RECOMMENDED METHODS OF ANALYSIS AND SAMPLING CODEX STAN 234-19991 PART A METHODS OF ANALYSIS BY COMMODITY CATEGORIES AND NAMES PART B METHODS OF SAMPLING BY COMMODITY CATEGORIES AND NAMES Table of Contents All Foods Cereals, Pulses and Legumes and Derived Products Cocoa Products and Chocolate Fats and Oils and Related Products Fish and Fishery Products Foods for Special Dietary Uses Fruit Juices Milk and Milk Products Natural Mineral Waters Processed Fruits and Vegetables Processed Meat and Poultry Products and Soups and Broths Quick Frozen Fruits and Vegetables Sugars and Honey Miscellaneous Products 1 The most updated version of the method should be used, in application of ISO/IEC 17025: 1999. The present list of methods reflects the amendments adopted by the 34th Session of the Codex Alimentarius Commission in 2011. 1 PART A METHODS OF ANALYSIS BY COMMODITY CATEGORIES AND NAMES Commodity All Foods All foods All foods All foods All foods All Foods (see also meat products) Individual Foods2 Individual Foods3 Provision Method Principle High performance liquid chromatography High performance liquid chromatography Spectrophotometry High performance liquid chromatography Part 1- General considerations Part 1: Optimized Monier-Williams method Part 2: Enzymatic method Type II II III III N/A III III Acesulfame K, Aspartame Cyclamate Cyclamate Saccharin Nitrates and/or Nitrites Sulphites Sulphites EN 12856 : 1999-04 EN 12857 : 1999-04 NMKL 123 (1998) EN 12856 : 1999-04 EN 12014-1:1997-04 EN 1988-1 : 1998-02 AOAC 990.28 EN 1988-2:1998 -02 NMKL 135 (1990) ISO 665:1977 (confirmed 1995) AOAC 923.03 ISO 2171:1993 ICC Method No 104/1 (1990) AOAC 945.38F; 920.39C Cereals, Pulses and Legumes and Derived Products Certain pulses Degermed maize (corn) meal and maize (corn) grits Degermed maize (corn) meal and maize (corn) grits Moisture Ash Gravimetry Gravimetry I I Fat, crude Gravimetry (ether extraction) I 2 3 Hominy, fruit juice, sea food Wine, dried apples, lemon juice, potato flakes, sultanas, beer 2 Degermed maize (corn) meal and maize (corn) grits Degermed maize (corn) meal and maize (corn) grits Degermed maize (corn) meal and maize (corn) grits Durum wheat semolina and durum wheat flour Durum wheat semolina and durum wheat flour Durum wheat semolina and durum wheat flour Instant Noodles Instant Noodles Instant Noodles Maize (corn) Peanuts (raw) Peanuts (raw) Peanuts (intended for further processing) Peanuts (Cereals, shell-fruits and derived products ( including peanuts)) Peanuts (intended for further processing) Pearl millet flour Moisture Particle size (granularity) Protein Ash (semolina) Moisture Protein (N x 5.7) Extraction of oil from instant noodles Acid Value Moisture Moisture Aflatoxins, total Aflatoxins, total Aflatoxins, total Sum of aflatoxins B1, B2, G1 and G2 Aflatoxins, total Ash ISO 712:1998 ICC Method No 110/1 (1986) AOAC 965.22 ICC Method No 105/1 (1986) AOAC 923.03 ISO 2171:1993 ISO 712:1998 ICC Method 110/1 (1986) ICC Method No 105/1 described in the standard described in the standard described in the standard ISO 6540:1980 (confirmed 1994) AOAC 991.31 AOAC 993.17 AOAC 975.36 EN 12955 : 1999-07 ISO 16050:2003 AOAC 979.18 AOAC 923.03 Gravimetry Sieving Titrimetry, Kjeldahl digestion Gravimetry Gravimetry Titrimetry, Kjeldahl digestion Gravimetry Titrimetry Gravimetry Gravimetry Immunoaffinity column (Aflatest) Thin layer chromatography Romer minicolmn HPLC with post column derivatization and immunoaffinity column clean up Holaday-Velasco minicolumn Gravimetry I I I I I I I I I I II III III III III I 3 Pearl millet flour Colour Modern Cereal Chemistry, 6th Ed., D.W. Kent-Jones and A.J. Amos (Ed.), pp. 605612, Food Trade Press Ltd, London, 1969. AOAC 945.38F; 920.39C ISO 5498:1981 (B.5 Separation) ISO 712:1998 ICC Method No 110/1 (1986) AOAC 920.87 AOAC 923.03 ISO 2171:1993 ICC Method No 104/1 (1990) Modern Cereal Chemistry, 6th Ed., D.W. Kent-Jones and A.J. Amos (Ed.), pp. 605612, Food Trade Press Ltd, London, 1969. AOAC 945.38F; 920.39C ICC Method No 113 (1972) ISO 6541:1981 (confirmed 1996) ISO 712:1998 ICC Method No 110/1 (1986) AOAC 965.22 ICC Method No 105/1 (1986) ISO 9648:1988 (confirmed 1994) AOAC 923.03 ISO 2171:1993 ICC Method No 104/1 (1990) AOAC 945.38F, 920.39C ISO 6540:1980 (confirmed 1994) ICC Method No 105/1 (1986) Colorimetry using specific colour grader Gravimetry (ether extraction) Gravimetry Gravimetry Titrimetry, Kjeldahl digestion Gravimetry IV Pearl millet flour Pearl millet flour Pearl millet flour Pearl millet flour Sorghum flour Fat, crude Fibre, crude Moisture Protein Ash I I I I I Sorghum flour Colour Colorimetry using specific colour grader Gravimetry (ether extraction) Gravimetry