Fatty Acids and Glycerides

1 Separation and Determination of Structure of Fatty Acids.- 1.1. Introduction.- 1.2. Preparation and Isolation of Fatty Acids.- 1.2.1. Generation of Free Acids.- 1.2.2. Preparation of Derivatives.- 1.2.3. Transesterification of Bound Acids.- 1.3. Preliminary Fractionation of Fatty Acids.- 1.3.1. Conventional TLC.- 1.3.2. Argentation TLC.- 1.3.3. Other Methods.- 1.4. Identification of Fatty Acids by Gas—Liquid Chromatography.- 1.4.1. Short-Chain Acids.- 1.4.2. Saturated Normal- and Branched-Chain and Cyclic Acids.- 1.4.3. Ethylenic Acids.- 1.4.4. Acetylenic Acids.- 1.4.5. Oxygenated Acids.- 1.5. Identification of Fatty Acids by Gas Chromatography/Mass Spectrometry.- 1.5.1. Saturated Normal- and Branched-Chain and Cyclic Acids.- 1.5.2. Ethylenic and Acetylenic Acids.- 1.5.3. Oxygenated Acids.- 1.5.4. Stable Isotope-Labeled Fatty Acids.- 1.6. Ancillary Methods of Fatty Acid Identification.- 1.6.1. Chemical Methods.- 1.6.2. Spectrometric Methods.- 1.6.3. Enzymatic Methods.- 1.7. Summary and Conclusions.- 1.8. References.- 2 Synthesis and Analysis of Stable Isotope- and Radioisotope-Labeled Fatty Acids.- 2.1. Introduction.- 2.2. Synthesis of Stable Isotope-Labeled Fatty Acids.- 2.2.1. Experimental Techniques.- 2.2.2. Saturated Fatty Acids.- 2.2.3. Monounsaturated Fatty Acids.- 2.2.4. Polyunsaturated Fatty Acids.- 2.2.5. Perdeutero Fatty Acids.- 2.2.6. 13C-Labeled Fatty Acids.- 2.3. Synthesis of Radioisotope-Labeled Fatty Acids.- 2.3.1. 14C-Labeled Fatty Acids.- 2.3.2. 3H-Labeled Fatty Acids.- 2.3.3. Biosynthesis of 14C- and 3H-Labeled Lipids.- 2.4. Analysis of Radioisotope-Labeled Fats.- 2.4.1. Liquid Scintillation Counting.- 2.4.2. Radio-Gas Chromatography.- 2.4.3. Thin-Layer Chromatography.- 2.4.4. Determination of Radiochemical Purity.- 2.4.5. Chemical Degradation Methods.- 2.4.6. Storage of Radioisotope-Labeled Fatty Acids.- 2.5. Analysis of Stable Isotope-Labeled Fatty Acids.- 2.5.1. Mass Spectrometry.- 2.5.2. Infrared Spectrometry.- 2.5.3. Nuclear Magnetic Resonance.- 2.6. Summary.- 2.7. References.- 3 Separation and Determination of the Structure of Acylglycerols and Their Ether Analogues.- 3.1. Introduction.- 3.2. Nomenclature.- 3.3. Selected Methods for Modifying or Degrading Glycerolipids.- 3.3.1. Methanolysis.- 3.3.2. Saponification.- 3.3.3. Phospholipase C Hydrolysis.- 3.3.4. Acetolysis.- 3.3.5. Thermal Cleavage.- 3.3.6. Reductive Hydrogenolysis.- 3.3.7. Grignard Hydrolysis.- 3.3.8. Oxidative Cleavage of Double Bonds.- 3.3.9. Hydrogenation.- 3.4. Derivatization Procedures.- 3.4.1. Acetates.- 3.4.2. Isopropylidenes.- 3.4.3. Trimethylsilyl Ethers.- 3.4.4. Tertiarybutyldimethylsilyl Ethers.- 3.4.5. Other Derivatives.- 3.5. Determination of the Structural Components of Glycerolipids.- 3.5.1. Glycerol.- 3.5.2. Fatty Alcohols.- 3.5.3. Monoalkyl- and Monoalk-1-enylglycerols.- 3.6. Extraction and Purification of Lipids.- 3.7. Thin-Layer Chromatography.- 3.7.1. Plate Preparation.- 3.7.2. Detection of Neutral Lipids.- 3.7.3. Recovery of Lipids from Silica Gel.- 3.7.4. Separations on Silica Gel.- 3.7.5. Separations on Silica Gel Impregnated with Borate or Arsenite.- 3.7.6. Separations on Silica Gel Impregnated with Silver Nitrate.- 3.7.7. Reversed-Phase TLC.- 3.7.8. Quantitation.- 3.8. Liquid Chromatography.- 3.9. Gas Chromatography.- 3.9.1. Column Packings.- 3.9.2. General Separation Characteristics.- 3.9.3. Equivalent Chain Length.