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Contents
1. Lipid Chemistry

LIPIDS AND 2.

3.
Lipoprotein Structure
Lipoprotein Physiology and Metabolism

LIPOPROTEINS Lipid Disorders

4.

5. Lipid and Lipoprotein Analyses

Lipid Chemistry Lipid Chemistry

Introduction 1. Fatty Acids
Composed of mostly carbons-hydrogen (C-H) bonds Linear chains of C-H bonds that terminated with -COOH
Classification: Classifications:
1. Fatty acids unesterified bound to albumin
2. Triglycerides esterified constituent of triglycerides or phospholipids
3. Cholesterol
short (4-6), medium (8-12), or long chain (>12)
4. Phospholipids
saturated (no double bonds), monosaturated (one double
Transported by lipoproteins (VLDL, LDL, HDL)
bond) or polyunsaturated (≥double bonds)

Lipid Chemistry Lipid Chemistry

1. Fatty Acids 2. Triglyceride
Linear chains of C-H bonds that terminated with -COOH Contain three fatty acids attached to one molecule of glycerol
Contain saturated fatty acids or unsaturated fatty acids
No charged groups, water insoluble, neutral lipid

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Lipid Chemistry Lipid Chemistry

3. Phospholipids 3. Phospholipids
Contain two fatty acids attached to one molecule of glycerol Contain two fatty
Third position contain phospholipid head groups acids attached to one
molecule of glycerol
Amphipathic (contain hydrophilic and hydrophobic head groups)
Third position contain
phospholipid head
groups
Amphipathic
contain hydrophilic
and hydrophobic
head groups)
Lecithin

Lipid Chemistry Lipid Chemistry

4. Cholesterol 4. Cholesterol
Unsaturated steroid alcohol contain four rings Converted to:
Amphipathic (contain hydrophilic and hydrophobic head groups) Bile salts (promote fat absorption in the intestine)
Classifications: Steroid hormones (glucocorticoids, mineralocorticoid, estrogen)
unesterified free cholesterol (amphipathic) Vitamin D and Cell membrane
esterified cholesteryl ester (neutral lipid)

Lipid Chemistry Contents

Chemical Structure of Lipids 1. Lipid Chemistry
2. Lipoprotein Structure
3. Lipoprotein Physiology and Metabolism
4. Lipid Disorders
5. Lipid and Lipoprotein Analyses

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Lipoprotein Structure Lipoprotein Structure

COMPONENTS CHARACTERISTICS
Composed of both lipids and Functions of Apolipoproteins
proteins (apolipoproteins) • Maintain structural integrity
Composition: • Ligands for cell receptor
Free cholesterol, • Activators and inhibitors of
phospholipids are found enzymes
on the surface • Amphipathic helix
Triglycerides and
cholesteryl esters are
found in the core regions

Lipoprotein Structure Lipoprotein Structure

Characteristics of the Major Human Apolipoproteins
Apolipoprotein Major LPP Location Function  Apo A-1: Major protein of HDL
Apo A-I HDL LCAT activator, ABCA1 lipid acceptor  Apo B: LDL, VLDL, chylomicrons
Apo A-II HDL inactivates LCAT Apo B-100: ligand for LDL receptor; covalently linked
Apo A-IV Chylos, VLDL, HDL to Apo (a)
Apo B-100 LDL, VLDL LDL receptor ligand Apo B-48: in chylomicrons
Apo B-48 Chylos Remnant receptor ligand Half of Apo B
Apo C-I Chylos, VLDL, HDL  Apo E: ligand for LDL receptor and chylomicron
Apo C-II Chylos, VLDL, HDL LPL cofactor
remnant receptor
Apo C-III Chylos, VLDL, HDL LPL inhibitor
Apo E2, E3, E4: affect lipoprotein metabolism
Apo E VLDL, HDL LDL receptor Ligand
Apo(a) Lp(a) plasminogen inhibitor

