主题：【资料】COMBINED COMPENDIUM OF FOOD ADDITIVE SPECIFICATIONS

COMBINED COMPENDIUM OF FOOD ADDITIVE SPECIFICATIONS
Joint FAO/WHO Expert Committee on Food Additives
All specifications monographs from the
1st to the 65th meeting (1956–2005)
Analytical methods, test procedures and laboratory solutions used by and referenced in the food additive specifications
Volume 4

CONTENTS
GENERAL INFORMATION....................................................................................................1
Revision of Food and Nutrition Paper No. 5 ................................................................ 1
Criteria for Replacement of Older Methods ................................................................. 2
Food Chemicals Codex................................................................................................. 2
Hazardous Reagents and Solvents ................................................................................ 2
Method Validation ........................................................................................................ 2
ANALYTICAL TECHNIQUES............................................................................................... 3
Chromatography ........................................................................................................... 3
Introduction....................................................................................................... 3
Chromatography Principles .............................................................................. 4
Chromatography Techniques ............................................................................ 4
Column Chromatography...................................................................... 4
Paper Chromatography ......................................................................... 5
Thin-Layer Chromatography ................................................................ 7
Gas Chromatography ........................................................................................ 9
Headspace-Gas Chromatography.....................................................................15
Analysis of Samples – General Method Instructions.......................................15
Gas Chromatography – Mass Spectrometry (GC-MS)....................................17
High Performance Liquid Chromatography ....................................................20
Spectrophotometry and Spectroscopy..........................................................................25
Definitions........................................................................................................25
Terminology.....................................................................................................25
Theory and Formulas for Calculation ..............................................................27
Apparatus .........................................................................................................29
UV/Visible Spectrophotometers ..........................................................29
Atomic Absorption Spectrophotometers .............................................29
Inductively Coupled Plasma – Atomic Emission Spectrophotometers30
SpectrofluorimetersInfrared Spectrometers...............................31
Nuclear Magnetic Resonance Spectrometers (NMR)..........................31
Procedures........................................................32
Applications ................33

GENERAL METHODS...........................................................................................................35
Appearance and Physical Properties............................................................................35
Boiling Point and Distillation Range ...............................................................35
Determination of pH (Potentiometric Method)................................................36
Melting Range/Melting Point ..........................................................................38
Refractive Index...............................................................................................38
Solidification Point ..........................................................................................39
Solubility..........................................................................................................41
Specific Gravity ...............................................................................................41
Specific Rotation..............................................................................................43
Identification Tests.......................................................................................................45
Inorganic Ions ..................................................................................................45
Organic Ions and Compounds..........................................................................48
Inorganic Components ................................................................................................51
Acid-Insoluble Matter .....................................................................................51
Arsenic Limit Test ..........................................................................................51
Ash ..................................................................................................................53
Total Ash..............................................................................................53
Acid-insoluble Ash ..............................................................................54
Sulfated Ash ........................................................................................54
Chlorides Limit Test .......................................................................................54
Chromium Limit Test ......................................................................................55
Cyclic Phosphate Determination......................................................................55
Fluoride Limit Test .........................................................................................57
Method I – Thorium Nitrate Colorimetric Method .............................57
Method II – Ion-Selective Electrode Method, Method A ...................58
Method III – Ion-Selective Electrode Method, Method B ..................59
Method IV – Ion-Selective Electrode Method, Method C...................59
Iron Limit Test ................................................................................................61
Loss on Drying.................................................................................................61
Loss on Ignition ..............................................................................................61
Metallic Impurities ..........................................................................................62
Measurement of Antimony, Barium, Cadmium, Chromium, Copper,
Lead and Zinc by Atomic Absorption Flame Technique...............65
Measurement of Antimony, Barium, Cadmium, Chromium, Copper,
Lead and Zinc by Inductively Coupled Plasma Technique ...........66
Measurement of Lead and Cadmium by Atomic Absorption
Electro-Thermal (Furnace) Atomization Technique......................67
Measurement of Arsenic and Antimony by Atomic Absorption
Hydride Technique...............................................................................68
Determination of Mercury by Cold Vapour Atomic Absorption
Technique.......................................................................................68

