RECOMMENDED METHODS OF ANALYSIS AND SAMPLING CODEX STAN PART A METHODS OF ANALYSIS BY COMMODITY CATEGORIES AND NAMES PART B

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1 RECOMMENDED METHODS OF ANALYSS AND SAMPLNG CODEX STAN PART A METHODS OF ANALYSS BY COMMODTY CATEGORES AND NAMES PART B METHODS OF SAMPLNG BY COMMODTY CATEGORES 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 SO/EC 17025: The present list of methods reflects the amendments adopted by the 34 th Session of the Codex Alimentarius Commission in

2 PART A METHODS OF ANALYSS BY COMMODTY CATEGORES AND NAMES Commodity Provision Method Principle Type All Foods All foods Acesulfame K, Aspartame EN : All foods Cyclamate EN : High performance liquid chromatography High performance liquid chromatography All foods Cyclamate NMKL 123 (1998) Spectrophotometry All foods Saccharin EN : High performance liquid chromatography All Foods (see also meat products) Nitrates and/or Nitrites EN : Part 1- General considerations N/A ndividual Foods 2 ndividual Foods 3 Sulphites Sulphites Cereals, Pulses and Legumes and Derived Products EN : AOAC EN : NMKL 135 (1990) Part 1: Optimized Monier-Williams method Part 2: Enzymatic method Certain pulses Moisture SO 665:1977 (confirmed 1995) Degermed maize (corn) meal and maize (corn) grits Degermed maize (corn) meal and maize (corn) grits Ash AOAC SO 2171:1993 CC Method No 104/1 (1990) Fat, crude AOAC F; C (ether extraction) 2 3 Hominy, fruit juice, sea food Wine, dried apples, lemon juice, potato flakes, sultanas, beer 2

3 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 Moisture SO 712:1998 CC Method No 110/1 (1986) 3 Particle size (granularity) AOAC Sieving Protein CC Method No 105/1 (1986) Titrimetry, Kjeldahl digestion Ash (semolina) AOAC SO 2171:1993 Moisture SO 712:1998 CC Method 110/1 (1986) Protein (N x 5.7) CC Method No 105/1 Titrimetry, Kjeldahl digestion nstant Noodles Extraction of oil from instant described in the standard noodles nstant Noodles Acid Value described in the standard Titrimetry nstant Noodles Moisture described in the standard Maize (corn) Moisture SO 6540:1980 (confirmed 1994) Peanuts (raw) Aflatoxins, total AOAC mmunoaffinity column (Aflatest) Peanuts (raw) Aflatoxins, total AOAC Thin layer chromatography Peanuts (intended for further processing) Peanuts (Cereals, shell-fruits and derived products ( including peanuts)) Peanuts (intended for further processing) Aflatoxins, total AOAC Romer minicolmn Sum of aflatoxins B 1, B 2, G 1 and G 2 EN : SO 16050:2003 HPLC with post column derivatization and immunoaffinity column clean up Aflatoxins, total AOAC Holaday-Velasco minicolumn Pearl millet flour Ash AOAC Pearl millet flour Colour Modern Cereal Chemistry, 6th Ed., D.W. Kent-Jones and A.J. Amos (Ed.), pp , Food Trade Press Ltd, London, Colorimetry using specific colour grader V

4 Pearl millet flour Fat, crude AOAC F; C (ether extraction) Pearl millet flour Fibre, crude SO 5498:1981 (B.5 Separation) Pearl millet flour Moisture SO 712:1998 CC Method No 110/1 (1986) 4 Pearl millet flour Protein AOAC Titrimetry, Kjeldahl digestion Sorghum flour Ash AOAC SO 2171:1993 CC Method No 104/1 (1990) Sorghum flour Colour Modern Cereal Chemistry, 6th Ed., D.W. Kent-Jones and A.J. Amos (Ed.), pp , Food Trade Press Ltd, London, Colorimetry using specific colour grader Sorghum flour Fat, crude AOAC F; C (ether extraction) Sorghum flour Fibre, crude CC Method No 113 (1972) SO 6541:1981 (confirmed 1996) Sorghum flour Moisture SO 712:1998 CC Method No 110/1 (1986) Sorghum flour Particle size (granularity) AOAC Sieving Sorghum flour Protein CC Method No 105/1 (1986) Titrimetry, Kjeldahl digestion Sorghum flour Tannins SO 9648:1988 (confirmed 1994) Spectrophotometry Sorghum grains Ash AOAC SO 2171:1993 CC Method No 104/1 (1990) Sorghum grains Fat, crude AOAC F, C (ether extraction) Sorghum grains Moisture SO 6540:1980 (confirmed 1994) Sorghum grains Protein CC Method No 105/1 (1986) Titrimetry, Kjeldahl digestion Sorghum grains Tannins SO 9648:1988 (confirmed 1994) Spectrophotometry Soy protein products Ash AOAC SO 2171:1993 (Method B) V

5 Soy protein products Fat CAC/RM Method 1 (extraction) Soy protein products Fibre, crude SO 5498:1981 Soy protein products Moisture AOAC (vacuum oven) Soy protein products Protein AOAC D (using factor 6.25) Titrimetry, Kjeldahl digestion Vegetable protein products Ash AOAC SO 2171:1993 (Method B), Direct Vegetable protein products Fat CAC/RM Method 1 (extraction) Vegetable protein products Fibre, crude AACC (1982) Ceramic fiber filteration Vegetable protein products Moisture AOAC (vacuum oven) Vegetable protein products Protein AOAC D (using factor 6.25) Titrimetry, Kjeldahl digestion Wheat flour Ash AOAC SO 2171:1993 CC Method No 104/1 (1990) Wheat flour Fat acidity AOAC Titrimetry Wheat flour Moisture SO 712:1998 CC Method No 110/1 (1986) Wheat flour Particle size (granularity) AOAC Sieving Wheat flour Protein CC Method No 105/1 (1986) Titrimetry, Kjeldahl digestion Wheat protein products including wheat gluten Wheat protein products including Wheat gluten Protein Vital wheat gluten and devitalized wheat gluten AOAC (wheat protein in grain Nx5.7) Solubilized wheat protein AOAC (wheat protein in flour Nx5.7) Kjeldahl Kjeldahl Fibre, crude AOAC Ceramic fiber filteration 5