Lipoprotein Structure Lipoprotein Structure

Characteristics of the Major Human Lipoproteins
MAJOR TYPES
Characteristics Chylomicrons VLDL LDL HDL
1. Chylomicrons
2. VLVL Density (g/mL) <0.93 0.93-1.006 1.019-1.063 1.063-1.21
3. LDL Diameter (nm) 80-1,200 30-80 18-30 5-12
4. HDL
Total lipid (% by
98 89-96 77 50
weight)
Triglyceride (%
84 44-60 11 3
by weight
Total cholesterol
7 16-22 62 19
(% by weight)
Major Protein Apo B-48 Apo B-100 Apo B-100 Apo A-1

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Lipoprotein Structure Lipoprotein Structure

MAJOR TYPES MAJOR TYPES
1. Chylomicrons 2. Very Low density Lipoproteins
• Largest and least dense • Also known as pre-beta lipoprotein
• Produced in the intestine • Produced in the Liver
• Delivery of dietary lipids to hepatic and peripheral cells • Transfer triglycerides from the liver to peripheral tissue

Tube of turbid
Tube of turbid plasma plasma left
left overnight in a overnight in a
refrigerator at 4OC refrigerator at
4OC

Lipoprotein Structure Lipoprotein Structure

MAJOR TYPES
3. Intermediate Density Lipoproteins (IDL)
• Also known as VLDL remnants
• Exist transiently during VLDL and LDL conversion
• Found in patients with Type III Hyperlipoproteinemia
MAJOR TYPES
4. Low Density Lipoproteins (LDL)
• Also known as Beta lipoprotein or bad cholesterol
• Formed from lipolysis of VLDL to IDL then to LDL
• Transfer dietary cholesterol to peripheral tissues

Lipoprotein Structure Lipoprotein Structure

MAJOR TYPES MINOR TYPES
5. High Density Lipoproteins (HDL) 6. Lipoprotein(a)
• Also known as Alpha lipoprotein or good cholesterol • LDL lipoprotein like particle
• Produced in the Liver and the Intestine • ↑ Confers increased risk for premature coronary heart
• Transfer cholesterol from peripheral cells back to the liver disease and stroke. Competes with plasminogen for fibrin

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Lipoprotein Structure Lipoprotein Structure

Adult reference ranges for Lipids
MINOR TYPES Conversion
7. Lipoprotein(X) Analyte Reference Range
Factor (mmol/L)
• Abnormal lipoprotein in biliary cirrhosis or cholestasis Total cholesterol 140-200 mg/dL 0.026
paitents; mutations in LCAT
HDL cholesterol 40-75 mg/dL
• Lacks Apo B 100; Removed by the RES
LDL cholesterol 50-130 mg/dL
Triglyceride 60-150 mg/dL 0.011

Contents Lipoprotein Structure

1. Lipid Chemistry LIPOPROTEIN METABOLISM
2. Lipoprotein Structure 1. Lipid Absorption

3. Lipoprotein Physiology and Metabolism 2. Exogenous Pathway

3. Endogenous Pathway
4. Lipid Disorders
4. Reverse Cholesterol Transport Pathway
5. Lipid and Lipoprotein Analyses

Lipoprotein Physiology and Metabolism Lipoprotein Physiology and Metabolism

Dietary Lipids
LIPOPROTEIN METABOLISM 4. Reverse Cholesterol
Transport pathway
1. Lipid Absorption
Conversion of dietary lipids into more polar (amphipathic) 1. Absorption Bile
compounds by Pancreatic Lipase Pathway

• Triglyceride Monoglycerides 2. Exogenous

Pathway
Pe r iphe r al

Cells
• Cholesterol esters Free cholesterol
• Phospholipids Lysophospholipids
Stool 3. Endogenous
Sitosterolemia: defective transporter, predisposition to Pathway
atherosclerosis

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Lipoprotein Physiology and Metabolism Lipoprotein Physiology and Metabolism

Dietary Lipids
LIPOPROTEIN METABOLISM 4. Reverse Cholesterol
Transport pathway
2. Exogenous Pathway
• Chylomicrons are synthesized in the intestine, carrying Bile
1. Absorption
dietary lipids to the circulation Pathway

