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MTAP 2 > CC2 (LIPID AND LIPOPROTEIN ANALYSES) > Flashcards

CC2 (LIPID AND LIPOPROTEIN ANALYSES) Flashcards

1
Q

SAMPLE for cholesterol measurement

A

plasma or serum

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2
Q

PATIENT PREP for cholesterol measurement

A

usual diet 2 weeks prior testing

May be tested from nonfasting spx (little effect on TC)

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3
Q

May be tested from nonfasting spx

A

CHOLESTEROL

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4
Q

PRECAUTIONS for cholesterol measurement

A
  • Avoid hemolyzed sample (free Hgb – false ↑)
  • Avoid icteric sample
  • No water contamination (dehydration is included in chole measurement)
  • Precise and accurate timing for color development
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5
Q

COLOR DEVELOPER MIXTURE FOR CHOLE:

A
  • Glacial acetic acid
  • Acetic anhydride
  • Conc. sulfuric acid
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6
Q

These makes it possible for the cholesterol to be carried out in colorimetric assay

A

Double bonds and hydroxyl group in the sterol structure

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7
Q

CDC Reference method for CHOLESTEROL

A

ABELL-LEVY-BRODIE-KENDALL METHOD

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8
Q

Used for cholesterol extraction

A

Hexane

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9
Q

Used for cholesterol hydrolysis

A

Alc. KOH

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10
Q

General steps for cholesterol method

A

Cholesteryl esters: hydrolyzed with alc. KOH

Unesterified cholesterol: extracted with hexane

Measured using: LIEBERMANN – BURCHARD reaction or SALKOWSKI reaction

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11
Q

NON-ENZYMATIC / CHEMICAL METHOD for CHOLESTEROL

A

ABELL KANDALL

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12
Q

Principle: Cholesterol dehydration & oxidation to form a colored compound (measured colorimetrically)

A

NON-ENZYMATIC / CHEMICAL METHODS (ABELL KANDALL)

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13
Q

Involves colorimetry only

A

One-step method

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14
Q

Involves extraction, colorimetry

A

Two-step method

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15
Q

Involves saponification, extraction, colorimetry

A

Three-step method

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16
Q

Involves precipitation, saponification, extraction, colorimetry

A

Four-step method

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17
Q

Chemical reaction after hydrolysis and hexane extraction of cholesterol

A

LIEBERMANN – BURCHARD reaction or SALKOWSKI reaction

Cholesterol + Sulfuric acid + Acetic anhydride –>
LIEBERMANN – BURCHARD: Green soln. (Cholestadienyl monosulfonic acid)

SALKOWSKI: Red soln. (Cholestadienyl disulfonic acid)

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18
Q

Product and solution color in LIEBERMANN – BURCHARD reaction for cholesterol

A

Cholestadienyl monosulfonic acid
Green soln.

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19
Q

Product and solution color in SALKOWSKI reaction for cholesterol

A

Cholestadienyl disulfonic acid
Red soln.

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20
Q

T/F

Color intensity absorbance = Cholesterol concentration

A

T

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21
Q

ENZYMATIC METHOD for CHOLESTEROL

A

CHOLESTEROL OXIDASE

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22
Q

Chemical equation for ENZYMATIC METHOD (CHOLESTEROL OXIDASE)

A

TC (cholesteryl ester and cholesterol) + H2O ▬Cholesteryl esterase▬► Cholesterol + FA

Cholesterol oxidase
Cholesterol + O2 ▬Cholesterol oxidase▬► 4-cholestenone + H2O2

H2O2 + Dye ▬Peroxidase▬► Color (Quinoneimine dye – violet/pink)

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23
Q

End product & color of enzymatic method (Cholesterol oxidase)

