Lipid Analysis

Isolation, Separation, Identification and Structural Analysis of Lipids

1st Edition - June 5, 2014
Author: William W. Christie
Language: English
eBook ISBN:
9 7 8 - 1 - 4 8 3 1 - 8 7 1 7 - 4

Lipid Analysis: Isolation, Separation, Identification, and Structural Analysis of Lipids focuses on the methods, principles, and approaches involved in the isolation,… Read more

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Lipid Analysis: Isolation, Separation, Identification, and Structural Analysis of Lipids focuses on the methods, principles, and approaches involved in the isolation, identification, separation, and structural analysis of lipids. The publication first elaborates on the structure, chemistry and occurrence of lipids, isolation of lipids from tissues, and chromatographic and spectroscopic analysis of lipids. Discussions focus on equipping a laboratory for lipid analysis, spectroscopy, chromatographic procedures, general principles of solvent extraction procedures, structural features of lipids important in analyses, simple and complex lipids, and fatty acids. The text then examines the preparation of volatile derivatives of lipids and analysis of fatty acids and simple lipid classes. Topics include alkyl diglycerides and neutral plasmalogens, analysis of the hydrolysis products of simple lipids, preparation of large quantities of pure fatty acids, physical characterization of fatty acids, location of double bonds in fatty acid chains, and saponification of lipids. The manuscript covers the analysis and radioassay of isotopically-labelled lipids, enzymatic hydrolysis of lipids, and analysis of molecular species of lipids. The publication is a primary reference for researchers interested in lipid analysis.

