RECOMMENDED METHODS OF ANALYSIS AND SAMPLING

CXS 234-1999

Adopted in 1999.
This document is amended yearly. Last amendment: 2024*

* The most updated version of the method should be used, in application of ISO/IEC 17025. The present list of methods reflects the
amendments adopted by the Forty-seventh Session of the Codex Alimentarius Commission in 2024.
 CXS 234-1999 2

Contents

1. PART A – METHODS OF ANALYSIS BY COMMODITY CATEGORIES AND NAMES

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

Spices and culinary herbs

Sugars and honey

Miscellaneous products
Irradiated foods

2. PART B – METHODS OF SAMPLING BY COMMODITY CATEGORIES AND NAMES

CXS 234-1999 3

PART A – METHODS OF ANALYSIS BY COMMODITY CATEGORIES AND NAMES

All foods
Commodity Provision Method Principle Type
High performance liquid II
All foods Acesulfame K, Aspartame EN 12856
chromatography
High performance liquid II
All foods Cyclamate EN 12857
chromatography
All foods Cyclamate NMKL 123 Spectrophotometry III
High performance liquid
All foods Saccharin EN 12856 III
chromatography
All foods (see also meat products) Nitrates and/or nitrites EN 12014-1 Part 1 – General considerations N/A
EN 1988-1 Part 1 – Optimized Monier-Williams III
Individual foodsi Sulphites
AOAC 990.28 method
EN 1988-2 III
Individual foodsii Sulphites Part 2: Enzymatic method
NMKL 135

Cereals, pulses and legumes and derived products

Commodity Provision Method Principle Type
Certain pulses (soybeans) Moisture ISO 665 Gravimetry (oven drying at 103 oC) I
Certain pulses except soybeans Moisture ISO 24557/AACC 44-17.01 Gravimetry (oven drying at 130 oC)
Degermed maize (corn) meal and Ash AOAC 923.03 Gravimetry I
maize (corn) grits ISO 2171
ICC 104/1
Degermed maize (corn) meal and Fat, crude AOAC 945.38F and 920.39C and ICC 11/1 Calculation from moisture and I
maize (corn) grits Gravimetry (ether extraction)

i
Hominy, fruit juice, seafood.
ii
Wine, dried apples, lemon juice, potato flakes, sultanas, beer.
CXS 234-1999 4

Cereals, pulses and legumes and derived products

Commodity Provision Method Principle Type
Degermed maize (corn) meal and Moisture ICC 110/1 Gravimetry I
maize (corn) grits (oven drying at 130 ºC – 133 oC)
Degermed maize (corn) meal and Particle size (granularity) AOAC 965.22iii Gravimetry (sieving) I
maize (corn) grits
Degermed maize (corn) meal and Protein ICC 105/2 and ICC 110/1 Calculation from moisture and I
maize (corn) grits titrimetry (Kjeldahl digestion)
Durum wheat semolina and durum Ash AOAC 923.03 / ISO 2171 and ISO 712 / Calculation from moisture and I
wheat flour ICC 110/1 gravimetry (incineration at 550 oC)
Durum wheat semolina and durum Moisture ISO 712 / ICC 110/1 Gravimetry (oven drying at 130 ºC – I
wheat flour 133 oC)
Durum wheat semolina and durum Protein ICC 105/2 and ISO 712 / ICC 110/1 Calculation from moisture and I
wheat flour titrimetry (Kjeldahl digestion)
Instant noodles Extraction of oil from instant See Appendix I, Part A Gravimetry I
noodles
Instant noodles Acid value See Appendix I, Part B Titrimetry (ether extraction) I

Instant noodles Moisture See Appendix I, Part C Gravimetry (oven drying at 105 oC) I
Maize (corn) Moisture ISO 6540 / ICC 110/1 Gravimetry (oven drying at 130 ºC – I
133 oC)
Pearl millet flour Ash AOAC 923.03 / ISO 2171 and ISO 712 / Calculation from moisture and I
ICC 110/1 gravimetry (incineration at 550 oC)
Pearl millet flour Colour ISO 16624 Diffuse reflectance colorimetry I
(specific colour grader)
Pearl millet flour Fat, crude AOAC 945.38F and 920.39C and ISO 712 / Calculation I
ICC 110/1 from moisture and gravimetry (ether
extraction)
Pearl millet flour Fibre, crude ISO 5498 and ISO 712 / ICC 110/1 Calculation from moisture and I
gravimetry (extraction and filtration)

iii
Sieve specifications as in ISO 3310/1.
CXS 234-1999 5

Cereals, pulses and legumes and derived products

Commodity Provision Method Principle Type
Pearl millet flour Moisture ISO 712 / ICC 110/1 Gravimetry (oven drying at 130 ºC– I
133 oC)
Pearl millet flour Protein ISO 20483 and ISO 712 / ICC 110/1 Calculation from moisture and I
titrimetry (Kjeldahl digestion)
Quinoa Moisture ISO 712 Gravimetry (oven drying) I
Quinoa Protein ISO 1871 Titrimetry (Kjeldahl digestion) IV
Sorghum flour Ash AOAC 923.03 Gravimetry I
ISO 2171
ICC 104/1
Sorghum flour Colour ISO 16624 Diffuse reflectance colorimetry I
(specific colour grader)
Sorghum flour Fat, crude AOAC 945.38F and 920.39C and ISO 712 / Calculation from moisture and I
ICC 110/1 gravimetry (ether extraction)
Sorghum flour Fibre, crude ICC 113 / ISO 6541 and ISO 712 / ICC Calculation from moisture and I
110/1 gravimetry (separation, incineration)
Sorghum flour Moisture ISO 712 / ICC 110/1 Gravimetry (oven drying at I
130 ºC – 133 oC)

Sorghum flour Particle size (granularity) AOAC 965.22iv Sieving I

Sorghum flour Protein ICC 105/2 and ISO 712 / ICC 110/1 Calculation from moisture and I
titrimetry (Kjeldahl digestion)
Sorghum flour Tannins ISO 9648 and ISO 712 / ICC 110/1 Calculation from moisture and I
spectrophotometry
Sorghum grains Ash AOAC 923.03 Gravimetry I
ISO 2171
ICC 104/1
Sorghum grains Fat, crude AOAC 945.38F and 920.39C and ISO 6540 Calculation from moisture and I
gravimetry (ether extraction)

iv
Sieve specifications as in ISO 3310/1.
CXS 234-1999 6

Cereals, pulses and legumes and derived products

Commodity Provision Method Principle Type
Sorghum grains Moisture ISO 6540 Gravimetry (oven drying at 130 ºC – I
133 oC)
Sorghum grains Protein ICC 105/2 and ISO 6540 Titrimetry, Kjeldahl digestion I
Sorghum grains Tannins ISO 9648 and ISO 6540 Calculation from moisture and I
spectrophotometry
Soy protein products Ash AOAC 923.03 Gravimetry I
ISO 2171: (Method B)
Soy protein products Fat ISO 734 Gravimetry (extraction) I
Soy protein products Fibre, crude ISO 5498 and AOAC 925.09 Calculation from moisture and I
gravimetry (extraction and filtration)
Soy protein products Moisture AOAC 925.09 Gravimetry (vacuum oven 98 ºC – I
100 oC)
Soy protein products Crude protein: excluding AOCS Ba 4f-00 Gravimetry (combustion) IV
added vitamins, minerals,
AACCI 46.30 IV
amino acids and food additives
ISO 16634-1 IV
Vegetable protein products Ash AOAC 923.03 Calculation from moisture and I
ISO 2171 and AOAC 925.09 gravimetry (incineration at 550 oC)
Vegetable protein products Fat ISO 734 Gravimetry (extraction) I
Vegetable protein products Fibre, crude AACC 32-10.01 and AOAC 925.09 Calculation from moisture and I
gravimetry (ceramic fibre filtration)
Vegetable protein products Moisture AOAC 925.09 Gravimetry (vacuum oven at 98 C – I
100 oC)
Vegetable protein products Crude protein: excluding AOCS Ba 4f-00 Gravimetry (combustion) IV
added vitamins, minerals,
AACCI 46.30 IV
amino acids and food additives
ISO 16634-1 IV
CXS 234-1999 7

Cereals, pulses and legumes and derived products

Commodity Provision Method Principle Type
Wheat flour Ash AOAC 923.03 Gravimetry I
ISO 2171
ICC 104/1
Wheat flour Fat acidity ISO 7305 and ISO 712 / ICC 110/1 Calculation from moisture and I
titrimetry (extraction)
Wheat flour Moisture ISO 712: Gravimetry (oven drying at 130 ºC I
ICC 110/1 and 133 oC)
Wheat flour Particle size (granularity) AOAC 965.22v Gravimetry (sieving) I
Wheat flour Protein ICC 105/2 and ISO 712 / ICC 110/1 Calculation from moisture and I
titrimetry (Kjeldahl digestion)
Wheat protein products including Crude protein excluding added Vital wheat gluten and devitalized wheat Calculation from moisture and I
wheat gluten vitamins, minerals, amino gluten titrimetry (Kjeldahl digestion)
acids and optional ingredients ISO 20483 and AOAC 925.09
Solubilized wheat protein Calculation from moisture and I
ISO 20483 and AOAC 925.09 titrimetry (Kjeldahl digestion)
Wheat protein products including Fibre, crude AOAC 962.09 and AOAC 925.09 Calculation from moisture and I
Wheat gluten gravimetry (ceramic fibre filtration)
Wheat protein products including Moisture AOAC 925.09 Gravimetry (vacuum oven at 98 oC – I
wheat gluten 100 oC)
Wheat protein products including Ash AOAC 923.03 Calculation from moisture and I
Wheat gluten ISO 2171 and AOAC 925.09 gravimetry (incineration at 550 oC)

Whole and decorticated pearl millet Ash AOAC 923.03 / ISO 2171 and ISO 712/ ICC Calculation from moisture and I
grains 110/1 gravimetry (incineration at 550 oC)
Whole and decorticated pearl millet Fat, crude AOAC 945.38F and 920.39C and ISO 712/ Calculation from moisture and I
grains ICC 110/1 gravimetry (ether extraction)

v
Sieve specifications as in ISO 3310/1.
CXS 234-1999 8

Cereals, pulses and legumes and derived products

Commodity Provision Method Principle Type
Whole and decorticated pearl millet Fibre, crude ISO 5498 and ISO 712 / ICC 110/1 Calculation from moisture and I
grains gravimetry (filtration through filter
paper)
Whole and decorticated pearl millet Moisture ISO 712 Gravimetry (oven drying 130 ºC – I
grains ICC 110/1 133 oC)

Whole and decorticated pearl millet Protein ISO 20483 and ISO 712 / ICC 110/1 Calculation from moisture and I
grains titrimetry (Kjeldahl digestion)
Whole maize (corn) meal Ash AOAC 923.03 Gravimetry I
ISO 2171
ICC 104/1
Whole maize (corn) meal Fat, crude AOAC 945.38Fand 920.39C and ICC 110/1 Calculation from moisture and I
gravimetry (ether extraction)
Whole maize (corn) meal Moisture ICC 110/1 Gravimetry (oven drying 130 ºC – I
ISO 6540 133 oC)
Whole maize (corn) meal Particle size (granularity) AOAC 965.22vi Sieving I
Whole maize (corn) meal Protein ICC 105/2 and ICC 110/1 Calculation from moisture and I
titrimetry (Kjeldahl digestion)

vi
Sieve specifications as in ISO 3310/1.
CXS 234-1999 9

Cocoa products and chocolate

Commodity Provision Method Principle Type

Chocolate and chocolate products Cocoa butter AOAC 963.15 Gravimetry (Soxhlet extraction) I
IOCCC 14
Chocolate and chocolate products Fat-free cocoa solids AOAC 931.05 Oven evaporation and factor I
Chocolate and chocolate products Fat-free milk solids IOCCC 17 or AOAC 939.02 Titrimetry, Kjeldahl digestion; after II
extraction of milk proteins
Chocolate and chocolate products Fat, total AOAC 963.15 Gravimetry (Soxhlet extraction) I
Chocolate and chocolate products Milkfat IOCCC 5 Titrimetry/Distillation I
AOAC 945.34; 925.41B; 920.80
Chocolate and chocolate products Moisture IOCCC 26 or AOAC 977.10 (Karl Fischer Gravimetry I
method); or AOAC 931.04 or
IOCCC 1
Chocolate and chocolate products Non-cocoa butter vegetable fat AOCS Ce 10/02 and described in the Described in the standard I
standard
Cocoa (cacao) mass or cocoa/ Cocoa shell AOAC 968.10 and 970.23 Spiral vessel count, stone cell count I
chocolate liquor, and cocoa cake
Cocoa (cacao) mass or cocoa/ Fat AOAC 963.15 or IOCCC 14 Gravimetry (Soxhlet extraction) I
chocolate liquor, and cocoa cake
Cocoa butter Free fatty acids ISO 660 or AOCS Cd 3d-63 Titrimetry I
Cocoa butter Unsaponifiable matter ISO 3596 or ISO 18609 or AOCS Ca 6b-53 Titrimetry after extraction with diethyl I
ether

Cocoa powders (cocoa) and dry Moisture IOCCC 26 or AOAC 977.10 (Karl Fischer Gravimetry I
cocoa-sugar mixtures method)
CXS 234-1999 10

Fats and oils and related products

Commodity Provision Method Principle Type

Fats and oils (all) Arsenic AOAC 963.21 and AOAC 942.17 Kjeldahl flask digestion and III
colorimetry (molybdenum blue)
Fats and oils (all) Arsenic AOAC 963.21 and AOAC 952.13 Kjeldahl flask digestion and III
colorimetry (diethyldithiorcarbamate)
Fats and oils (all) Arsenic AOAC 986.15 Atomic absorption spectrophotometry II
(hydride generation)
Fats and oils (all) Insoluble impurities ISO 663 Calculation from total insoluble I
content in n-hexane or light
petroleum. Gravimetry,
drying at 103 0C
Fats and oils (all) Moisture and volatile matter ISO 662 Gravimetry, drying at 103 0C I
Fats and oils (all) Soap content ISO 10539 / AOCS Cc 17-95 Titrimetry (colorimetric) I
Fats and oils Synthetic phenolic antioxidants AOCS Ce 6a-2021 Liquid chromatography II
Fats and oils Synthetic phenolic antioxidants AOAC 983.15 Liquid chromatography III
Fats and oils not covered by individual Acidity: acid value ISO 660 / AOCS Cd 3d-63 Titrimetry I
standards
Fats and oils not covered by individual Copper and iron AOAC 990.05 / ISO 8294 / AOCS Ca 18b- Atomic absorption spectrophotometry II
standards 91 (direct graphite furnace)
Fats and oils not covered by individual Peroxide value AOCS Cd 8b-90 Titrimetry (colorimetric) I
standards ISO 3960 / NMKL 158

Fat spreads and blended spreads Total fat ISO 17189 | IDF 194 Gravimetry, direct determination of fat I
using solvent extraction
Fish oils Fatty acid composition AOCS Ce 2c-66 and AOCS Ce 1i-07 / Preparation of methyl esters and gas III
AOCS Ce 1j-07 chromatography
Fish oils Fatty acid composition ISO 12966-2 and ISO 12966-4 Preparation of methyl esters and gas III
chromatography
CXS 234-1999 11
Fats and oils and related products