Gravimetry Sieving Titrimetry, Kjeldahl digestion Spectrophotometry Gravimetry IV Sorghum flour Sorghum flour Sorghum flour Sorghum flour Sorghum flour Sorghum flour Sorghum grains Fat, crude Fibre, crude Moisture Particle size (granularity) Protein Tannins Ash I I I I I I I Sorghum grains Sorghum grains Sorghum grains Fat, crude Moisture Protein Gravimetry (ether extraction) Gravimetry Titrimetry, Kjeldahl digestion I I I 4 Sorghum grains Soy protein products Soy protein products Soy protein products Soy protein products Soy protein products Vegetable protein products Vegetable protein products Vegetable protein products Vegetable protein products Vegetable protein products Wheat flour Tannins Ash Fat Fibre, crude Moisture Protein Ash Fat Fibre, crude Moisture Protein Ash ISO 9648:1988 (confirmed 1994) AOAC 923.03 ISO 2171:1993 (Method B) CAC/RM 55-1976 - Method 1 ISO 5498:1981 AOAC 925.09 AOAC 955.04D (using factor 6.25) AOAC 923.03 ISO 2171:1993 (Method B) CAC/RM 55-1976 - Method 1 AACC (1982) 32-17 AOAC 925.09 AOAC 955.04D (using factor 6.25) AOAC 923.03 ISO 2171:1993 ICC Method No 104/1 (1990) AOAC 939.05 ISO 712:1998 ICC Method No 110/1 (1986) AOAC 965.22 ICC Method No 105/1 (1986) Vital wheat gluten and devitalized wheat gluten AOAC 979.09 (wheat protein in grain Nx5.7) Solubilized wheat protein AOAC 920.87 (wheat protein in flour Nx5.7) Spectrophotometry Gravimetry Gravimetry (extraction) Gravimetry Gravimetry (vacuum oven) Titrimetry , Kjeldahl digestion Gravimetry, Direct Gravimetry (extraction) Ceramic fiber filteration Gravimetry (vacuum oven) Titrimetry, Kjeldahl digestion Gravimetry I I I I I II I I I I II I Wheat flour Wheat flour Wheat flour Wheat flour Wheat protein products including wheat gluten Fat acidity Moisture Particle size (granularity) Protein Protein Titrimetry Gravimetry Sieving Titrimetry, Kjeldahl digestion Kjeldahl I I I I I Kjeldahl I 5 Wheat protein products including Wheat gluten Wheat protein products including Wheat gluten Whole and decorticated pearl millet grains Whole and decorticated pearl millet grains Whole and decorticated pearl millet grains Whole and decorticated pearl millet grains Whole and decorticated pearl millet grains Whole maize (corn) meal Fibre, crude Ash AOAC 962.09 AOAC 923.03 ISO 2171:1980, method B AOAC 923.03 AOAC 945.38F; 920.39C ISO 5498:1981 (B.5 Separation) ISO 712:1998 ICC Method No 110/1 (1986) AOAC 920.87 AOAC 923.03 ISO 2171:1993 ICC Method No 104/1 (1990) AOAC 945.38F; 920.39C ISO 712:1998 ICC Method No 110/1 (1986) AOAC 965.22 ICC Method No 105/1 (1986) Ceramic fiber filteration Gravimetry I I Ash Fat, crude Fibre, crude Moisture Protein Ash Gravimetry Gravimetry (ether extraction) Gravimetry Gravimetry Titrimetry, Kjeldahl digestion Gravimetry I I I I I I Whole maize (corn) meal Whole maize (corn) meal Whole maize (corn) meal Whole maize (corn) meal Cocoa Products and Chocolate Chocolate and chocolate products Chocolate and chocolate products Chocolate and chocolate products Fat, crude Moisture Particle size (granularity) Protein Gravimetry (ether extraction) Gravimetry Sieving Titrimetry, Kjeldahl digestion I I I I Cocoa butter Fat-free cocoa solids Fat-free milk solids AOAC 963.15 IOCCC 14-1972 AOAC 931.05 IOCCC 17-1973 or AOAC 939.02 Gravimetry (Soxhlet extraction) Oven evaporation and factor Titrimetry, Kjeldahl digestion; after extraction of milk proteins I I II 6 Chocolate and chocolate products Chocolate and chocolate products Chocolate and chocolate products Fat, total Milkfat Moisture AOAC 963.15 IOCCC 5-1962 AOAC 945.34; 925.41B; 920.80 IOCCC 26-1988 or AOAC 977.10 (Karl Fischer method); or AOAC 931.04 or IOCCC 1-1952 Gravimetry (Soxhlet extraction) Titrimetry/Distillation Gravimetry I I I Chocolate and chocolate products Cocoa (Cacao) Mass or Cocoa/ Chocolate Liquor, and Cocoa Cake Cocoa (Cacao) Mass or Cocoa/ Chocolate Liquor, and Cocoa Cake Cocoa butter Cocoa butter Non-cocoa butter vegetable fat AOCS Ce 10/02 and described in the Standard Cocoa shell Fat Free fatty acids Unsaponifiable matter AOAC 968.10 and 970.23 AOAC 963.15 or IOCCC 14 (1972) ISO660:1996 amended 2003; or AOCS Cd 3d-63 (03) ISO 3596:2000 or ISO 18609: 2000; or AOCS Ca 6b-53 (01) IOCCC 26-1988 or AOAC 977.10 (Karl Fischer method) Described in the Standard Spiral vessel count, Stone cell count Gravimetry (Soxhlet extraction) Titrimetry Titrimetry after extraction with diethyl ether Gravimetry I I I I I I Cocoa powders (cocoa) and dry cocoa- Moisture sugar mixtures Fats and Oils and Related Products Fats and Oils (all) Fats and Oils (all) Fats and Oils (all) Fats and oils Arsenic Arsenic Arsenic Butylhydroxyanisole, butylhydroxytoluene, tertbutylhydroquinone, & propyl