Preface

Acknowledgements

1. The Structure, Chemistry and Occurrence of Lipids

A. Introduction

B. The Fatty Acids

1. Saturated Fatty Acids

2. Monoenoic Fatty Acids

3. Non-Conjugated Polyunsaturated Fatty Acids

4. Branched-Chain and Cyclopropane Fatty Acids

5. Unusual Fatty Acids of Plant Origin

C. Simple Lipids

1. Triglycerides and Partial Glycerides

2. Alkyl Diglycerides and Neutral Plasmalogens

3. Cholesterol and Cholesteryl

4. Wax Esters and Other Minor Components

D. Complex Lipids

1. Phosphoglycerides

2. Glycosyl Diglycerides and Related Compounds

3. Sphingolipids

E. Structural Features of Lipids Important in Analyses

2. The Isolation of Lipids From Tissues

A. General Principles of Solvent Extraction Procedures

1. Storage of Tissues

2. the Solubility of Lipids in Organic Solvents

3. Removal of Non-Lipid Contaminants

4. Artefacts of Extraction Procedures

5. Some Practical Considerations

B. Recommended Procedures

1. Extraction of Large Amounts of Tissue

2. Chloroform-Methanol (2:1, v/v) Extraction and "Folch" Wash

3. Sephadex G-25 Columns for Removing Non-Lipid Contaminants

3. Chromatographic and Spectroscopic Analysis of Lipids. General Principles

A. A Statement of the Problem

B. Chromatographic Procedures

1. Gas-Liquid Chromatography

2. Adsorption Chromatography

3. Diethylaminoethyl (DEAE) Cellulose Chromatography

4. Partition Chromatography

5. Chromatography on Adsorbents Containing Complexing Agents

C. Some Practical Considerations

1. Autoxidation of Lipids

2. Solvents

3. Contaminants

D. Spectroscopy

1. Infrared Absorption Spectroscopy

2. Ultraviolet Absorption Spectroscopy

3. Nuclear Magnetic Resonance Spectroscopy

4. Mass Spectrometry

5. Optical Rotary Dispersion

E. Equipping a Laboratory for Lipid Analysis

4. The Preparation of Volatile Derivatives of Lipids

A. Saponification of Lipids

B. The Preparation of Methyl Esters of Fatty Acids

1. Acid-Catalysed Estenfication and Transesterification

2. Base-Catalysed Transesterification

3. Diazomethane

4. Special Cases

C. Derivatives of Free Hydroxyl Groups

1. Acetylation

2. Trifluoroacetates

3. Trimethyhilyl Ethers

4. Isopropylidene Compounds

5. N-Butylboronate Derìvatives

D. Derivatives of Fatty Aldehydes

1. Hydrazone Derivatives

2. Acetals

E. Derivatives of Double Bonds

1. Mercuric Acetate Derivatives

2. Hydroxylation

3. Epoxidation

4. Deuterohydrazine Reduction

5. The Analysis of Fatty Acids

A. Introduction

B. Analytical Gas-Liquid Chromatography

1. Fatty Acids of Animal Origin

2. Unusual Fatty Acids

3. Spurious Peaks on Recorder Traces

C. Isolation of Individual Fatty Acids for Structural Analysis

1. Adsorption Chromatography

2. Silver Nitrate Chromatography

3. Preparative Gas Chromatography

4. Liquid-Liquid Partition Chromatography

D. Spectroscopy of Fatty Acids

1. Infrared Absorption Spectroscopy

2. Ultraviolet Spectroscopy

3. Nuclear Magnetic Resonance Spectroscopy

4. Mass Spectrometry

E. Hydrogenation

F. Location of Double Bonds in Fatty Acid Claims

1. Permanganate-Periodate Oxidation

2. Ozonolysis and Reductive Cleavage

3. Oxidation After Partial Reduction of Polyunsaturated

G. Location of Other Functional Groups in Fatty Adds

H. Physical Characterisation of Fatty Acids

I. Preparation of Large Quantities of Pure Fatty Acids

1. Saturated Fatty Acids

2. Polyunsaturated Fatty Acids

6. The Analysis of Simple Lipid Classes

A. Chromatographic Separation of the Common Simple Lipid Classes

1. Thin-Layer Chromatography

2. Column Chromatography

3. Gas-Liquid Chromatography

4. Separation of Isomeric Mono- and Diglycerides

B. Analysis of the Hydrolysis Products of Simple Lipids

1. Fatty Acids

2. Glycerol

3. Cholesterol

C. Alkyl Diglycerides and Neutral Plasmalogens

1. Separation and Identification

2. Isolation and Identification of Aldehydes from Plasmalogens

3. Isolation and Identification of I-Alkyl Glycerols

D. Wax Esters and Related Compounds

7. The Analysis of Complex Lipids

A. Preliminary Separation and Preparation of Lipid Samples

1. Simple Group Separations

2. Cations Associated with Complex Lipids

B. Column Chromatography

1. Silicic Acid

2. Florisil

3. Alumina

4. DEAE and TEAE Cellulose Chromatography

C. Thin-Layer Chromatography

1. Single-Dimensional TLC Systems

2. Two-Dimensional TLC Systems

3. Location and Identification of Complex Lipids on TLC Plates

D. Determination of Complex Lipids Separated by Chromatographic Procedures

E. Analysis of Complex Lipids as their Partial Hydrolysis Products

F. Sphingolipids

1. Cerebrosides and Ceramide Polyhexosides

2. Gangliosides

3. Long-Chain Bases

G. Alkyl- and Alkenyl-Ether Derivatives of Complex Lipids

H. Phosphonolipids

I. Determination of Water-Soluble Hydrolysis Products of Complex Lipids

1. Phosphorus

2. Glycerol

3. Carbohydrates, N-Acetylneuraminic Acid and Related Compounds

4. Nitrogenous Bases of Phospholipids

8. The Analysis of Molecular Species of Lipids

A. Introduction

B. General Methods of Analysis

1. Liquid-Solid and Liquid-Liquid Chromatography

2. Gas-Liquid Chromatography of Intact Lipids

C. Molecular Species of Simple Lipids

1. Cholesteryl Esters

2. Monoglycerides

3. Diglycerides

4. Triglycerides

5 . Wax Esters

D. Molecular Species of Complex Lipia3

1. General Approaches to the Problem

2. Phosphatidyl Choline

3. Phosphatidyl Ethanolamine

4. Other Glycerophosphatides

5. Glycosyl Diglycerides

6. Sphingomyelin and Glycosphingolipids

9. Enzymatic Hydrolysis of Lipids

A. Introduction

B. Positional Distribution of Fatty Acids in Triglycerides

1. Pancreatic Lipase Hydrolysis

2. Stereospecific Analysis of Triglycerides

C. Enzymatic Hydrolysis of Complex Lipid

1. Phospholipase A2

2. Pancreatic Lipase and Related Enzymes

3. Phospholipase C

4. Phospholipase D

D. The Use of Enzymatic Hydrolysis in the Determination of Molecular Species of Lipids

10. The Analysis and Radioassay of Isotopically-Labelled Lipids

A. Introduction

B. Thin-Layer Chromatography

C. Gas-Liquid Chromatography

1. Radioassay After Preparative GLC

2. Continuous Monitoring of the Column Effluent

D. Location of 14C in Aliphatic Chains

11. A Summary

A. Introduction

B. Extraction of Lipids from Tissues

C. Fatty Acid Composition

D. the Analysis of Simple Lipids

E. the Analysis of Complex Lipids

F. Structural Analysis of Lipids

Appendix A: Commercial Sources of Lipid Standards

Appendix B: Sources of Information

References

Index

Life Sciences

Physical Sciences & Engineering

Social Sciences & Humanities

Health

Lipid Analysis

Isolation, Separation, Identification and Structural Analysis of Lipids

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