Commodity Provision Method Principle Type

Fish oils Acidity: acid value AOCS Ca 5a-40 / AOCS Cd 3d-63 / ISO Titrimetry I
660 / NMKL 38
Fish oils Peroxide value AOCS Cd 8b-90 / ISO 3960 / NMKL 158 / Titrimetry (colorimetric) I
European Pharmacopoeia 2.5.5
Fish oils Phospholipids USP-FCC 12 2S (Krill oil – phospholipids), Nuclear magnetic resonance I
spectroscopy
Fish oils P-Anisidine value European Pharmacopoeia 2.5.36/ Spectrophotometry I
AOCS Cd 18-90/
ISO 6885
Fish oils Triglycerides AOCS Cd 11d-96 Liquid chromatography with II
evaporative light scattering detection
Fish oils Triglycerides European Pharmacopoeia 1352 Liquid chromatography with III
refractive index detection
Fish oils Triglycerides USP 40 NF37 Liquid chromatography with III
refractive index detection
Fish oils Vitamin Avii European Pharmacopoeia Monograph on Liquid chromatography III
Cod Liver Oil (Type A), Monograph
01/2005:1192, with LC end-point 2.2.29
Fish oils Vitamin Avii EN 12823-1 Liquid chromatography II
Fish oils Vitamin D EN 12821 (Determination of vitamin D by Liquid chromatography III
high performance liquid chromatography –
Measurement of cholecalciferol (D3) or
ergocalciferol (D2))
Fish oils Vitamin Dviii NMKL 167 / EN 12821 Liquid chromatography II
Fish oils (calanus oil) Wax content AOCS Ch 8-02 Gas chromatography (FID) IV

vii
The respective Standard for fish oils (CXS 329-2017) states that vitamin A is expressed as 'retinol equivalents' (RE) where RE takes into account the fact that different vitamers of vitamin A differ in activity.
ISO/TR 23304:2021 "Food products – Guidance on how to express vitamins and their vitamers".
viii
The provisions account for vitamins D2 and D3.
CXS 234-1999 12
Fats and oils and related products

Commodity Provision Method Principle Type

Named animal fats Acidity: acid value ISO 660 / AOCS Cd 3d-63 Titrimetry I
Named animal fats Fatty acid composition ISO 12966-2 and ISO 12966-4 Preparation of methyl esters and gas III
chromatography
Named animal fats Copper and iron AOAC 990.05 / ISO 8294 / AOCS Ca 18b- Atomic absorption spectrophotometry II
91 (direct graphite furnace)
Named animal fats Iodine value (IV) ISO 3961 / AOAC 993.20 / AOCS Cd 1d-92 Titrimetry (Wijs) I
/ NMKL 39
Named animal fats Peroxide value AOCS Cd 8b-90 / ISO 3960 / NMKL 158 Titrimetry (colorimetric) I
Named animal fats Relative density ISO 6883, with the appropriate conversion Pycnometry I
factor / AOCS Cc 10c-95
Named animal fats Refractive index ISO 6320 / AOCS Cc 7-25 Refractometry II
Named animal fats Saponification value ISO 3657 / AOCS Cd 3-25 Titrimetry (colorimetric) I
Named animal fats Unsaponifiable matter ISO 3596 / ISO 18609 / AOCS Ca 6b-53 Gravimetry, drying at 103 0C and I
titrimetry (colorimetry)
Named animal fats Titre ISO 935 Thermometry I
Named animal fats Titre AOCS Cc 12-59ix Thermometry IV

Named vegetable oils Acidity: ISO 660 / AOCS Cd 3d-63 / AOCS Ca 5a- Titrimetry I
40
Acid value
Named vegetable oils Free fatty acids ISO 660 / AOCS Cd 3d-63 / AOCS Ca 5a- Titrimetry I
40
Named vegetable oils Apparent density ISO 6883 / AOCS Cc 10c-95 Pycnometry I

Named vegetable oils Baudouin test (modified AOCS Cb 2-40 Colour reaction I
Villavecchia or sesame seed
oil test)

ix
AOCS Cc 12-59 is the preferred method in certain regions. Due to difference in practical application of AOCS Cc 12-59 compared to ISO 935, it is listed as a Type IV method.
CXS 234-1999 13
Fats and oils and related products

Commodity Provision Method Principle Type

Named vegetable oils Carotenoids, total BS 684-2.20 Spectrophotometry II
Named vegetable oils Copper and iron AOAC 990.05 / ISO 8294 / AOCS Ca 18b- Atomic absorption spectrophotometry II
91 (direct graphite furnace)

Named vegetable oils Crismer value AOCS Cb 4-35 and AOCS Ca 5a-40 Calculation from individual fatty acid I
composition (gas chromatography of
methyl esters) and turbidity

Named vegetable oils Fatty acid composition ISO 12966-2 and ISO 12966-4 / AOCS Ce Gas chromatography of methyl esters II
2-66 and AOCS Ce 1h-05
Named vegetable oils Halphen test AOCS Cb 1-25 Colorimetry I
Named vegetable oils Insoluble impurities ISO 663 Calculation from total insoluble I
content in n-hexane or light
petroleum.
Gravimetry, drying at 103 0C

Named vegetable oils Iodine value ISO 3961 / AOAC 993.20 / AOCS Cd 1d-92 Titrimetry (Wijs) I
/ NMKL 39
Named vegetable oils Moisture and volatile matter ISO 662 Gravimetry, drying at 103 0C I

Named vegetable oils Peroxide value (PV) AOCS Cd 8b-90 / ISO 3960 / NMKL 158 Titrimetry (colorimetric) I

Named vegetable oils Refractive index ISO 6320 / AOCS Cc 7-25 Refractometry II

Named vegetable oils Reichert-Meissi value and AOCS Cd 5-40 Calculation from soluble and insoluble I
Polenske value volatile fatty acids. Titrimetry
(colorimetric)

Named vegetable oils Relative density ISO 6883 / AOCS Cc 10c-95 Pycnometry I
Named vegetable oils Saponification value (SV) ISO 3657 / AOCS Cd 3-25 Titrimetry (colorimetric) I

Named vegetable oils Slip point ISO 6321 / AOCS Cc 3b-92 for all oils,
Open ended capillary tube I
except palm oils
or AOCS Cc 3-25 for palm oils only
Named vegetable oils Sterol composition and total ISO 12228-1 / AOCS Ch 6-91 Thin-layer chromatography and gas II
sterols chromatography
CXS 234-1999 14
Fats and oils and related products

Commodity Provision Method Principle Type

Named vegetable oils Tocopherol content ISO 9936 / AOCS Ce 8-89 Liquid chromatography with II
fluorescence detection

Named vegetable oils Unsaponifiable matter ISO 3596 / AOCS Ca 6b-53 Diethyl ether extraction and I
gravimetry, drying at 103 0C and
titrimetry (colorimetry) and correction
for free fatty acids titrimetry
(colorimetry)

Named vegetable oils Unsaponifiable matter ISO 18609x Hexane extraction and gravimetry, IV
drying at 103 ºC and titrimetry
(colorimetry) and correction for free
fatty acids titrimetry (colorimetry) xi
4𝛼-desmethylsterol and total Separation and Gas chromatography
Olive oils and olive pomace oils COI/T.20/Doc. No 26 II
sterol content (FID)
4𝛼-desmethylsterol and total Separation and Gas chromatography
Olive oils and olive pomace oils ISO 12228-2 (part 2) III
sterol content (FID)
4𝛼-desmethylsterol and total Separation and Gas chromatography
Olive oils and olive pomace oils AOCS Ch 6-91 III
sterol content (FID)
Olive oils and olive pomace oils Absorbency in ultraviolet COI/T.20/Doc. No. 19 / ISO 3656 / AOCS Absorption in ultraviolet I
Ch 5-91
Olive oils and olive pomace oils Acidity, free (acid value) ISO 660 (Section 9.1) / AOCS Cd 3d-63 / Titrimetry I
COI/T.20/Doc. No. 34
Olive oils and olive pomace oils Alpha-tocopherol ISO 9936 HPLC (UV or fluorescence) II
Olive oils and olive pomace oils Alpha-tocopherol AOCS Ce 8-89 HPLC (UV or fluorescence) III
Olive oils and olive pomace oils Difference between the actual COI/T.20/Doc. No. 20 and COI/T.20/Doc. Analysis of triglycerides by HPLC I
and theoretical ECN 42 No. 33 and fatty acids by GC followed by
triglyceride content calculation
Olive oils and olive pomace oils Erythrodiol + uvaol COI/T.20/Doc. No. 26 Separation and gas chromatography II
(FID)
Olive oils and olive pomace oils Fatty acid composition COI/T.20/Doc. No. 33 Gas chromatography (FID) of methyl II
esters

x
Results obtained from ISO 18609 are systematically lower. In case of limitations due to climate or regulations that prohibit the use of diethyl ether, ISO 18609 can be used instead of the Type I method.
xi
The technique in ISO 18609 is gravimetric. The correction by titration and colorimetry is only when it is necessary to correct for free fatty acids.
CXS 234-1999 15
Fats and oils and related products

Commodity Provision Method Principle Type

Olive oils and olive pomace oils Fatty acid composition AOCS Ce 2-66 and AOCS Ch 2-91 / Ce 1h- Gas chromatography (FID) of methyl III
05 esters
Olive oils and olive pomace oils Fatty acid composition ISO 12966-2 and ISO 12966-4 Gas chromatography (FID) of methyl III
esters
Olive oils and olive pomace oils 2-glyceryl monopalmitate COI/T.20/Doc. No. 23 Hydrolysis and derivatization gas II
percentage chromatography (FID)
Olive oils and olive pomace oils Fatty acid ethyl ester content COI/T.20/Doc. No. 28 Gas chromatography (FID) II

Olive oils and olive pomace oils Halogenated solvents, traces ISO 16035 Headspace gas chromatography II
(ECD)
Olive oils and olive pomace oils Insoluble impurities in light ISO 663 Gravimetry I
petroleum
Olive oils and olive pomace oils Iodine value ISO 3961 / AOAC 993.20 / AOCS Cd 1d-92 Wijs-titrimetry I
/ NMKL 39
Olive oils and olive pomace oils Moisture and volatile matter ISO 662 Gravimetry I

Olive oils and olive pomace oils Organoleptic characteristics COI/T.20/Doc. No. 15 Sensory Panel test I

Olive oils and olive pomace oils Peroxide value ISO 3960 / AOCS Cd 8b-90 / NMKL 158 Titrimetry I

Olive oils and olive pomace oils Relative density ISO 6883 / AOCS Cc 10c-95 Pycnometry I

Olive oils and olive pomace oils Refractive index ISO 6320 / AOCS Cc 7-25 Refractometry II

Olive oils and olive pomace oils Saponification value ISO 3657 / AOCS Cd 3-25 Titrimetry I

Olive oils and olive pomace oils Stigmastadienes COl/T.20/Doc. no. 11 Gas chromatography (FID) II

Olive oils and olive pomace oils Stigmastadienes AOCS Cd 26-96 Gas chromatography (FID) III

Olive oils and olive pomace oils Stigmastadienes ISO 15788-2 HPLC III

Olive oils and olive pomace oils Stigmastadienes ISO 15788-1 Gas chromatography (FID) III

Olive oils and olive pomace oils Trans fatty acids content COI/T.20/Doc. No. 33 Gas chromatography (FID) of methyl II
esters
Olive oils and olive pomace oils Trans fatty acids content ISO 12966-2 and ISO 12966-4 Gas chromatography (FID) of methyl III
esters
CXS 234-1999 16

Fats and oils and related products

Commodity Provision Method Principle Type
Olive oils and olive pomace oils Trans fatty acids content AOCS Ce 2-66 and AOCS Ce 1h-05 Gas chromatography (FID) of III
methyl esters
Olive oils and olive pomace oils Unsaponifiable matter ISO 3596 / AOCS Ca 6b-53 Gravimetry
I

Olive oils and olive pomace oils Wax content COI/T.20/Doc. No. 28 Gas chromatograph (FID) II

Olive oils and olive pomace oils Wax content AOCS Ch 8-02 Gas chromatography (FID)
III

Table 1. Method performance criteria for iron and copper in olive oils and olive pomace oils

Method performance criteria Example

of
ML Minimum Limit of Limit of methods
Commodity Provision Principle
(mg/kg) applicable detection quantification Recovery that
Precision (RSDR) (%) no more than meet the
range (LOD) (LOQ) (%)
(mg/kg) (mg/kg) (mg/kg) criteria

ISO 8294
AOAC
990.05 GF-AAS
Olive oils
AOCS GF-AAS
and olive Iron 3.0 1.8–4.2 0.3 0.6 27 80–110
Ca 17a- ICP-OES
pomace oils
18 ICP-OES
ISO
21033

ISO 8294
AOAC
Olive oils GF-AAS
990.05
and olive Copper 0.1 0.03–0.17 0.01 0.02 44 80–110 GF-AAS
AOCS
pomace oils ICP-OES
Ca 17a-
18
CXS 234-1999 17

Fish and fishery products

Commodity Provision Method Principle Type

Canned tuna and bonito Determination of presentation See Appendix II
Crackers from marine and freshwater Crude protein AOAC 2001.11 Titrimetry (Kjeldahl digestion) IV
fish, crustacean and molluscan
shellfish
Crackers from marine and freshwater Crude protein Described in the standard
fish, crustacean and molluscan
shellfish
Crackers from marine and freshwater Moisture Described in the standard
fish, crustacean and molluscan
shellfish
Fish and fishery products, except raw Drained weight See Appendix III Gravimetry I
bivalve molluscs (shucked)
Raw bivalve molluscs (shucked) Drained weight Described in the standard
Fish and fishery products, except Net weight See Appendix III Gravimetry I
frozen abalone (covered by glaze),
raw fresh chilled or frozen abalone,
quick-frozen raw scallop products, raw
bivalve molluscs
Fish and fishery products Washed drained weight See Appendix III Gravimetry I
Fish and fishery products Sensory and physical Described in the standard and see Sensory analysis, visual inspections, I
determinations Appendix IV and CXG 31-1999 counting
Fish sauce Total nitrogen AOAC 978.02 Titrimetry (Kjeldahl digestion) I
Fish sauce Amino acid nitrogen AOAC 920.04 and AOAC 920.03 Determining formaldehyde titration I
method
Subtracting by ammoniacal nitrogen
(magnesium oxide method)
Fish sauce pH NMKL 179 Potentiometry II
CXS 234-1999 18
Fish and fishery products

Commodity Provision Method Principle Type

Fish sauce pH AOAC 981.12 Potentiometry IV
The pH shall be measured in a sample of
fish sauce diluted with water to 1:10 using a
pH metre. The dilution of fish sauce is
necessary because of the high ionic
strength in the undiluted sauce
Frozen abalone (covered by glaze) Net weight AOAC 963.18 Gravimetry I
Frozen fish and fishery products Thawing and cooking See Appendix V Thawing and heating I
procedures
CXS 234-1999 19
Fish and fishery products

Commodity Provision Method Principle Type

Quick-frozen blocks of fish fillet, Proportion of fish fillet and AOAC 988.09 Physical separation I
minced fish flesh and mixtures of fillets minced fish
and minced fish flesh
Quick-frozen fish fillets Gelatinous condition AOAC 983.18 and AOAC 950.46A Gravimetry I
(determined as moisture)
Quick-frozen finfish, uneviscerated Gelatinous condition AOAC 983.18 and AOAC 950.46A Gravimetry I
and eviscerated (determined as moisture)
Quick-frozen blocks of fish fillet, Gelatinous condition AOAC 983.18 and AOAC 950.46A Gravimetry I
minced fish flesh and mixtures of fillets (determined as moisture)
and minced fish flesh
Quick-frozen fish sticks (fish fingers) Fish content (declaration) AOAC 996.15 and calculation (See Gravimetry I
and fish portions – breaded or in batter Appendix VI)
Quick-frozen fish sticks (fish fingers) Proportion of fish fillet and WEFTA Method – See Appendix VII Gravimetry I
and fish portions-breaded and in batter minced fish
Quick-frozen fish sticks (fish fingers), Gelatinous condition AOAC 983.18 and AOAC 950.46A Gravimetry I
fish portions and fish fillets – breaded (determined as moisture)
or in batter
Quick-frozen raw scallop products Net weight AOAC 963.18 and Appendix III Gravimetry I
Raw bivalve molluscs Net weight AOAC 963.18 and Appendix III Gravimetry I
Raw fresh chilled or frozen abalone Net weight AOAC 963.18 Gravimetry I
Salted Atlantic herring and salted sprat Water content (determined as AOAC 950.46B a) Gravimetry I
moisture)
Salted Atlantic herring and salted sprat Determination of salt content See Appendix VIII
and sturgeon caviar
Live and raw bivalve molluscs Determination of Escherichia See Appendix IX
coli
Smoked fish, smoke-flavoured fish and Determination of Listeria See Appendix IX
smoke-dried fish monocytogenes
Smoked fish, smoke-flavoured fish and Determination of Clostridium See Appendix IX
smoke-dried fish botulinum
CXS 234-1999 20
Fish and fishery products