Nickel in Polyols .............................................................................................71
Nickel Limit Test ............................................................................................72
Nitrogen Determination (Kjeldahl Method) ....................................................72
Method I ..............................................................................................72
Method II (Semi-Micro) ......................................................................73
Non-Volatile Residue.......................................................................................73
Phosphate Determination as P2O5 ...................................................................73
Method I ..............................................................................................73
Method II .............................................................................................74
Selenium Limit Test.........................................................................................74
Sulfates Limit Test ..........................................................................................75
Water-Insoluble Matter....................................................................................75
Water Determination (Karl Fischer Titrimetric Method) ................................75
Organic Components ...................................................................................................79
Chlorinated Organic Compounds Limit Test...................................................79
Cyclohexylamine in Cyclamates......................................................................79
Dicyclohexylamine in Cyclamates ..................................................................79
1,4-Dioxane and Ethylene Oxide.....................................................................80
Fumaric and Maleic Acid.................................................................................82
Gum Constituents Identification......................................................................83
Norbixin ...........................................................................................................83
Oxalate Limit Test ...........................................................................................84
Polyols: Thin Layer Chromatography .............................................................84
Pyrrolidone Carboxylic Acid ...........................................................................85
Readily Carbonizable Substances ....................................................................85
Reducing Substances (as Glucose) ..................................................................87
Method I (Volumetric).........................................................................87
Method II (Gravimetric) ......................................................................87
Related Foreign Substances in Flavour Enhancers..........................................87
Residual Solvents.............................................................................................87
Method I (Determination carried out in water)....................................88
Method II (Determination carried out in methanol) ............................88
Toluenesulfonamides in Saccharines...............................................................90
Triphenylphosphine Oxide...............................................................................91
Microbiological Analyses ............................................................................................93
Total (Aerobic) Plate Count.............................................................................93
Spiral Plate Count Method (Alternative Method)............................................95
Coliforms and E-Coli.......................................................................................98
MPN Method (Alternative Method) ..............................................................100
Salmonella......................................................................................................101
Staphylococcus aureus...................................................................................112
Direct Plate Count Method ................................................................112
MPN Method .....................................................................................115
Enumeration of Yeasts and Moulds...............................................................115

