Cholesterol: Methods for Analysis Flashcards
(28 cards)
hydrolysis of cholesterol esters
saponification
reagent used in saponification
KOH (alcoholic potassium hydroxide)
removal of protein interference
extraction
reagent used for extraction
Bloor’s rgt
(3 ethanol : 1 ether)
precipitation of free cholesterol
purification
reagent used for purification
digitonin
formation of colored compounds
colorimetry
reagents used for colorimetry
- sulfuric acid (H2SO4)
- acetic anhydride (CH3CO)2O
- ferric iron
cholestadienyl MONOSULFONIC acid and end product
Liebermann-Burchardt
green
cholestadienyl DISULFONIC acid and end product
Salkowski
red
CDC reference method
Modified Abell-Kendall
Sample + reagent → incubate → measure the colored product using colorimeters
one-step method or
Pearson, Stern & Mac Gavack
colorimetry and extraction of cholesterol
two-step method
Bloor’s
colorimetry + extraction + saponification
three-step method
what test uses a three-step method involving: colorimetry + extraction + saponification
Modified Abell-Kendall
reagents used in Modified Abell-Kendall steps:
saponification - _____
extraction - _____
colorimetry - _____
saponification - alcoholic KOH
extraction - n-hexane/ petroleum ether
colorimetry - Liebermann-Burchardt
colorimetry + extraction + saponification + purification
Four-step method or
Schoenheimer/Sperry
Enzymatic method involves
Cholesteryl esterase
Cholesterol oxidase
Peroxidase
Enzymatic method color is measured at
500 nm
Reference Value:
<200mg/dL
desirable
Reference Value:
200-239 mg/dL
borderline high
Reference Value:
≥240 mg/dL
high
age (years) : 2-19
moderate risk:
high risk for CHD:
moderate risk: ≥170 mg/dL
high risk for CHD: ≥185 mg/dL
age (years) : 20-29
moderate risk:
high risk for CHD:
moderate risk: ≥200mg/dL
high risk for CHD: ≥220mg/dL