Commodity Provision Method Principle Type

Salted fish and dried salted fish of the Moisture AOAC 937.07 and AOAC 950.46B (air- Gravimetry I
Gadidae family of fishes drying (a))
Salted fish and dried salted fish of the Salt saturation See equation in footnotexii Calculation I
Gadidae family of fishes
Salted fish and dried salted fish of the Water content in the whole See Appendix VIII Gravimetry I
Gadidae family of fishes fish
Smoked fish, smoke-flavoured fish Water phase salt (salt AOAC 952.08 and sodium chloride see Gravimetry and titrimetry and I
and smoke-dried fish determined as chloride method criteria in Table 5 calculation
expressed as sodium chloride) Water phase salt = (% salt x 100) / (%
water + % salt)

Smoked fish, smoke-flavoured fish Water activity NMKL 168 | Electrometry II

and smoke-dried fish ISO 18787
Live and raw bivalve molluscs Paralytic shellfish toxicity AOAC 959.08 Mouse bioassay IV
Live and raw bivalve molluscs Paralytic shellfish toxicity AOAC 2011.27 Receptor binding assay IV

Table 2. Method performance criteria for histamine for fish and fishery products
Provision ML Minimum LOD LOQ RSDR (%) Recovery (%) Examples of Principle
(mg/100 g) applicable range (mg/100 g) (mg/100 g) no more applicable methods
(mg/100 g) than that meet the criteria
Histamine 10 (average) 8–12 1 2 16 90–107 AOAC 977.13 / NMKL Fluorometry,
99, HPLC-UV, HPLC-
NMKL 196, ISO 19343 UV, HPLC-FLD
Histamine 20 16–24 2 4 14 90–107 AOAC 977.13 / NMKL Fluorometry,
(each unit) 99, HPLC-UV, HPLC-
NMKL 196, ISO 19343 UV, HPLC-FLD

xii
The % salt saturation is calculated as follows:
1. % salt in water = (% salt content / (% salt content + % moisture)) x 100%
2. % salt saturation = (% salt in water / 26.4 %*) x 100%
* The solubility of sodium chloride in water is 36 g per 100 g water, and the constant is calculated as follows: 36 g sodium chloride / (100 g water + 36 g sodium chloride) x 100% = 26.4%
CXS 234-1999 21
Determination of biotoxins in live and raw bivalve molluscs, live abalone and raw fresh chilled or frozen abalone
The method selected should be chosen on the basis of practicability and preference should be given to methods which have applicability for routine use.
Method performance criteria for determination of toxin analogues by chemical methods
Methods shall meet the numerical criteria listed in Table 3 and may either meet the minimum applicable range, or LOD and LOQ criteria listed.

Table 3. Method performance criteria for determination of toxin analogues by chemical methods
Toxin group Toxin Minimum LOD LOQ Precision (RSDR) Recovery (%) Examples of applicable
applicable (mg/kg) (mg/kg) (%) methods that meet the
range (mg/kg) no more than criteria
STX group Saxitoxin 0.05–0.2 0.01 0.02 44 50–130 AOAC 2005.06 (HPLC-FLD)
(STX) NMKL 182 (HPLC-FLD), EN
NEO 0.05–0.2 0.01 0.02 44 50–130 14526 (HPLC-FLD)
dcSTX 0.05–0.2 0.01 0.02 44 50–130 AOAC 2011.02 (HPLC-FLD)
GTX1 0.05–0.2 0.01 0.02 44 50–130 NMKL 197 (HPLC-FLD),
GTX2 0.1–0.5 0.03 0.06 38 50–130 Turner et al. (2020)
J.AOAC Int. Vol. 103, No. 2,
GTX3 0.1–0.5 0.03 0.06 38 50–130 p533-62 (uHPLC-MSMS)
GTX4 0.05–0.2 0.01 0.02 44 50–130
GTX5 0.1–0.5 0.03 0.06 38 50–130
GTX6 0.1–0.5 0.03 0.06 38 50–130
dcGTX2 0.1–0.5 0.03 0.06 38 50–130
dcGTX3 0.1–0.5 0.03 0.06 38 50–130
C1 0.1–0.5 0.03 0.06 38 50–130
C2 0.1–0.5 0.03 0.06 38 50–130
C3 0.5–1.5 0.1 0.2 32 50–130
C4 0.5–1.5 0.1 0.2 32 50–130
OA group OA 0.03–0.2 0.01 0.02 44 EU-harmonized SOP using
70–130 HPLC-MSMS – see reference
DTX1 0.03–0.2 0.01 0.02 44 60–115 below*
70–130 For other methods see
references **
DTX2 0.1–0.5 0.03 0.06 38 60–115
70–130
Domoic aid DA 14–26 2 4 20 70–130 EN 14176 (HPLC-UV)
AOAC 991.26 (HPLC-UV)
CXS 234-1999 22
Toxin group Toxin Minimum LOD LOQ Precision (RSDR) Recovery (%) Examples of applicable
applicable (mg/kg) (mg/kg) (%) methods that meet the
range (mg/kg) no more than criteria
AZA group AZA1 0.03–0.2 0.01 0.02 44 EU-harmonized SOP using
70–130 HPLC-MSMS – see reference
AZA2 0.03–0.2 0.01 0.02 44 70–130 below*
AZA3 0.03–0.2 0.01 0.02 44 70–130 For other methods see
references**
* https://www.aesan.gob.es/en/CRLMB/docs/docs/metodos_analiticos_de_desarrollo/EU-Harmonized-SOP-LIPO-LCMSMS_Version5.pdf
** H.J. van den Top, A. Gerssen, P. McCarron, H.P. van Egmond. Quantitative determination of marine lipophilic toxins in mussels, oysters and cockles using liquid chromatography-mass spectrometry: inter-
laboratory validation study. Food Additives & Contaminants: Part A, 2011, Vol. 28, Iss. 12.
Total toxicity is estimated as the sum of the molar concentrations of detected analogues multiplied by the relevant specific toxicity equivalency factors (TEFs). Internationally scientifically validated TEFs must
be used. The science behind TEFs is developing. Current internationally validated TEFs are available on the FAO website.
Methods should be validated and used for the relevant toxin analogues that may contribute to total toxicity. Currently known toxin analogues to consider are listed in Table 3.
Where toxin analogues that are not listed in Table 3 are determined the competent authority must assess the contribution of these analogues to total toxicity while conducting further investigations.

Table 4. Method performance criteria for screening and for determination of methylmercury*
Commodity Provision ML Min appl. LOD LOQ Precision Recovery Examples of Principle
range (%) (%) applicable
(mg/kg) (mg/kg) (mg/kg)
methods that
(mg/kg) not more
meet the criteria
than
Tuna (all methylmercury* 1.2 0.64–1.8 0.12 0.24 31 80–110 EN 16801 / GC-ICP/MS
species) NMKL 202
AAS-flame
AOAC 977.15**
ICP-MS
NMKL 186** /
ICP-MS
AOAC 2013.06**
ICP-MS
/ EN 15763**
Alfonsino methylmercury* 1.5 0.82–2.2 0.15 0.30 30 80–110 AOAC 988.11 GC-electron capture
EN 16801 / GC-ICP/MS
NMKL 202
AAS-flame
AOAC 977.15**
ICP-MS
NMKL 186** /
ICP-MS
AOAC 2013.06**
ICP-MS
/ EN 15763**
CXS 234-1999 23

Marlin (all methylmercury* 1.7 0.95–2.5 0.17 0.34 30 80–110 AOAC 988.11 GC-electron capture
species) GC-ICP/MS
EN 16801 /
NMKL 202 AAS-flame
AOAC 977.15** ICP-MS
NMKL 186** / ICP-MS
AOAC 2013.06** ICP-MS
/ EN 15763**
Shark (all methylmercury* 1.6 0.88–2.3 0.16 0.32 30 8–110 AOAC 988.11 GC-electron capture
species) GC-ICP/MS
EN 16801 /
NMKL 202 AAS-flame
AOAC 977.15** ICP-MS
NMKL 186** / ICP-MS
AOAC 2013.06** ICP-MS
/ EN 15763**
Orange Methylmercury* 0.8 0.40–1.2 0.08 0.16 33 80–110 AOAC 988.11 GC-electron capture
roughy EN 16801 / GC-ICP/MS
NMKL 202
AAS-flame
AOAC 977.15**
ICP-MS
NMKL 186** /
ICP-MS
AOAC 2013.06**
ICP-MS
/ EN 15763**
Pink cusk eel Methylmercury* 1.0 0.52–1.5 0.10 0.20 32 80–110 AOAC 988.11 GC-electron capture
EN 16801 / GC-ICP/MS
NMKL 202
AAS-flame
AOAC 977.15**
ICP-MS
NMKL 186** /
ICP-MS
AOAC 2013.06**
ICP-MS
/ EN 15763**
* Countries or importers may decide to use their own screening when applying the ML for methylmercury in fish by analysing total mercury in fish. If the total mercury concentration is below or equal to the ML
for methylmercury, no further testing is required, and the sample is determined to be compliant with the ML. If the total mercury concentration is above the ML for methylmercury, follow-up testing shall be
conducted to determine if the methylmercury concentration is above the ML. The ML also applies to fresh or frozen fish intended for further processing.
**Method applicable for determination of mercury and can be used for screening of methyl mercury, see *
CXS 234-1999 24
Table 5. Method performance criteria for sodium chloride and for salt determined as chloride expressed as sodium chloride
Commodity Provision ML Min. appl. LOD LOQ Precision Recovery Examples of Principle
(%) range (%) (%) (RSDR) (%) applicable
(%) (%) methods that
no more meet the
than criteria
Boiled dried Sodium chloride 15 (NaCl) 13.8–16.2 1.5 3.0 5.3 98–102 NMKL 178 Potentiometric
salted and salt titration
anchovies determined as 9.1 (Cl-) 8.3–9.9 0.91 1.8 5.7 98–102 AOAC 971.27 Potentiometric
chloride titration
expressed as AOAC 937.09 Titration
sodium chloride

Fish sauce Salt determined 20 (NaCl) 18 2.0 4.0 5.1 98–102 NMKL 178 Potentiometric
as chloride Minimum titration
expressed as limit AOAC 971.27 Potentiometric
sodium chloride titration
12 (Cl-) AOAC 976.18 Titration
11 1.2 2.4 5.5 98–102 AOAC 937.09 Titration
CXS 234-1999 25
Foods for special dietary uses

Commodity Provision Method Principle Type

Special foods Ash AOAC 942.05 Gravimetry I
Special foods Calcium AOAC 984.27 ICP emission spectrometry III
Special foods Calories by calculation Method described in CAC/VOL IX-Ed.1, Calculation method III
Part III
Special foods Carbohydrates Method described in CAC/VOL IX-Ed.1, Calculation III
Part III
Special foods Chloride AOAC 971.27 (Codex general method) Potentiometry II
Special foods Dietary fibre, total AOAC 985.29 Gravimetry (enzymatic I
digestion)
Special foods Fat CAC/RM 55 Gravimetry (extraction) I
Special foods Fat in foods not containing starch, meat or CAC/RM 1, B-2 Gravimetry I
vegetable products
Special foods Fill of containers CAC/RM 46 (See Appendix X) Weighing I
Special foods Folic acid AOAC 944.12 Microbioassay II
Special foods Linoleate (in the form of glycerides) AOAC 922.06; 969.33; 963.22 Acid hydrolysis, preparation II
of methyl esters and gas
chromatography
Special foods Linoleate (in the form of glycerides) AOAC 922.06; 979.19 Acid hydrolysis and III
spectrophotometry
Special foods Loss on drying (milk-based) AOAC 925.23 Gravimetry I
ISO 6731 | IDF 21

Special foods Nicotinamide for foods not based on milk AOAC 961.14 Colorimetry II
Special foods Nicotinamide for milk-based foods AOAC 944.13 Microbioassay II
Special foods Pantothenic acid/enriched foods AOAC 945.74 Microbioassay II
Special foods Pantothenic acid/non-enriched foods The Analyst 89 (1964):1, 3-6, ibid. 232 Microbioassay IV
US Dept Agr., Agr. Handbook 97 (1965)
CXS 234-1999 26
Foods for special dietary uses

Commodity Provision Method Principle Type

Special foods Phosphorous AOAC 986.24 Colorimetry II
(molybdovanadate)
Special foods Protein efficiency ratio (PER) AOAC 960.48 Rat bioassay I
Special foods Protein, crude Method described in CAC/Vol IX-Ed. 1, Part III Titrimetry, Kjeldahl digestion I
Special foods Riboflavin AOAC 970.65 Fluorometry II
Special foods Sodium and potassium ISO 8070 | IDF 119 Flame atomic absorption
II
spectrometry
Special foods Sodium and potassium AOAC 984.27 ICP emission spectrometry III
Special foods Vitamin A AOAC 974.29 Colorimetry IV
Special foods Vitamin A in foods in which carotenes have AOAC 941.15 Spectrophotometry III
been added as a source of vitamin A
Special foods Vitamin B12 AOAC 952.20 Microbioassay II
Special foods Vitamin B6 AOAC 961.15 Microbioassay II
Special foods Vitamin C AOAC 967.22 Microfluorometry II
Special foods Vitamin C AOAC 967.21 Colorimetry III
(dichloroindophenol)
Special foods Vitamin D (D3, milk-based infant formula) AOAC 992.26 Liquid chromatography II
Special foods Vitamin E AOAC 971.30 Colorimetry IV
Special foods Vitamin E (milk-based infant formula) AOAC 992.03 Liquid chromatography II
Special foods Sodium and potassium ISO 8070 | IDF 119 Flame atomic absorption II
spectrometry
Follow-up formula Dietary fibre, total AOAC 991.43 Gravimetry (enzymatic I
digestion)
Follow-up formula Iodine (milk-based formula) AOAC 992.24 Ion-selective potentiometry II
CXS 234-1999 27
Foods for special dietary uses

Commodity Provision Method Principle Type

Follow-up formula Pantothenic acid AOAC 992.07 Microbioassay II
Measures total pantothenate
(free pantothenic acid + CoA- + ACP-bound)
and measured as D-pantothenic acid (or
calcium D-pantothenate)
Follow-up formula Vitamin A AOAC 974.29 Colorimetry IV
Follow-up formula Vitamin A (retinol isomers) AOAC 992.04 HPLC II
Follow-up formula Vitamin A (retinol) (above 500 IU/l milk AOAC 992.06 HPLC III
after reconstitution)
Follow-up formula Vitamin K AOAC 2015.09 / ISO 21446 HPLC-FLD II
CXS 234-1999 28
Foods for special dietary uses

Commodity Provision Method Principle Type

Foods with low-sodium Iodine AOAC 925.56 Titrimetry II
content (including salt
substitutes)
Foods with low-sodium Silica (colloidal, calcium silicate) AOAC 950.85N Gravimetry IV
content (including salt
substitutes)
Enzyme-Linked Immunoassay R5 Mendez
Gluten-free foods Gluten Immunoassay I
(ELISA) Method
Eur J Gastroenterol Hepatol 2003; 15: 465-474

Infant formula Biotin AOAC 2016.02 / ISO 23305 HPLC-UV II

Infant formula Biotin EN 15607 (d-biotin) HPLC- FLD III
(Measures total D-biotin [free + D-biocytin])