6 Wheat protein products including Wheat gluten Ash AOAC SO 2171:1980, method B 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 Ash AOAC Fat, crude AOAC F; C (ether extraction) Fibre, crude SO 5498:1981 (B.5 Separation) Moisture SO 712:1998 CC Method No 110/1 (1986) Protein AOAC Titrimetry, Kjeldahl digestion Whole maize (corn) meal Ash AOAC SO 2171:1993 CC Method No 104/1 (1990) Whole maize (corn) meal Fat, crude AOAC F; C (ether extraction) Whole maize (corn) meal Moisture SO 712:1998 CC Method No 110/1 (1986) Whole maize (corn) meal Particle size (granularity) AOAC Sieving Whole maize (corn) meal Protein CC Method No 105/1 (1986) Titrimetry, Kjeldahl digestion Cocoa Products and Chocolate Chocolate and chocolate products Cocoa butter AOAC OCCC (Soxhlet extraction) Chocolate and chocolate products Fat-free cocoa solids AOAC Oven evaporation and factor Chocolate and chocolate products Fat-free milk solids OCCC or AOAC Titrimetry, Kjeldahl digestion; after extraction of milk proteins Chocolate and chocolate products Fat, total AOAC (Soxhlet extraction) 6

7 Chocolate and chocolate products Milkfat OCCC AOAC ; B; Chocolate and chocolate products Moisture OCCC or AOAC (Karl Fischer method); or AOAC or OCCC Titrimetry/Distillation Chocolate and chocolate products Non-cocoa butter vegetable fat AOCS Ce 10/02 and described in the Standard Described in the Standard Cocoa (Cacao) Mass or Cocoa/ Chocolate Liquor, and Cocoa Cake Cocoa shell AOAC and Spiral vessel count, Stone cell count Cocoa (Cacao) Mass or Cocoa/ Chocolate Liquor, and Cocoa Cake Fat AOAC or OCCC 14 (1972) (Soxhlet extraction) Cocoa butter Free fatty acids SO660:1996 amended 2003; or AOCS Cd 3d-63 (03) Titrimetry Cocoa butter Unsaponifiable matter SO 3596:2000 or SO 18609: 2000; or AOCS Ca 6b-53 (01) Titrimetry after extraction with diethyl ether Cocoa powders (cocoa) and dry cocoasugar mixtures Moisture OCCC or AOAC (Karl Fischer method) Fats and Oils and Related Products Fats and Oils (all) Arsenic AOAC (Codex general method) Colorimetry (diethyldithiocarbamate) Fats and Oils (all) Arsenic AOAC (Codex general method) Colorimetry (molybdenum blue) Fats and Oils (all) Arsenic AOAC (Codex general method) Atomic absorption spectrophotometry Fats and oils Butylhydroxyanisole, butylhydroxytoluene, tertbutylhydroquinone, & propyl gallate AOAC ; or AOCS Ce-6-86 (09) Liquid chromatography Fats and Oils (all) nsoluble impurities SO 663:2007 7

8 Fats and Oils (all) Lead AOAC SO 12193:2004 (Codex general method) or AOCS Ca 18c-91 (09) Atomic absorption spectrophotometry (direct graphite furnace) Fats and Oils (all) Matter volatile at 105 C SO 662:1998 (open-drying) Fats and Oils (all) Soap content BS 684 Section 2.5; or AOCS Cc (09) Fats and oils not covered by individual standards Fats and oils not covered by individual standards Fats and oils not covered by individual standards Acid Value SO 660:1996; or AOCS Cd 3d-63 (09) Titrimetry Copper and ron AOAC SO 8294:1994 or AOCS Ca 18b-91 (09) (Codex general method) Peroxide value AOCS Cd 8b-90 (09) SO 3960:2007 Atomic absorption Spectrophotometry (direct graphite furnace) Titrimetry using iso-octane Fat spreads and blended spreads Fat content SO DF 194: 2003 Named Animal Fats Acidity SO 660:2009; or AOCS Cd 3d-63 (09) Titrimetry Named Animal Fats Copper and ron AOAC SO 8294:1994; or AOCS Ca 18b-91 (09) (Codex general method) Named Animal Fats GLC ranges of fatty acid composition SO 5508 :1990 and SO :2011 or AOCS Ce 2-66 (09) and Ce 1e-91 (09) or Ce 1f-96 (09) Named Animal Fats odine value (V) SO 3961: 2009; or AOAC ; or AOCS Cd 1d-1992 (09) Atomic absorption Spectrophotometry (direct graphite furnace) Gas chromatography of methyl esters Wijs-Titrimetry Named Animal Fats Peroxide value AOCS Cd 8b-90 (09) SO 3960:2007 Named Animal Fats Relative density SO/AOCS method for apparent density to be inserted Named Animal Fats Refractive index SO 6320:2000 and corr 2006; or AOCS Cc 7-25 (09) 8 Titrimetry using iso-octane Pycnometry Refractometry Named Animal Fats Saponification value SO 3657:2002; or AOCS Cd 3-25 (09) Titrimetry

9 Named Animal Fats Unsaponifiable matter SO 3596:2000 or SO 18609: 2000; or AOCS Ca 6b-53 (09) Titrimetry after extraction with diethyl ether Named Animal Fats Titre SO 935:1988; or AOCS Cc (09) Thermometry Named Vegetable Oils Acidity SO 660:2009; or AOCS Cd 3d-63 (09) Titrimetry Named Vegetable Oils Apparent density SO 6883: 2007, with the appropriate conversion factor; or AOCS Cc 10c-95 (09) Named Vegetable Oils Baudouin test (modified Villavecchia or sesameseed oil test) Pycnometry AOCS Cb 2-40 (09) Colour reaction Named Vegetable Oils Carotenoids, total BS 684 Section 2.20 Spectrophotometry Named Vegetable Oils Copper and iron SO 8294: 1994; or AOAC ; or AOCS Ca 18b-91 (09) Named Vegetable Oils Crismer value AOCS Cb 4-35 (97) and AOCS Ca 5a-40 Named Vegetable Oils GLC ranges of fatty acid composition (09) SO 5508:1990 and SO ; or AOCS Ce 2-66 (09) and Ce 1e-91 (09) or Ce 1f-96 (09) AAS Turbidity Gas chromatography of methyl esters Named Vegetable Oils Halphen test AOCS Cb 1-25 (09) Colorimetry Named Vegetable Oils nsoluble impurities SO 663: 2000 Named Vegetable Oils odine value (V) Wijs - SO 3961: 1996; or AOAC ; or AOCS Cd 1d-1992 (97); or NMKL 39 (2003) Wijs-Titrimetry 4 Named Vegetable Oils Lead AOAC ; or SO 12193: 2004; or AOCS Ca 18c-91 (03) Atomic Absorption Named Vegetable Oils Moisture & volatile matter at 105 C SO 662: 1998 Named Vegetable Oils Peroxide value (PV) AOCS Cd 8b-90 (09); or SO 3960: 2007 Titrimetry 4 t is possible to calculate the odine Value from fatty acid composition data obtained by gas chromatography e.g. using AOCS Cd 1b-87 (97) 9

