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Contents

LIPIDS AND 1.

2.

3.
Lipid Chemistry
Lipoprotein Structure
Lipoprotein Physiology and Metabolism

LIPOPROTEINS 4.

5.
Lipid Disorders
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 colesterol (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
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 MAJOR TYPES
Apo A-I HDL LCAT activator, ABCA1 lipid acceptor 1. Chylomicrons
Apo A-II HDL inactivates LCAT 2. VLVL
Apo A-IV Chylos, VLDL, HDL 3. LDL
Apo B-100 LDL, VLDL LDL receptor ligand 4. HDL
Apo B-48 Chylos Remnant receptor ligand
Apo C-I Chylos, VLDL, HDL
Apo C-II Chylos, VLDL, HDL LPL cofactor
Apo C-III Chylos, VLDL, HDL LPL inhibitor
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
Density (g/mL) <0.93 0.93-1.006 1.019-1.063 1.063-1.21 • Largest and least dense
Diameter (nm) 80-1,200 30-80 18-30 5-12 • Produced in the intestine
• Delivery of dietary lipids to hepatic and peripheral cells
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
2. Very Low density Lipoproteins 3. Low Density Lipoproteins (LDL)
• Also known as pre-beta lipoprotein • Also known as Beta lipoprotein or bad cholesterol
• Produced in the Liver • Formed from lypolysis of VLDL to IDL then to LDL
• Transfer triglycerides from the liver to peripheral tissue • Transfer dietery cholesterol to peripheral tissues

Lipoprotein Structure Lipoprotein Structure

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

Lipoprotein Structure Contents

Adult reference ranges for Lipids
1. Lipid Chemistry
Conversion
Analyte Reference Range Lipoprotein Structure
Factor (mmol/L) 2.

Total cholesterol 140-200 mg/dL 0.026 3. Lipoprotein Physiology and Metabolism

HDL cholesterol 40-75 mg/dL 4. Lipid Disorders
LDL cholesterol 50-130 mg/dL 5. Lipid and Lipoprotein Analyses
Triglyceride 60-150 mg/dL 0.011

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Lipoprotein Structure Lipoprotein Physiology and Metabolism

LIPOPROTEIN METABOLISM LIPOPROTEIN METABOLISM

1. Lipid Absorption 1. Lipid Absorption
2. Exogenous Pathway Conversion of dietery lipids into more polar (amphipathic)
3. Endogenous Pathway compounds by Pancreatic Lipase
4. Reverse Cholesterol Transport Pathway • Triglyceride  Monoglycerides
• Cholesterol esters  Free cholesterol
• Phospholipids  Lysophospholipids

Lipoprotein Physiology and Metabolism Lipoprotein Physiology and Metabolism

Dietary Lipids
4. Reverse Cholesterol LIPOPROTEIN METABOLISM
Transport pathway
2. Exogenous Pathway
1. Absorption Bile • Chylomicrons are synthesized in the intestine, carrying
Pathway dietery lipids to the circulation
2. Exogenous
Pathway Peripheral • Lipoprotein Lipase (LPL) hydrolyzes triglycerides in the
Cells
chylomicrons into FA and glycerol or reesterified for long
term storage in the hepatic cells
Stool 3. Endogenous • Chylomicrons  Chylomicron remnant particles
Pathway

Lipoprotein Physiology and Metabolism Lipoprotein Physiology and Metabolism

Dietary Lipids
4. Reverse Cholesterol LIPOPROTEIN METABOLISM
Transport pathway
3. Endogenous Pathway
1. Absorption Bile • Triglycerides in the liver are packaged into VLDL, carrying
Pathway lipids to the circulation
2. Exogenous
Pathway Peripheral • VLDL is converted into VLDL remnants by action of LPL and
Cells
taken up by liver
• Half of VLDL is transformed into LDL for delivery of
Stool 3. Endogenous LPL exogenous cholesterol to peripheral cells.
Pathway

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Lipoprotein Physiology and Metabolism Lipoprotein Physiology and Metabolism

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

Stool 3. Endogenous LPL

Pathway

Lipoprotein Physiology and Metabolism Contents

Dietary Lipids
4. Reverse Cholesterol 1. Lipid Chemistry
Transport pathway
2. Lipoprotein Structure
1. Absorption Bile 3. Lipoprotein Physiology and Metabolism
Pathway
2. Exogenous 4. Lipid Disorders
Pathway Peripheral
Cells
5. Lipid and Lipoprotein Analyses