gallate Insoluble impurities AOAC 952.13 (Codex general method) AOAC 942.17 (Codex general method) AOAC 985.16 (Codex general method) AOAC 983.15; or AOCS Ce-6-86 (09) Colorimetry (diethyldithiocarbamate) Colorimetry (molybdenum blue) Atomic absorption spectrophotometry Liquid chromatography II III III II Fats and Oils (all) Gravimetry ISO 663:2007 I 7 Fats and Oils (all) Lead AOAC 994.02 ISO 12193:2004 (Codex general method) or AOCS Ca 18c-91 (09) ISO 662:1998 ISO 660:1996; or AOCS Cd 3d-63 (09) AOAC 990.05 ISO 8294:1994 or AOCS Ca 18b-91 (09) (Codex general method) AOCS Cd 8b-90 (09) ISO 3960:2007 ISO 17189 | IDF 194: 2003 ISO 660:2009; or AOCS Cd 3d-63 (09) AOAC 990.05 ISO 8294:1994; or AOCS Ca 18b-91 (09) (Codex general method) ISO 5508 :1990 and ISO 12966-2:2011 or AOCS Ce 2-66 (09) and Ce 1e-91 (09) or Ce 1f-96 (09) ISO 3961: 2009; or AOAC 993.20; or AOCS Cd 1d-1992 (09) AOCS Cd 8b-90 (09) ISO 3960:2007 ISO/AOCS method for apparent density to be inserted ISO 6320:2000 and corr 2006; or AOCS Cc 7-25 (09) ISO 3657:2002; or AOCS Cd 3-25 (09) Atomic absorption spectrophotometry (direct graphite furnace) Gravimetry (open-drying) Titrimetry Atomic absorption Spectrophotometry (direct graphite furnace) Titrimetry using iso-octane Gravimetry Titrimetry Atomic absorption Spectrophotometry (direct graphite furnace) Gas chromatography of methyl esters Wijs-Titrimetry II Fats and Oils (all) Fats and Oils (all) Matter volatile at 105°C Soap content I I I II BS 684 Section 2.5; or AOCS Cc 17-95 (09) Gravimetry Fats and oils not covered by individual Acid Value standards Fats and oils not covered by individual Copper and Iron standards Fats and oils not covered by individual Peroxide value standards Fat spreads and blended spreads Named Animal Fats Named Animal Fats Fat content Acidity Copper and Iron I I I II Named Animal Fats Named Animal Fats GLC ranges of fatty acid composition Iodine value (IV) II I Named Animal Fats Named Animal Fats Named Animal Fats Named Animal Fats Peroxide value Relative density Refractive index Saponification value Titrimetry using iso-octane Pycnometry Refractometry Titrimetry I II II I 8 Named Animal Fats Named Animal Fats Named Vegetable Oils Named Vegetable Oils Named Vegetable Oils Unsaponifiable matter Titre Acidity Apparent density ISO 3596:2000 or ISO 18609: 2000; or AOCS Ca 6b-53 (09) ISO 935:1988; or AOCS Cc 12-59 (09) ISO 660:2009; or AOCS Cd 3d-63 (09) ISO 6883: 2007, with the appropriate conversion factor; or AOCS Cc 10c-95 (09) Titrimetry after extraction with diethyl ether Thermometry Titrimetry Pycnometry Colour reaction I I I I I Named Vegetable Oils Named Vegetable Oils Named Vegetable Oils Named Vegetable Oils Named Vegetable Oils Named Vegetable Oils Named Vegetable Oils AOCS Cb 2-40 (09) Baudouin test (modified Villavecchia or sesameseed oil test) BS 684 Section 2.20 Carotenoids, total Copper and iron Crismer value GLC ranges of fatty acid composition Halphen test Insoluble impurities Iodine value (IV) ISO 8294: 1994; or AOAC 990.05; or AOCS Ca 18b-91 (09) AOCS Cb 4-35 (97) and AOCS Ca 5a-40 Turbidity (09) ISO 5508:1990 and ISO 12966-2; or AOCS Gas chromatography of methyl esters Ce 2-66 (09) and Ce 1e-91 (09) or Ce 1f-96 (09) AOCS Cb 1-25 (09) Colorimetry ISO 663: 2000 Gravimetry Wijs - ISO 3961: 1996; or AOAC 993.20; Wijs-Titrimetry 4 or AOCS Cd 1d-1992 (97); or NMKL 39 (2003) AOAC 994.02 ; or ISO 12193: 2004; or AOCS Ca 18c-91 (03) ISO 662: 1998 AOCS Cd 8b-90 (09); or ISO 3960: 2007 Atomic Absorption Gravimetry Titrimetry Spectrophotometry AAS II II I II I I I Named Vegetable Oils Named Vegetable Oils Named Vegetable Oils 4 Lead Moisture & volatile matter at 105°C Peroxide value (PV) II I I It is possible to calculate the Iodine Value from fatty acid composition data obtained by gas chromatography e.g. using AOCS Cd 1b-87 (97) 9 Named Vegetable Oils Named Vegetable Oils Named Vegetable Oils Refractive index Reichert value and Polenske value Relative density ISO 6320: 2000 and corr 2006; or AOCS Cc Refractometry 7-25 (09) AOCS Cd 5-40 (09) Titrimetry IUPAC 2.101 with the appropriate conversion factor See comment above (Named Animal Fats) 5 ISO 3657: 2002; or AOCS Cd 3-25 (09) Pycnometry II I I Named Vegetable Oils Named Vegetable Oils Saponification value (SV) Slip point Titrimetry I I ISO 6321:2002 for all oils; Open ended capillary tube AOCS Cc 3b-92 (09) for all oils except palm oils; AOCS Cc 3-25 (09) for palm oils only BS 684 Section 2.5; or AOCS Cc 17-95 (09) Gravimetry ISO 12228: 1999; or AOCS Ch 6-91 (09) ISO 9936:2006 and corrigendum 2008 or AOCS Ce 8-89 (09) ISO 3596:2000; or ISO 18609: 2000; or AOCS Ca 6b-53 (09) COI/T.20/Doc. No. 19 or ISO 3656:2011 or