LIPIDS AND LIPOPROTEINS

Contents
1.

2.
3. 4. 5.

Lipid Chemistry Lipoprotein Structure Lipoprotein Physiology and Metabolism Lipid Disorders Lipid and Lipoprotein Analyses

Lipid Disorders
Lipid Disorder

1. Arteriosclerosis 2. Hyperlipoproteinemia i. Hypercholesterolenemia ii. Hypercholesterolenemia iii. Combined hyperlipidemia 3. Hypolipoporoteinemia

Lipid Disorders
1. Arteriosclerosis
Deposition of esterified cholesterol in artery walls
Coronary Artery disease CAD heart Peripheral vascular disease PVD arteries in arms/legs

Cerebrovascular disease CVD vessels of the brain

Lipid Disorders
1. Arteriosclerosis
Deposition of esterified cholesterol in artery walls
Coronary Artery disease CAD heart Peripheral vascular disease PVD arteries in arms/legs Cerebrovascular disease CVD vessels of the brain

Lipid Disorders
2. Hyperlipoproteinemia i. Hypercholesterolenemia Cholesterol LDL, receptors

ii. Hypertriglyceridemia Triglycerides LPL or Apo C-II VLDL -/VLDL remnants Chylos -/ Chylos remnants iii. Combined Hyperlipoprotenemia Triglycerides, Cholesterol VLDL and Chylos remnants Presence of apo E2/2

Lipid Disorders
Lipid Disorder 1. Arteriosclerosis 2. Hyperlipoproteinemia i. Hypercholesterolenemia ii. Hypertriglyceridemia iii. Combined hyperlipidemia 3. Liporotein (a) Elevation 4. Hypolipoporoteinemia

Contents
1.

2.
3. 4. 5.

Lipid Chemistry Lipoprotein Structure Lipoprotein Physiology and Metabolism Lipid Disorders Lipid and Lipoprotein Analyses

Lipid and Lipoprotein Analysis

LIPID MEASUREMENT 1. Cholesterol Measurement 2. Triglyceride Measurement 3. Lipoprotein Measurement i. High-Density Lipoprotein Methods
ii. Low-Density Lipoprotein Methods
iii. Apolipoprotein Methods

Lipoprotein Physiology and Metabolism

LIPID MEASUREMENT 1. Cholesterol Measurement Non-Enzymatic Methods (Abell Kendall)
i. ii. Cholesteryl esters are hydrolyzed with alcoholic KOH Unesterified cholesterol is extracted with hexane Cholesterol + sulfuric acid + acetic anhydride Green Sol.

iii. Measured using Liebermann Burchard reaction

Lipoprotein Physiology and Metabolism

LIPID MEASUREMENT 1. Cholesterol Measurement Enzymatic Method (Cholesteryl oxidase)
i. Total Cholesterol (cholesteryl ester and cholesterol) + H2O Cholesteryl esterase Cholesterol + Fatty Acid

ii.

Cholesterol + O2 Cholesterol oxidase 4-cholestenone + H2O2

iii. H2O2 + Dye Peroxidase Color

Lipid and Lipoprotein Analysis

LIPID MEASUREMENT 1. Cholesterol Measurement 2. Triglyceride Measurement 3. Lipoprotein Measurement i. High-Density Lipoprotein Methods
ii. Low-Density Lipoprotein Methods
iii. Apolipoprotein Methods

Lipoprotein Physiology and Metabolism

LIPID MEASUREMENT 2. Triglyceride Measurement Non-enzymatic Method
i. ii. Triglyceride Alcoholic KOH Glycerol + Fatty Acid Glycerol + Periodic acid Formaldehyde

a. Van Handel and Zilversmith iii. Formaldehyde + Chromotropic acid Blue Solution b. Hantzch Condensation (Fluorometric Method) iii. Formaldehyde + Diacetyl acetone + NH3 Yellow Soln

Lipoprotein Physiology and Metabolism

LIPID MEASUREMENT 2. Triglyceride Measurement Enzymatic Methods (Glycerol Kinase)
i. ii. Triglyceride Lipase Glycerol + Fatty Acid Glycerol + ATP Glycerokinase Glycerophosphate +ADP

a.

Pyruvate Kinase
iii. ADP + PEP Pyruvate kinase ATP + Pyruvate iv. Pyruvate + NADH + H+ LD lactate + NAD+ (340 nm)

b.

Glycerol-Phosphate Oxidase iii. Glycerophosphate + O2 GO Dihydroxyacetone + H2O2 iv. H2O2 + Dye Peroxidase Color

PEP = phosphoenol pyruvate; LD = Lactate dehydrogenase; GO = Glycerophosphate oxidase

Lipid and Lipoprotein Analysis

LIPID MEASUREMENT 1. Cholesterol Measurement 2. Triglyceride Measurement 3. Lipoprotein Measurement i. High-Density Lipoprotein Methods
ii. Low-Density Lipoprotein Methods
iii. Apolipoprotein Methods

Lipid and Lipoprotein Analysis

LIPID MEASUREMENT 3. Lipoprotein Measurement i. General methods ii. High-Density Lipoprotein Methods
iii. Low-Density Lipoprotein Methods

iv. Apolipoprotein Methods

Lipid and Lipoprotein Analysis

LIPID MEASUREMENT 3. Lipoprotein Measurement i. General methods Ultracentrifugation
Based on the molecular density Order from the lightest to heaviest i. Chylomicrons ii. VLDL iii. LDL iv. HDL

Lipid and Lipoprotein Analysis

LIPID MEASUREMENT 3. Lipoprotein Measurement i. General methods Electrophoresis
Based on migration in an electric field

Lipid stains (Oil red O, Fat Red 7B and Sudan Black)

4 bands with fat stain i. ii. iii. iv. Alpha-lipoprotein fastest Pre- VLDL, LP(a) Beta LDL Chylomicrons stationary at origin

Lipid and Lipoprotein Analysis

LIPID MEASUREMENT 3. Lipoprotein Measurement ii. High-Density Lipoprotein (HDL) Methods
Polyanion Precipitation Lipoproteins (LDL, VLDL) are precipitated with polyanions (e.g. Heparin, dextran sulfate or Na phosphotungstate) in the 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

Lipid and Lipoprotein Analysis

LIPID MEASUREMENT 3. Lipoprotein Measurement iii. Low-Density Lipoprotein (LDL) Methods a. LDL = Total cholesterol (HDL + VLDL)
Friedewald:

VLDL

= Triglyceride / 5 = mg/dL = Triglyceride / 2.175 = mmol/L

b. Ultracentrifugation of serum at native density gradient of 1.006 g/L to float VLDL

Lipid and Lipoprotein Analysis

LIPID MEASUREMENT 3. Lipoprotein Measurement iv. Apolipoprotein Methods Apo B for determination of LDL and VLDL conc.
Apo A-1 for determination of HDL conc.

Lipid and Lipoprotein Analysis

LIPID MEASUREMENT 3. Lipoprotein Measurement i. General methods ii. High-Density Lipoprotein Methods
iii. Low-Density Lipoprotein Methods

iv. Apolipoprotein Methods