- 3.9.4. Separation of Neutral Lipids.- 3.9.5. Quantitative Gas Chromatography.- 3.10. Mass Spectrometry.- 3.10.1. Monoacylglycerols.- 3.10.2. Monoalkylglycerols.- 3.10.3. Monoalk-1-enylglycerols.- 3.10.4. Diacylglycerols.- 3.10.5. Alkylacylglycerols.- 3.10.6. Triacylglycerols.- 3.10.7. Other Modes of Ionization.- 3.11. Other Techniques.- 3.11.1. Infrared Spectroscopy.- 3.11.2. Nuclear Magnetic Resonance Spectroscopy.- 3.11.3. Optical Rotation and Stereospecific Analysis.- 3.11.4. Calculation Methods.- 3.12. Combined Techniques.- 3.13. Conclusions.- 3.14. References.- 4 Stereospecific Analysis of Triacylglycerols.- 4.1. Introduction.- 4.2. Physical and Chemical Methods for the Determination of Stereochemical Distribution of Fatty Acids in Triacylglycerols.- 4.3. Enzymatic Methods for Determination of Stereochemical Distribution of Fatty Acids in Triacylglycerols.- 4.3.1. Generation of Diacylglycerols.- 4.3.2. Stereospecific Selection of Diacylglycerols or Their Derivatives.- 4.3.3. Calculations and Checks on Accuracy.- 4.3.4. Studies with Radioactive Acylglycerols.- 4.4. Positional Distribution of Fatty Acids in Natural Triacylglycerols.- 4.4.1. Animal Fats.- 4.4.2. Human Tissues.- 4.4.3. Interorgan and Interspecies Comparisons.- 4.4.4. Plant Oils.- 4.4.5. Yeast and Microorganisms.- 4.5. Concluding Remarks.- 4.6. References.- 5 Stereospecific Synthesis of Enantiomeric Acylglycerols.- 5.1. Introduction.- 5.2. General Procedures for Specification of Absolute Configuration of Enantiomeric sn-Glycerol Derivatives.- 5.3. Sources and Practical Methods for Preparation of Enantiomeric sn-Glycerol Derivatives.- 5.3.1. d-Mannitol and l-Mannitol.- 5.3.2. Preparation of 1,2:5,6-Diisopropylidene-d- and -l-mannitols.- 5.3.3. Preparation of 1,2- and 2,3-Isopropylidene-sn-glycerols.- 5.3.4. Preparation of 1- and 3-Benzyl-sn-glycerols.- 5.3.5. Preparation of Enantiomeric Alkyl- and Alkenyl-sn-glycerols.- 5.4. Methods of Configurational Conversion and Inversion of sn-Glycerol Derivatives.- 5.4.1. Conversion of 1,2-Isopropylidene-sn-glycerol into 2,3-Isopropylidene-sn-glycerol.- 5.4.2. Inversion of 3-Benzyl-sn-glycerol into 1-Benzyl-sn-glycerol.- 5.4.3. Preparation of 1-Benzyl-sn-glycerol from d-Mannitol via l,3:2,5:4,6-O-Trimethylene-d-mannitol.- 5.5. Selection of Protective Groups for Stereospecific Synthesis of Acyl-, Alkyl-, and Alkylacyl-sn-glycerols.- 5.5.1. Isopropylidene Group.- 5.5.2. Benzylidene Group.- 5.5.3. Triphenylmethyl (Trityl) Group.- 5.5.4. Benzyl Group.- 5.5.5. 2,2,2-Trichloroethoxycarbonyl Group.- 5.5.6. Other Protective Groups.- 5.6. Stereospecific Acylation of Enantiomeric sn-Glycerol Derivatives.- 5.6.1. Selection of Acylating Reagents.- 5.6.2. Synthesis of Enantiomeric 1- and 3-Monoacyl-sn-glycerols.- 5.6.3. Synthesis of 2-Monoacyl-sn-glycerols.- 5.6.4. Synthesis of Enantiomeric 1,2- and 2,3-Diacyl-sn-glycerols.- 5.6.5. Synthesis of Enantiomeric 1,3-Diacyl-sn-glycerols.- 5.6.6. Synthesis of Enantiomeric Mixed-Acid Triacyl-sn-glycerols.- 5.7. Synthesis of Enantiomeric Isotope-Labeled Acyl- and Alkylacyl-sn-glycerols.- 5.7.1. Synthesis of Enantiomeric Isotope-Labeled Acyl-sn-glycerols.- 5.7.2. Synthesis of Enantiomeric Isotope-Labeled Alkylacyl-sn-glycerols.- 5.8. Methods of Isolation and Purification of Synthetic Acyl-sn-glycerols.- 5.8.1. Selection of Solvents.- 5.8.2. Distillation and Crystallization.- 5.8.3. Chromatography.