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FINALS CC 1 > TESTS FOR LIVER SYNTHETIC FUNCTION > Flashcards

TESTS FOR LIVER SYNTHETIC FUNCTION Flashcards

1
Q

Measures the totality of protein fractions in the plasma (albumin + globulin)

A

TOTAL PROTEIN

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2
Q

Assessment of nutritional status and presence of severe disease involving the
liver, kidney and bone marrow

A

TOTAL PROTEIN

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3
Q

Total Protein specimen

A

SERUM

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4
Q

Why you should never use plasma as specimen in total protein?

A

it contains fibrinogen (cause false increase by 0.2-0.4 g/dl)

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5
Q

Total Protein Interference

A

hemolysis, ictericia

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6
Q

Total Protein Reference value (SERUM)

A

6.5-8.3 g/dL (CF to g/L: 10)

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7
Q

10 METHODS FOR TOTAL PROTEIN

A
  1. KJELDAHL METHOD
  2. BIURET METHOD
  3. FOLIN-CIOCALTEU (LOWRY) METHOD
  4. UV ABSORPTION METHOD
  5. REFRACTOMETRY
  6. TURBIDIMETRY AND NEPHELOMETRY
  7. SALT FRACTIONATION
  8. COOMASIE BRILLIANT BLUE DYE
  9. NINHYDRIN
  10. SERUM PROTEIN ELECTROPHORESIS (SPE)
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8
Q

Classical method for protein quantitation

A

KJELDAHL METHOD

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9
Q

Reference method but NOT ROUTINELY USED (time consuming)

A

KJELDAHL METHOD

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10
Q

KJELDAHL METHOD Measures the amount of _________ in speciment

A

NITROGEN

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11
Q

KJELDAHL METHOD assumes that nitrogen content of protein is

A

16% (15.1 – 16.8%)

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12
Q

1 g of nitrogen is equal to

A

6.54 g of protein

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13
Q

3 STEPS of KJELDAHL METHOD

A

DIGESTION → DISTILLATION → TITRATION

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14
Q

Most widely used method for total protein determination

A

BIURET METHOD

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15
Q

A colorimetric non-enzymatic method that reads absorbance of color at ________

A

540 nm
BIURET METHOD

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16
Q

Biuret reagents:

A

o Alkaline Copper sulfate
o NaK tartrate (Rochelle salt) → prevents precipitation of copper
o NaOH
o KI → stabilizer

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17
Q

PRINCIPLE OF BIURET METHOD

A

peptide bonds present in protein forms a bond with cupric ions
forming a violet-colored chelate. (↑ bonds = ↑ purple color intensity = ↑ CHON)

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18
Q

Biuret Method Interference

A

Lipemic Sample

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19
Q

Has the highest analytical sensitivity (can measure very small conc. of protein)

A

FOLIN-CIOCALTEU (LOWRY) METHOD

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20
Q

FOLIN-CIOCALTEU (LOWRY) METHOD CAN BE MEASURED

A

SPECTROPHOTOMETRICALLY

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21
Q

FOLIN-CIOCALTEU (LOWRY) METHOD PRINCIPLE

A

Oxidation of phenolic compounds such as tyrosine, tryptophan, and histidine → reduces main reagent → to give a deep blue color

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22
Q

FOLIN-CIOCALTEU (LOWRY) MAIN REAGENT

A

Phospotngstic-molybdic acid or phenol reagent

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23
Q

FOLIN-CIOCALTEU (LOWRY) COLOR ENHANCER

A

Biuret reagent

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24
Q

UV ABSORPTION METHOD PRINCIPLE

A

The absorbance of proteins at 210nm (due to the absorbance of the
peptide bonds at this specific wavelength, EXCEPT aromatic AA)

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25
Q

Absorption @ 280nm

A

aromatic AA (tryptophan, tyrosine, and phenylalanine)

26
Q

Based on measurement of refractive index of serum total proteins

A

REFRACTOMETRY

27
Q

↑ changes in refractive index = ↑ protein

A

REFRACTOMETRY

28
Q

Measurement depends on formation of a uniform fine precipitate which scatters incident light in suspension ____________ or block light ____________