Infant formula Calories (by calculation) Method described in CAC/Vol IX-Ed.1, Calculation I
Part IIIxiii
Infant formula Calcium AOAC 2015.06/ISO 21424 | IDF 243 ICP-MS II
Infant formula Calcium AOAC 2011.14/ISO 15151 | IDF 229 ICP emission spectroscopy III
Infant formula Calcium ISO 8070 │ IDF 119 Flame atomic absorption III
spectrophotometry
Infant formula Calcium AOAC 985.35 Flame atomic absorption III
spectroscopy

xiii 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 may be 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.
CXS 234-1999 29
Foods for special dietary uses

Commodity Provision Method Principle Type

Infant formula Carnitine AOAC 2015.10/ISO 21468 UHPLC-MS/MS II
Infant formula Chloride AOAC 986.26 Potentiometry III
Infant formula Chloride AOAC 2016.03/ISO 21422 | IDF 242 Potentiometry II
Infant formula Choline AOAC 2015.10/ISO 21468 UHPLC-MS/MS II
Infant formula Choline AOAC 999.14 Enzymatic colorimetric method III
with limitations on applicability
due to choline and ascorbate
concentration
Infant formula Copper AOAC 2015.06/ISO 21424 | IDF 243 ICP-MS II
Infant formula Copper AOAC 985.35 Flame atomic absorption III
spectroscopy
Infant formula Copper AOAC 2011/14/ISO 15151 | IDF 229 ICP emission spectroscopy III
Infant formula Chromium (Section B of CXS 72-1981 only) EN 14082 Graphite furnace atomic III
absorption after dry ashing
Infant formula Chromium (Section B of CXS 72-1981 only) EN 14083 Graphite furnace AAS after III
pressure digestion
Infant formula Chromium (Section B of CXS 72-1981 only) AOAC 2006.03 ICP emission spectroscopy III
Infant formula Chromium (Section B of CXS 72-1981 only) AOAC 2011.19/ISO 20649 |IDF 235 ICP-MS II
Infant formula Crude proteinxiv ISO 8968-1 | IDF 20-1 Titrimetry (Kjeldahl) I
Infant formula Fatty acids (including trans fatty acid) AOAC 996.06 Gas chromatography III

xiv 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.
CXS 234-1999 30
Foods for special dietary uses

Commodity Provision Method Principle Type

Infant formula Fatty acids (including trans fatty acid) AOCS Ce 1i-07 Gas chromatography III
Infant formula Folic acid AOAC 992.05 Microbioassay III
(Measures free folic acid + free, unbound
natural folates, aggregated, and measured as
folic acid)
EN 14131
(Total folate (free + bound), aggregated and
measured as folic acid)
Infant formula Folic acid AOAC 2011.06 LC-MS/MS II
Infant formula Iodine AOAC 2012.15 / ISO 20647 | IDF 234 ICP-MS II
(for milk-based formula)
Infant formula Iron AOAC 2015.06 / ICP-MS II
ISO 21424 | IDF 243
Infant formula Iron AOAC 2011.14 / ISO 15151 | IDF 229 ICP emission spectroscopy III

Infant formula Ironxv AOAC 985.35 Flame atomic absorption III

spectrophotometry
Infant formula Iron AOAC 999.11│NMKL139 AAS after dry ashing II
Infant formula Magnesium AOAC 2015.06 / ICP-MS II
ISO 21424 | IDF 243
Infant formula Magnesium AOAC 2011.14 / ISO 15151 | IDF 229 ICP emission spectroscopy III
Infant formula Magnesium ISO 8070 │ IDF 119 Flame atomic absorption III
spectrophotometry
Infant formula Magnesium AOAC 985.35 Flame atomic absorption III
spectroscopy

xv
General Codex methods are also available.
CXS 234-1999 31
Foods for special dietary uses

Commodity Provision Method Principle Type

Infant formula Manganese AOAC 2015.06 / ICP-MS II
ISO 21424 | 243
Infant formula Manganese AOAC 2011.14 / ISO 15151 | IDF 229 ICP emission spectroscopy III
Infant formula Manganese AOAC 985.35 Flame atomic absorption III
spectrophotometry
Infant formula Melamine ISO/TS 15495 | IDF/RM 230 LC-MS/MS IV
Infant formula Molybdenum (Section B of CXS 72-1981 EN 14083 Graphite furnace AAS after III
only) pressure digestion
Infant formula Molybdenum (Section B of CXS 72-1981 AOAC 2006.03 ICP emission spectroscopy III
only)
Infant formula Molybdenum (Section B of CXS 72-1981 AOAC 2011.19 / ISO 20649 | IDF 235 ICP-MS II
only)
Infant formula Myo-Inositol AOAC 2011.18 / ISO 20637 LC-pulsed amperometry II
Infant formula Niacin AOAC 2015.14 / ISO 21470 Enzymatic digestion and II
UHPLC-MS/MS
Infant formula Niacin AOAC 985.34 (niacin (preformed) and Microbioassay and turbidimetry III
nicotinamide)
Infant formula Niacin EN 15652 HPLC IIIxvi
(Free and bound and phosphorylated forms
measured either as aggregate of nicotinic
acid + nicotinamide, or as individual forms)
Infant formula Pantothenic acid AOAC 2012.16 UHPLC-MS/MS II
ISO 20639
Infant formula Phosphorus AOAC 2015.06 / ICP-MS II
ISO 21424 | IDF 243
Infant formula Phosphorus AOAC 2011.14 / ISO 15151 | IDF 229 ICP emission spectroscopy III

xvi When published as EN method.

CXS 234-1999 32
Foods for special dietary uses

Commodity Provision Method Principle Type

Infant formula Phosphorus AOAC 986.24 Spectrophotometry III
(molybdovanadate)
Infant formula Riboflavin AOAC 2015.14 / ISO 21470 Enzymatic digestion and II
UHPLC-MS/MS
Infant formula Riboflavin AOAC 985.31xvii Fluorimetry III
Infant formula Riboflavin EN 14152 (Measures natural and HPLC III
supplemental forms, free, bound and
phosphorylated (FMN and FAD) aggregated
and measured as riboflavin)
Infant formula Selenium AOAC 996.16 or AOAC 996.17 Continuous hydride generation III
flame atomic absorption
spectrometry (HGAAS)
Infant formula Selenium EN 14627 Hydride generation atomic III
absorption spectrometry
(HGAAS)
Infant formula Selenium AOAC 2006.03 ICP emission spectroscopy III
Selenium AOAC 2011.19 / ISO 20649 | IDF 235 ICP-MS II
Infant formula Sodium and potassium AOAC 2015.06 / ICP-MS II
ISO 21424 | 243
Infant formula Sodium and potassium AOAC 2011.14 / ISO 15151 | IDF 229 ICP emission spectroscopy III
Infant formula Sodium and potassium ISO 8070 | IDF 119 Flame atomic absorption III
spectrophotometry
Infant formula Thiamine AOAC 2015.14 / ISO 21470 Enzymatic digestion and II
UHPLC-MS/MS
Infant formula Thiamine AOAC 986.27xvii Fluorimetry III

xvii
Care should be taken in the application of the method due to spectral interference.
CXS 234-1999 33
Foods for special dietary uses

Commodity Provision Method Principle Type

Infant formula Thiamine EN 14122 HPLC with pre-or post-column III
(Measures all vitamin B1 forms (natural and derivatization to thiochrom
added free, bound and phosphorylated)
following extraction and conversion to
thiamine)
Infant formula Total amino acids (excluding taurine and AOAC 2018.06 / ISO 4214 | IDF 254 / AACC UHPLC-UV II
tryptophan) 07-50.01
for use according to Section 3.1.3 (a)
notes 2) and 3) of CXS 72-1981
Infant formula Total carbohydrates AOAC 986.25 Determination by difference I
AOAC 990.19 or
Moisture/total solids AOAC 990.20
Gravimetry
ISO 6731|IDF 21

Ash AOAC 942.05 Gravimetry

Infant formula Total fat AOAC 989.05 Gravimetry (Röse-Gottlieb) I
ISO 23318 | IDF 249
Infant formula Total fat ISO 8262-1 |IDF 124-1 Gravimetry (Weibull-Berntrop) I
for milk-based infant formula (products not
completely soluble in ammonia)
Infant formula Total fatty acids AOAC 996.06 Gas chromatography III
Commodity Provision Method Principle Type
Infant formula Total fatty acids AOAC 2012.13 / ISO 16958 | IDF231 Gas chromatography II

Infant formula Total nucleotides AOAC 2011.20 LC II

ISO 20638
Infant formula Total phospholipids AOCS Ja7b-91 Gas chromatography with III
suitable extraction and
preparation procedures
CXS 234-1999 34
Foods for special dietary uses

Commodity Provision Method Principle Type

Infant formula Tryptophan AOAC 2017.03 HPLC II

For use according to Section 3.1.3 (a)
notes 2 and 3 of CXS 72-1981
Infant formula Vitamin A EN 12823-1 (all-trans-retinol and 13-cis- HPLC III
retinol)
Vitamin A (both natural + supplemental ester
forms) aggregated and quantified as
individual retinol isomers (13-cis and all-trans)
Infant formula Vitamin A palmitate AOAC 2012.10 HPLC II
(retinyl palmitate), ISO 20633
vitamin A acetate
(retinyl acetate)
Infant formula Vitamin B12 AOAC 2014.02 LC-UV III
Infant formula Vitamin C AOAC 2012.22 / ISO/DIS 20635 HPLC-UV II
Infant formula Vitamin D EN 12821 HPLC-UV III
(D2 and/or D3 measured as single
components. Hydroxylated forms not
measured)
NMKL 167
Infant formula Vitamin D AOAC 995.05 HPLC-UV III
D2 and D3 measured
Infant formula Vitamin D AOAC 2016.05 / ISO 20636 LC-MS II
Commodity Provision Method Principle Type
Infant formula Vitamin E AOAC 992.03 HPLC III
Measures all rac-vitamin E (both natural +
supplemental ester forms) aggregated and
quantified as α-congeners
 CXS 234-1999 35
Foods for special dietary uses

Commodity Provision Method Principle Type

Infant formula Vitamin E EN 12822 HPLC II
(Measures vitamin E (both natural +
supplemental ester forms) aggregated and
quantified as individual tocopherol congeners
(α, β, γ, δ)
Infant formula Vitamin E AOAC 2012.10 / ISO 20633 HPLC II
Infant formula Vitamin B6 AOAC 2015.14 / ISO 21470 Enzymatic digestion and UHPLC- II
MS/MS
Infant formula Vitamin B6 AOAC 985.32 Microbioassay III
Infant formula Vitamin B6 EN 14166 Microbioassay III
(Aggregates free and bound pyridoxal,
pyridoxine and pyridoxamine and measures as
pyridoxine)
Infant formula Vitamin B6 AOAC 2004.07 III
EN 14164 HPLC
(Free and bound phosphorylated forms
(pyridoxal, pyridoxine and pyridoxamine)
converted and measured as pyridoxine)
Infant formula Vitamin B12 AOAC 986.23 Turbidimetric method
(Measures total vitamin B12 as cyanocobalamin)
III
Infant formula Vitamin B12 AOAC 2011.10 / ISO 20634 HPLC II
Infant formula Vitamin K AOAC 2015.09 / ISO 21446 HPLC-FLD II
Infant formula Zinc AOAC 2015.06 / ICP-MS II
ISO 21424 | IDF 243
Infant formula Zinc AOAC 2011.14 / ISO 15151 | IDF 229 ICP emission spectroscopy III
Infant formula Zinc AOAC 985.35 Flame atomic absorption III
spectroscopy
CXS 234-1999 36
Table 6. Methods of analysis for dietary fibre: Guidelines for use of nutrition and health claims (CXG 23-1997): 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 (1) Method applicable for determining dietary fibres that do not include AOAC 985.29 Enzymatic gravimetry Type I
the lower molecular weight fraction (4) AACC Intl 32-05.01
All foods (1) Method applicable for determining dietary fibres that do not include AOAC 991.43 Enzymatic gravimetry Type I
the lower molecular weight fraction and also includes determination AACC Intl 32-07.01
for soluble and insoluble dietary fibres (4)
NMKL 129
All foods (1) Method applicable for determining dietary fibres that do not include AOAC 993.21 Gravimetry Type I
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)
All foods (1) Method applicable for determining dietary fibres that do not include AOAC 994.13 Enzymatic GC/ Type I
the lower molecular weight fraction. Provides sugar residue AACC Intl 32- 25.01 colorimetry gravimetry
composition of dietary fibre polysaccharides, as well as content of
Klason lignin (4) NMKL 162
All foods (1) Insoluble dietary fibres in food and food products (4) AOAC 991.42 Enzymatic gravimetry Type I
(Specific for insoluble fibre)
AACC Intl 32-20.01
All foods (1) Soluble dietary fibres in food and food products (4) AOAC 993.19 Enzymatic gravimetry Type I
(Specific for soluble fibre)
General methods that measure both the higher (monomeric units > 9) and the lower molecular weight fraction (monomeric units <=9) (2)
All foods (1) Method applicable for determining the content of dietary fibres of AOAC 2001.03 Enzymatic gravimetry Type I
higher and lower molecular weight, in food where resistant starches AACC Intl 32-41.01 and Liquid
are not present chromatography
All foods (1) Method applicable for determining the content of dietary fibres of ICC Standard No. 185 / AOAC Enzymatic gravimetry Type I
higher and lower molecular weight. The method is applicable in food 2017.16 / AACC 32-60-01 High pressure liquid
that may, or may not, contain resistant starch chromatography
All foods (1) Method applicable for determining the content of insoluble and AACC Intl 32-50.01 Enzymatic gravimetry Type I
soluble dietary fibres of higher and lower molecular weight. The High Pressure Liquid
AOAC 2011.25
method is applicable in food that may, or may not, contain resistant Chromatography
starches
CXS 234-1999 37

Standard Provisions Method Principle Type

General methods that do not measure the lower molecular weight fraction (i.e. monomeric units < = 9)(2)
Methods that measure individual specific components (monomeric units: the whole range for each type of components is covered)(2)
All foods (1) (13)(14) Beta-D-Glucans AOAC 995.16 Enzymatic Type II
AACC Intl 32-23.01
All foods (1) Fructans (oligofructoses, inulin, hydrolysed inulin, polyfructoses, AOAC 997.08 Enzymatic & HPAEC- Type II
fructooligosaccharides) AACC Intl 32-31.01 PAD
(applicable to added fructans)
All foods (1) Fructans (oligofructoses, inulin, hydrolysed inulin, polyfructoses, AOAC 999.03 Enzymatic & Type III
fructooligosaccharides) AACC Intl 32-32.01 colorimetric
(not applicable highly depolymerized fructans)
All foods (1) Polydextrose AOAC 2000.11 HPAEC-PAD Type II
AACC Intl 32-28.01
All foods (1) Trans-galacto-oligo saccharides AOAC 2001.02 HPAEC-PAD Type II
AACC Intl 32-33.01
All foods (1) Resistant starch (Recommended for RS3) AOAC 2002.02 Enzymatic Type II
AACC Intl 32-40.01

Other methods(2) that have not been subjected to interlaboratory evaluation under AOAC international guidelines
Yeast Insoluble glucans and mannans of yeast cell wall (for yeast cell Eurasyp (European association for Chemical & Type IV
cell wall wall only) specialty yeast product) – LM HPAEC-PAD
Bonanno. Biospringer. 2004 – online
version:
http://www.eurasyp.org/public.techniq
ue.home.screen
All foods Fructo-oligosaccharides (monomeric units < 5) Ouarné et al. 1999 in Complex HPAEC-PAD Type IV
Carbohydrates in Foods. Edited by S.
Sungsoo, L. Prosky & M. Dreher.
Marcel Dekker Inc, New York
CXS 234-1999 38
All foods Non-starch polysaccharides (NSP) (3) Englyst H.N., Quigley M.E., Hudson Gas-liquid Type IV
G. 1994. Determination of dietary fibre chromatography
as non-starch polysaccharides with
gas-liquid chromatographic high-
performance liquid chromatographic
or spectrophotometric measurement
of constituent sugars – Analyst 119,
1497–1509
(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 International.
(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 on nutrition labelling (CXG 2- 1985).
(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.

