MIDTERM LAB: LIPID PROFILE Flashcards
Requires at least _____ of fasting
10-16 hours
OPTIMAL FASTING:
12 HOURS
samples are not accepted
Hemolyzed and icteric
Specimen:
Serum or plasma EDTA
Not affected by fasting:
CHOLESTEROL determination AND HDL
Lipid Profile TEST:
Triglyceride determination
Total Cholesterol determination
HDL determination
LDL determination (computed)
VLDL determination (computed)
Friedewald Equation
TC = HDL + LDL + VLDL
LDL = TC – HDL – TAG/5 (mg/dL)
LDL = TC – HDL – TAG/2.175 (mmol/L)
Friedewald Equation FOR mg/dl:
LDL = TC – HDL – TAG/5 (mg/dL
Friedewald Equation FOR mmol/l:
LDL = TC – HDL – TAG/2.175 (mmol/L)
De Long Equation:
TC = HDL + LDL + VLDL
LDL = TC – HDL – TAG/ 6.5 (mmol/L)
LDL = TC – HDL – TAG/2.825 (mmol/L)
De Long Equation FOR mg/dl:
LDL = TC – HDL – TAG/ 6.5 (mmol/L)
De Long Equation FOR mmol/L:
LDL = TC – HDL – TAG/2.825 (mmol/L)
Cholesterol Measurement:
- Abell-Kendall Method
- Abell, Levey, Brody Method
- Sperry Method
- Salkowski Method
- Enzymatic Method
- Isotope Dilution/Mass Spectrometry (IDMS)
OLD reference method
Abell-Kendall Method
3-step method
Abell-Kendall Method
Abell-Kendall Method reagent:
Libermann-Burchard reagent: gHAc, acetic anhydride,
concentrated H2SO4
Abell-Kendall Method 3-step method:
Saponification -> Extraction -> Colorimetry
Current Reference Method
Abell, Levey, Brody Method
Abell-Kendall Method End Color:
GREEN
Abell-Kendall Method End Product:
cholestadienyl monosulfuric acid
Abell, Levey, Brody Method Reagents:
Libermann-Burchard rgt + KOH and alcoholic hexane
Abell, Levey, Brody Method End Color:
GREEN
Sperry Method 4-step method:
Saponification -> Extraction -> Colorimetry
-> Purification with digitonide
4-step method
Sperry Method
Salkowski Method End Product:
cholestadienyl disulfuric acid
Salkowski Method End Color:
RED
most commonly used
Enzymatic Method
Enzymatic Method interferences:
ascorbate, hemoglobin, bilirubin
Definitive method
Isotope Dilution/Mass Spectrometry (IDMS)
GOLD STANDARD FOR CHOLESTEROL
Isotope Dilution/Mass Spectrometry (IDMS)
Triglyceride Measurement:
- Van Handel & Zilversmit Method
- Modified Van Handel & Zilversmit Method
- Hantzsch Method
Colorimetric Method
Van Handel & Zilversmit Method
Van Handel & Zilversmit Method Reagent:
chromotropic acid
Van Handel & Zilversmit Method End Color:
BLUE
Modified Van Handel & Zilversmit Method End Color:
PINK
Fluorometric method
Hantzsch Method
Modified Van Handel & Zilversmit Method Reagent:
sulphuric acid, salicylic acid
Colorimetric Method and Reference Method
Modified Van Handel & Zilversmit Method
Hantzsch Method End Color:
YELLOW
Hantzsch Method Reagent:
acetylacetone (diacetyl acetone)
Reference method; Measured in Svedverg units
Ultracentrifugation
Lipoprotein Analysis:
- Ultracentrifugation
- Electrophoresis
Ultracentrifugation Reagent:
potassium bromide w/ 1.063 density
Fluorometric method
Ultracentrifugation
Floating layer:
Chylo, VLDL
Sinking layer:
LDL, HDL
Electrophoresis
- Preferred gel:
agarose gel
From origin to the most anodal:
Chylo, LDL, VLDL, HDL