- 5.9. Determination of Structure and Absolute Configuration of Synthetic and Natural Acyl-sn-glycerols.- 5.9.1. Chemical and Enzymatic Methods.- 5.9.2. Physical Methods.- 5.10. Summary and Conclusions.- 5.11. References.- 6 Metabolic Studies with Natural and Synthetic Fatty Acids and Enantiomeric Acylglycerols.- 6.1. Introduction.- 6.2. Activation of Fatty Acids.- 6.3. Biosynthesis of Phosphatidic Acid.- 6.3.1. Acylation of sn-Glycerol-3-phosphate.- 6.3.2. Acylation of Dihydroxyacetone Phosphate.- 6.3.3. Synthesis of Phosphatidic Acid from Monoacylglycerols and Diacylglycerols.- 6.3.4. Relative Contributions of the Pathways of Phosphatidic Acid Synthesis.- 6.4. Biosynthesis of Diacylglycerols.- 6.4.1. Synthesis of Diacylglycerols from Monoacylglycerols.- 6.4.2. Synthesis of Diacylglycerols from Phosphatidic Acid.- 6.4.3. Synthesis of Other Diacylglycerols.- 6.5. Biosynthesis of Triacylglycerols.- 6.5.1. Synthesis of Triacylglycerols from Diacylglycerols.- 6.5.2. Triacylglycerol Synthetase.- 6.5.3. Relative Contributions of the Major Pathways of Triacylglycerol Synthesis.- 6.6. Factors Influencing the Biosynthesis of Acylglycerols.- 6.6.1. Nutritional Factors.- 6.6.2. Hormonal Factors.- 6.6.3. Pharmacological Factors.- 6.7. Methods Employed in Studying the Biosynthesis of Acylglycerols.- 6.8. Kinetic Problems Encountered in Enzyme Assays Involving Lipid Substrates.- 6.9. Enzymatic Lipolysis of Acylglycerols.- 6.9.1. Pharyngeal Lipases.- 6.9.2. Pancreatic Lipase.- 6.9.3. Lipoprotein Lipase.- 6.9.4. Hormone-Sensitive Lipase.- 6.9.5. Monoacylglycerol Lipase.- 6.10. Summary and Conclusions.- 6.11. References.- 7 Composition of Selected Dietary Fats, Oils, Margarines, and Butter.- 7.1. Introduction.- 7.2. Methodology.- 7.2.1. Extraction Techniques.- 7.2.2. Esterification Techniques for Fatty Acids.- 7.2.3. Gas Chromatographic Analysis of Fatty Acid Methyl Esters.- 7.2.4. Sterol Derivatization.- 7.2.5. Gas Chromatographic Analysis of Sterols.- 7.2.6. Lipoxidase:cis,cis-Polyunsaturated Fatty Acid Analysis.- 7.3. Basic Processing.- 7.3.1. General Discussion.- 7.3.2. Additives.- 7.3.3. Mayonnaise and Salad Dressing.- 7.4. Source Oils.- 7.4.1. Soybean.- 7.4.2. Cottonseed.- 7.4.3. Peanut.- 7.4.4. Corn.- 7.4.5. Safflower.- 7.4.6. Sunflower.- 7.4.7. Rapeseed.- 7.4.8. Other Cruciferae.- 7.4.9. Olive.- 7.4.10. Sesame.- 7.4.11. Cocoa.- 7.4.12. Palm and Palm Kernel.- 7.4.13. Coconut.- 7.5. Confectionery Products.- 7.6. Nuts.- 7.7. Cooking Oils.- 7.8. Shortenings.- 7.9. Margarines.- 7.10. Dairy Products and Creamers.- 7.11. Animal Fats.- 7.12. Marine Products.- 7.13. Summary and Trends.- 7.14. References.- 8 Fatty Acid Composition of Glycerolipids of Animal Tissues.- 8.1. Introduction.- 8.2. Methods of Analysis.- 8.2.1. Isolation and Preparation of Fatty Acids.- 8.2.2. Quantitative Determination of Fatty Acids.- 8.2.3. Quality Control.- 8.3. Fatty Acids of Different Animal Species and Man.- 8.3.1. Differences among Homologous Tissues.- 8.3.2. Differences among Homologous Cell Types.- 8.3.3. Differences among Homologous Subcellular Fractions.- 8.3.4. Differences among Homologous Lipoproteins.- 8.4. Fatty Acids of Individual Animal Species and Man.- 8.4.1. Tissue Differences.- 8.4.2. Differences among Cell Types.- 8.4.3. Variations among Subcellular Fractions.- 8.4.4. Discrepancies among Plasma Lipoprotein Classes.- 8.4.5. Characteristics of Skin Surface and Fecal Lipids.- 8.5. Summary and Conclusions.- 8.6. References.