A

NEPHELOMETRY, TURBIDIMETRY

29
Q

Used for detection of proteins as little as 1ug

A

COOMASIE BRILLIANT BLUE DYE

30
Q

Develops violet color by reacting with primary amines widely used for detection of peptides and amino acids after paper chromatography

A

NINHYDRIN

31
Q

Separation technique based on differences in migration of charge

A

SERUM PROTEIN ELECTROPHORESIS (SPE)

32
Q

Migration of charged particles in an electric field

A

SERUM PROTEIN ELECTROPHORESIS (SPE)

33
Q

Consists of ______ (positive charge) and ______ (negative charge)

A

ANODE, CATHODE
SPE

34
Q

Either positive or negative depending on pH condition

A

Amphoteric
↑ pH: negative; ↓pH: positive

35
Q

What buffer is used in Amphoteric

A

Barbital (Veronal) → pH 8.6

36
Q

Isoelectric Property of Proteins

A

Amphoteric
No charge at isoelectric point
The acidic and basic amino acids content of proteins determines its net charge.

37
Q

Albumin band Reference Value

A

3.5-5.0 g/dl (53-65%)

38
Q

α1 Globulin band
(90% α1-antitrypsin) Reference Value

A

0.1-0.3 g/dl (2.5-5%)

39
Q

α2 Globulin band Reference Value

A

0.6-1.0 g/dl (7-13%)

40
Q

β Globulin band Reference Value

A

0.7-1.1 g/dl (8-14%)

41
Q

γ Globulin band (immunoglobulins) Reference Value

A

0.8-1.6 g/dl (12-22%)

42
Q

Most to least anodal

A
  1. Albumin band
  2. α1 Globulin band (90% α1-antitrypsin)
  3. α2 Globulin band
  4. β Globulin band
  5. γ Globulin band (immunoglobulins)
43
Q

The liver synthesizes _________ of albumin

A

9-12 g/day

44
Q

Concentration is __________ to the severity of hepatic disease.

A

directly proportional

45
Q

Low albumin is seen due to __________

A

decreased synthesis

46
Q

Globulins are precipitated which removes globulin from total protein

A

SALT PRECIPITATION

47
Q

Charges protein positively so that the following anionic dyes would bind

A

DYE BINDING

48
Q

Most commonly used and sensitive in dye binding

A

Bromcresol Green (BCG)

49
Q

Most specific dye in dye binding

A

Bromcresol purple (BCP)

50
Q

Dye that is not specific for albumin

A

Methyl Orange

51
Q

More specific dye to albumin than methyl orange

A

2,4’-hydroxyazobenzenebenzoic acid (HABA)

52
Q

2,4’-hydroxyazobenzenebenzoic acid (HABA) interferences

A

conjugated bilirubin, aspirin, penicillin, sulfonamides

53
Q

Validates if globulin is truly higher than albumin (for checking)

A

ALBUMIN/GLOBULIN RATIO

54
Q

In inverted A/G ratio:

A

Globulin is higher than albumin

55
Q

inverted A/G ratio is seen in

A

n cirrhosis, multiple myeloma, waldenstrom’s macroglobulinemia

56
Q

A/G ratio reference value

A

1.3-3.1 g/dL (CF to g/L: 10)

57
Q

Aka: Vitamin K Response Test

A

PROTHROMBIN TEST

58
Q

Prior to testing, in prothrombin test patient is administered first with _______ for _______

A

10mg of vitamin K for 1-3 days (intramuscularly)

59
Q

It differentiates intrahepatic disorder from extrahepatic disorder

A

PROTHROMBIN TEST

60
Q

If PT is still prolonged after vit. K admin

A

indicates intrahepatic disorder

61
Q

If PT becomes normal after vit. K admin

A

indicates extrahepatic disorder

62
Q

prolonged PT signifies massive cellular damage

A

Acute/Chronic Hepatitis

FINALS CC 1 (6 decks)

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