Fruit juices and nectars

Commodity Provisions Method Principle Type
Fruit juices and nectars Ascorbic acid-L (additives) IFUMA 17A HPLC II
Fruit juices and nectars Ascorbic acid-L (additives) ISO 6557-1 Fluorescence spectrometry IV
Fruit juices and nectars Ascorbic acid-L (additives) AOAC 967.21 Indophenol method III
IFUMA 17
ISO 6557-2
Fruit juices and nectars Carbon dioxide (additives and IFUMA 42 Titrimetry IV
processing aids) (back-titration after precipitation)
Fruit juices and nectars Cellobiose IFUMA 4 Capillary gas chromatography IV
Fruit juices and nectars Citric acidxviii (additives) AOAC 986.13 HPLC II

xviii All juices except citrus based juices.

CXS 234-1999 39
Fruit juices and nectars
Commodity Provisions Method Principle Type
xix
Fruit juices and nectars Citric acid (additives) EN 1137 Enzymatic determination III
IFUMA 22
Fruit juices and nectars Glucose and fructose EN 12630 HPLC III
(permitted ingredients) IFUMA 67 NMKL 148

Fruit juices and nectars Glucose-D and fructose-D EN 1140 Enzymatic determination II
(permitted ingredients) IFUMA 55

Fruit juices and nectars HFCS and HIS in apple juice Determination of HFCS and HIS by CAP GC method IV
(permitted ingredients) Capillary GC method
JAOAC 84, 486 (2001)
Fruit juices and nectars Malic acid (additives) AOAC 993.05 Enzymatic determination and HPLC III
Fruit juices and nectars Malic acid-D EN 12138 Enzymatic determination II
IFUMA 64
Fruit juices and nectars Malic acid-D in apple juice AOAC 995.06 HPLC II
Fruit juices and nectars Malic acid-L EN 1138 Enzymatic determination II
IFUMA 21
Fruit juices and nectars Pectin (additives) IFUMA 26 Precipitation/photometry I
Fruit juices and nectars Benzoic acid and its salts; IFUMA 63 HPLC II
sorbic acid and its salts
NMKL 124
Fruit juices and nectars Benzoic acid and its salts ISO 5518, ISO 6560 Spectrometry III

xix
All juices except citrus based juices.
CXS 234-1999 40
Fruit juices and nectars

Commodity Provisions Method Principle Type

Fruit juices and nectars Preservatives in fruit juices ISO 5519 Spectrometry III
(sorbic acid and its salts)
Fruit juices and nectars Quinic, malic and citric acid Determination of quinic, malic and citric acid in HPLC III
in cranberry juice cocktail cranberry juice cocktail and apple juice
and apple juice (permitted AOAC 986.13
ingredients and additives)
Fruit juices and nectars Saccharin NMKL 122 Liquid chromatography II
Fruit juices and nectars Soluble solids AOAC 983.17 Indirect by refractometry I
EN 12143 IFUMA 8
ISO 2173
Fruit juices and nectars Sucrose (permitted EN 12146 Enzymatic determination III
ingredients) IFUMA 56
Fruit juices and nectars Sucrose (permitted EN 12630 HPLC II
ingredients) IFUMA 67 NMKL 148
Fruit juices and nectars Sulphur dioxide (additives) Optimized Monier-Williams AOAC 990.28 Titrimetry after distillation II
IFUMA 7A NMKL 132
Fruit juices and nectars Sulphur dioxide (additives) NMKL 135 Enzymatic determination III
Fruit juices and nectars Sulphur dioxide (additives) ISO 5522, ISO 5523 Titrimetry after distillation III
Fruit juices and nectars Tartaric acid in grape juice EN 12137 HPLC II
(additives) IFUMA 65
Fruit juices and nectars Total nitrogen EN 12135 IFUMA 28 Digestion/titration I
CXS 234-1999 41
Fruit juices and nectars

Commodity Provisions Method Principle Type

Fruit juices and nectars Sections 3.2 Quality criteria Determination of acetic acid Enzymatic determination II
and 3.3 Authenticity of EN 12632; IFUMA 66
CXS 247-2005xx
Fruit juices and nectars Determination of alcohol (ethanol) Enzymatic determination II
IFUMA 52
Fruit juices and nectars Detection of anthocyanins HPLC I
IFUMA 71
Fruit juices and nectars Determination of ash in fruit products Gravimetry I
AOAC 940.26; EN 1135; IFUMA 9
Fruit juices and nectars Detection of beet sugar in fruit juices Deuterium NMR II
AOAC 995.17
Fruit juices and nectars Determination of benzoic acid as a marker in HPLC III
orange juice
AOAC 994.11
Fruit juices and nectars Determination of C13/C12 ratio of ethanol derived Stable isotope mass spectrometry II
from fruit juices
JAOAC 79, No. 1, 1996, 62-72
Fruit juices and nectars Determination of carbon stable isotope ratio of Stable isotope mass spectrometry II
apple juice
AOAC 981.09– JAOAC 64, 85 (1981)
Fruit juices and nectars Determination of carbon stable isotope ratio of Stable isotope mass spectrometry II
orange juice
AOAC 982.21

xx
3.4 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.
CXS 234-1999 42
Fruit juices and nectars

Commodity Provisions Method Principle Type

Fruit juices and nectars Determination of carotenoid, total/individual Spectrophotometry I
groups
EN 12136; IFUMA 59
Fruit juices and nectars Determination of centrifugable pulp Centrifugation/% value I
EN 12134; IFUMA 60

Fruit juices and nectars Determination of chloride (expressed as Electrochemical titrimetry III
sodium chloride)
EN 12133
IFUMA 37
Fruit juices and nectars Determination of chloride in vegetable juice Titration II
AOAC 971.27 (Codex general method)
ISO 3634
Fruit juices and nectars Determination of essential oils (Scott titration) (Scott) distillation, titration I
AOAC 968.20 - IFUMA 45xxi
Fruit juices and nectars Determination of essential oils (in citrus fruit) Distillation and direct reading of the I
(volume determination)xxi volume determination
ISO 1955
Fruit juices and nectars Determination of fermentability Microbiological method I
IFUMA 18
Fruit juices and nectars Determination of formol number Potentiometric titration I
EN 1133
IFUMA 30
Fruit juices and nectars Determination of free amino acids Liquid chromatography II
EN 12742 IFUMA 57
Fruit juices and nectars Determination of fumaric acid HPLC II
IFUMA 72
Fruit juices and nectars Determination of glucose fructose and HPLC II
saccharose
EN 12630 IFUMA 67 NMKL 148
Fruit juices and nectars Determination of gluconic acid Enzymatic determination II
IFUMA 76

xxi
Because there is no numerical value in the standard, duplicate Type I methods have been included which may lead to different results.
CXS 234-1999 43
Fruit juices and nectars
Commodity Provisions Method Principle Type
Fruit juices and nectars Determination of glycerol Enzymatic determination II
IFUMA 77
Fruit juices and nectars Determination of hesperidin and naringin HPLC II
EN 12148 IFUMA 58
Fruit juices and nectars Determination of hydroxymethylfurfural HPLC II
IFUMA 69
Fruit juices and nectars Determination of hydroxymethylfurfural Spectrometry III
ISO 7466
Fruit juices and nectars Determination of isocitric acid-D Enzymatic determination II
IFUMA 54
Fruit juices and nectars Determination of Lactic acid- D and L Enzymatic determination II
EN 12631
IFUMA 53
Fruit juices and nectars Determination of L-malic/total malic acid ratio Enzymatic determination and HPLC II
in apple juice
AOAC 993.05
Fruit juices and nectars Determination of naringin and neohesperidin HPLC III
in orange juice
AOAC 999.05
Fruit juices and nectars Determination of pH value Potentiometry
NMKL 179 II
EN 1132 IFUMA 11 ISO 1842 IV
Fruit juices and nectars Determination of phosphorus/phosphate Photometric determination II
EN 1136
IFUMA No 50
Fruit juices and nectars Determination of proline by photometry – non- Photometry I
specific determination
EN 1141 IFUMA 49
Fruit juices and nectars Determination of relative density Pycnometry II
EN 1131 (1993); IFUMA 01 & IFU Method No
General sheet (1971)
Fruit juices and nectars Determination of relative density Densitometry III
IFUMA 01A
Fruit juices and nectars Determination of sodium, potassium, calcium, Atomic absorption spectroscopy II
magnesium in fruit juices
EN 1134 IFUMA 33
Fruit juices and nectars Determination of sorbitol-D Enzymatic determination II
IFUMA62
CXS 234-1999 44
Fruit juices and nectars
Commodity Provisions Method Principle Type
Fruit juices and nectars Determination of stable carbon isotope ratio Stable isotope mass spectrometry II
in the pulp of fruit juices
ENV 13070
Analytica Chimica Acta 340 (1997)
Fruit juices and nectars Determination of stable carbon isotope ratio of Stable isotope mass spectrometry II
sugars from fruit juices
ENV 12140
Analytica Chimica Acta 271 (1993)
Fruit juices and nectars Determination of stable hydrogen isotope ratio Stable isotope mass spectrometry II
of water from fruit juices
ENV 12142
Fruit juices and nectars Determination of stable oxygen isotope ratio in Stable isotope mass spectrometry II
fruit juice water
ENV 12141
Fruit juices and nectars Detection of starch Colorimetric I
AOAC 925.38 IFUMA 73
Fruit juices and nectars Determination of sugar beet derived syrups Oxygen isotope ratio analysis I
in frozen concentrated orange juice δ18O
Measurements in water
AOAC 992.09
Fruit juices and nectars Determination of titrable acids, total Titrimetry I
EN 12147
IFUMA 03
ISO 750
Fruit juices and nectars Determination of total dry matter (vacuum Gravimetric determination I
oven drying at 70 °C)xxii
EN 12145
IFUMA 61
Fruit juices and nectars Determination of total solids (microwave oven Gravimetric determination I
drying)xxii AOAC 985.26
Fruit juices and nectars Determination of vitamin C (dehydro-ascorbic Microfluorometry III
acid and ascorbic acid)
AOAC 967.22

xxii
Because there is no numerical value in the standard, duplicate Type I methods have been included which may lead to different results.
CXS 234-1999 45
Milk and milk products
Provisions Method Principle Type
Commodity
Milk and milk products Melamine ISO 23970 | IDF 252 LC-MS/MS II
Blend of evaporated skimmed milk Total fat ISO 23318 | IDF 249 Gravimetry (Röse-Gottlieb) I
and vegetable fat
Blend of evaporated skimmed milk Milk solids-not-fat (MSNF)xxiii ISO 6731 | IDF 21 Calculation from total solids content I
and vegetable fat and ISO 23318 | IDF 249 and fat content,
gravimetry, drying at 102 oC and
gravimetry (Röse-Gottlieb)
Blend of evaporated skimmed milk Milk protein in MSNFxxiiiiii ISO 6731 | IDF 21 and Calculation from total solids content, IV
and vegetable fat ISO 23318 | IDF 249 and fat content and protein content,
ISO 8968-1 | IDF 20-1 gravimetry, drying at 102 oC and
gravimetry (Röse-Gottlieb) and
titrimetry (Kjeldahl)
Blend of evaporated skimmed milk Milk protein in MSNFxxiiiiii ISO 6731 | IDF 21 and Calculation from total solids content, IV
and vegetable fat ISO 23318 | IDF 249 and fat content and protein content,
AOAC 991.20 gravimetry, drying at 102 oC and
gravimetry (Röse-Gottlieb) and
titrimetry (Kjeldahl)
Reduced fat blend of evaporated Total fat ISO 23318| IDF 249 Gravimetry (Röse-Gottlieb) I
skimmed milk and vegetable fat

xxiii
Milk total solids and milk solids-not-fat (MSNF) content include water of crystallization of lactose.
CXS 234-1999 46
Milk and milk products
Provisions Method Principle Type
Commodity
Reduced fat blend of evaporated Milk solids-not-fat (MSNF)xxiv ISO 6731 | IDF 21 Calculation from total solids content I
skimmed milk and vegetable fat and ISO 23318 | IDF 249 and fat content,
gravimetry, drying at 102 oC and
gravimetry (Röse-Gottlieb)
Reduced fat blend of evaporated Milk protein in MSNFxxiv ISO 6731 | IDF 21 and Calculation from total solids content, IV
skimmed milk and vegetable fat ISO 23318 | IDF 249 and fat content and protein content,
ISO 8968-1 | IDF 20-1 gravimetry, drying at 102 oC and
gravimetry (Röse-Gottlieb) and
titrimetry (Kjeldahl)
Reduced fat blend of evaporated Milk protein in MSNFxxiv ISO 6731 | IDF 21 and Calculation from total solids content, IV
skimmed milk and vegetable fat ISO 23318 | IDF 249 and fat content and protein content,
AOAC 991.20 gravimetry, drying at 102 oC and
gravimetry (Röse-Gottlieb) and
titrimetry (Kjeldahl)
Blend of skimmed milk and vegetable Total fat ISO 23318 | IDF 249 Gravimetry (Röse-Gottlieb) I
fat in powdered form
Blend of skimmed milk and vegetable Waterxxv (moisture) Described in Appendix XIxxvi Gravimetry, drying at 102 oC IV
fat in powdered form
Blend of skimmed milk and vegetable Waterxxv (moisture) ISO 5537 | IDF 26 Gravimetry, drying at 87 °C I
fat in powdered form
Blend of skimmed milk and vegetable Milk protein in MSNFxxiv ISO 5537 | IDF 26 and Calculation from total solids content, IV
fat in powdered form ISO 23318 | IDF 249 and fat content and protein content,
ISO 8968-1 | IDF 20-1 gravimetry, drying at 87 oC and
gravimetry (Röse-Gottlieb) and
titrimetry (Kjeldahl)

xxiv
Milk total solids and milk solids-not-fat (MSNF) content include water of crystallization of lactose.
xxv
Water content excluding the crystallized water bound to lactose (generally known as moisture content).
xxvi
Due to accessibility to equipment and calibration of the method ISO 5537 | IDF 26, the method described in Appendix XI is listed as Type IV.
CXS 234-1999 47
Milk and milk products
Provisions Method Principle Type
Commodity
Blend of skimmed milk and vegetable Milk protein in MSNFxxvii Described in Appendix XIxxviii and Calculation from total solids content, IV
fat in powdered form ISO 23318 | IDF 249 and fat content and protein content,
ISO 8968-1 | IDF 20-1 gravimetry, drying at 102 oC and
gravimetry (Röse-Gottlieb) and
titrimetry (Kjeldahl)
Blend of skimmed milk and vegetable Milk protein in MSNFxxvii ISO 5537 | IDF 26 and Calculation from total solids content, IV
fat in powdered form and fat content and protein content,
AOAC 991.20 gravimetry, drying at 87 oC and
gravimetry (Röse-Gottlieb) and
titrimetry (Kjeldahl)
Blend of skimmed milk and vegetable Milk protein in MSNFxxvii Described in Appendix XIxxviiiiii and Calculation from total solids content, IV
fat in powdered form ISO 23318 |IDF 249 and AOAC 991.20 fat content and protein content,
gravimetry, drying at 102 oC and
gravimetry (Röse-Gottlieb) and
titrimetry (Kjeldahl)
Reduced fat blend of skimmed milk Total fat ISO 23318 | IDF 249 Gravimetry (Röse-Gottlieb) I
powder and vegetable fat in powdered
form
Reduced fat blend of skimmed milk Waterxxix (moisture) ISO 5537 | IDF 26 Gravimetry, drying at 87 °C I
powder and vegetable fat in powdered
form
Reduced fat blend of skimmed milk Waterxxix (moisture) Described in Appendix XIxxviii Gravimetry, drying at 102 oC IV
powder and vegetable fat in powdered
form
Reduced fat blend of skimmed milk Milk protein in MSNFxxvii ISO 5537 | IDF 26 and Calculation from total solids content, IV
powder and vegetable fat in powdered ISO 1736 | IDF 9 and fat content and protein content,
form ISO 8968-1 | IDF 20-1 gravimetry, drying at 87 oC and
gravimetry (Röse-Gottlieb) and
titrimetry (Kjeldahl)