Stool 3. Endogenous LPL

Pathway

Lipid Disorders Lipid Disorders

Lipid Disorder 1. Arteriosclerosis
1. Arteriosclerosis • Deposition of esterified
2. Hyperlipoproteinemia cholesterol, in artery walls
i. Hypercholesterolenemia • Increased smooth muscle cells,
ii. Hypertriglyceridemia extracellular lipid, calcification,
iii. Combined hyperlipidemia fibrous tissue, macrophages,
3. Hypolipoporoteinemia lymphocytes and platelets
(plaque)
• Rupture or erosion can caused
thrombosis that block the
circulation
• E.g. CAD, PVD, CVD

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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
Lipid Disorder
1. Arteriosclerosis
2. Hyperlipoproteinemia
i. Hypercholesterolenemia
ii. Hypertriglyceridemia
iii. Combined hyperlipidemia
3. Liporotein (a) Elevation
4. Hypolipoporoteinemia
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
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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
Contents
1. Lipid Chemistry
2. Lipoprotein Structure
3. Lipoprotein Physiology and Metabolism
4. Lipid Disorders
5. Lipid and Lipoprotein Analyses
Lipoprotein Physiology and Metabolism
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.
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Lipoprotein Physiology and Metabolism Lipid and Lipoprotein Analysis

LIPID MEASUREMENT LIPID MEASUREMENT
1. Cholesterol Measurement 1. Cholesterol Measurement
Enzymatic Method (Cholesteryl oxidase) 2. Triglyceride Measurement
i. Total Cholesterol (cholesteryl ester and cholesterol) + H2O 3. Lipoprotein Measurement
–Cholesteryl esterase Cholesterol + Fatty Acid i. High-Density Lipoprotein Methods
ii. Cholesterol + O2 –Cholesterol oxidase
ii. Low-Density Lipoprotein Methods
4-cholestenone + H2O2
iii. H2O2 + Dye –Peroxidase Color iii. Apolipoprotein Methods

Lipoprotein Physiology and Metabolism Lipoprotein Physiology and Metabolism

LIPID MEASUREMENT LIPID MEASUREMENT

2. Triglyceride Measurement
Non-enzymatic Method
i. Triglyceride –Alcoholic KOH Glycerol + Fatty Acid
ii. 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 Sol’n
PEP = phosphoenol pyruvate; LD = Lactate dehydrogenase; GO = Glycerophosphate oxidase
2. Triglyceride Measurement
Enzymatic Methods (Glycerol Kinase)
i. Triglyceride –Lipase Glycerol + Fatty Acid
ii. 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

Lipid and Lipoprotein Analysis Lipid and Lipoprotein Analysis

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

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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
§ Ultracentrifugation § Electrophoresis
§ Based on the molecular § Based on migration in an electric field
density § Lipid stains (Oil red O, Fat Red 7B and Sudan Black)
§ Order from the lightest § 4 bands with fat stain
to heaviest
i. Chylomicrons i. Alpha-lipoprotein –fastest
ii. VLDL ii. Pre-β – VLDL, LP(a)
iii. LDL iii. Beta – LDL
iv. HDL iv. Chylomicrons – stationary at origin

Lipid and Lipoprotein Analysis Lipid and Lipoprotein Analysis

LIPID MEASUREMENT LIPID MEASUREMENT
3. Lipoprotein Measurement 3. Lipoprotein Measurement
ii. High-Density Lipoprotein (HDL) Methods iii. Low-Density Lipoprotein (LDL) Methods
§ Polyanion Precipitation a. LDL = Total cholesterol – (HDL + VLDL)
§ Lipoproteins (LDL, VLDL) are precipitated with § Friedewald:
polyanions (e.g. Heparin, dextran sulfate or Na = Triglyceride / 5 = mg/dL
phosphotungstate) in the presence of divalent VLDL = Triglyceride / 2.175 = mmol/L
cations (e.g. Mg or Mn), which are sediment by
centrifugation (10-30 min. for 10,000g or 3 min. b. Ultracentrifugation of serum at native density gradient
of 1.006 g/L to float VLDL
for 15,000g
§ HDL is quantified in the supernate by Abell-Kendal

Lipid and Lipoprotein Analysis Lipid and Lipoprotein Analysis

LIPID MEASUREMENT LIPID MEASUREMENT
3. Lipoprotein Measurement 3. Lipoprotein Measurement
iv. Apolipoprotein Methods i. General methods
§ Apo B – for determination of LDL and VLDL conc. ii. High-Density Lipoprotein Methods
§ Apo A-1 – for determination of HDL conc. iii. Low-Density Lipoprotein Methods
iv. Apolipoprotein Methods

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Lipid and LPP Population Distributions Lipid and LPP Population Distributions

POSITIVE RISK FACTOR NEGATIVE RISK FACTOR

Age: ≥45 years for men; ≥55 years or premature menopause for women HDL cholesterol concentration ≥60mg/dL
Family history of premature CHD LDL cholesterol <100 mg/dL
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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