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CC1 > LIPIDS LAB > Flashcards

LIPIDS LAB Flashcards

1
Q

preferred sample

A

serum collected in SST

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2
Q

preferred for LPP assays

A

edta plasma

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3
Q

fasting hours required

A

12-14 hour

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4
Q

Turbidity of the chylomicrons can interfere with the
spectrophotometric methods of lipid profile by?

A

falsely increased result

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5
Q

specimen type for total cholesterol and HDL

A

non-fasting specimen

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6
Q

specimen type for TAG and LDL

A

fasting specimens

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7
Q
  • can lead to decreased plasma volume in relation to volume of pack RBC
  • can interfere with the
    result
A

hemoconcentration

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8
Q

long period storage of
serum/plasma

A

-70C or lower

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9
Q

short-term storage of serum/plasma (up to a month or two)

A

-20C

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10
Q

cholesterol measurement methods

A

can be chemical or enzymatic

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11
Q

fasting requirement for cholesterol measurement?

A

8-12hours

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12
Q

specimen type for cholesterol measurement

A

fasting or non-fasting specimen

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13
Q

chemical methods for measuring cholesterol

A
  • Libermann-Burchardt Rxn
  • Salkowski Rxn
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14
Q

end product of Libermann-Burchardt Rxn

A

Cholestadienyl Monosulfonic Acid

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15
Q

end color of Libermann-Burchardt Rxn

A

green

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16
Q

color developer mixture of Libermann-Burchardt Rxn

A

a. glacial acetic acid
b. acetic anhydride
c. concentrated sulfuric acid

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17
Q

end product of salkowski rxn

A

Cholestadienyl Disulfonic Acid

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18
Q

end color of salkowski rxn

A

red

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19
Q

general methods for measuring cholesterol

A
  1. one-step method
  2. two-step method
  3. three-step method
  4. four-step method
20
Q

one-step method

A

colorimetry (Pearson, Stern, and Mac Gavack)

21
Q

two-step method

A

extraction + colorimetry (Bloors)

22
Q

three-step method

A

saponification + extraction + colorimetry (Abell-Kendal)

23
Q

four-step method

A

saponification + extraction + colorimetry + precipitation (Schoenheimer, Sperry, Parekh, and Jung)

24
Q

what are the reagents for step methods?

A
  • Liebermann-Burchardt (colorimetry)
  • Petroleum ether (saponification)
  • Alcoholic potassium hydroxide
  • Digitonin (precipitation)
25
Q

CDC REFERENCE METHOD FOR CHOLESTEROL
MEASUREMENT

A

Gas Chromatography Mass Spectrophotometry (GC-MS)

26
Q

TRADITIONAL REFERENCE METHOD FOR
CHOLESTEROL

A

Abell, Levy and Brodie Method

27
Q

enzymatic methods for measuring cholesterol

A

cholesterol oxidase rxn

28
Q

interference for enzymatic method of measuring total cholesterol

A
  • ascorbic acid, bilirubin, hgb
  • ascorbic acid: ↑ascorbic acid ↓ TC
  • bilirubin: oxidized by H2O2
  • hemoglobin: has pseudo-peroxidase activity that diminish H2O2 produced in the cholesterol oxidase rxn
29
Q
  • Uses hexane extraction / petroleum ether after
    hydrolysis with alcoholic KOH followed by reaction with
    Liebermann-Burchardt color reagent
  • EC: green
A

Abell, Levy and Brodie Method

30
Q

gold standard method for cholesterol

A

ISOTOPE DILUTION MASS SPECTROMETRY

31
Q

fasting hours requirement for TAG measurement

A

10-12 hours

32
Q

specimen for TAG measurement

A

either plasma or serum

33
Q

CDC Reference Method for TAG Measurement

A
  • Saponification: alcoholic KOH
  • Extraction: choloroform
  • Treatment w/ silicilic acid: to remove phospholipids
  • EP: pink chromophore
34
Q

interference for TAG measurement

A

HAB
- hemolysis
- ascorbic acid
- bilirubin

35
Q

methods for measuring TAG

A
  • chemical method
  • enzyme method
36
Q

chemical methods for measuring TAG

A
  1. colorimetric method - Van Handel and Zilversmith
  2. fluorometric method - Hantzsch Condensation
37
Q

Van Handel and Zilversmith
(colorimetric)

A
  • TAG + KOH → Glycerol + Fatty Acid
  • Glycerol + Periodate → Formic Acid + Formaldehyde
  • Formaldehyde + Chromotropic Acid → Chromogen
38
Q

Hantzsch Condensation
(fluorometric)

A
  • TAG + KOH → Fatty Acids + Glycerol
  • Glycerol + Periodate → Formic acid + Formaldehyde
  • HCHO + Diethyl acetone + NH3 → Diacetyl Lutidine Compound
39
Q

Involves liberation of fatty acids from cholesterol, from
triglycerides and KOH

A

Hantzsch Condensation

40
Q

enzymatic methods for measuring TAG

A
  • Glycerol Kinase Method
41
Q

Reaction A for Glycerol Kinase Method

A
  • TAG + Lipase → Glycerol + Fatty acid
  • Glycerol + ATP + Glycerol Kinase → Glycerolphosphate +ADP
  • ADP + Phosphoenol pyruvate + Pyruvate Kinase → ATP + Pyruvate
  • Pyruvate + NADH + H ion + Lactate dehydrogenase → Lactate + NAD
42
Q

Reaction B for Glycerol KInase Method

A
  • TAG + Lipase → Glycerol + Fatty Acid
  • Glycerol + ATP + Glycerol Kinase → GlycerolPO4 + ADP
  • Glycerol PO4 + NAD + GlycerolPO4dehydrogenase → DihydroxyacetonePO4 + NADH
  • NADH + Tetrazolium dye + Diaphorase → Formasan + NAD+
43
Q

Conditions where there is increased TAG

A
  • Hypothyroidism
  • Alcoholism
  • Nephrotic Syndrome
  • Hyperlipoproteinemia tyeps I, IIb, III, IV, V
  • Pancreatitis
44
Q

Conditions where there is decreased TAG

HMM… B-aba ng tag

A
  • Hyperthyroidism
  • Malabsorption syndrome
  • Malnutrition burns
  • Brain infection
45
Q
A

CC1 (3 decks)

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