• Lipoprotein Lipase (LPL) hydrolyzes triglycerides in the 2. Exogenous

Pathway
Pe r iphe r al

Cells
chylomicrons into FA and glycerol or reesterified for long
term storage in the hepatic cells
• Chylomicrons Chylomicron remnant particles Stool 3. Endogenous LPL
Pathway

Lipoprotein Physiology and Metabolism Lipoprotein Physiology and Metabolism

Dietary Lipids
LIPOPROTEIN METABOLISM 4. Reverse Cholesterol
Transport pathway
3. Endogenous Pathway
• Triglycerides in the liver are packaged into VLDL, carrying Bile
1. Absorption
lipids to the circulation Pathway

• VLDL is converted into VLDL remnants by action of LPL and 2. Exogenous

Pathway
Pe r iphe r al

Cells
taken up by liver
• Half of VLDL is transformed into LDL for delivery of
exogenous cholesterol to peripheral cells. Stool 3. Endogenous LPL
Pathway

Lipoprotein Physiology and Metabolism Lipoprotein Physiology and Metabolism

Dietary Lipids
LIPOPROTEIN METABOLISM 4. Reverse Cholesterol
Transport pathway
4. Reverse Cholesterol Transport Pathway
• HDL remove excess cholesterol transport pathway deliver Bile
1. Absorption
cholesterol to the liver Pathway

• Aqueous Diffusion Pathway 2. Exogenous Pe r iphe r al

Pathway Cells
• ABCA1 transporter

Stool 3. Endogenous LPL

Pathway

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Contents
1. Lipid Chemistry
2. Lipoprotein Structure
3. Lipoprotein Physiology and Metabolism
4. Lipid Disorders
5. Lipid and Lipoprotein Analyses
Lipid Disorders
1. Arteriosclerosis
• Deposition of esterified
cholesterol, in artery walls
• Increased smooth muscle cells,
extracellular lipid, calcification,
fibrous tissue, macrophages,
lymphocytes and platelets
(plaque)
• Rupture or erosion can caused
thrombosis that block the
circulation
• E.g. CAD, PVD, CVD
Lipid Disorders
1. Arteriosclerosis
• Treatment:
1. HMG-COA reductase drug
inhibitors, “statins”
• Block intracellular cholesterol
synthesis by inhibiting HMG-
COA reductase
• Increases expression of LDL
receptor
• Myositis and hepatotoxic effect
• Fish oil products
• Decrease hepatic TG synthesis
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Lipid Disorders
Lipid Disorder
1. Arteriosclerosis
2. Hyperlipoproteinemia
i. Hypercholesterolenemia
ii. Hypertriglyceridemia
iii. Combined hyperlipidemia
3. Hypolipoporoteinemia
Lipid Disorders
1. Arteriosclerosis
• Coronary Artery disease
(CAD) heart
(angina and MI)
• Peripheral vascular disease
(PVD) 
arteries in arms or legs
• Cerebrovascular disease
(CVD) 
vessels of the brain (stroke)
Lipid Disorders
2. Hyperlipoproteinemia
2. Niacin or nicotinic Acid i. Hypercholesterolenemia
Reduce LDL-C (5-25%) and raise • ↑ Cholesterol
HDL-C by 15-35% • ↑ LDL, ↓ receptors
• Inhibits lipolysis of TG ii. Hypertriglyceridemia
3. Fibric Acid derivatives • ↑ Triglycerides
• Dec TG by 20-50% and • ↓ LPL or Apo C-II
inc HDL by 10-20% • VLDL -/VLDL remnants
• Chylos -/Chylos remnants
4. Ezetimibe
• Inhibits cholesterol iii. Combined Hyperlipoprotenemia
absorption • ↑ Triglycerides, Cholesterol
• Used with statins • ↑VLDL and Chylos remnants
• Presence of apo E2/2
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Lipid Disorders Lipid Disorders