AOCS Ch 5-91 (09). ISO 660:2009 or AOCS Cd 3d-63 (09) ISO 9936:1997 COI/T.20/Doc. no. 20 or AOCS Ce 5b-89 (09) IUPAC 2.4316 ISO 6800:1997 or AOCS Ch 3-91 (09) Gas chromatography HPLC Gravimetry Absorption in ultra violet Titrimetry HPLC Analysis of triglycerides of HPLC and calculation Gas chromatography Gas chromatography Named Vegetable Oils Named Vegetable Oils Named Vegetable Oils Named Vegetable Oils Olive Oils and Olive Pomace Oils Olive Oils and Olive Pomace Oils Olive Oils and Olive Pomace Oils Olive Oils and Olive Pomace Oils Olive Oils and Olive Pomace Oils Olive Oils and Olive Pomace Oils Soap content Sterol content Tocopherol content Unsaponifiable matter Absorbency in ultra-violet Acidity, free (acid value) Alpha-tocopherol Difference between the actual and theoretical ECN 42 triglyceride content Erythrodiol + uvaol content Fatty acids in the 2-position of the triglycerides I II II I II I II I II I 6 The method is no longer available. The method is no longer available. 5 10 Olive Oils and Olive Pomace Oils Olive Oils and Olive Pomace Oils Olive Oils and Olive Pomace Oils Olive Oils and Olive Pomace Oils Olive Oils and Olive Pomace Oils Olive Oils and Olive Pomace Oils Olive Oils and Olive Pomace Oils Olive Oils and Olive Pomace Oils Olive Oils and Olive Pomace Oils Olive Oils and Olive Pomace Oils Olive Oils and Olive Pomace Oils Olive Oils and Olive Pomace Oils Olive Oils and Olive Pomace Oils Olive Oils and Olive Pomace Oils Olive Oils and Olive Pomace Oils Olive Oils and Olive Pomace Oils Olive Oils and Olive Pomace Oils Margarine Margarine Margarine Margarine Halogenated solvents, traces Insoluble impurities in light petroleum Iodine value Iron and copper Lead Moisture and volatile matter Organoleptic characteristics Peroxide value Relative density Refractive index Saponification value Sterol composition and total sterols Stigmastadienes Stigmastadienes Trans fatty acids content Unsaponifiable matter Wax content Fat Milkfat Sodium chloride Vitamin A COI/T.20/Doc. no. 8 ISO 663:2000 ISO 3961:2009 or AOAC 993.20 or AOCS Cd 1d-92 (09) or NMKL 39 (2003) ISO 8294:1994 or AOAC 990.05 AOAC 994.02 or ISO 12193:2004 or AOCS Ca 18c-91(09) ISO 662:1998 COI/T.20/Doc. no. 15 ISO 3960:2007 or AOCS Cd 8b-90 (09) IUPAC 2.101, with the appropriate conversion factor. See comment above6 ISO 6320:2000 and corr 2006 or AOCS Cc 7-25 (09) ISO 3657:2002 or AOCS Cd 3-25 (09) COI/T.20/Doc. no. 10 or ISO 12228:1999 or AOCS Ch 6-91 (09). COl/T.20/Doc. no. 11 or ISO 157881:1999 or AOCS Cd 26-96 (09). ISO 15788-2: 2003 COI/T.20/Doc no. 17 or ISO 15304:2002 or AOCS Ce 1f-96 (09) ISO 3596:2000 or ISO 18609:2000 or AOCS Ca 6b-53 (09) COI/T.20/Doc. no. 18 or AOCS Ch 8-02 (09) IUPAC 2.801 CAC/RM 15-1969 AOAC 971.27 (Codex general method) AOAC 960.45 Gas chromatography Gravimetry Wijs-Titrimetry AAS AAS Gravimetry Panel test Titrimetry Pycnometry Refractometry Titrimetry Gas chromatography Gas chromatography HPLC Gas chromatography of methyl esters Gravimetry Gas chromatography Gravimetry Titrimetry Potentiometry Spectrophotometry II I I II II I I I I II I II II III II I II I I II II 11 Margarine Margarine Margarine Minarine Minarine Minarine Minarine Minarine Minarine Minarine Fish and Fishery Products Fish and fishery products Fish and fishery products Fish and fishery products: canned products Fish and fishery products: canned products Boiled Dried Salted Anchovies Canned shrimps or prawns Frozen fish and fishery products Vitamin D Vitamin E Water Fat Milkfat Sodium chloride Vitamin A Vitamin D Vitamin E Water AOAC 936.14 IUPAC 2.411 CAC/RM 17-1969 (described in the Standard) IUPAC 2.801 CAC/RM 15-1969 (described in the Standard) AOAC 971.27 (Codex general method) AOAC 960.45 AOAC 936.14 IUPAC 2.411 CAC/RM 17-1969 Bioassay TLC followed by spectrophotometry or GLC Gravimetry Gravimetry Titrimetry Potentiometry Spectrophotometry Bioassay TLC followed by spectrophotometry or GLC Gravimetry II II I I I II II II II I Histamine Mercury Drained weight Net weight Sodium Chloride (chloride expressed as sodium chloride) Size, determination of Thawing and cooking procedures AOAC 977.13 AOAC 977.15 Described in the Standard Described in the Standard AOAC 937.09 Described in the Standard Described in the Standards Fluorimetry Flameless atomic absorption spectrophotometry Weighing Weighing Titrimetry Number per 100 g Thawing and heating II III I I II I I 12 Quick frozen blocks of fish fillet, minced fish flesh and mixtures of fillets and minced fish flesh Quick frozen blocks of fish fillet, minced fish flesh and mixtures of fillets and minced fish flesh Quick frozen blocks of fish fillet, minced fish flesh and mixtures of fillets and minced