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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
Absorption @ 280nm
aromatic AA (tryptophan, tyrosine, and phenylalanine)
26
Based on measurement of refractive index of serum total proteins
REFRACTOMETRY
27
↑ changes in refractive index = ↑ protein
REFRACTOMETRY
28
Measurement depends on formation of a uniform fine precipitate which scatters incident light in suspension ____________ or block light ____________
NEPHELOMETRY, TURBIDIMETRY
29
Used for detection of proteins as little as 1ug
COOMASIE BRILLIANT BLUE DYE
30
Develops violet color by reacting with primary amines widely used for detection of peptides and amino acids after paper chromatography
NINHYDRIN
31
Separation technique based on differences in migration of charge
SERUM PROTEIN ELECTROPHORESIS (SPE)
32
Migration of charged particles in an electric field
SERUM PROTEIN ELECTROPHORESIS (SPE)
33
Consists of ______ (positive charge) and ______ (negative charge)
ANODE, CATHODE SPE
34
Either positive or negative depending on pH condition
Amphoteric ↑ pH: negative; ↓pH: positive
35
What buffer is used in Amphoteric
Barbital (Veronal) → pH 8.6
36
Isoelectric Property of Proteins
Amphoteric No charge at isoelectric point The acidic and basic amino acids content of proteins determines its net charge.
37
Albumin band Reference Value
3.5-5.0 g/dl (53-65%)
38
α1 Globulin band (90% α1-antitrypsin) Reference Value
0.1-0.3 g/dl (2.5-5%)
39
α2 Globulin band Reference Value
0.6-1.0 g/dl (7-13%)
40
β Globulin band Reference Value
0.7-1.1 g/dl (8-14%)
41
γ Globulin band (immunoglobulins) Reference Value
0.8-1.6 g/dl (12-22%)
42
Most to least anodal
1. Albumin band 2. α1 Globulin band (90% α1-antitrypsin) 3. α2 Globulin band 4. β Globulin band 5. γ Globulin band (immunoglobulins)
43
The liver synthesizes _________ of albumin
9-12 g/day
44
Concentration is __________ to the severity of hepatic disease.
directly proportional
45
Low albumin is seen due to __________
decreased synthesis
46
Globulins are precipitated which removes globulin from total protein
SALT PRECIPITATION
47
Charges protein positively so that the following anionic dyes would bind
DYE BINDING
48
Most commonly used and sensitive in dye binding
Bromcresol Green (BCG)
49
Most specific dye in dye binding
Bromcresol purple (BCP)
50
Dye that is not specific for albumin
Methyl Orange
51
More specific dye to albumin than methyl orange
2,4’-hydroxyazobenzenebenzoic acid (HABA)
52
2,4’-hydroxyazobenzenebenzoic acid (HABA) interferences
conjugated bilirubin, aspirin, penicillin, sulfonamides
53
Validates if globulin is truly higher than albumin (for checking)
ALBUMIN/GLOBULIN RATIO
54
In inverted A/G ratio:
Globulin is higher than albumin
55
inverted A/G ratio is seen in
n cirrhosis, multiple myeloma, waldenstrom’s macroglobulinemia
56
A/G ratio reference value
1.3-3.1 g/dL (CF to g/L: 10)
57
Aka: Vitamin K Response Test
PROTHROMBIN TEST
58
Prior to testing, in prothrombin test patient is administered first with _______ for _______
10mg of vitamin K for 1-3 days (intramuscularly)
59
It differentiates intrahepatic disorder from extrahepatic disorder
PROTHROMBIN TEST
60
If PT is still prolonged after vit. K admin
indicates intrahepatic disorder
61
If PT becomes normal after vit. K admin
indicates extrahepatic disorder
62
prolonged PT signifies massive cellular damage
Acute/Chronic Hepatitis

FINALS CC 1 (6 decks)

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