10 Named Vegetable Oils Refractive index SO 6320: 2000 and corr 2006; or AOCS Cc Refractometry 7-25 (09) Named Vegetable Oils Reichert value and Polenske AOCS Cd 5-40 (09) Titrimetry value Named Vegetable Oils Relative density UPAC with the appropriate conversion factor See comment above (Named Animal Fats) 5 Pycnometry Named Vegetable Oils Saponification value (SV) SO 3657: 2002; or AOCS Cd 3-25 (09) Titrimetry Named Vegetable Oils Slip point SO 6321:2002 for all oils; AOCS Cc 3b-92 (09) for all oils except palm oils; AOCS Cc 3-25 (09) for palm oils only Open ended capillary tube Named Vegetable Oils Soap content BS 684 Section 2.5; or AOCS Cc (09) Named Vegetable Oils Sterol content SO 12228: 1999; or AOCS Ch 6-91 (09) Gas chromatography Named Vegetable Oils Tocopherol content SO 9936:2006 and corrigendum 2008 HPLC or AOCS Ce 8-89 (09) Named Vegetable Oils Unsaponifiable matter SO 3596:2000; or SO 18609: 2000; or AOCS Ca 6b-53 (09) Olive Oils and Olive Pomace Oils Absorbency in ultra-violet CO/T.20/Doc. No. 19 or SO 3656:2011 Absorption in ultra violet or AOCS Ch 5-91 (09). Olive Oils and Olive Pomace Oils Acidity, free (acid value) SO 660:2009 Titrimetry or AOCS Cd 3d-63 (09) Olive Oils and Olive Pomace Oils Alpha-tocopherol SO 9936:1997 HPLC Olive Oils and Olive Pomace Oils Difference between the actual CO/T.20/Doc. no. 20 or AOCS Ce 5b-89 Analysis of triglycerides of HPLC and theoretical ECN 42 (09) and calculation triglyceride content Olive Oils and Olive Pomace Oils Erythrodiol + uvaol content UPAC Gas chromatography Olive Oils and Olive Pomace Oils Fatty acids in the 2-position of the triglycerides SO 6800:1997 or AOCS Ch 3-91 (09) Gas chromatography 5 The method is no longer available. 6 The method is no longer available. 10

11 Olive Oils and Olive Pomace Oils Halogenated solvents, traces CO/T.20/Doc. no. 8 Gas chromatography Olive Oils and Olive Pomace Oils nsoluble impurities in light SO 663:2000 petroleum Olive Oils and Olive Pomace Oils odine value SO 3961:2009 or AOAC or Wijs-Titrimetry AOCS Cd 1d-92 (09) or NMKL 39 (2003) Olive Oils and Olive Pomace Oils ron and copper SO 8294:1994 or AOAC AAS Olive Oils and Olive Pomace Oils Lead AOAC or SO 12193:2004 or AAS AOCS Ca 18c-91(09) Olive Oils and Olive Pomace Oils Moisture and volatile matter SO 662:1998 Olive Oils and Olive Pomace Oils Organoleptic characteristics CO/T.20/Doc. no. 15 Panel test Olive Oils and Olive Pomace Oils Peroxide value SO 3960:2007 or AOCS Cd 8b-90 (09) Titrimetry Olive Oils and Olive Pomace Oils Relative density UPAC 2.101, with the appropriate Pycnometry conversion factor. See comment above 6 Olive Oils and Olive Pomace Oils Refractive index SO 6320:2000 and corr 2006 or AOCS Refractometry Cc 7-25 (09) Olive Oils and Olive Pomace Oils Saponification value SO 3657:2002 or AOCS Cd 3-25 (09) Titrimetry Olive Oils and Olive Pomace Oils Sterol composition and total sterols CO/T.20/Doc. no. 10 or SO 12228:1999 or AOCS Ch 6-91 (09). Olive Oils and Olive Pomace Oils Stigmastadienes COl/T.20/Doc. no. 11 or SO :1999 or AOCS Cd (09). 11 Gas chromatography Gas chromatography Olive Oils and Olive Pomace Oils Stigmastadienes SO : 2003 HPLC Olive Oils and Olive Pomace Oils Trans fatty acids content CO/T.20/Doc no. 17 or SO 15304:2002 or AOCS Ce 1f-96 (09) Olive Oils and Olive Pomace Oils Unsaponifiable matter SO 3596:2000 or SO 18609:2000 or AOCS Ca 6b-53 (09) Olive Oils and Olive Pomace Oils Wax content CO/T.20/Doc. no. 18 or AOCS Ch 8-02 (09) Gas chromatography of methyl esters Gas chromatography Margarine Fat UPAC Margarine Milkfat CAC/RM Titrimetry Margarine Sodium chloride AOAC (Codex general method) Potentiometry Margarine Vitamin A AOAC Spectrophotometry

12 Margarine Vitamin D AOAC Bioassay Margarine Vitamin E UPAC TLC followed by spectrophotometry or GLC Margarine Water CAC/RM (described in the Standard) Minarine Fat UPAC Minarine Milkfat CAC/RM (described in the Standard) Titrimetry Minarine Sodium chloride AOAC (Codex general method) Potentiometry Minarine Vitamin A AOAC Spectrophotometry Minarine Vitamin D AOAC Bioassay Minarine Vitamin E UPAC TLC followed by spectrophotometry or GLC Minarine Water CAC/RM Fish and Fishery Products Fish and fishery products Histamine AOAC Fluorimetry Fish and fishery products Mercury AOAC Flameless atomic absorption spectrophotometry Fish and fishery products: canned products Drained weight Described in the Standard Weighing Fish and fishery products: canned products Boiled Dried Salted Anchovies Net weight Described in the Standard Weighing Sodium Chloride (chloride expressed as sodium chloride) AOAC Titrimetry Canned shrimps or prawns Size, determination of Described in the Standard Number per 100 g Frozen fish and fishery products Thawing and cooking procedures Described in the Standards Thawing and heating 12