xxvii
Milk total solids and milk solids-not-fat (MSNF) content include water of crystallization of lactose.
xxviii
Due to accessibility to equipment and calibration of the method ISO 5537 | IDF 26, the method as described in Appendix XI is listed as Type IV.
xxix
Water content excluding the crystallized water bound to lactose (generally known as moisture content).
CXS 234-1999 48
Milk and milk products
Provisions Method Principle Type
Commodity
Reduced fat blend of skimmed milk Milk protein in MSNFxxx ISO 5537 | IDF 26 and Calculation from total solids content, IV
powder and vegetable fat in powdered ISO 23318 | IDF 249 and fat content and protein content,
form AOAC 991.20 gravimetry, drying at 87 oC and
gravimetry (Röse-Gottlieb) and
titrimetry (Kjeldahl)
Reduced fat blend of skimmed milk Milk protein in MSNFxxx Described in Appendix XIxxxi and Calculation from total solids content, IV
powder and vegetable fat in powdered ISO 23318 |IDF 249 and fat content and protein content,
form AOAC 991.20 gravimetry, drying at 102 oC and
gravimetry (Röse-Gottlieb) and
titrimetry (Kjeldahl)
Blend of sweetened condensed Total fat ISO 23318 | IDF 249 Gravimetry (Röse-Gottlieb) I
skimmed milk and vegetable fat
Blend of sweetened condensed Sucrose ISO 2911 | IDF 35 Polarimetry IV
skimmed milk and vegetable fat
Blend of sweetened condensed Milk solids-not-fat (MSNF)xxx ISO 6734 | IDF 15 and Calculation from total solids content, IV
skimmed milk and vegetable fat (for ISO 23318 | IDF 249 and fat content and sucrose content,
products sweetened with sucrose ISO 2911 | IDF 35 gravimetry, drying at 102 °C and
only) gravimetry (Röse-Gottlieb) and
polarimetry
Blend of sweetened condensed Milk protein in MSNFxxx ISO 6734 | IDF 15 and Calculation from total solids content, IV
skimmed milk and vegetable fat (for ISO 23318 | IDF 249 and fat content, sucrose content and
products sweetened with sucrose ISO 2911 | IDF 35 and protein content,
only) ISO 8968-1 | IDF 20-1 gravimetry, drying at 102 °C and
gravimetry (Röse-Gottlieb) and
polarimetry and
titrimetry (Kjeldahl)

xxx
Milk total solids and milk solids-not-fat (MSNF) content include water of crystallization of lactose.
xxxi Due to accessibility to equipment and calibration of the method ISO 5537 | IDF 26, the method as described in Appendix XI is listed as Type IV.
CXS 234-1999 49
Milk and milk products
Provisions Method Principle Type

Blend of sweetened condensed Milk protein in MSNFxxxii ISO 6734 | IDF 15 and Calculation from total solids content, IV
skimmed milk and vegetable fat (for ISO 23318 | IDF 249 and fat content, sucrose content and
products sweetened with sucrose ISO 2911 | IDF 35 and protein content,
only) AOAC 991.20 gravimetry, drying at 102 °C and
gravimetry (Röse-Gottlieb) and
polarimetry and
titrimetry (Kjeldahl)
Reduced fat blend of sweetened Total fat ISO 23318 | IDF 249 Gravimetry (Röse-Gottlieb) I
condensed skimmed milk and
vegetable fat
Reduced fat blend of sweetened Milk solids-not-fat (MSNF)xxxii ISO 6734 | IDF 15 and Calculation from total solids content, IV
condensed skimmed milk and ISO 23318 | IDF 249 and fat content and sucrose content,
vegetable fat (for products sweetened ISO 2911 | IDF 35 gravimetry, drying at 102 °C and
with sucrose only) gravimetry (Röse-Gottlieb) and
polarimetry
Reduced fat blend of sweetened Milk protein in MSNFxxxii ISO 6734 | IDF 15 and Calculation from total solids content, IV
condensed skimmed milk and ISO 23318 | IDF 249 and fat content, sucrose content and
vegetable fat (for products sweetened ISO 2911 | IDF 35 and protein content,
with sucrose only) ISO 8968-1 | IDF 20-1 gravimetry, drying at 102 °C and
gravimetry (Röse-Gottlieb) and
polarimetry and
titrimetry (Kjeldahl)
Reduced fat blend of sweetened Milk protein in MSNFxxxii ISO 6734 | IDF 15 and Calculation from total solids content, IV
condensed skimmed milk and ISO 23318 | IDF 249 and fat content, sucrose content and
vegetable fat (for products sweetened ISO 2911 | IDF 35 and protein content,
with sucrose only) AOAC 991.20 gravimetry, drying at 102 °C and
gravimetry (Röse-Gottlieb) and
polarimetry and
titrimetry (Kjeldahl)
Butter Milk solids-not-fat (MSNF)xxxii ISO 3727-2 | IDF 80-2 Gravimetry I

xxxii
Milk total solids and milk solids-not-fat (MSNF) content include water of crystallization of lactose.
CXS 234-1999 50
Milk and milk products
Provisions Method Principle Type
Commodity

Butter Milkfat (total fat) ISO 17189 | IDF 194 Gravimetry (direct determination of I
fat using solvent extraction)
Butter Milk fat purity ISO 17678 | IDF 202 Calculation from determination of I
triglycerides by gas
chromatography– FID
Butter Salt ISO 1738 | IDF 12/ Titrimetry (Mohr: determination of III
chloride, expressed as sodium
AOAC 960.29
chloride)
Butter Salt ISO 15648 | IDF 179 Potentiometry (determination of II
chloride, expressed as sodium
chloride)
Butter Waterxxxiii ISO 3727-1 | IDF 80-1 Gravimetry I
Cheese Milkfat ISO 23319 | IDF 250 Gravimetry (Schmid-Bondzynski- I
Ratzlaff)
Cheese Moisture ISO 5534 | IDF 4 Gravimetry, drying at 102 °C I
Cheese (and cheese rind) Natamycin ISO 9233-1 | IDF 140-1 Molecular absorption III
spectrophotometry
Cheese (and cheese rind) Natamycin ISO 9233-2 | IDF 140-2 HPLC-UV II

Cheese Propionic acid ISO/TS 19046-1I IDF/RM 233-1 Gas chromatography– FID IV

Cheese Propionic acid ISO/TS 19046-2I IDF/RM 233-2 Ion exchange chromatography-UV IV

Cheese Sodium chloride ISO 5943 | IDF 88 Potentiometry (determination of II

chloride, expressed as sodium
chloride)
Cheeses, individual Dry matter (total solids)xxxiv ISO 5534 | IDF 4 Gravimetry, drying at 102 °C I

xxxiii
Water content excluding the crystallized water bound to lactose (generally known as moisture content).
xxxiv
Milk total solids and milk solids-not-fat (MSNF) content include water of crystallization of lactose.
CXS 234-1999 51
Milk and milk products
Provisions Method Principle Type
Commodity
Cheeses, individual Milkfat in dry matter ISO 5534 | IDF 4 Calculation from dry matter content I
ISO 23319 | IDF 250 and fat content,
gravimetry, drying at 102 °C and
gravimetry (Schmid-Bondzynski-
Ratzlaff)
Cheeses in brine Milkfat in dry matter ISO 5534 | IDF 4 Calculation from dry matter content I
ISO 23319 | IDF 250 and fat content,
gravimetry, drying at 102 °C and
gravimetry (Schmid-Bondzynski-
Ratzlaff)
Cottage cheese Fat-free dry matter ISO 5534 | IDF 4 and Calculation from dry matter content I
ISO 23319 | IDF 250 and fat content,
gravimetry, drying at 102 °C and
gravimetry (Schmid-Bondzynski-
Ratzlaff)
Calculation from dry matter content
Cottage cheese (for samples and fat content,
ISO 5534 | IDF 4 and
containing lactose over 5% or with Milkfat in dry matter I
ISO 8262-3 | IDF 124-3 gravimetry, drying at 102 °C and
non-dairy ingredients)
gravimetry (Weibull-Berntrop)
Calculation from dry matter content
and fat content,
Cottage cheese (for samples ISO 5534 | IDF 4 and
Milkfat in dry matter gravimetry, drying at 102 °C and I
containing lactose up to 5%) ISO 23319 | IDF 250
gravimetry (Schmid-Bondzynski-
Ratzlaff)
Cottage cheese (for samples Gravimetry (Schmid-Bondzynski-
Milkfat ISO 23319 | IDF 250 I
containing lactose up to 5%) Ratzlaff)
Cottage cheese (for samples Milkfat ISO 8262-3 | IDF 124-3 Gravimetry (Weibull-Berntrop) I
containing lactose over 5% or with
non-dairy ingredients)
Cheese, unripened, including fresh Milk protein ISO 8968-1 | IDF 20-1 Titrimetry, Kjeldahl I
cheese
Cream and prepared creams Milk protein ISO 8968-1 | IDF 20-1 Titrimetry (Kjeldahl) I
CXS 234-1999 52
Milk and milk products
Provisions Method Principle Type
Commodity

Cream Milkfat ISO 23318 | IDF 249 Gravimetry (Röse-Gottlieb) I

Cream Solidsxxxv ISO 6731 | IDF 21 Gravimetry (drying at 102 °C) I
Creams lowered in milkfat content Milkfat ISO 23318 | IDF 249 / AOAC 995.19 Gravimetry (Röse-Gottlieb) I
Creams, whipped creams and Milk solids-not-fat (MSNF)xxxv ISO 3727-2 | IDF 80-2 Gravimetry I
fermented creams
I
Cream cheese Dry matter ISO 5534 | IDF 4 Gravimetry drying at 102 °C (forced air
oven)
Cream cheese Moisture on fat-free basis ISO 5534 | IDF 4 Calculation from fat content and I
moisture content,
ISO 23319 | IDF 250 gravimetry drying at 102 °C (forced air
oven),
gravimetry (Schmid-Bondzynski-
Ratzlaff)
Dairy fat spreads Milk fat purity ISO 17678 | IDF 202 Calculation from determination of I
triglycerides by gas chromatography–
FID
Dairy fat spreads Milkfat (total fat) ISO 17189 | IDF 194 Gravimetry I
Gravimetry (direct determination of fat
using solvent extraction)
Dairy permeate powders Lactose ISO 22662 | IDF 198 High performance liquid II
chromatography
Dairy permeate powders Milkfat ISO 23318 | IDF 249 Gravimetry (Röse-Gottlieb) I
Dairy permeate powders Nitrogen ISO 8968-1 | IDF 20-1 Titrimetry (Kjeldahl) I
Dairy permeate powders Moisturexxxvi ISO 5537 | IDF 26 Gravimetry (drying at 87 °C) I
Dairy permeate powders Ash NMKL 173 / AOAC 930.30 Gravimetry (ashing at 550 °C) I

Edible casein products Free acidity ISO 5547 | IDF 91 Titrimetry (aqueous extract) I

xxxv
Milk total solids and milk solids-not-fat (MSNF) content include water of crystallization of lactose.
xxxvi
Moisture content excluding the water of crystallization of lactose.
CXS 234-1999 53
Milk and milk products
Provisions Method Principle Type
Commodity
Edible casein products (caseins Ash (including P2O5) ISO 5545 | IDF 90 I
Gravimetry (ashing at 825 °C)
obtained by rennet precipitation and of
caseinates, with the exception of
ammonium caseinate)
CXS 234-1999 54
Milk and milk products
Provisions Method Principle Type
Commodity
Edible casein products (acid caseins, Ash (including P2O5) ISO 5544 | IDF 89 I
Gravimetry (ashing at 825 °C)
of ammonium caseinates, of their
mixtures with rennet casein and with
caseinates, and of caseins of
unknown type)
Edible casein products Lactose ISO 5548 | IDF 106 Photometry (phenol and H2SO4) IV
Edible casein products Milkfat (total fat) ISO 23319 | IDF 250 Gravimetry (Schmid-Bondzynski- I
Ratslaff)
Edible casein products pH ISO 5546 | IDF 115 Electrometry II
Edible casein products Milk protein (total N x 6.38 in ISO 5550 | IDF 78 and Calculation from dry matter content I
dry matter) and protein content
ISO 8968-1 | IDF 20-1
Gravimetry, drying at 102 oC and
titrimetry (Kjeldahl)
Edible casein products Sediment (scorched particles) ISO 5739 | IDF 107 Visual comparison with standard discs, IV
after filtration
Edible casein products Waterxxxvii ISO 5550 | IDF 78 Gravimetry (drying at 102 °C) I
Emmental Calcium ISO 8070 | IDF 119 Flame atomic absorption III
> = 800 mg/100 g
Emmental Calcium AOAC 2015.06 / ISO 21424 |IDF 243 ICP mass spectrometry II
> = 800 mg/100 g
Emmental Calcium AOAC 2011.14 / ISO 15151 | IDF 229 ICP emission spectroscopy III
> = 800 mg/100 g
Emmental Propionic acid ISO/TS 19046-1I IDF/RM 233-1 Gas Chromatography -FID IV
Emmental Propionic acid ISO/TS 19046-2I IDF/RM 233-2 Ion exchange chromatography - UV IV

Evaporated milks Milkfat ISO 23318 | IDF 249 Gravimetry (Röse-Gottlieb) I

xxxvii
Water content excluding the crystallized water bound to lactose (generally known as moisture content).
CXS 234-1999 55
Milk and milk products
Provisions Method Principle Type
Commodity

Calculation from total solids content, I

Evaporated milks Milk protein in MSNFxxxviii ISO 6731 | IDF 21 and
ISO 23318 | IDF 249 and fat content and protein content,
ISO 8968-1 | IDF 20-1 gravimetry, drying at 102 oC and
gravimetry (Röse-Gottlieb) and
titrimetry (Kjeldahl)
Evaporated milks Milk solidsxxxviiiError! Bookmark not ISO 6731 | IDF 21 Gravimetry, drying at 102 °C I
defined.

