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1. CC 1 Midterms > Laboratory Methodology (TOPIC 1) > Flashcards

Laboratory Methodology (TOPIC 1) Flashcards

1
Q

PARAMETERS USED IN THE DIAGNOSIS OF LIPOPROTEIN DISORDERS

A
  1. Lipid Profile test
    a. Triglycerides
    b. Total Cholesterol
    i.HDL-cholesterol determination
    ii. LDL-cholesterol determination
    2.Plasma Appearance Test
    3.Lipoprotein electrophoresis
    4.Ultracentrifugation
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2
Q

neutral fats because fatty acids are tied up in ester linkages

A

Triglycerides

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3
Q

depot fat or storage material of adipose tissue and main form of lipid storage in man

A

Triglycerides

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4
Q

major constituents of chylomicrons and prebeta lipoprotein

A

Triglycerides

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5
Q

higher value in serum than in plasma

A

Triglycerides

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6
Q

also known as “triacyl glycerol”

A

Triglycerides

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7
Q

Triglycerides also known as

A

“triacyl glycerol”

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8
Q

Time for fasting for triglycerides

A

12-14 hours

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9
Q

what are the Types of measurement of Triglycerides

A

Chemical Method
Hantzsh/Lutidine reaction
Van Handel and Zilversmit

Enzymatic Method
Weiland Method
Trinder’s Method
Eggstan and Kreutz Method

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10
Q

Chemical Method
Step 1: Extraction

What is the process

A

Removal of lipid from protein by treatment of organic solvent such as CHLOROFORM, ISOPROPANOL, ETHER

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11
Q

Chemical Method
Step 2: Purification by adsorption

What is the process

A

Removal of
PHOSPHOLIPID
MONOGLYCERIDES
DIGLYCERIDES
GLUCOSE
BILIRUBIN

by addition of
SILICIC ACID
ZEOLITE
FLOROSIL

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12
Q

Chemical Method
Step 3: Hydrolysis or saponification

What is the process

A

Breaking down of
TRIGLYCERIDES into GLYCEROL and FATTY ACIDS

by addition of
KOH or SODIUM METHOXIDE

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13
Q

Chemical Method
Step 4: Oxidation

What is the process

A

Conversion of
GLYCEROL to FORMALDEHYDE and FORMIC ACID (measurable compound)

by SODIUM PERIODATE

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14
Q

Chemical Method
Step 5: Colorimetric measurement

What is the process

A

FORMALDEHYDE is added with color reagents and absorbance is measured in spectrophotometer.

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15
Q

Mostt common chemical method

A

Hantzsh/Lutidine Reaction

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16
Q

Hantzsh/Lutidine Reaction principle

A

Formaldehye + Acetylacetone + NH4 — 3,5 diacetyl-1,4-dihydrolutidine (YELLOW at 410nm)

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17
Q

Van Handel and Zilversmit principle

A

Formaldehyde + H2SO4 + CTA (chromotropic acid) — pink chromopore

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18
Q

Triglyceride chemical method color reagents

A

Chromotrophic acid and sulfuric acid
Diphenylhydrazone
Acetylacetone and ammonia

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19
Q

Chromotrophic acid and sulfuric acid

Color and measured at?

A

pink color

500-600nm

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20
Q

Diphenylhydrazone

measured at?

A

500-600 nm

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21
Q

Acetylacetone and ammonia

Color and measured at?

A

yellow color

412nm

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22
Q

Triglyceride Enzymatic method
Initial Reaction

A

Triglyceride –(Lipase)–> glycerol + 3 fatty acids

Glycerol + ATP –(Glycerol Kinase)–> Glycerol phosphate + ADP

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23
Q

Weiland Method Principle

A

Glycerol phosphate + NAD –(Glycerophosphate dehydrogenase)–> dihydroacetone phosphate + NADH + H

NADH + tetrazolium dye –(Diaphorase)–> formazan + NAD

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24
Q

Weiland Method: The absorbance of NADH can be measured at ___ nm after GCPD reaction.