13 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 Quick Frozen Fish sticks (fish fingers) and fish portions - breaded or in batter Quick frozen fish sticks (fish fingers) and fish portions - breaded or in batter Quick Frozen Fish Sticks (fish fingers) and Fish Portions-Breaded and in Batter (except for certain fish species with soft flesh) Quick frozen fish sticks (fish fingers) and fish portions - breaded or in batter Salted Atlantic Herring and Salted Sprat Proportion of fish fillet and minced fish Net content of frozen fish blocks covered by glaze AOAC Physical separation Described in the Standard Sodium chloride AOAC (Codex general method) Potentiometry Net weight of products covered by glaze Fish content (declaration) Described in the Standard Water spraying and sieving AOAC and calculation (described in the standard) Net weight Described in the Standard Weighing Proportion of fish fillet and minced fish WEFTA Method (described in the Stnadard) Sodium chloride AOAC (Codex general method) Potentiometry Water content AOAC B air drying Salted Fish of the Gadidae Family Salt Described in CODEX STAN Titrimetry (Mohr) Salt determined as chloride expressed as sodium chloride Salted Fish and Dried Salted Fish of the Gadidae Family of Fishes Salt Content Water content Sampling and method described in the Standard Sturgeon Caviar Salt content Described in CODEX STAN Titrimetry (Mohr) Salt determined as chloride expressed as sodium chloride 13

14 Foods for Special Dietary Uses Special foods Ash AOAC Special foods Calcium AOAC CP emission spectrometry Special foods Calories by calculation Method described in CAC/VOL X-Ed.1, Part Special foods Carbohydrates Method described in CAC/VOL X-Ed.1, Part Calculation method Calculation Special foods Chloride AOAC (Codex general method) Potentiometry Special foods Dietary fibre, total AOAC (enzymatic digestion) Special foods Fat CAC/RM (extraction) Special foods Fat in foods not containing starch, meat or vegetable products CAC/RM , B-2 Special foods Fill of containers CAC/RM Weighing Special foods Folic acid AOAC Microbioassay Special foods Special foods Linoleate (in the form of glycerides) Linoleate (in the form of glycerides) AOAC ; ; AOAC ; Special foods Loss on drying AOAC AOAC Special foods Loss on drying (milk based) AOAC DF Standard 21B:1987 SO 6731:1989 Special foods Nicotinamide for foods not based on milk Acid hydrolysis, preparation of methyl esters and gas chromatography Acid hydrolysis and spectrophotometry AOAC Colorimetry 14

15 Special foods Special foods Special foods Nicotinamide for milk-based foods Pantothenic acid/enriched foods Pantothenic acid/non-enriched foods AOAC Microbioassay AOAC Microbioassay The Analyst 89 (1964):1, 3-6, ibid. 232 US Dept Agr., Agr. Handbook 97 (1965) 15 Microbioassay Special foods Phosphorous AOAC Colorimetry (molybdovanadate) Special foods Protein efficiency ratio (PER) AOAC Rat bioassay Special foods Protein, crude Method described in CAC/VOL X-Ed. 1, Part Titrimetry, Kjeldahl digestion Special foods Riboflavin AOAC Fluorometry Special foods Sodium and potassium SO 8070:1987 (confirmed 1992) DF Standard 119A:1987 Flame emission spectrophotometry Special foods Sodium and potassium AOAC CP emission spectrometry Special foods Vitamin A AOAC Colorimetry V Special foods Vitamin A in foods in which carotenes have been added as a source of vitamin A AOAC Spectrophotometry Special foods Vitamin B 12 AOAC Microbioassay Special foods Vitamin B 6 AOAC Microbioassay Special foods Vitamin C AOAC Microfluorometry Special foods Vitamin C AOAC Colorimetry (dichloroindophenol) Special foods Vitamin D AOAC Rat bioassay V Special foods Vitamin D (D 3, milk based infant formula) AOAC Liquid chromatography Special foods Vitamin E AOAC Colorimetry V Special foods Vitamin E (milk based infant formula) AOAC Liquid chromatography V

16 Follow-up formula Dietary fibre, total AOAC (enzymatic digestion) follow-up formula odine (milk based formula) AOAC on-selective potentiometry Follow-up formula Pantothenic acid AOAC (Measures total pantothenate (free pantothenic acid + CoA- + ACP-bound) and measured as D-pantothenic acid (or calcium D-pantothenate) Microbioassay Follow-up formula Vitamin A AOAC Colorimetry V Follow-up formula Vitamin A (retinol isomers) AOAC HPLC Follow-up formula Vitamin A (retinol) (above 500 U/l milk after reconstitution) Follow-up formula Vitamin K AOAC EN 14148:2003 (vitamin K 1 ) (Measures either aggregated cis + trans K 1 or can measure individual cis and trans forms depending on LC column.) AOAC HPLC HPLC with C30 column to separate the cis- and the trans- K vitamins Foods with low-sodium content (including salt substitutes) odine AOAC Titrimetry Foods with low-sodium content (including salt substitutes) Gluten-free foods Silica (colloidal, calcium silicate) Gluten AOAC N V Enzyme-Linked mmunoassay R5 Mendez (ELSA) Method Eur J Gastroenterol Hepatol 2003; 15: mmunoassay nfant formula Biotin EN 15607:2008 (d-biotin) (Measures total D-biotin (free + D-biocytin) HPLC 16

17 nfant formula Calories (by calculation) Method described in CAC/Vol X-Ed.1, Part 7 Calculation nfant formula Calcium SO 8070 DF 119: 2007 Flame atomic absorption spectrophotometry nfant formula Calcium AOAC Flame atomic absorption spectroscopy nfant formula Calcium AOAC CP emission spectroscopy nfant formula Chloride AOAC Potentiometry nfant formula Choline AOAC Enzymatic Colorimetric Method with limitations on applicability due to choline and ascorbate concentration. nfant formula Copper AOAC Flame atomic absorption spectroscopy nfant formula Copper AOAC CP emission spectroscopy nfant formula nfant formula nfant formula Chromium (Section B of STAN 72 only) Chromium (Section B of STAN 72 only) Chromium (Section B of STAN 72 only) EN 14082:2003 EN 14083:2003 Graphite furnace atomic absorption after dry ashing Graphite furnace AAS after pressure digestion AOAC CP emission spectroscopy 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) (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 17