fish flesh Quick frozen fish fillets Proportion of fish fillet and minced fish Net content of frozen fish blocks covered by glaze Sodium chloride AOAC 988.09 Physical separation I Described in the Standard Gravimetry I AOAC 971.21 (Codex general method) Potentiometry II Net weight of products Described in the Standard covered by glaze Quick Frozen Fish sticks (fish fingers) Fish content (declaration) AOAC 996.15 and calculation (described in and fish portions - breaded or in batter the standard) Quick frozen fish sticks (fish fingers) Net weight Described in the Standard and fish portions - breaded or in batter Quick Frozen Fish Sticks (fish fingers) Proportion of fish fillet and WEFTA Method (described in the Stnadard) and Fish Portions-Breaded and in minced fish Batter (except for certain fish species with soft flesh) Quick frozen fish sticks (fish fingers) Sodium chloride AOAC 971.27 (Codex general method) and fish portions - breaded or in batter Salted Atlantic Herring and Salted Water content AOAC 950.46B Sprat Salted Fish of the Gadidae Family Salted Fish and Dried Salted Fish of the Gadidae Family of Fishes Sturgeon Caviar Salt Salt Content Water content Salt content Described in CODEX STAN 167-1989 Sampling and method described in the Standard Described in CODEX STAN 167-1989 Water spraying and sieving Gravimetry Weighing Gravimetry I I I I Potentiometry air drying Titrimetry (Mohr) Salt determined as chloride expressed as sodium chloride Gravimetry Titrimetry (Mohr) Salt determined as chloride expressed as sodium chloride II I I I I 13 Foods for Special Dietary Uses Special foods Special foods Special foods Special foods Special foods Special foods Special foods Special foods Ash Calcium Calories by calculation Carbohydrates Chloride Dietary fibre, total Fat Fat in foods not containing starch, meat or vegetable products Fill of containers Folic acid Linoleate (in the form of glycerides) Linoleate (in the form of glycerides) Loss on drying Loss on drying (milk based) AOAC 942.05 AOAC 984.27 Method described in CAC/VOL IX-Ed.1, Part III Method described in CAC/VOL IX-Ed.1, Part III AOAC 971.27 (Codex general method) AOAC 985.29 CAC/RM 55-1976 CAC/RM 1-1973, B-2 Gravimetry ICP emission spectrometry Calculation method Calculation Potentiometry Gravimetry (enzymatic digestion) Gravimetry (extraction) Gravimetry I III III III II I I I Special foods Special foods Special foods CAC/RM 46-1972 AOAC 944.12 AOAC 922.06; 969.33; 963.22 Weighing Microbioassay Acid hydrolysis, preparation of methyl esters and gas chromatography Acid hydrolysis and spectrophotometry Gravimetry Gravimetry I II II Special foods Special foods Special foods AOAC 922.06; 979.19 AOAC 934.01 AOAC 925.23 AOAC 925.23 IDF Standard 21B:1987 ISO 6731:1989 AOAC 961.14 III I I Special foods Nicotinamide for foods not based on milk Colorimetry II 14 Special foods Special foods Special foods Special foods Special foods Special foods Special foods Special foods Special foods Special foods Special foods Nicotinamide for milk-based foods Pantothenic acid/enriched foods Pantothenic acid/non-enriched foods Phosphorous Protein efficiency ratio (PER) Protein, crude Riboflavin Sodium and potassium Sodium and potassium Vitamin A AOAC 944.13 AOAC 945.74 The Analyst 89 (1964):1, 3-6, ibid. 232 US Dept Agr., Agr. Handbook 97 (1965) AOAC 986.24 AOAC 960.48 Method described in CAC/VOL IX-Ed. 1, Part III AOAC 970.65 ISO 8070:1987 (confirmed 1992) IDF Standard 119A:1987 AOAC 984.27 AOAC 974.29 Microbioassay Microbioassay Microbioassay Colorimetry (molybdovanadate) Rat bioassay Titrimetry, Kjeldahl digestion Fluorometry Flame emission spectrophotometry ICP emission spectrometry Colorimetry Spectrophotometry II II IV II I I II II III IV III Vitamin A in foods in which AOAC 941.15 carotenes have been added as a source of vitamin A Vitamin B12 Vitamin B6 Vitamin C Vitamin C Vitamin D Vitamin D (D3, milk based infant formula) Vitamin E Vitamin E (milk based infant formula) AOAC 952.20 AOAC 961.15 AOAC 967.22 AOAC 967.21 AOAC 936.14 AOAC 992.26 AOAC 971.30 AOAC 992.03 Special foods Special foods Special foods Special foods Special foods Special foods Special foods Special foods Microbioassay Microbioassay Microfluorometry Colorimetry (dichloroindophenol) Rat bioassay Liquid chromatography Colorimetry Liquid chromatography II II II III IV II IV II 15 Follow-up formula follow-up formula Follow-up formula Dietary fibre, total Iodine (milk based formula) Pantothenic acid AOAC 991.43 AOAC 992.24 AOAC 992.07 (Measures total pantothenate (free pantothenic acid + CoA- + ACP-bound) and measured as D-pantothenic acid (or calcium D-pantothenate) Gravimetry (enzymatic digestion) Ion-selective