The absorbance of formazan can be measure at ___ nm

A

340

500-600

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25
Q

Trinders Method Principle
Triglyceride enzymatic method

A

Glycerophosphate + O2 –(Glycerophosphate oxidase)–> dihydroacetone + H2O2

H2O2 + phenol + 4-aminoantipyrine –(Peroxidase)–> quinoneimine dye

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26
Q

Trinders Method: Quinonimine dye is measured at ___ nm

A

500-505

27
Q

EGGSTAN AND KREUTZ METHOD
Principle

A

ADP + phosphoenol pyruvate –(Pyruvate Kinase)–> ATP + Pyruvate

Pyruvate + NADH + H –(Lactate Dehydrogenase)–> Lactate + NAD

28
Q

EGGSTAN AND KREUTZ METHOD: NAD is measured at __ nm

A

340

29
Q

Normal Values of Triglycerides

Acceptable
Borderline
High Risk

and Coversion factor to mmol/l

A

<250 mg/dl
250-500 mg/dl
>500 mg/dl

0.011

30
Q

Cholesterol Determination

Chemical Methods

A

Zlatkis, Zak, Boyle
Carr-Drekter
Abell-Kendall
Babson
Schoenheimer-Sperry
Lieberman-Burchard reaction
Salkowski reaction

31
Q

Cholesterol Determination

Enzymatic Method

A

Trinders reaction
Oxygen consumption

32
Q

General Steps in Cholesterol Chemical Method

A

Extraction - cholesterol is seperated from protein
Saponification - cholesterol ester is broken down to free cholesterol and fatty acids
Purification - cholesterol is precipitated
Colorimetry - addition of color reagent to be measure in spectrophotometer

33
Q

Cholesterol Chemical Method

What is the Procedure for the Method

Zlatkis, Zak, Boyle

A

One step method
(colorimetry)

34
Q

Cholesterol Chemical Method

What is the Procedure for the Method

Carr-Drekter

A

Two step method
Extraction-Colorimetric

35
Q

Cholesterol Chemical Method

What is the Procedure for the Method

Abell-Kendall Babson

A

Three step method
(Saponification-Extraction-Colorimetric)

36
Q

Cholesterol Chemical Method

What is the Procedure for the Method

Schoenheimer-Sperry

A

Four-Step Method
(Hydrolysis-Extraction-Precipitation-Colorimetric)

37
Q

Cholesterol :One step method (colorimetric)

What are the 2 reactions

A

Lieberman-Burchard Reaction
Salkowski Reaction

38
Q

Lieberman-Burchard reaction

Principle

Result

Stability

A

Cholesterol(from chloroform extract of serum) + acetic anhydride + sulfuric acid

Cholestapolyene sulfonic acid (GREEN)

Less Stable

39
Q

Salkowski Reaction

Principle

Result

Stability

A

chole + HAc + FE(III)

Cholesapolyene carbonium ion (RED PURPLE)

More Stable

40
Q

Cholesterol Enzymatic Method

Trinders Reaction Principle

Measure at ___ nm

A

Cholesterol ester + H2O –(Cholesteryl ester hydrolase)–> free cholesterol + fatty acids

Free cholesterol + O2 –(Cholesterol Oxidase)–> Cholestene-3-one + H2O2

H2O2 + Phenol + aminoantipyrine –(peroxidase)–> quinoneimine dye (Pink)

500

41
Q

Cholesterol Enzymatic Method

Oxygen Consumption Principle

A

Cholesterol ester + H2O –(Cholesteryl ester hydrolase)–> free cholesterol + fatty acids

Free cholesterol + O2 –(cholesterol oxidase)–> Cholestene-3-one + H2O2

H2O2 + Peroxidase –(Peroxidase)–> O2 + H2O

O2 electrode the O2 releases from H2O2 decomposition

42
Q

Trinders Reaction CHOLESTEROL

A
  1. Cholesterol ester + H2O (CHOLESTEROL ESTER HYDROLASE) = Free cholesterol + Fatty acids
  2. Free Cholesterol + O2 (CHOLESTEROL OXIDASE) = cholestene-3-one + H2O2
  3. H2O2 + phenol + aminoantipyrine (PEROXIDASE) = Quinoneimine Dye (PINK)
  4. Color produces measured at 500nm
43
Q