18 nfant formula Crude protein 8 AOAC SO /2 DF 20-1/2: 2001 Titrimetry (Kjeldahl) nfant formula Fatty acids (including trans AOAC Gas chromatography fatty acid) nfant formula nfant formula Fatty acids (including trans fatty acid) Folic acid AOCS Ce 1h-05 (09) Gas chromatography AOAC (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) nfant formula Folic acid J AOAC nt. 2000:83; (Measures free folic acid + proportion of free, natural folate) nfant formula Folic acid J Chromatogr. A., 928, 77-90, 2001 (Measures total folates after conversion to, and measurement as 5-Me-H4PteGlu) nfant formula odine (for milk-based formula) Microbioassay Optical Biosensor mmunoassay HPLC, incorporating immunoaffinity clean-up and conversion to 5- methyltetrahydrofolate AOAC on-selective potentiometry nfant formula ron 9 AOAC Flame atomic absorption spectrophotometry nfant formula ron AOAC CP emission spectroscopy nfant formula ron AOAC NMKL139:1991 AAS after dry ashing V V 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 9 General Codex methods are also available 18

19 nfant formula Magnesium SO 8070 DF 119: 2007 Flame atomic absorption spectrophotometry nfant formula Magnesium AOAC Flame atomic absorption spectroscopy nfant formula Magnesium AOAC CP emission spectroscopy nfant formula Manganese AOAC Flame atomic absorption spectrophotometry nfant formula Manganese AOAC CP emission spectroscopy nfant formula Molybdenum (Section B of STAN 72 only) EN 14083:2003 Graphite furnace AAS after pressure digestion nfant formula Melamine SO/TS DF/RM 230:2010 LC-MS/MS V nfant formula Molybdenum (Section B of STAN 72 only) nfant formula Niacin AOAC (niacin (preformed) and nicotinamide) nfant formula Niacin pren 15652:2009 (Free and bound and phosphorylated forms measured either as aggregate of nicotinic acid + nicotinamide, or as individual forms) AOAC CP emission spectroscopy Microbioassay and turbidimetry nfant formula Phosphorus AOAC Spectrophotometry (molybdovanadate) HPLC 10 nfant formula Phosphorus AOAC CP emission spectroscopy nfant formula Riboflavin AOAC Fluorimetry nfant formula Riboflavin EN 14152:2003 (Measures natural and supplemental forms, free, bound and phosphorylated (FMN and FAD) aggregated and measured as riboflavin.) HPLC 10 when published as EN method 11 Care should be taken in the application of the method due to spectral interference 19

20 nfant formula Selenium AOAC or AOAC Continuous hydride generation Flame atomic absorption spectrometry (HGAAS) nfant formula Selenium EN 14627:2005 Hydride generation atomic absorption spectrometry (HGAAS) nfant formula Selenium AOAC CP emission spectroscopy nfant formula Sodium and potassium AOAC CP emission spectrometry nfant formula Sodium and potassium SO 8070 DF 119:2007 Flame atomic absorption spectrophotometry nfant formula Thiamine AOAC Fluorimetry nfant formula Thiamine EN 14122:2003 (Measures all vitamin B 1 forms (natural and added free, bound and phosphorylated) following extraction and conversion to thiamine) nfant formula Total carbohydrates Moisture/Total Solids Ash AOAC nfant formula Total fat AOAC SO 8381 DF 123:2008 nfant formula Total fat for milk-based infant formula (Products not completely soluble in ammonia) AOAC AOAC or AOAC DF 21B:1987 or SO 6731:1989 HPLC with pre-or post column derivatization to thiochrom Determination by difference (Röse-Gottlieb) SO DF 124-1: 2005 (Weibull-Berntrop) 12 Care should be taken in the application of the method due to spectral interference 20

21 nfant formula Total phospholipids AOCS Ja7b-91 Gas chromatography with suitable extraction and preparation procedures nfant formula Vitamin A EN :2000 (all-trans-retinol and 13- cis-retinol) Vitamin A (both natural + supplemental ester forms) aggregated and quantified as individual retinol isomers (13 - cis and alltrans) nfant formula Vitamin D AOAC D 3 measured nfant formula Vitamin D EN 12821:2000 (D2 and/or D3 measured as single components. Hydroxylated forms not measured.) NMKL 167: 2000 nfant formula Vitamin D AOAC D2 and D3 measured nfant formula Vitamin E AOAC Measures all rac-vitamin E (both natural + supplemental ester forms) aggregated and quantified as α-congeners nfant formula Vitamin E EN 12822: 2000 (Measures Vitamin E (both natural + supplemental ester forms) aggregated and quantified as individual tocopherol. HPLC HPLC HPLC HPLC HPLC HPLC nfant formula Vitamin B 6 AOAC Microbioassay nfant formula Vitamin B 6 EN 14166:2008 (Aggregates free and bound pyridoxal, pyridoxine and pyridoxamine and measures as pyridoxine) Microbioassay 21

22 nfant formula Vitamin B 6 AOAC EN 14164:2008 (Free and bound phosphorylated forms (pyridoxal, pyridoxine and pyridoxamine) converted and measured as pyridoxine) nfant formula Vitamin B 6 EN 14663:2005 (includes glycosylated forms) (Free and bound phosphorylated and glycosylated forms measured as the individual forms pyridoxal, pyridoxine and pyridoxamine) nfant formula Vitamin B 12 AOAC (Measures total vitamin B 12 as cyanocobalamin) nfant formula Vitamin C EN 14130:2003 (Measures ascorbic acid + dehydroascorbic acid) 22 HPLC HPLC Turbidimetric Method nfant formula Zinc AOAC Flame atomic absorption spectroscopy nfant formula Zinc AOAC CP emission spectroscopy HPLC Methods of analysis for dietary fibre: Guidelines for Use of Nutrition and Health Claims: Table of Conditions for Claims Standard Provisions Method Principle Type General methods that do not measure the lower molecular weight fraction (i.e. monomeric units<=9) (2) All foods Method applicable for determining dietary fibres that do not include AOAC Enzymatic Type (1) the lower molecular weight fraction. (4) AACC ntl (1991,1999) gravimetry All foods (1) Method applicable for determining dietary fibres that do not include the lower molecular weight fraction and also includes determination AOAC AACC ntl (1999, 1991) Enzymatic gravimetry Type All foods (1) for soluble and insoluble dietary fibres (4) Method applicable for determining dietary fibres that do not include the lower molecular weight fraction, in foods and food products containing more than 10% dietary fibres and less than 2% starch (e.g. fruits) (4) NMKL 129, 2003 AOAC gravimetry Type

23 Standard Provisions Method Principle Type All foods (1) Enzymatic GC/ Type Method applicable for determining dietary fibres that do not include the lower molecular weight fraction. Provides sugar residue composition of dietary fibre polysaccharides, as well as content of Klason lignin (4). AOAC AACC ntl (1999, 1994) NMKL 162, 1998 colorimetry gravimetry All foods (1) All foods (1) nsoluble dietary fibres in food and food products (4) Soluble dietary fibres in food and food products (4) AOAC (Specific for insoluble fibre) AACC ntl (1999, 1982) AOAC (Specific for soluble fibre) Enzymatic gravimetry Enzymatic gravimetry Type Type 23