potentiometry Microbioassay I II II Follow-up formula Follow-up formula Follow-up formula Follow-up formula Vitamin A Vitamin A (retinol isomers) AOAC 974.29 AOAC 992.04 Colorimetry HPLC HPLC HPLC with C30 column to separate the cis- and the trans- K vitamins Titrimetry Gravimetry IV II III II Vitamin A (retinol) (above 500 AOAC 992.06 IU/l milk after reconstitution) AOAC 999.15 Vitamin K EN 14148:2003 (vitamin K1) (Measures either aggregated cis + trans K1 or can measure individual cis and trans forms depending on LC column.) Iodine Silica (colloidal, calcium silicate) Biotin Calories (by calculation) AOAC 925.56 AOAC 950.85N Foods with low-sodium content (including salt substitutes) Foods with low-sodium content (including salt substitutes) Infant formula Infant formula II IV II I EN 15607:2008 (d-biotin) HPLC (Measures total D-biotin (free + D-biocytin) Method described in CAC/Vol IX-Ed.1, Calculation Part III7 7 Section 9. Calories by calculation – Section 9.2 Conversion Factors (a) protein 4 kcal per g (b) carbohydrate 4 kcal per g (c) fat 9 kcal per g (d) monosaccharides 3.75 kcal per g (e) specific food ingredients See “Energy and Protein Requirements”(FAO Nutrition Meeting Report Series No. 52 or WHO Technical Report Series No. 522) 16 Infant formula Infant formula Infant formula Infant formula Infant formula Calcium Calcium Calcium Chloride Choline ISO 8070 │ IDF 119: 2007 AOAC 985.35 AOAC 984.27 AOAC 986.26 AOAC 999.14 Flame atomic absorption spectrophotometry Flame atomic absorption spectroscopy ICP emission spectroscopy Potentiometry Enzymatic Colorimetric Method with limitations on applicability due to choline and ascorbate concentration. Flame atomic absorption spectroscopy ICP emission spectroscopy Graphite furnace atomic absorption after dry ashing Graphite furnace AAS after pressure digestion ICP emission spectroscopy Titrimetry (Kjeldahl) Gas chromatography Gas chromatography II III III III II Infant formula Infant formula Infant formula Infant formula Infant formula Infant formula Infant formula Infant formula Copper Copper Chromium (Section B of STAN 72 only) Chromium (Section B of STAN 72 only) Chromium (Section B of STAN 72 only) Crude protein8 Fatty acids (including trans fatty acid) Fatty acids (including trans fatty acid) AOAC 985.35 AOAC 984.27 EN 14082:2003 EN 14083:2003 AOAC 2006.03 AOAC 991.20 ISO 8968-1/2 | IDF 20-1/2: 2001 AOAC 996.06 AOCS Ce 1h-05 (09) II III II III III I II III (f) other specific calorie conversion factors maybe used where the formulation of the food and the nutrient content are known and where such specific conversion factors are physiologically more meaningful than the factors listed above 8 Determination of Crude Protein The calculation of the protein content of infant formulas prepared ready for consumption may be based on N x 6.25, unless a scientific justification is provided for the use of a different conversion factor for a particular product. The value of 6.38 is generally established as a specific factor appropriate for conversion of nitrogen to protein in other milk products, and the value of 5.71 as a specific factor for conversion of nitrogen to protein in other soy products 17 Infant formula Folic acid Infant formula Folic acid Infant formula Folic acid AOAC 992.05 (Measures free folic acid + free, unbound natural folates, aggregated and measured as folic acid) EN 14131:2003 (Total folate (free + bound), aggregated and measured as folic acid) J AOAC Int. 2000:83; 1141-1148 (Measures free folic acid + proportion of free, natural folate) J Chromatogr. A., 928, 77-90, 2001 (Measures total folates after conversion to, and measurement as 5-Me-H4PteGlu) AOAC 992.24 AOAC 985.35 AOAC 984.27 AOAC 999.11│NMKL139:1991 ISO 8070 │ IDF 119: 2007 AOAC 985.35 AOAC 984.27 AOAC 985.35 AOAC 984.27 EN 14083:2003 Microbioassay II Optical Biosensor Immunoassay HPLC, incorporating immunoaffinity clean-up and conversion to 5methyltetrahydrofolate Ion-selective potentiometry Flame atomic absorption spectrophotometry ICP emission spectroscopy AAS after dry ashing Flame atomic absorption spectrophotometry Flame atomic absorption spectroscopy ICP emission spectroscopy Flame atomic absorption spectrophotometry ICP emission spectroscopy Graphite furnace AAS after pressure digestion IV IV Infant formula Infant formula Infant formula Infant formula Infant formula Infant formula Infant formula Infant formula Infant formula Infant formula Iodine (for milk-based formula) Iron9 Iron Iron Magnesium Magnesium Magnesium Manganese Manganese Molybdenum (Section B of STAN 72 only) II III III II II III III II III II 9 General Codex methods are also available 18 Infant formula Infant formula