Trinders Reaction TRIGLYCERIDES

complete

A
  1. Triglycerids (LIPASE) = Glycerol + 3 Fatty acids
  2. Glycerol + ATP (GLYCEROL KINASE) = glycerol phosphates + ADP
  3. Glycerophosphate + O2 (GLYCEROPHOSPHATE OXIDASE)= dihydroacetone + H2O2
  4. H2O2 + phenol + 4-aminoantipyrine (PEROXIDASE) = quinoneimine dye + 2H2O2
  5. Dye measured at 500-505nm
44
Q

Cholesterol Values

Desirable
Borderline
High Risk

A

<200 mg/dl
200-239 mg/dl
>240 mg/dl

45
Q

Methods for HDL cholesterol

A

Precipitation method
Immunoassays

46
Q

Precipitation method for HDL cholesterol (DEFINITION)

A

Precipitating reagent is added to precipitate VLDL and LDL, supernatant is for HDL

47
Q

Immunoassays for HDL cholesterol

A

ELISA
Immunonephelometric

48
Q

HDL-cholesterol values

Male
Female
High risk of heart disease

A

28-62
37-77
<35 mg/dl

49
Q

Friedwald Formula

A

LDL-C mg/dl = TC - HDL - C (Triglyceride/5)

50
Q

Gold standard of LDL quantification

A

Ultracentrifugation

51
Q

Normal Values for LDL-Cholesterol

Desirable
Borderline High-Risk
High Risk

A

<130 mg/dl
130-159 mg/dl
>160 mg/dl

52
Q

Phospholipids Chemical Method

A

Extraction
Oxidation- phospholipid phosphorus is converted to inorganic phosphorus

Colorimetry - inorganic phosphorus + molybdate blue = molybdenum blue

Total phosphorus X 25= phospholipid mass

53
Q

Phospholipids Enzymatic Method

A

Phospholipase D- phospholipid is hydrolyzed to choline

Choline oxidase- choline is oxidized to betaine and H2O2

Peroxidase –H2O2 is added with phenol and 4-aminoantipyrine = quinonimine dye

Color produced is measured at 500 nm

54
Q

Fatty Acids Normal Value

Adult
Children and obese

A

0.3-0.9 mmol/l
>1.1 mmol/l

55
Q

Fatty acids methods

A

Gas-layer chromatography (GLC)
Dole titration method

56
Q

GUIDELINES TO REDUCE RISK OF CORONARY HEART DISEASE(From the National Cholesterol Education Program)

Cholesterol (mg/dl)
Desirable
Borderline
High Risk

A

<200

200-239

> or = 240

57
Q

GUIDELINES TO REDUCE RISK OF CORONARY HEART DISEASE(From the National Cholesterol Education Program)

Triglycerides (mg/dl)
Desirable
Borderline
High Risk

A

<200

200-499

> or = 500

58
Q

GUIDELINES TO REDUCE RISK OF CORONARY HEART DISEASE(From the National Cholesterol Education Program)

Chol/HDL Ratio
Desirable
Borderline
High Risk

A

<3.9

4.0-4.9

> or = 5.0

59
Q

GUIDELINES TO REDUCE RISK OF CORONARY HEART DISEASE(From the National Cholesterol Education Program)

LDL-Chol (mg/dl)
Desirable
Borderline
High Risk

A

<130

130-159

> or = 160

59
Q

GUIDELINES TO REDUCE RISK OF CORONARY HEART DISEASE(From the National Cholesterol Education Program)

HDL-Chol (mg/dl)
Desirable
Borderline
High Risk

A

> 40

35-40

<35

60
Q

used to diagnose dysbetalipoproteinemia and familial combined hyperlipidemia.

A

Apolipoprotein B analysis

60
Q

Requested in addition to lipid profile test

A

Apolipoprotein B analysis

61
Q

Specimen for Apolipoprotein B analysis

A

Non-Fasting Serum

62
Q

Apolipoprotein B analysis
Value for high risk of Cardiovascular Disease (CVD)

A

> or = 130 mg/dl

1. CC 1 Midterms (14 decks)

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