

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7
Q

What minor source organs that can we found amylase?

A

Small intestine, skeletal muscle, liver and fallopian tube

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8
Q

What is the significant diagnosis for amylase?

A

Acute pancreatitis and salivary gland lesions.

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

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9
Q

What is the substrate and product of amylase determination?

A

SUBSTRATE: 2-chloro-4-nitrophenyl-maltotrioside

it reacts directly with alpha-amylase in the sample.

PRODUCT: 2-chloro-4-nitrophenol

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10
Q

Does amylase determination method requires ancillary enzymes?

A

False

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11
Q

What are the activators of Amylase determination?

A

Calcium and Chloride

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12
Q

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

A

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

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13
Q

What are the isoenzymes of Amylase?

A

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

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

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14
Q

What amylase isoenzyme markedly increases in acute pancreatitis?

A

P3

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15
Q

Which amylase isoenzyme migrates faster?

A

S-type

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16
Q

What is the inhibitor of S-type amylase isoenzyme?

A

Wheat germ lectin

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17
Q

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

A

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

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18
Q

What are the Amylase Methods? Define each method.

A

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

19
Q

Amyloclastic is required to reach it time at?

A

Chromic point: fast disappearance (high amylase)

20
Q

Another term for amyloclastic?

A

Idiometric

21
Q

What indicator is used in amyloclastic?

A

Iodine (forming dark blue color)

22
Q

What is the end-product amyloclastic method?

A

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

23
Q

The disappearance of blue color result into?

A

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

24
Q

In Saccharogenic method, it is reported in what units?

A

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

25
Q

How Saccharogenic method processed?

A

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

26
Q

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

A

Chromolytic/ Chromogen Labeled Substrate Method/Colorimetric

27
Q

What AMS method measures colorimetrically?

A

Chromolytic/Chromogen labelled substrate method/Chromogenic

28
Q

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

A

Coupled enzyme reaction/Continuous Monitoring Method

29
Q

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

A

340 nm; 6.9 pH

30
Q

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

A

Substrate: maltotriose/ maltopentose/ maltopentaose

Indicator reaction: conversion of NAD to NADH

31
Q

In the Amylase determination reaction, what substrate is used?

A

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

32
Q

In the Amylase determination reaction, what product is released?

A

2-chloro-4-nitrophenol (CNP)

33
Q

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

A

405 nm at 37 Celsius

34
Q

In the Amylase determination reaction, what reagents is used?

A

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

35
Q

In the Amylase determination reaction, what buffer is used?

A

MES: Morpholino Ethane Sulfonic Acid (6 pH)

36
Q

When can we add the working reagent?

A

It can be added once the spectrophotometer is ready.

37
Q

In AMS Determination it is allowed to used hemolyzed samples.

A

No.

38
Q

What inhibits amylase activity?

A

Triglycerides, especially in Acute pancreatitis patient with hyperlipidemia.

39
Q

What factors that causes false decrease in AMS activity?

A

Triglycerides, Oxalates, Citrates

40
Q

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

A

Heparinized plasma

40
Q

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

A

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

41
Q

What is the reference range of AMS concentration in serum?

A

28-100 unit per liter

42
Q

What is the reference range of AMS concentration in urine?

A

1 to 15 U/hour

Clinical Chemistry 2 (4 decks)

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