Infant formula Infant formula Melamine Molybdenum (Section B of STAN 72 only) Niacin Niacin ISO/TS 15495 | IDF/RM 230:2010 AOAC 2006.03 AOAC 985.34 (niacin (preformed) and nicotinamide) prEN 15652:2009 (Free and bound and phosphorylated forms measured either as aggregate of nicotinic acid + nicotinamide, or as individual forms) AOAC 986.24 AOAC 984.27 AOAC 985.31 11 LC-MS/MS ICP emission spectroscopy Microbioassay and turbidimetry HPLC IV III III II10 Infant formula Infant formula Infant formula Infant formula Phosphorus Phosphorus Riboflavin Riboflavin Spectrophotometry (molybdovanadate) ICP emission spectroscopy Fluorimetry HPLC II III III II Infant formula Selenium EN 14152:2003 (Measures natural and supplemental forms, free, bound and phosphorylated (FMN and FAD) aggregated and measured as riboflavin.) AOAC 996.16 or AOAC 996. 17 Infant formula Infant formula Infant formula Infant formula Infant formula 10 11 12 Selenium Selenium Sodium and potassium Sodium and potassium Thiamine EN 14627:2005 AOAC 2006.03 AOAC 984.27 ISO 8070 | IDF 119:2007 AOAC 986.2712 Continuous hydride generation Flame atomic absorption spectrometry (HGAAS) Hydride generation atomic absorption spectrometry (HGAAS) ICP emission spectroscopy ICP emission spectrometry Flame atomic absorption spectrophotometry Fluorimetry III II III III II III when published as EN method Care should be taken in the application of the method due to spectral interference Care should be taken in the application of the method due to spectral interference 19 Infant formula Thiamine Infant formula Total carbohydrates Moisture/Total Solids EN 14122:2003 (Measures all vitamin B1 forms (natural and added free, bound and phosphorylated) following extraction and conversion to thiamine) AOAC 986.25 AOAC 990.19 or AOAC 990.20 IDF 21B:1987 or ISO 6731:1989 AOAC 942.05 AOAC 989.05 ISO 8381|IDF 123:2008 ISO 8262-1 |IDF 124-1: 2005 HPLC with pre-or post column derivatization to thiochrom II Determination by difference Gravimetry I Infant formula Infant formula Ash Total fat Total fat for milk-based infant formula (Products not completely soluble in ammonia) Total phospholipids Gravimetry Gravimetry (Röse-Gottlieb) Gravimetry (Weibull-Berntrop) I I Infant formula AOCS Ja7b-91 Infant formula Vitamin A Infant formula Vitamin D EN 12823-1:2000 (all-trans-retinol and 13cis-retinol) Vitamin A (both natural + supplemental ester forms) aggregated and quantified as individual retinol isomers (13 - cis and alltrans) AOAC 992.26 D3 measured Gas chromatography with suitable extraction and preparation procedures HPLC III III HPLC III 20 Infant formula Vitamin D Infant formula Infant formula Vitamin D Vitamin E Infant formula Vitamin E Infant formula Infant formula Vitamin B6 Vitamin B6 EN 12821:2000 (D2 and/or D3 measured as single components. Hydroxylated forms not measured.) NMKL 167: 2000 AOAC 995.05 D2 and D3 measured AOAC 992.03 Measures all rac-vitamin E (both natural + supplemental ester forms) aggregated and quantified as α-congeners EN 12822: 2000 (Measures Vitamin E (both natural + supplemental ester forms) aggregated and quantified as individual tocopherol congeners (α, β, γ, δ). AOAC 985.32 EN 14166:2008 (Aggregates free and bound pyridoxal, pyridoxine and pyridoxamine and measures as pyridoxine) AOAC 2004.07 EN 14164:2008 (Free and bound phosphorylated forms (pyridoxal, pyridoxine and pyridoxamine) converted and measured as pyridoxine) EN 14663:2005 (includes glycosylated forms) (Free and bound phosphorylated and glycosylated forms measured as the individual forms pyridoxal, pyridoxine and pyridoxamine) HPLC II HPLC HPLC III III HPLC II Microbioassay Microbioassay III III Infant formula Vitamin B6 II HPLC Infant formula Vitamin B6 HPLC III 21 Infant formula Vitamin B12 Infant formula Vitamin C Infant formula Infant formula Zinc Zinc AOAC 986.23 (Measures total vitamin B12 as cyanocobalamin) EN 14130:2003 (Measures ascorbic acid + dehydroascorbic acid) AOAC 985.35 AOAC 984.27 Turbidimetric Method II HPLC II Flame atomic absorption spectroscopy ICP emission spectroscopy II III Methods of analysis for dietary fibre: Guidelines for Use of Nutrition and Health Claims: Table of Conditions for Claims Standard Provisions Method General methods that do not measure the lower molecular weight fraction (i.e. monomeric units 9) and the lower molecular weight fraction (monomeric units = 107 cfu/g Streptococcus thermophilus & Lactobacillus delbrueckii subsp. bulgaricus >= 107 cfu/g ISO 7889/IDF 117: 2003 Colony count at 37°C Test for identification: morphological , cultural and biochemical characteristics I ISO 9232/IDF 146:2003 I Natural Mineral Waters Natural mineral waters Natural