Colony count at 25 °C IV
Fermented milks Colony-forming units of yeasts ISO 6611 | IDF 94
and/or moulds
Gravimetry, drying at 102 °C I
Fermented milks Dry matter (total solids)xxxviiixviii ISO 13580 | IDF 151
Potentiometry, titration to pH 8.30 IV
Fermented milks Total acidity expressed as ISO/TS 11869 | IDF/RM 150
percentage of lactic acid
Colony count at 37 °C I
Fermented milks Lactobacillus acidophilus ISO 20128 | IDF 192
Colony count at 37 °C I
Fermented milks Quantification of Lactobacillus ISO 7889 | IDF 117
delbrueckii subsp. bulgaricus
Yoghurt and yoghurt products and Streptococcus
thermophilus
I
Fermented milks Identification of Lactobacillus ISO 9232 | IDF 146 Test for strain identification
delbrueckii subsp. bulgaricus
Yoghurt and yoghurt products and Streptococcus
thermophilus
Sum of microorganisms ISO 27205 | IDF 149 (Annex A) Colony count at 25 °C, 30 °C, 37 °C
constituting the starter culture and 45 °C according to the starter
Fermented milks (bacteria in fermented milk organism in question I
deriving (or originating) from
starter culture)
I
Fermented milks Milkfat ISO 23318 | IDF 249 Gravimetry (Röse-Gottlieb)

xxxviii
Milk total solids and milk solids-not-fat (MSNF) content include water of crystallization of lactose.
CXS 234-1999 56
Milk and milk products
Provisions Method Principle Type
Commodity
I
Fermented milks Milk protein ISO 8968-1 | IDF 20-1 Titrimetry (Kjeldahl)
Milk powders and cream powders Acidity, titratable ISO 6091 | IDF 86 Titrimetry, titration to pH 8.4 I
Milk powders and cream powders Milkfat ISO 23318 | IDF 249 Gravimetry (Röse-Gottlieb) I
Milk powders and cream powders Milk protein ISO 8968-1 | IDF 20-1 Titrimetry (Kjeldahl) I
Visual comparison with standard discs,
Milk powders and cream powders Scorched particles ISO 5739 | IDF 107 IV
after filtration
Visual comparison with standard discs,
Milk powders and cream powders Scorched particles ADPI Scorched Particles, 2016 IV
after filtration
Milk powders and cream powders Solubility index ISO 8156 | IDF 129 Centrifugation I
Milk powders and cream powders Waterxxxix (moisture) ISO 5537 | IDF 26 Gravimetry (drying at 87 °C) I
Milk powders and cream powders Waterxxxix (moisture) Described in Appendix XIxl Gravimetry (drying at 102 °C) IV
Milk fat products Fatty acids, free (expressed as ISO 1740 | IDF 6 Titrimetry I
oleic acid)
Milk fat products Milkfat purity ISO 17678 | IDF 202 Calculation from determination of I
triglycerides by gas chromatography–
FID
Milk fat products (anhydrous milkfat) Peroxide value (expressed as ISO 3976 | IDF 74 Photometry I
meq. of oxygen/kg fat)
Milk fat products Waterxxxix ISO 5536 | IDF 23 Titrimetry (Karl Fischer) II
Mozzarella Milkfat in dry matter – with ISO 5534 | IDF 4 and Calculation from dry matter content I
high moisture ISO 23319 | IDF 250 and fat content,
gravimetry, drying at 102 oC and
gravimetry (Schmid-Bondzynski-
Ratzlaff)
Mozzarella Milkfat in dry matter – with low ISO 5534 | IDF 4 and Calculation from dry matter content I
moisture ISO 23319 | IDF 250 and fat content,
gravimetry, drying at 102 oC and
gravimetry (Schmid-Bondzynski-
Ratzlaff)
Sweetened condensed milk Milkfat ISO 23318 | IDF 249 Gravimetry (Röse-Gottlieb) I

xxxix
Water content excluding the crystallized water bound to lactose (generally known as moisture content).
xl
Due to accessibility to equipment and calibration of the method ISO 5537 | IDF 26, the method described in Appendix XI is listed as Type IV.
CXS 234-1999 57
Milk and milk products
Provisions Method Principle Type
Commodity
Sweetened condensed milks (for Milk protein in MSNFxli ISO 6734 | IDF 15 and Calculation from total solids content, I
products sweetened with sucrose ISO 23318 | IDF 249 and fat content, sucrose and protein
only) ISO 2911 | IDF 35 and content,
ISO 8968-1 | IDF 20-1 gravimetry, drying at 102 °C and
polarimetry,
gravimetry (Röse-Gottlieb),
titrimetry (Kjeldahl)
Sweetened condensed milks Solidsxli ISO 6734 | IDF 15 Gravimetry, drying at 102 °C I
Whey cheeses by coagulation Milkfat ISO 23319 | IDF 250 Gravimetry (Schmid-Bondzynski- I
Ratzlaff)
Whey cheeses by coagulation Milkfat in dry matter ISO 23319 | IDF 250 and Calculation from fat content and dry I
ISO 5534 | IDF 4 matter content,
gravimetry (Schmid-Bondzynski-Ratzlaff),
gravimetry, drying at 102 °C
Whey cheeses by concentration Milkfat (total fat) ISO 23318 | IDF 59249 Gravimetry (Röse-Gottlieb) I
(carbohydrate contents below 5%)
Whey cheeses by concentration (does Milkfat (total fat) ISO 8262-3 |IDF 124-3 Gravimetry (Weibull-Berntrop) I
not dissolve completely in the
ammonia, contains fats and fatty acid
(FFA) in significant quantities or
carbohydrate content > 5%)
Whey cheeses by concentration Milkfat in dry matter ISO 23318 | IDF 249 and Calculation from fat content and dry I
(for carbohydrate content under 5%) (total fat in dry matter) ISO 2920 | IDF 58 matter content,
gravimetry (Röse-Gottlieb)
gravimetry, drying at 88 °C
Whey cheeses by concentration (does Milkfat in dry matter ISO 8262-3 | IDF 124-3 and Calculation from fat content and dry I
no dissolve completely in the (total fat in dry matter) ISO 2920 | IDF 58 matter contents,
ammonia, contains FFA in significant gravimetry (Weibull-Berntrop)
quantities, or carbohydrate content gravimetry, drying at 88 °C
>5%)

xli
Milk total solids and milk solids-not-fat (MSNF) content include water of crystallization of lactose.
CXS 234-1999 58
Milk and milk products
Provisions Method Principle Type
Commodity

Whey powders Ash ISO 5545 | IDF 90 Gravimetry (ashing at 825 M °C) IV
Whey powders Lactose ISO 5765-1/2 | IDF 79-1/2 Enzymatic method: Part 1– Glucose II
moiety or Part 2– Galactose moiety
Whey powders Milkfat ISO 23318 | IDF 249 Gravimetry (Röse-Gottlieb) I
Milk protein
Whey powders ISO 8968-1 | IDF 20-1 Titrimetry (Kjeldahl) I

Whey powders Waterxlii (moisture) ISO 5537 | IDF 26 Gravimetry (drying at 87 °C) I

xlii
Water content excluding the crystallized water bound to lactose (generally known as moisture content).
CXS 234-1999 59
Table 7. Numeric performance criteria for methods of analysis for copper and iron in milk fat products
Minimum applicable Examples of
ML LOD LOQ range applicable
Commodity Provision RSDR (%) Recovery Principle
(mg/kg) (mg/kg) (mg/kg) methods that
Minimum Maximum
meet the criteria
AOAC 2015.06 /
ISO 21424 | IDF ICP mass spectrometry
243
Milk fat
Copper 0.05 0.010 0.020 44.0 60–115% 0.028 0.072 Photometry,
products ISO 5738 | IDF 76
(diethyldithiocarbamate)
Photometry,
AOAC 960.40
(diethyldithiocarbamate)
AOAC 2015.06 /
Milk fat
Iron 0.2 0.020 0.040 40.8 80–110% 0.08 0.32 ISO 21424 | IDF ICP mass spectrometry
products
243
CXS 234-1999 60
Table 8. Numeric performance criteria for copper and iron in edible casein products
Examples of
Minimum applicable
ML LOD LOQ Recovery
Commodity Provision RSDR (%) applicable range methods that Principle
(mg/kg) (mg/kg) (mg/kg) (%)
Minimum Maximum meet the
criteria
ICP mass spectrometry
AOAC 2015.06 / ICP emission
ISO 21424 | IDF spectroscopy
243
AOAC 2011.14 /
ISO 15151 | IDF
Edible casein 229
Copper 5 0.50 1.0 25.1 80–110 3.1 6.9
products
AOAC 985.35 Atomic absorption
spectrophotometry

ISO 5738 | IDF Colorimetry

76 (diethyldiethiocarbamate)

AOAC 2015.06 /
ISO 21424 | IDF
ICP mass spectrometry
243
Iron 20 2.0 4.0 20.4 80–110 13,9 26.1 ICP emission
AOAC 2011.14 /
spectroscopy
ISO 15151 | IDF
Edible casein 229
products AOAC 2015.06 /
ISO 21424 | IDF
Iron (in ICP mass spectrometry
243
roller dried 50 5.0 10.0 17.8 90–107 36.7 63.3 ICP emission
AOAC 2011.14 /
caseinates) spectroscopy
ISO 15151 | IDF
229
CXS 234-1999 61

Natural mineral waters

Provisions Method Principle Type
Commodity

Natural mineral waters Calcium ISO 7980 Atomic absorption III

spectrophotometry
Natural mineral waters Chloride Examination of Water Pollution Control. II
WHO Pergamon Press (1982) Vol. 2,
pp. 205-208
Natural mineral waters Chloride AOAC 973.51 Titrimetry (mercuric nitrate) III
Natural mineral waters Chloride ISO 9297 Titrimetry III
Natural mineral waters Iron, dissolved ISO 6332 Spectrophotometry II
Natural mineral waters Magnesium ISO 6059 Titrimetry II
Natural mineral waters Magnesium ISO 7980 Atomic absorption III
spectrophotometry
Natural mineral waters Phenols ISO 6439 Spectrophotometry I
Natural mineral waters Potassium Examination of Water Pollution Control. II
WHO Pergamon Press (1982) Vol. 2,
pp. 142-145
Natural mineral waters Sodium Examination of Water Pollution Control. II
WHO Pergamon Press (1982) Vol. 2
pp. 148-151
Natural mineral waters Sodium Examination of Water Pollution Control. III
WHO Pergamon Press (1982) Vol. 2,
pp. 151-152
Natural mineral waters Sulphates ISO 9280 Gravimetry III
Natural mineral waters Sulphide Handb. Spurenanal. 1974 IV
CXS 234-1999 62
Table 9a. Criteria applicable to health-related substances in the Standard for natural mineral waters (CXS 108-1981)1

Provision ML Min. applicable LOD LOQ Precision RSDR (%) Recovery Suggested methods Principle
(mg/L) range (mg/L) (mg/L) (mg/L) no more than (%) meeting the criteria

Antimony 0.005 0.0028 0.001 0.002 44 80–110 ISO 17294-2 ICP-MS

ISO 15586 GF-AAS
EPA 200.8 ICP-MS
Arsenic 0.01 0.0056 0.002 0.004 44 90–107 ISO 17294-2 ICP-MS
ISO 15586 GF-AAS
ISO 11969 AAS (Hydride)
EPA 200.8 ICP-MS
Barium 0.7 0.35 0.07 0.14 34 95–105 ISO 11885 ICP-OES
ISO 17294-2 ICP-MS
EPA 200.8
ICP-MS
Borate 5 3.1 0.5 1 25 97–103 ISO 9390 Spectrophotometry
ISO 11885 ICP-OESxliii
ISO 17294-2
ICP-MSxliii
Cadmium 0.003 0.0017–0.0043 0.0006 0.0012 44 40–120 ISO 17294-2 ICP-MS
EPA 200.8 ICP-MS
EN 17851 ICP-MS
EN 14083 GF-AAS
Chromium 0.05 0.028 0.01 0.02 44 90–107 ISO 11885 ICP-OES
ISO 17294-2 ICP-MS
ISO 15586ISO 18412 (Cr GF-AAS
VI) Photometric
ISO 23913 (Cr VI) CIA, spectrophotometry
ISO 9174 (Section 4) AAS
EPA 200.8 ICP-MS
Copper 1 0.52 0.1 0.2 32 97–103 ISO 11885 ICP-OES
ISO 17294-2 ICP-MS

xliii
Total boron is determined.
CXS 234-1999 63
Provision ML Min. applicable LOD LOQ Precision RSDR (%) Recovery Suggested methods Principle
(mg/L) range (mg/L) (mg/L) (mg/L) no more than (%) meeting the criteria

ISO 15586 GF-AAS

ISO 8288 Flame-AAS
EPA 200.8 ICP-MS
Cyanide 0.07 0.039 0.014 0.028 44 90–107 ISO 14403 CFA
ISO 6703-1 Photometric, trimetric
Fluoride 1.0 0.52 0.1 0.2 32 97–103 ISO 10304-1 LC of ions
ISO 10359-1 (dissolved Electrochemical probe
fluoride)
ISO 10359-2 (inorganic Digestion, distillation
bound)

Lead 0.01 0.006–0.014 0.002 0.004 44 60–115 ISO 17294-2 ICP-MS

ISO 15586 GF-AAS
EPA 200.8 ICP-MS
Manganese 0.4 0.18 0.04 0.08 37 95–105 ISO 11885I ICP-OES
SO 17294-2 ICP-MS
ISO 15586 GF-AAS
EPA 200.8 ICP-MS
Mercury 0.001 0.00056 0.0002 0.0004 44 80–110 EN 1483 AAS
ISO 17852 Enrichment by amalgamation (III)
ISO 5666 AFS
ISO 16590 AAS after tin (II) chloride
EPA 200.8 reduction
Enrichment by amalgamation (III)
ICP-MS
Nickel 0.02 0.011 0.004 0.008 44 90–107 ISO 17294-2 ICP-MS
ISO 15586 GF-AAS
EPA 200.8 ICP-MS
Nitrate 50 37 5 10 18 98–102 ISO 10304-1 LC of ions
ISO 13395 CFA, FIA, spectrophotometry
ISO 7890-3
CXS 234-1999 64
Provision ML Min. applicable LOD LOQ Precision RSDR (%) Recovery Suggested methods Principle
(mg/L) range (mg/L) (mg/L) (mg/L) no more than (%) meeting the criteria

Nitrite 0.1 0.03 0.01 0.02 44 95–105 ISO 10304-1 LC of ions UV

ISO 13395 CFA, FIA, spectrophotometry
ISO 6777
Selenium 0.01 0.0056 0.002 0.004 44 90–107 ISO 17294-2 ICP-MS
ISO 15586 GF-AAS
ISO 9965 AAS (hydride)
EPA 200.8 ICP-MS

Table 9b. Performance characteristics of suggested methods

Provision ML Applicable range-from: LOD RSDR (%) Recovery (%) Suggested methods Principle

Surface active agents - 0.05–5.0 mg/L 0.05 mg/l < 44 70–100 ISO 16265 CFA
Mineral oil - >0.1 mg/L < 41 71–102 ISO 9377-2 GC
(hydrocarbon index)
PCB - >15 ng/L < 20 70–130 AOAC 990.06 GC ECD
Pesticide (organochlorine) - > 15 ng/L < 20 70–130 AOAC 990.06 GC ECD
PAH - 0.005 μg/L < 10 80–110 ISO 17993 HPLC FD
0.04 μg/L < 18 80–110 ISO 7981-1 TLC
0.005 μg/L < 19 80–100 ISO 7981-2 HPLC

Processed fruits and vegetables

Commodity Provision Method Principle Type

Processed fruits and vegetables
(except jams, jellies, marmalades,
Benzoic acid NMKL 124 Liquid chromatography (UV) II
pickled cucumbers, mango chutney,
coconut milk and coconut cream)
CXS 234-1999 65
Processed fruits and vegetables

Commodity Provision Method Principle Type

Processed fruits and vegetables

(except jams, jellies, marmalades,
Benzoic acid AOAC 983.16 Gas chromatography III
pickled cucumbers, mango chutney,
coconut milk and coconut cream)

Processed fruits and vegetables

(canned strawberries, pickled
cucumbers, preserved tomatoes, Calcium AOAC 968.31 Complexometry/Titrimetry II
canned citrus fruits, certain canned
vegetables)
AOAC 968.30
Processed fruits and vegetables Drained weight Gravimetry (sieving) I
(Codex general method)
Processed fruits and vegetables Fill of glass containers ISO 8106 Gravimetry I
CXS 234-1999 66

Processed fruits and vegetables

Commodity Provision Method Principle Type

Processed fruits and Fill of metal ISO 90-1 Gravimetry I

vegetables containers

Packing
I
medium
Processed fruits and canned berry AOAC 932.12
Refractometry
vegetables fruits ISO 2173
(raspberry,
strawberry)
Processed fruits and ISO 1842 Potentiometry IV
pH
vegetables (pickled
cucumbers, table
olives, processed
tomato concentrates,
preserved tomatoes,
mango chutney and
aqueous coconut
products)
Canned bamboo AOAC 981.12 Potentiometry IV
pH
shoots
Processed fruits and AOAC 981.12 Potentiometry III
pH
vegetables
(pickled cucumbers,
table olives, processed
tomato concentrates,
preserved tomatoes,
mango chutney and
aqueous coconut
products)
CXS 234-1999 67