SPECIFIC METHODS..........................................................................................................119
Enzyme Preparations .................................................................................................119
_-Amylase Activity (Bacterial)......................................................................119
_-Amylase Activity (Fungal) .........................................................................120
Antibacterial Activity.....................................................................................122
Catalase Activity............................................................................................123
Cellulase Activity...........................................................................................124
Ethylenimine Limit Test ................................................................................126
_-Galactocidase (Lactase) Activity................................................................128
Glucoamylase Activity (Amyloglucosidase Activity) ...................................130
_-Glucanase Activity .....................................................................................132
Glucose Isomerase Activity ...........................................................................133
Glucose Oxidase Activity ..............................................................................136
Gluteraldehyde Limit Test .............................................................................137
Gluteraldehyde Determination in High Fructose Corn Syrup (High
Fructose Glucose Syrup)..........................................................................138
Hemicellulase Activity...................................................................................139
Invertase Activity...........................................................................................142
Milk Clotting Activity....................................................................................143
Protease Activity (Viscometer Method) ........................................................144
Proteolytic Activity, Bacterial (PC)...............................................................147
Proteolytic Activity, Fungal (HUT)...............................................................148
Proteolytic Activity, Fungal (SAP)................................................................150
Proteolytic Activity, Plant..............................................................................152
Pullulanase Activity .......................................................................................154
Xylanase Activity (Method 1) .......................................................................156
Xylanase Activity (Method 2) .......................................................................158
Fats, Oils and Hydrocarbons......................................................................................161
Acid Value .....................................................................................................161
Aromatic Hydrocarbons Determination.........................................................161
Average Molecular Weight............................................................................163
Carbon Number at 5% Distillation Point .......................................................166
Congealing Range..........................................................................................167
Free Fatty Acids.............................................................................................167
Hydroxyl Value..............................................................................................168
Identification Tests for Functional Groups....................................................169
Test A – Methyl Esters of Fatty Acids...............................................169
Test B – Acetic Acid..........................................................................171
Test C – Succinic Acid ......................................................................171
Test D – Fumaric Acid.......................................................................171
Test E – Tartaric Acid........................................................................171
Test F – Citric Acid............................................................................171
Test G – Lactic Acid..........................................................................171
Test H – Glycerol...............................................................................172
Iodine Value (Modified Wijs Method) ..........................................................172
1-Monoglyceride and Free Glycerol Contents...............................................173
Polycyclic Aromatic Hydrocarbons...............................................................174
Polyglycerol Determination in Polyglycerol Esters.......................................180
Polyols............................................................................................................181
Propylene Glycol Dimer and Trimer Determination .....................................182
Saponification ................................................................................................184
Saponification Value......................................................................................184
Sorbitan Ester Content ...................................................................................185
Sulfur..............................................................................................................186
Viscosity, 100°...............................................................................................188
Flavouring Agents......................................................................................................191
Acid Value .....................................................................................................191
Assay..............................................................................................................191

Boiling Point ..................................................................................................191
Ester Determination .......................................................................................191
Melting Point (Melting Range)......................................................................191
Refractive Index.............................................................................................191
Solubility in Ethanol ......................................................................................191
Solubility in General ......................................................................................192
Specific Gravity .............................................................................................192
Food Colours..............................................................................................................193
Chloride as Sodium Chloride.........................................................................193
Chloroform-insoluble Matter .........................................................................193
Colouring Matters ..........................................................................................194
Identification by Chromatography.....................................................194
Identification by Spectrophotometry .................................................198
Colouring Matters Content by Spectrophotometry............................199
Colouring Matters Content of Water-soluble Colouring Matters ......200
Colouring Matters Content of Organic Solvent-soluble Colouring
Matters ...............................................................................................200
Colouring Matters Content of Lakes..................................................201
Colouring Matters Content by Titration with Titanous Chloride ......201
Subsidiary Colouring Matter Content............................................................205
Ether-extractable Matter ................................................................................208
Hydrochloric acid-insoluble Matter in Lakes ................................................209
Leuco Base in Sulfonated Triarylmethane Colours .......................................210
Organic Compounds other than Colouring Matters.......................................211
Sulfate as Sodium Sulfate ..............................................................................214
Unsulfonated Primary Aromatic Amines.......................................................215
Water Content (Loss on Drying)....................................................................216
Water-insoluble Matter ..................................................................................217
Water-soluble Chlorides and Sulfates in Aluminum Lakes...........................217
ASSAY METHODS..............................................................................................................219
Alginates Assay (Carbon Dioxide Determination by Decarboxylation) ...................219
Cellulose Derivatives Assay (Ethoxyl and Methoxyl Group Determination) ...........220
Polysorbates Assay (Oxyethylene Group Determination).........................................221
MEDIA, REAGENTS AND SOLUTIONS...........................................................................225
Media ........................................................................................................................225
Reagents.....................................................................................................................239
Buffer Solutions .........................................................................................................243
Buffer Test Solutions .....................................................................................243
Standard Buffer Solutions..............................................................................244
Standard Solutions .....................................................................................................247
Test Solutions (TS) ....................................................................................................251
Volumetric Solutions .................................................................................................285
Normal Solutions (N).....................................................................................285
Molar Solutions (M) ......................................................................................285
Preparation and Methods of Standardization.................................................285