24 General methods that measure both the higher (monomeric units > 9) and the lower molecular weight fraction (monomeric units <=9) (2) All foods AOAC Enzymatic Type (1) Method applicable for determining the content of dietary fibres of AACC ntl (2002) gravimetry and higher and lower molecular weight, in food where resistant starches Liquid are not present chromatography All foods (1) Method applicable for determining the content of dietary fibres of higher and lower molecular weight. The method is applicable in AOAC AACC ntl (2009) Enzymatic- Type food that may, or may not, contain resistant starches. High Pressure Liquid Chromatography Methods that measure individual specific components (monomeric units: the whole range for each type of components is covered) (2) All foods (1 3)(1 4) Beta-D-Glucans AOAC Enzymatic Type (1) AACC ntl (1999, 1995) All foods Fructans (oligofructoses, inulin, hydrolyzed inulin, polyfructoses, AOAC Enzymatic & Type (1) fructooligosaccharides) (applicable to added fructans) AACC ntl (2001) HPAEC-PAD All foods Fructans (oligofructoses, inulin, hydrolyzed inulin, polyfructoses, AOAC Enzymatic & Type (1) fructooligosaccharides) (not applicable highly depolymerised fructans) AACC ntl (2001) colorimetric All foods Polydextrose AOAC HPAEC-PAD Type (1) AACC ntl (2001) All foods Trans-galacto-oligo saccharides AOAC HPAEC-PAD Type (1) AACC ntl (2001) All foods (1) Resistant starch (Recommended for RS3) AOAC AACC ntl (2002) Enzymatic Type 24

25 Other methods (2) that have not been subjected to interlaboratory evaluation under AOAC international guidelines All foods nsoluble glucans and mannans of yeast cell wall (for yeast cell Eurasyp (European association for wall only) specialty yeast product) LM Bonanno. Biospringer online version : ue.home.screen. All foods Fructo-oligosaccharides (monomeric units<5) Ouarné et al in Complex Carbohydrates in Foods. Edited by S. Sungsoo, L. Prosky & M. Dreher. Marcel Dekker nc, New York All foods Non-starch polysaccharides (NSP) (3) Englyst H.N, Quigley M.E., Hudson G. (1994) Determination of dietary fibre as non-starch polysaccharides with gas-liquid chromatographic high performance liquid chromatographic or spectrophotometric measurement of constituent sugars Analyst 119, Chemical HPAEC-PAD HPAEC-PAD & Gas-Liquid Chromatography Type V Type V Type V (1) Users should consult the description of each method for the food matrices that were the subject of interlaboratory study in the Official methods of Analysis of AOAC nternational. (2) Two issues are left for national authorities: to include monomeric units 3-9 and which isolated or synthetic compounds have physiological benefit. (Refer to the Guidelines for Nutrition Labelling (CAC/GL ), as revised in (3) Quantitation lost for resistant starch. Refer to specific methods. (4) Quantitation lost for inulin, resistant starch, polydextrose and resistant maltodextrins. Refer to specific methods. 25

26 Commodity Provisions Method Principle Type Ascorbic acid-l (additives) FU Method No 17a (1995) HPLC Ascorbic acid-l (additives) SO : 1986 Fluorescence spectrometry V Ascorbic acid-l (additives) AOAC ndophenol method FU Method No 17 SO : 1984 Carbon dioxide (additives and FU Method No 42 (1976) Titrimetry V processing aids) (back-titration after precipitation) Cellobiose FU Recommendation No.4 October 2000 Capillary gas chromatography V Citric acid 13 (additives) AOAC HPLC Citric acid 5 (additives) EN 1137: 1994 Enzymatic determination FU Method No 22 (1985) Glucose and fructose EN HPLC (permitted ingredients) FU Method No 67 (1996) NMKL 148 (1993) Glucose-D and fructose-d EN 1140 Enzymatic determination (permitted ingredients) FU Method No 55 (1985) HFCS & HS in apple juice Determination of HFCS & HS by Capillary CAP GC Method V (permitted ingredients) GC method JAOAC 84, 486 (2001) Malic acid (additives) AOAC Enzymatic determination and HPLC Malic acid-d EN Enzymatic determination FU Method No 64 (1995) Malic acid-d in apple juice AOAC HPLC Malic acid-l EN 1138 (1994) FU Method No 21 (1985) Enzymatic determination 13 All juices except citrus based juices 26

27 Pectin (additives) FU Method No 26 (1964/1996) Precipitation/photometry Benzoic acid and its salts; FU Method No 63 (1995) HPLC sorbic acid and its salts NMKL 124 (1997) Benzoic acid and its salts SO 5518:1978 Spectrometry SO 6560: 1983 Preservatives in fruit juices SO 5519: 1978 Spectrometry (sorbic acid and its salts) Quinic, malic & citric acid in cranberry juice cocktail and apple juice (permitted ingredients and additives) Determination of quinic, malic and citric acid in cranberry juice cocktail and apple juice AOAC Saccharin NMKL 122 (1997) Liquid chromatography Soluble solids AOAC EN (1996) FU Method No 8 (1991) SO 2173: 2003 Sucrose (permitted ingredients) Sucrose (permitted ingredients) EN (1996) FU Method No 56 (1985/1998) EN FU Method No 67 (1996) NMKL 148 (1993) HPLC ndirect by refractometry Enzymatic determination Sulphur dioxide (additives) Optimized Monier Williams AOAC Titrimetry after distillation FU method No. 7A (2000) NMKL 132 (1989) Sulphur dioxide (additives) NMKL 135 (1990) Enzymatic determination Sulphur dioxide (additives) SO 5522:1981 SO 5523:1981 Tartaric acid in grape juice EN (1997) (additives) FU Method No 65 (1995) Total nitrogen EN (1997) FU Method No 28 (1991) HPLC Titrimetry after distillation HPLC Digestion/titration 27