A

Quinoneimine dye – violet/pink

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24
Q

SAMPLE for TAG

A

plasma or serum

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25
PATIENT PREP for TAG
Strict fasting (12 hours) If + FBS: 10 hours
26
Transfer of phosphate group
Kinase
27
CDC Reference method for TAG
MODIFIED VAN HANDEL AND ZILVERSMITH
28
Chemical equation for NON-ENZYMATIC/CHEMICAL METHOD of TAG
HYDROLYSIS: TAG ▬Alcoholic KOH▬► Glycerol + FA Glycerol + Periodic acid ▬▬► Formaldehyde Formaldehyde + Chromotropic acid → Blue soln (Van Handel and Zilversmith) or Formaldehyde + Diacetyl acetone + NH3 → Yellow soln (Hantzch Condensation)
29
2 methods included in NON-ENZYMATIC/CHEMICAL METHOD for TAG
Van Handel and Zilversmith (Colorimetric) Hantzch Condensation (Fluorometric)
30
Equation of Van Handel and Zilversmith (Colorimetric)
Formaldehyde + Chromotropic acid → Blue soln
31
Equation of Hantzch Condensation (Fluorometric)
Formaldehyde + Diacetyl acetone + NH3 → Yellow soln
32
T/F Color intensity absorbance = TAG concentration
T
33
Part of TAG structure utilized in analysis
Glycerol
34
Colorimetric method for TAG Color of solution
Van Handel and Zilversmith Blue
35
Fluorometric method for TAG Color of solution
Hantzch Condensation Yellow
36
ENZYMATIC METHOD for TAG
GLYCEROL KINASE
37
Equation for TAG ENZYMATIC METHODS (GLYCEROL KINASE)
TAG ▬Lipase▬► Glycerol + FA Glycerol + ATP ▬Glycerokinase▬► Glycerophosphate + ADP > PK > Glycerol-Phosphate Oxidase
38
Pyruvate Kinase equation for TAG (enzymatic) utilizing ADP
ADP + PEP ▬PK▬► ATP + Pyruvate Pyruvate + NADH + H+ ▬LD▬►Lactate + NAD+ (340 nm; decreased absorbance)
39
Glycerol-Phosphate Oxidase equation for TAG (enzymatic) utilizing Glycerophosphate
Glycerophosphate + O2 ▬GO▬► Dihydroxyacetone + H2O2 H2O2 + Dye ▬Peroxidase▬► Color
40
SAMPLE for LIPOPROTEIN
Serum from SST (laboratory) EDTA plasma (research studies)
41
PATIENT PREP for LIPOPROTEIN
Fasting (12 hours) If nonfasting sample – only test HDL and TC
42
Indicates hyperlipidemia; must be treated using ultracentrifugation or enzymatic cleavage
Lipemic (fasting sample)
43
GENERAL METHODS for LIPOPROTEIN
ULTRACENTRIFUGATION ELECTROPHORESIS
44
Reference method for LPP quantitation
ULTRACENTRIFUGATION
45
ULTRACENTRIFUGATION of LPP is based on
molecular density
46
solution applied during ULTRACENTRIFUGATION of LPP & its SG
Potassium bromide (SG 1.063)
47
Method wherein plasma is centrifuged for 24 hrs
ULTRACENTRIFUGATION of LPP
48
Resulting layers after ULTRACENTRIFUGATION of LPP
Lightest to heaviest: 1) CM 2) VLDL 3) LDL 4) HDL
49
LPP ELECTROPHORESIS is based on
migration in an electric field (depends on protein constituent)
50
pH of proteins
8.6 *proteins are negative charge
51
Lipid stains used in LPP ELECTROPHORESIS
Oil red O Fat Red 7B Sudan Black
52
4 bands obtained after fat stain
HDL (α-lipoprotein) VLDL, LP(a) LDL CM
53
Fastest (most anodic) lipoprotein
HDL (α-lipoprotein)
54
Pre-β lipoprotein
VLDL LP(a)
55
β lipoprotein
LDL
56
Lipoprotein stationary at origin (cathode)
CM
57
PLASMA PROTEINS
A. Albumin B. Globulin * α1-globulins * α2-globulins * β-globulins * y-globulins
58
HIGH-DENSITY LIPOPROTEIN (HDL) METHOD
POLYANION PRECIPITATION