2. Hyperlipoproteinemia 2. Hyperlipoproteinemia

i. Hypercholesterolenemia ii. Familial hypertriglyceridemia or secondary causes

• Familial Hypercholesterolenemia (FH)
• Homozygotes: levels 800-1,0000mg/dL
• Heterozygotes: autosomal codominant; 300-600mg/dL
• Increase in LDL-C, deficient in LDL receptors
• Class 1: no LDL receptor
• Class 2: membrane protein synthesized but not transported
• Class 3: LDL receptor cannot bind LDL
• Class 4: cannot internalize LDL
• Class 5: do not recycle
• Homozygotes: LDL pheresis, Heterozygote: statins
• DM: increased shunting of glucose into pentose pathway,
increased FA synthesis
• Renal failure: depresses removal of large MW constituents
• Can cause acute and recurrent pancreatitis
Severe hypertriglyceridemia
• Deficiency of LPL located on chromosome 8 or deficiency in
Apo C II

Lipid Disorders Lipid Disorders

2. Hyperlipoproteinemia 3. Lipoprotein (a) elevation
• Seen in CHD patients
iii. Combined Hyperlipoproteinemia • Competes with plasminogen for fibrin binding sites; increases
• Elevated levels of TG and cholesterol plaque formation
• Familial Combined Hyperlipoproteinemia (FCH)
4. Non- HDL Cholesterol
• Excessive Hepatic synthesis of apoprotein Binc. VLDL secretion
inc. production of LDL • Reflects TC minus HDL-C
• High risk for CHD • Measured in a non-fasting state
• Independent predictor for CVD and diabetes patients
• Familial dysbetalipoproteinemia / Type III hyperlipoproteinemia
• Accumulation of cholesterol rich VLDL and chylomicron remnants 5. Hypobetalipoproteinemia- Isolated low levels of LDL-C
• Presence of Apo E2/2
• Will migrate in the β region instead of in pre β region 6. Hypoalphalipoproteinemia- decrease in circulating HDL
• Tangier disease: 1-2 mg/dL

Contents Lipid and Lipoprotein Analysis

1. Lipid Chemistry LIPID MEASUREMENT
2. Lipoprotein Structure 1. Cholesterol Measurement

3. Lipoprotein Physiology and Metabolism 2. Triglyceride Measurement

3. Lipoprotein Measurement
4. Lipid Disorders
i. High-Density Lipoprotein Methods
5. Lipid and Lipoprotein Analyses
ii. Low-Density Lipoprotein Methods
iii. Apolipoprotein Methods

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Lipoprotein Physiology and Metabolism Lipoprotein Physiology and Metabolism

LIPID MEASUREMENT LIPID MEASUREMENT

1. Cholesterol Measurement
Non-Enzymatic Methods (Abell Kendall)
i. Cholesteryl esters are hydrolyzed with alcoholic KOH
ii. Unesterified cholesterol is extracted with hexane
iii. Measured using Liebermann – Burchard reaction
Cholesterol + sulfuric acid + acetic anhydride Green Sol.
1. Cholesterol Measurement
Enzymatic Method (Cholesterol oxidase)
i. Total Cholesterol (cholesteryl ester and cholesterol) + H2O
–Cholesteryl esterase Cholesterol + Fatty Acid
ii. Cholesterol + O2 –Cholesterol oxidase 
4-cholestenone + H 2O 2
iii. H2O 2 + Dye –Horseradish Peroxidase  Color

Lipid and Lipoprotein Analysis Lipoprotein Physiology and Metabolism

LIPID MEASUREMENT LIPID MEASUREMENT

1. Cholesterol Measurement 2. Triglyceride Measurement
2. Triglyceride Measurement Non-enzymatic Method
3. Lipoprotein Measurement i. Triglyceride –Alcoholic KOH  Glycerol + Fatty Acid
i. High-Density Lipoprotein Methods ii. Glycerol + Periodic acid  Formaldehyde
ii. Low-Density Lipoprotein Methods a. Van Handel and Zilversmith
iii. Apolipoprotein Methods iii. Formaldehyde + Chromotropic acid  Blue Solution
b. Hantzch Condensation (Fluorometric Method)
iii. Formaldehyde + Diacetyl acetone + NH 
3 Yellow Sol’n