mineral waters Calcium Chloride ISO 7980:1986 (confirmed 1995) Examination of Water Pollution Control. WHO Pergamon Press (1982) Vol. 2, pp. 205-208 AOAC 973.51 ISO 9297:1989 (confirmed 1994) ISO 9308-1:1990 Titrimetry (Mercuric nitrate) Titrimetry Membrane filtration Atomic absorption spectrophotometry III II Natural mineral waters Natural mineral waters Natural mineral waters Chloride Chloride Coliform organism, thermotolerant organism and presumptive Escherichia coli Faecal Streptococci Iron, dissolved Magnesium Magnesium Phenols Potassium III III I Natural mineral waters Natural mineral waters Natural mineral waters Natural mineral waters Natural mineral waters Natural mineral waters ISO 7899-2:1984 ISO 6332:1988 (confirmed 1995) ISO 6059:1984 (confirmed 1995) ISO 7980:1986 (confirmed 1995) ISO 6439:1990 (confirmed 1995) Examination of Water Pollution Control. WHO Pergamon Press (1982) Vol.2, pp. 142-145 Membrane filtration Spectrophotometry Titrimetry Atomic absorption spectrophotometry Spectrophotometry I II II III I II 41 Natural mineral waters Sodium Examination of Water Pollution Control. WHO Pergamon Press (1982) Vol.2 pp. 148-151 Examination of Water Pollution Control. WHO Pergamon Press (1982) Vol.2, pp. 151-152 ISO 6461-2:1986 (confirmed 1996) ISO 9280:1990 (confirmed 1995) Handb. Spurenanal. 1974 Membrane filtration Gravimetry II Natural mineral waters Sodium III Natural mineral waters Natural mineral waters Natural mineral waters Spores of sulphite-reducing anaerobes (Clostridia) Sulphates Sulphide I III IV Criteria applicable to health-related substances in the Standard for Natural Mineral Waters Provision Antimony Arsenic ML (mg/L) 0.005 0.01 Min. applicable range (mg/L) 0.0028 0.0056 LOD LOQ (mg/L) (mg/L) 0.001 0.002 0.002 0.004 Precision Recover RSDR (%) y No more than (%) 44 80-110 44 90-107 Suggested methods Principle meeting the criteria ISO 17294-2:2003 ISO 15586:2003 EPA 200.8 ISO 17294-2:2003 ISO 15586:2003 ISO 11969:1996 EPA 200.8 ISO 11885:2007 ISO 17294-2:2003 EPA 200.8 ICP-MS GF-AAS ICP-MS ICP-MS GF-AAS AAS (Hydride) ICP-MS ICP-OES ICP-MS ICP-MS Barium 0.7 0.35 0.07 0.14 34 95-105 42 Provision ML (mg/L) 5 0.003 Min. applicable range (mg/L) 3.1 0.0017 LOD LOQ (mg/L) (mg/L) 0.5 1 Precision Recover RSDR (%) y No more than (%) 25 44 97-103 80-110 Suggested methods Principle meeting the criteria ISO 9390:1990 ISO 11885:2007 ISO 17294-2:2003 ISO 11885:2007 ISO 17294-2:2003 ISO 15586:2003 ISO 5961:1994 (Section 3) EPA 200.8 ISO 11885:2007 ISO 17294-2:2003 ISO 15586:2003 ISO 18412:2005 (Cr VI) ISO 23913:2006 (Cr VI) ISO 9174:1998 (Section 4) EPA 200.8 ISO 11885:2007 ISO 17294-2:2003 ISO 15586:2003 ISO 8288:1986 EPA 200.8 ISO 14403:2002 ISO 6703-1:1998 ISO 10304-1:2007 ISO 10359-1:1992 (dissolved fluoride) ISO 10359-2:1994 (inorganic bound) ISO 17294-2:2003 Spectrophotometry ICP-OES21 ICP-MS21 ICP-OES ICP-MS GF-AAS AAS ICP-MS ICP-OES ICP-MS GF-AAS Photometric CIA, spectrophotometry AAS ICP-MS ICP-OES ICP-MS GF-AAS Flame-AAS ICP-MS CFA Photometric, trimetric HPLC Electrochemical probe Digestion, distillation ICP-MS Borate Cadmium 0.0006 0.0012 Chromium 0.05 0.028 0.01 0.02 44 90-107 Copper 1 0.52 0.1 0.2 32 97-103 Cyanide Fluoride 0.07 1.0 0.039 0.52 0.014 0.1 0.028 0.2 44 32 90-107 97-103 Lead 21 0.01 0.0056 0.002 0.004 44 90-107 Total Boron is determined 43 Provision ML (mg/L) Min. applicable range (mg/L) 0.18 LOD LOQ (mg/L) (mg/L) Precision Recover RSDR (%) y No more than (%) 37 95-105 Suggested methods Principle meeting the criteria ISO 15586:2003 EPA 200.8 ISO 11885:2007 ISO 17294-2:2003 ISO 15586:2003 EPA 200.8 EN 1483:2007 ISO 17852:2006 ISO 5666:1999 ISO 16590:2000 EPA 200.8 ISO 17294-2:2003 ISO 15586:2003 EPA 200.8 ISO 10304-1:2007 ISO 13395:1996 ISO 7890-3:1988 ISO 10304-1:2007 ISO 13395:1996 ISO 6777:1984 ISO 17294-2:2003 ISO 15586:2003 ISO 9965:1993 EPA 200.8 GF-AAS ICP-MS ICP-OES ICP-MS GF-AAS ICP-MS AAS Enrichment by amalgamation (III) AFS AAS after tin(II) chloride reduction Enrichment by amalgamation (III) ICP-MS ICP-MS GF-AAS ICP-MS LC of ions UV CFA, FIA, Spectrophotometry Spectrophotometry LC of ions UV CFA, FIA, Spectrophotometry Spectrophotometry ICP-MS GF-AAS AAS (Hydride) ICP-MS Manganese 0.4 0.04 0.08 Mercury 0.001 0.00056 0.0002 0.0004 44 80-110 Nickel Nitrate Nitrite 0.02 50 0.1 0.011 37 0.03 0.004 5 0.01 0.008 10 0.02 44 18 44 90-107 98-102 95-105 Selenium 0.01 0.0056 0.002 0.004 44 90-107 44 Performance characteristics of suggested methods Provision Surface active agents Mineral oil (hydrocarbon index) PCB Pesticide (organochlorine) PAH ML Applicable range- from: 0.05 – 5.0 mg/L >0.1 mg/L >15 ng/L > 15 ng/ L 0.005 µg/L 0.04 µg/L 0.005 µg/L LOD 0.05 mg/l RSDR (%) < 44 < 41