28 Sections 3.2 Quality Criteria and 3.3 Authenticity 14 Determination of acetic acid EN 12632; FU Method No 66 (1996) Determination of alcohol (ethanol) FU Method No 52 (1996) Detection of anthocyanins FU Method No 71 (1998) Determination of ash in fruit products AOAC ;EN 1135 (1994); FU Method No 9 (1989) Detection of beet sugar in fruit juices AOAC Determination of benzoic acid as a marker in orange juice AOAC Determination of C 13 /C 12 ratio of ethanol derived from fruit juices JAOAC 79, No. 1, 1996, Determination of carbon stable isotope ratio of apple juice AOAC JAOAC 64, 85 (1981) Determination of carbon stable isotope ratio of orange juice AOAC Determination of carotenoid, total/individual groups EN (1997); FU Method No 59 (1991) Enzymatic determination Enzymatic determination HPLC Deuterium NMR HPLC Stable isotope mass spectrometry Stable isotope mass spectrometry Stable isotope mass spectrometry Spectrophotometry Verification of Composition, Quality and Authenticity Fruit juices and nectars should be subject to testing for authenticity, composition, and quality where applicable and where required. The analytical methods used should be those found in Section 9, Methods of Analysis and Sampling. The verification of a sample s authenticity / quality can be assessed by comparison of data for the sample, generated using appropriate methods included in the standard, with that produced for fruit of the same type and from the same region, allowing for natural variations, seasonal changes and for variations occurring due to processing. 28

29 Determination of centrifugable pulp EN (1997) - FU Method No 60 (1991) Determination of chloride (expressed as sodium chloride) EN12133 (1997) FU Method No 37 (1991) Determination of chloride in vegetable juice AOAC (Codex general method) SO 3634:1979 Determination of essential oils (Scott titration AOAC FU 45b* Determination of essential oils (in citrus fruit) (volume determination)* SO 1955:1982 Determination of fermentability FU Method No 18 (1974) Determination of formol number EN 1133 (1994) FU Method No 30 (1984) Determination of free amino acids EN (xxxx) FU Method No 57 (1989) Determination of fumaric acid FU Method No 72 (1998) Determination of glucose fructose and saccharose EN FU Method No 67 (1996) NMKL 148 (1993) Determination of gluconic acid FU Method No 76 (2001) Determination of glycerol FU Method No 77 (2001) Determination of hesperidin and naringin EN (1996) - FU Method No 58 (1991) Centrifugation/% value Electrochemical titrimetry Titration (Scott) distillation, titration Distillation and direct reading of the volume determination Microbiological method Potentiometric titration Liquid Chromatography HPLC HPLC Enzymatic determination Enzymatic determination HPLC 29

30 Determination of hydroxymethylfurfural FU Method No 69 (1996) Determination of hydroxymethylfurfural SO 7466:1986 Determination of isocitric acid-d FU Method No 54 (1984) Determination of Lactic acid- D and L EN (1999) FU Method No 53 (1983/1996) Determination of L-malic/total malic acid ratio in apple juice AOAC Determination of naringin and neohesperidin in orange juice AOAC Determination of ph-value NMKL 179:2005 EN 1132 (1994);FU Method No 11 (1989);SO 1842: 1991 Determination of phosphorus/phosphate EN 1136 (1994) FU Method No 50 (1983) Determination of proline by photometry non-specific determination EN 1141 (1994); FU Method No 49 (1983) Determination of relative density EN 1131 (1993); FU Method No 1 (1989) & FU Method No General sheet (1971) Determination of Relative density FU Method No 1A Determination of sodium, potassium, calcium, magnesium in fruit juices EN 1134 (1994); FU Method No 33 (1984) Determination of sorbitol-d FU Method No 62 (1995) HPLC Spectrometry Enzymatic determination Enzymatic determination Enzymatic determination and HPLC HPLC Potentiometry Photometric determination Photometry Pycnometry Densitometry Atomic Absorption Spectroscopy Enzymatic determination V 30

31 Determination of stable carbon isotope ratio in the pulp of fruit juices ENV (1998) Analytica Chimica Acta 340 (1997) Determination of stable carbon isotope ratio of sugars from fruit juices ENV Analytica Chimica Acta.271 (1993) Determination of stable hydrogen isotope ratio of water from fruit juices ENV (1997) Determination of stable oxygen isotope ratio in fruit juice water ENV 12141(1997) Detection of starch AOAC (1925) FU Method No 73 (2000) Determination of sugar beet derived syrups in frozen concentrated orange juice δ 18 O Measurements in Water AOAC Determination of titrable acids, total EN (1995) FU Method No Method No 3, (1968) SO 750:1998 Determination of total dry matter (vacuumoven drying at 70 C)* EN (1996) FU Method No 61 (1991) Determination of total solids (Microwave oven drying)* AOAC Determination of Vitamin C (dehydroascorbic acid and ascorbic acid) AOAC Determination of Vitamin C EN : 2004 Stable isotope mass spectrometry Stable isotope mass spectrometry Stable isotope mass spectrometry Stable isotope mass spectrometry Colorimetric Oxygen isotope ratio analysis Titrimetry Gravimetric determination Gravimetric determination Microfluorometry HPLC 31

32 * Because there is no numerical value in the Standard duplicate Type methods have been included which may lead to different results. Milk and Milk Products Milk products ron NMKL 139 (1991) AOAC (Codex general method) Milk products ron NMKL 161 (1998) / AOAC Atomic absorption spectrophotometry Atomic spectrophotometry absorption Milk products ron AOAC nductively Coupled Plasma optical emission spectrophotometry Milk products ron SO 6732 DF 103:2010 Photometry (bathophenanthroline) V Milk and Milk Products Melamine SO/TS DF/RM 230:2010 LC-MS/MS V Milk products (products not completely soluble in ammonia) Blend of evaporated skimmed milk and vegetable fat Blend of evaporated skimmed milk and vegetable fat Blend of evaporated skimmed milk and vegetable fat Reduced fat blend of evaporated skimmed milk and vegetable fat Milkfat DF SO :2005 (Weibull-Berntrop) Total fat SO 1737 DF 13:2008 (Röse-Gottlieb) V Milk solids-not-fat (MSNF) SO 6731 DF 21:2010 and SO 1737 DF 13:2008 Calculation from total solids content and fat content (Röse-Gottlieb) Milk protein in MSNF 15 SO /2 DF 20-1/2:2001/ Titrimetry (Kjeldahl) V AOAC Total fat SO 1737 DF 13:2008 (Röse-Gottlieb) V Milk total solids and Milk solids-not-fat content include water of crystallization of lactose 15 Milk total solids and MSNF content include water of crystallization of lactose 32