Lipoprotein Physiology and Metabolism Lipid and Lipoprotein Analysis

LIPID MEASUREMENT LIPID MEASUREMENT
2. Triglyceride Measurement 1. Cholesterol Measurement
Enzymatic Methods (Glycerol Kinase) 2. Triglyceride Measurement
i. Triglyceride –LipaseGlycerol + Fatty Acid 3. Lipoprotein Measurement
ii. Glycerol + ATP –GlycerokinaseGlycerophosphate +ADP i. High-Density Lipoprotein Methods
a. Pyruvate Kinase ii. Low-Density Lipoprotein Methods
iii. ADP + PEP –Pyruvate kinaseATP + Pyruvate iii. Apolipoprotein Methods
+
iv. Pyruvate + NADH + H+ –LDlactate + NAD (340 nm)
b. Glycerol-Phosphate Oxidase
iii. Glycerophosphate + O2 –GODihydroxyacetone + H O2 2
iv. H2O 2 + Dye –PeroxidaseColor
PEP = phosphoenol pyruvate; LD = Lactate dehydrogenase; GO = Glycerophosphate oxidase

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Lipid and Lipoprotein Analysis Lipid and Lipoprotein Analysis

LIPID MEASUREMENT LIPID MEASUREMENT
3. Lipoprotein Measurement 3. Lipoprotein Measurement
i. General methods i. General methods
ii. High-Density Lipoprotein Methods  Ultracentrifugation
iii. Low-Density Lipoprotein Methods  Based on the molecular
iv. Apolipoprotein Methods density
 Order from the lightest
to heaviest
i. Chylomicrons
ii. VLDL
iii. LDL
iv. HDL

Lipid and Lipoprotein Analysis Lipid and Lipoprotein Analysis

LIPID MEASUREMENT LIPID MEASUREMENT
3. Lipoprotein Measurement 3. Lipoprotein Measurement
i. General methods ii. High-Density Lipoprotein (HDL) Methods
 Electrophoresis  Polyanion Precipitation
 Based on migration in an electric field  Lipoproteins (LDL, VLDL) are precipitated with
 Lipid stains (Oil red O, Fat Red 7B and Sudan Black) polyanions (e.g. Heparin, dextran sulfate or Na
phosphotungstate) in the presence of divalent
 4 bands with fat stain
cations (e.g. Mg or Mn), which are sediment by
i. Alpha-lipoprotein (HDL) –fastest centrifugation (10-30 min. for 10,000g or 3 min.
ii. Pre-β – VLDL, LP(a) for 15,000g
iii. Beta – LDL  HDL is quantified in the supernate by Abell-Kendal
iv. Chylomicrons – stationary at origin

Lipid and Lipoprotein Analysis Lipid and Lipoprotein Analysis

LIPID MEASUREMENT LIPID MEASUREMENT
3. Lipoprotein Measurement 3. Lipoprotein Measurement
iii. Low-Density Lipoprotein (LDL) Methods iv. Apolipoprotein Methods
a. LDL = Total cholesterol – (HDL + VLDL)  Apo B – for determination of LDL and VLDL conc.
 Friedewald:  Apo A-1 – for determination of HDL conc.
= Triglyceride / 5 = mg/dL
VLDL = Triglyceride / 2.175 = mmol/L
b. Ultracentrifugation of serum at native density gradient
of 1.006 g/L to float VLDL

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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 LPP Population Distributions
NEGATIVE RISK FACTOR
HDL cholesterol concentration ≥60mg/dL
LDL cholesterol <100 mg/dL
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Lipid and LPP Population Distributions
POSITIVE RISK FACTOR
Age: ≥45 years for men; ≥55 years or premature menopause for women
Family history of premature CHD
Current cigarette smoking
Hypertension (BP ≥140/90 mm Hg or taking antihypertensive medication
LDL cholesterol concentration ≥160mg/dL, with ≤1 risk factor
LDL cholesterol concentration ≥130mg/dL, with ≥2 risk factor
LDL cholesterol concentration ≥100mg/dL, with CHD or risk equivalent
HDL cholesterol conentration >40 mg/DL
Diabetes mellitus = CHD risk equivalent
Metabolic syndrome (multiple risk factor0
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