59
a two-step method including precipitation and Abell-Kendall
POLYANION PRECIPITATION (HDL measurement)
60
Polyanion added to serum (removes non-HDL lipoproteins)
Precipitation (POLYANION PRECIPITATION for HDL)
61
Used to quantify HDL
Abell-Kendal
62
Examples of Polyanions
Heparin Dextran sulfate Na phosphotungstate
63
Examples of divalent cations
Mg Mn
64
Lipoproteins (LDL, VLDL) are precipitated with polyanions in presence of divalent cations (e.g. Mg or Mn), which are sediment by centrifugation (10-30 min. for 10,000g or 3 min. for 15,000g) HDL is quantified in the supernate by Abell-Kendal (used in cholesterol assay)
POLYANION PRECIPITATION
65
Lipoproteins only derived values (computed) in the lab
LDL and VLDL
66
LDL computation
LDL= Total cholesterol – (HDL + VLDL)
67
VLDL computation
Friedewald: VLDL = TAG / 5 = mg/dL (conventional) VLDL= TAG / 2.175 = mmol/L (SI)
68
Ultracentrifugation of serum at native density gradient of ______ to float VLDL
1.006 g/L
69
Apolipoprotein used for determination of LDL and VLDL conc.
Apo B
70
Apolipoprotein used for determination of HDL conc.
Apo A-1
71
T/F Apolipoprotein conc. = Corresponding lipoprotein conc.
T
72
TESTS FOR APOLIPOPROTEIN:
Immunoassays (Immunonephelometric assay, Immunoturbidimetric assay, Immunochemical assay)
73
Involves specific antibodies (reagent); apolipoproteins (target)
Immunoassays (Immunonephelometric assay, Immunoturbidimetric assay, Immunochemical assay)
74
Target in immunoassays for apolipoprotein
Apo A-1
75
Reagent used in immunoassays for apolipoprotein
Anti-Apo A-1
76
triggers CHD
POSITIVE RISK FACTOR
77
POSITIVE RISK FACTOR for CHD: AGE
≥ 45 years for men ≥ 55 years or premature menopause for women
78
POSITIVE RISK FACTOR for CHD: Hypertension
BP ≥140/90 mmHg or taking antihypertensive medication
79
LDL cholesterol concentration with less than or equal to 1 risk factor
≥ 160 mg/dL
80
LDL cholesterol concentration with greater than or equal to 2 risk factor
≥ 130 mg/dL
81
LDL cholesterol concentration with CHD or risk equivalent
≥ 100 mg/dL
82
POSITIVE RISK FACTOR for CHD: HDL cholesterol concentration
<40 mg/dL
83
CHD risk equivalent (has active lipolysis – stored fats released in serum)
Diabetes mellitus
84
LIST OF POSITIVE RISK FACTOR – triggers CHD
* Age: ≥ 45 years for men; ≥ 55 years or premature menopause for women * Family history of premature CHD * Current cigarette smoking * Hypertension: (BP >140/90 mmHg or taking antihypertensive medication) * LDL cholesterol concentration: (> 160 mg/dL, with <1 risk factor) * LDL cholesterol concentration: (> 130 mg/dL, with >2 risk factor) * LDL cholesterol concentration: (> 100 mg/dL, with CHD or risk equivalent) * HDL cholesterol concentration: (<40 mg/dL) * Diabetes mellitus = CHD risk equivalent (has active lipolysis – stored fats released in serum) * Metabolic syndrome (multiple risk factor)
85
NEGATIVE RISK FACTOR for CHD
* HDL cholesterol concentration (> 60 mg/dL) * LDL cholesterol (<100 mg/dL)
86
Included in LIPID PROFILE TEST
1. Total cholesterol 2. TAG 3. HDL 4. LDL 5. VLDL
87
Lipids not routinely measured in the lab
Fatty Acids Phospholipids

MTAP 2 (23 decks)

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