33 Reduced fat blend of evaporated skimmed milk and vegetable fat Reduced fat blend of Evaporated skimmed milk and vegetable fat Blend of skimmed milk and vegetable fat in powdered form Blend of skimmed milk and vegetable fat in powdered form Blend of skimmed milk and vegetable fat in powdered form Reduced fat blend of skimmed milk powder and vegetable fat in powdered form Reduced fat blend of skimmed milk powder and vegetable fat in powdered form Reduced fat blend of skimmed milk powder and vegetable fat in powdered form Blend of sweetened condensed skimmed milk and vegetable fat Blend of sweetened condensed skimmed milk and vegetable fat Blend of sweetened condensed skimmed milk and vegetable fat Blend of sweetened condensed skimmed milk and vegetable fat MSNF 15 SO 6731 DF 21:2010 and SO 1737 DF 13:2008 Calculation from total solids content and fat content (Röse-Gottlieb) Titrimetry (Kjeldahl) Milk protein in MSNF 15 SO /2 DF 20-1/2:2001/ V AOAC Total fat SO 1736 DF 9:2008 (Röse-Gottlieb) Water 16 SO 5537 DF 26:2004, drying at 87 C Milk protein in MSNF 15 SO /2 DF 20-1/2:2001/ Titrimetry (Kjeldahl) V AOAC Total fat SO 1736 DF 9:2008 (Röse-Gottlieb) Water Error! Bookmark not defined. SO 5537 DF 26:2004, drying at 87 C Milk protein in MSNF 15 SO /2 DF 20-1/2:2001/ AOAC Titrimetry (Kjeldahl) Total fat SO 1737 DF 13:2008 (Röse-Gottlieb) Sucrose SO 2911 DF 35:2004 Polarimetry V Milk solids-not-fat (MSNF) 15 SO 6734 DF 15:2010 Calculation from total solids content V Milk protein in MSNF 15 SO /2 DF 20-1/2:2001/ AOAC Titrimetry (Kjeldahl) V V 16 Water content excluding the crystallized water bound to lactose (generally known as moisture content ) 33

34 Reduced fat blend of sweetened condensed skimmed milk and vegetable fat Reduced fat blend of sweetened condensed skimmed milk and vegetable fat Reduced fat blend of sweetened condensed skimmed milk and vegetable fat Total fat SO 1737 DF 13:2008 (Röse-Gottlieb) MSNF 15 SO 6734 DF 15:2010 Calculation from total solids content V Milk protein in MSNF 15 SO /2 DF 20-1/2:2001/ AOAC Butter Copper SO 5738 DF 76:2004 AOAC Titrimetry (Kjeldahl) Photometry, diethyldithiocarbamate Butter Lead AOAC (Codex general method) Atomic absorption spectrophotometry Butter Milk solids-not-fat DF 80-2 SO :2001 Butter Milkfat DF 80-3 SO :2003 Butter Milk fat purity SO DF 202:2010 Calculation from determination of triglycerides by gas chromatography Butter Salt SO 1738 DF 12:2004 / AOAC Titrimetry (Mohr: determination of chloride, expressed as sodium chloride) Butter Salt DF 179 SO 15648:2004 Potentiometry (determination of chloride, expressed as sodium chloride) Butter Vegetable fat (sterols) SO DF 159:2006 Gas chromatography Butter Vegetable fat (sterols) SO DF 200:2006 Gas chromatography Butter Water DF 80 SO 37271:2001 Cheese Citric acid SO/TS 2963 DF/RM 34:2006 Enzymatic method V Cheese Citric acid AOAC Photometry Cheese Milkfat SO 1735 DF 5:2004 (Schmid-Bondzynski- Ratslaff) V

35 Cheese Moisture SO 5534 DF 4:2004, drying at 102 C Cheese (and cheese rind) Natamycin SO DF 140-1:2007 Molecular absorption spectrophotometry Cheese Sodium chloride SO 5943 DF 88:2006 Potentiometry (determination of chloride, expressed as sodium chloride) Cheeses, individual Dry matter (Total solids) SO 5534/DF 4: 2004, drying at 102 C Cheeses in brine Milkfat in dry matter (FDM) DF 5 SO 1735:2004 (Schmid-Bondzynski- Ratzlaff) Cottage cheese Fat-free dry matter SO 5534 DF 4:2004 and SO 1735 DF 5:2004 Cottage cheese Milk fat SO 1735 DF 5:2004 Cottage cheese Cheese, Unripened ncluding Fresh Cheese Cream and Prepared Creams Milk fat in dry matter SO DF 124-3:2005 SO DF 124-3:2005 Protein SO /2 DF 20-1/2:2001/ AOAC and SO /2 DF 20-1/2:2001/ Milk protein AOAC Calculation from dry matter content and fat content, drying at 102 C (Schmid-Bondzynski- Ratzlaff) (Schmid-Bondzynski- Ratzlaff) (for samples containing lactose up to 5%) (Weibull-Berntrop) (for samples containing lactose over 5%) (Weibull-Berntrop) Titrimetry, Kjeldahl Titrimetry (Kjeldahl) Cream Milkfat SO 2450 DF 16:2008 (Röse-Gottlieb) Cream Solids SO 6731 DF 21:2010 (drying at 102 C) Creams Lowered in Milkfat Content Milkfat SO 2450 DF 16:2008 / AOAC Creams, Whipped Creams and Fermented Creams Milk solids-not-fat SO DF 80-2:2001 AOAC

36 Cream cheese Dry matter SO 5534 DF 4:2004 drying at 102 C (forced air oven) Cream cheese Moisture on fat free basis SO 5534 DF 4:2004 SO 1735 DF 5:2004 Calculation from fat content and moisture content drying at 102 C (forced air oven) (Schmid-Bondzynski- Ratzlaff) Dairy fat spreads Milk fat purity SO DF 202:2010 Calculation from determination of triglycerides by gas chromatography Dairy fat spreads Total fat SO DF 194:2003 Direct determination of fat using solvent extraction Dairy fat spreads Vegetable fat (sterols) SO DF 159:2006 Gas chromatography Dairy fat spreads Vegetable fat (sterols) SO DF 200:2006 Gas chromatography Edible casein products Acids, free SO 5547 DF 91:2008 Titrimetry (aqueous extract) V Edible casein products Ash (including P 2 O 5 ) SO 5545 DF 90:2008 or 17 (ashing at 825 C ) SO 5544 DF 89:2008 Edible Casein Products Casein in protein SO DF 29-1:2004 Titrimetry, Kjeldahl Edible casein products Copper AOAC Atomic absorption spectrophotometry Edible casein products Copper DF 76 SO 5738:2004 Colorimetry (diethyldiethiocarbamate) Edible casein products Lactose DF 106 SO 5548:2004 Photometry (phenol and H 2 SO 4 ) V 17 refer to scope of methods 36

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