Processed fruits and vegetables

Commodity Provision Method Principle Type
Processed fruits and NMKL 179 Potentiometry II
pH
vegetables
(pickled cucumbers,
table olives, processed
tomato concentrates,
preserved tomatoes,
mango chutney and
aqueous coconut
products)
Processed fruits and Soluble solids ISO 2173 Refractometry I
vegetables (packing
(pickled cucumbers, medium)
processed tomato
concentrates,
preserved tomatoes,
canned applesauce,
jams, jellies and
marmalades, mango
chutney and certain
canned fruit)
Processed fruits and Sorbates
vegetables
Gas chromatography
(except jams, jellies, AOAC 983.16 III
(Flame ionization)
marmalades, pickled
cucumbers)
Processed fruits and Sorbates
vegetables
Liquid chromatography
(except jams, jellies, NMKL 124 II
(UV)
marmalades, pickled
cucumbers)
CXS 234-1999 68

Processed fruits and vegetables

Commodity Provision Method Principle Type

Processed fruits and I

Total solids AOAC 920.151 Gravimetry
vegetables
Aqueous coconut Gravimetry (Röse- I
Total fats ISO 23318 | IDF 249
products Gottlieb)
Aqueous coconut I
Total solids ISO 6731 | IDF 21 Gravimetry
products
Calculation: I
Aqueous coconut
Gravimetry (Röse-
products Non-fat solids ISO 23318 | IDF249 and ISO 6731 | IDF 21
Gottlieb)
Gravimetry
Aqueous coconut I
Moisture ISO 6731 | IDF 21 Gravimetry
products
Canned apple sauce Fill of glass ISO 8106 Gravimetry I
containers
Canned apple sauce Fill of metal ISO 90-1 Gravimetry I
containers
Canned apple sauce Soluble solids Refractometry I
(packing ISO 2173 (Codex general method for processed fruits and vegetables)
medium)
Canned green beans Tough strings See Appendix XII Stretching I
and wax beans
Canned green peas Fill of glass ISO 8106 Gravimetry I
containers
Canned green peas Fill of metal ISO 90-1 Gravimetry I
containers
Canned green peas Types of peas, See Appendix XIII Visual examination I
distinguishing
Canned mangoes Soluble solids AOAC 932.14C Brix spindle method I
(packing (refractometry)
medium)
CXS 234-1999 69

Processed fruits and vegetables

Commodity Provision Method Principle Type
Canned mushrooms Drained weight AOAC 968.30 Gravimetry (sieving) I
Canned palmito Mineral ISO 762 Gravimetry I
impurities
AOAC 968.30 I
Canned stone fruits Drained weight Gravimetry (sieving)

Soluble solids I
Canned stone fruits (packing ISO 2173 Refractometry
medium)
Canned strawberries Calcium AOAC 968.31 Complexometric titrimetry II
Canned strawberries Mineral ISO 762 Gravimetry
impurities
Certain canned citrus Calcium Atomic absorption
fruits NMKL 153 spectrophotometry II
(flame)
Certain canned citrus Calcium
AOAC 968.31 Complexometry titrimetry III
fruits
Citrus marmalade Calcium AOAC 968.31 Complexometric titrimetry II
Dates Identification of See Appendix XIV Visual examination I
defects
Dates Moisture AOAC 934.06 Gravimetry (vacuum I
oven)
Desiccated coconut Total acidity of ISO 660 or AOCS Cd 3d-63 Titrimetry I
the extracted oil
Desiccated coconut Ash AOAC 950.49 Gravimetry (ashing) I
Desiccated coconut Extraneous Described in Appendix XV Counting extraneous IV
vegetable matter material with the naked
eye
CXS 234-1999 70

Processed fruits and vegetables

Commodity Provision Method Principle Type
Desiccated coconut Moisture AOAC 925.40 Gravimetry (loss on I
drying)
Desiccated coconut Oil content AOAC 948.22 Gravimetry I
Dried apricots Identification of See Appendix XVI Visual inspection I
defects (gravimetry)
Dried apricots Moisture AOAC 934.06 Gravimetry (vacuum I
oven)
Dried apricots Sulphur dioxide AOAC 963.20 Colorimetry II
Jams, jellies and ISO 8106 I
Fill of glass Gravimetry
marmalades containers

Jams, (fruit preserves) Soluble solids ISO 2173 Refractometry I

and jellies and
marmalades
Mango chutney Ash insoluble in ISO 763 Gravimetry I
HCl
Pickled cucumbers Acidity, total AOAC 942.15 Titrimetry I
Pickled cucumbers Drained weight AOAC 968.30 Gravimetry I
Pickled cucumbers Mineral ISO 762 Gravimetry I
impurities
Pickled cucumbers Salt (NaCl) AOAC 971.27 (Codex general method) Potentiometry II
Pickled cucumbers Volume fill by See Appendix XVII Displacement I
displacement
Preserved tomatoes Calcium AOAC 968.31 Complexometric titrimetry III
Preserved tomatoes Calcium Atomic absorption
NMKL 153 spectrophotometry II
(flame)
CXS 234-1999 71
Processed fruits and vegetables
Commodity Provision Method Principle Type
Preserved tomatoes Drained weight AOAC 968.30 Gravimetry (sieving) I
Note: Use a No. 14
screen instead of ‘7/16’
or No. 8
Preserved tomatoes Mould count AOAC 965.41 Howard mould count I
Processed tomato Lactic acid Spectrometry (enzymatic II
EN 12631
concentrates determination)
Processed tomato Mineral ISO 762 Gravimetry IV
concentrates impurities (sand)
Processed tomato Mould count AOAC 965.41 Howard mould count I
concentrates
Processed tomato Natural tomato AOAC 970.59 Refractometry I
concentrates soluble solids
Processed tomato Sodium chloride AOAC 971.27 (Codex general method) Potentiometry II
concentrates
Processed tomato Tomato soluble
AOAC 970.59 Refractometry I
concentrates solids
Raisins Mineral ISO 762 Ashing I
impurities
Raisins Mineral oil CAC/RM 52 Extraction and separation II
on alumina
Raisins Moisture AOAC 972.20 Electrical conductance I
Raisins Sorbitol AOAC 973.28 Gas chromatography II
(flame ionization)
Raisins Sulphur dioxide AOAC 963.20 Colorimetry II
Table olives Drained weight AOAC 968.30 (Codex general method for processed fruits and vegetables) Gravimetry (sieving) I
Table olives Fill of glass ISO 8106 Gravimetry I
containers
CXS 234-1999 72

Processed fruits and vegetables

Commodity Provision Method Principle Type
Table olives Fill of metal ISO 90-1 (for metal containers) Gravimetry I
containers (Codex general method for processed fruits and vegetables)
Table olives pH of brine NMKL 179 (Codex general method for processed fruits and vegetables) Potentiometry II
Table olives pH of brine AOAC 981.12 (Codex general method for processed fruits and vegetables) Potentiometry III
Table olives pH of brine ISO 1842 Potentiometry IV
Table olives Salt in brine AOAC 971.27 | NMKL 178 (Codex general method) Potentiometry II

Table 10. Numeric performance criteria for benzoic acid, calcium, sorbates and tin in processed fruits and vegetables

Limit of Precision
Minimum Limit of
ML detection (RSDR) Recovery Example of applicable methods
Commodity Provision applicable quantification
(mg/kg) (LOD) (%) no (%) that meet the criteria
range (mg/kg) (LOQ) (mg/kg)
(mg/kg) more than

Jams, jellies and Benzoic

1 000 830–1 170 100 200 11.3 95–105 ISO 5518, NMKL 124, AOAC 983.16
marmalades Acid

Pickled Benzoic
1 000 830–1 170 100 200 11.3 95–105 NMKL 124, AOAC 983.16
cucumbers Acid

Benzoic
Mango chutney 250 197–302 25 50 13.9 90–107 ISO 5518, NMKL 124, AOAC 983.16
Acid

Coconut milk and Benzoic

1 000 830–1 170 100 200 11.3 95–105 ISO 5518, NMKL 124, AOAC 983.16
coconut cream Acid

Jams, jellies and

Sorbates 1 000 830–1 170 100 200 11.3 95–105 NMKL 124, AOAC 983.16
marmalades

Pickled
Sorbates 1 000 830–1 170 100 200 11.3 95–105 NMKL 124, AOAC 983.16
cucumbers
CXS 234-1999 73

Limit of Precision
Minimum Limit of
ML detection (RSDR) Recovery Example of applicable methods
Commodity Provision applicable quantification
(mg/kg) (LOD) (%) no (%) that meet the criteria
range (mg/kg) (LOQ) (mg/kg)
(mg/kg) more than

Processed fruits
Tin 250 197–302 25 50 13.9 90–107 AOAC 980.19, NMKL 126, NMKL 191
and vegetables

NMKL 190, EN 15764, NMKL 126,

Table olives Tin 250 197–302 25 50 13.9 90–107
NMKL 191
CXS 234-1999 74

Processed meat and poultry products and soups and broths

Provisions Method Principle Type
Commodity
Spectrometric III
determination of nitrate and
Meat products Nitrates and/or nitrites EN 12014-3 nitrite content of meat
products after enzymatic
reduction of nitrate to nitrite
Ion exchange III
Meat products Nitrates and/or nitrites EN 12014-4 NMKL 165
chromatographic method
Processed meat and poultry products Fat ISO 1443 Gravimetry I
Processed meat and poultry products Lead AOAC 934.07 Colorimetry (dithizone) II
Processed meat and poultry products Nitrates ISO 3091 Colorimetry (cadmium II
reduction)
Processed meat and poultry products Nitrites ISO 2918 Colorimetry IV
Processed meat and poultry products Tin AOAC 985.16 (Codex general method) Atomic absorption II
spectrophotometry
Processed meat and poultry products Nitrogen/protein ISO 937 Titrimetry II
Bouillons and consommés Amino nitrogen AIIBP Method No 2/7 Volumetry (modified Van II
(soups and broths) Slyke)
Bouillons and consommés Creatinine AIIBP Method No 2/5 HPLC II
(soups and broths)
Bouillons and consommés Nitrogen, total AOAC 928.08 Kjeldahl II
(soups and broths)
Bouillons and consommés Sodium chloride AIIBP Method No 2/4 Potentiometric titration II
(soups and broths) (chloride expressed as
sodium chloride)
Canned corned beef Lead AOAC 972.25 (Codex general method) Atomic absorption II
spectrophotometry
Canned corned beef Nitrites, potassium AOAC 973.31 (Codex general method) Colorimetry II
and/or sodium salt
CXS 234-1999 75
Processed meat and poultry products and soups and broths
Provisions Method Principle Type
Commodity

Canned corned beef Nitrites, potassium ISO 2918 Colorimetry IV

and/or sodium salt
Canned corned beef Tin (products in AOAC 985.16 (Codex general method) Atomic absorption II
tinplate and other spectrophotometry
containers)
Cooked cured chopped meat Fat ISO 1443 Gravimetry (extraction) I
Cooked cured chopped meat Lead AOAC 972.25 (Codex general method) Atomic absorption II
spectrophotometry
Cooked cured chopped meat Nitrites AOAC 973.31 (Codex general method) Colorimetry II
Cooked cured chopped meat Nitrites ISO 2918 Colorimetry IV
Cooked cured chopped meat Tin AOAC 985.16 (Codex general method) Atomic absorption II
spectrophotometry
Cooked cured ham Fat ISO 1443 Gravimetry (extraction) I
Cooked cured ham Gelatin, added Described in the standard Calculation I
Cooked cured ham Lead AOAC 972.25 (Codex general method) Atomic absorption II
spectrophotometry
Cooked cured ham Nitrites AOAC 973.31 (Codex general method) Colorimetry II
Cooked cured ham Nitrites ISO 2918 Colorimetry IV
Cooked cured ham Protein (conversion ISO 937 Titrimetry, Kjeldahl II
factor 6.25) digestion
Cooked cured ham Tin AOAC 985.16 (Codex general method) Atomic absorption II
spectrophotometry
Cooked cured pork shoulder Fat ISO 1443 Gravimetry (extraction) I
Cooked cured pork shoulder Gelatin, added Described in the standard Calculation I
Cooked cured pork shoulder Lead AOAC 972.25 (Codex general method) Atomic absorption II
spectrophotometry
CXS 234-1999 76
Processed meat and poultry products and soups and broths
Provisions Method Principle Type
Commodity

Cooked cured pork shoulder Nitrites AOAC 973.31 (Codex general method) Colorimetry II
Cooked cured pork shoulder Nitrites ISO 2918 Colorimetry IV
Cooked cured pork shoulder Protein ISO 937 Titrimetry, Kjeldahl II
digestion
Cooked cured pork shoulder Tin AOAC 985.16 (Codex general method) Atomic absorption II
spectrophotometry
Luncheon meat Fat ISO 1443 Gravimetry (extraction) I
Luncheon meat Lead AOAC 972.25 (Codex general method) Atomic absorption II
spectrophotometry
Luncheon meat Nitrites, potassium AOAC 973.31 (Codex general method) Colorimetry II
and/or sodium salt
Luncheon meat Nitrites, potassium ISO 2918 Colorimetry IV
and/or sodium salt
Luncheon meat Tin AOAC 985.16 (Codex general method) Atomic absorption II
spectrophotometry

Quick-frozen fruits and vegetables

Commodity Provisions Method Principle Type

Quick-frozen fruits and Net weight AOAC 963.26 Weighing I

vegetables(non-glazed)
Quick-frozen fruits and vegetables Thawing procedure See Appendix XVIII Thawing I
Quick-frozen fruits and vegetables: Mineral impurities AOAC 971.33 Gravimetry I
berries, leek and carrot

Quick-frozen fruits and vegetables: Soluble solids, total AOAC 932.12 Refractometry I
berries, whole kernel corn and
corn-on-the-cob
CXS 234-1999 77

Quick-frozen fruits and vegetables

Provisions Method Principle Type
Commodity

Quick-frozen fruits and vegetables: Drained fruit/drained AOAC 953.15 Draining I

peaches and berries berries

Quick-frozen fruits and vegetables: Cooking procedure See Appendix XIX Cooking I
vegetables
Quick-frozen French-fried potatoes Moisture AOAC 984.25 Gravimetry (convection oven) I
Quick-frozen green and wax beans Tough strings See Appendix XII Stretching I
Quick-frozen peas Solids, alcohol See Appendix XX Gravimetry I
insoluble
Quick-frozen spinach Dry matter, sodium See Appendix XXI Weighing I
chloride-free

Spices and culinary herbs

Commodity Provisions Method Principle Type

Allspice, juniper berry and star anise Moisture ISO 939 Distillation I
Allspice, juniper berry and star anise Total ash on dry basis ISO 939 and ISO 928 Calculation from moisture and ash (at I
5500C), distillation and gravimetry
Allspice, juniper berry and star anise Acid-insoluble ash on dry basis ISO 939 and ISO 930 Calculation from moisture and ash (at I
5500C), distillation and gravimetry
Allspice, juniper berry and star anise Volatile oils on dry basis ISO 939 and ISO 6571 Calculation from moisture and volatile oils, I
distillation and distillation
Allspice, juniper berry and star anise Extraneous matter ISO 927 Visual examination followed by gravimetry I
Allspice, juniper berry and star anise Foreign matter ISO 927 Visual examination followed by gravimetry I
Allspice, juniper berry and star anise Mould visible ISO 927 Visual examination followed by gravimetry I
Allspice, juniper berry and star anise Mammalian and other excreta MPM V-8 Spices, Condiments, Visual examination followed by gravimetry IV
(whole spice) Flavors and Crude Drugs MPM:
V-8, Spices
CXS 234-1999 78
Spices and culinary herbs

Commodity Provisions Method Principle Type

Allspice, juniper berry and star anise Whole dead insects and live ISO 927 Visual examination (counting) I
insects
Allspice, juniper berry and star anise Insect fragments (for whole ISO 927 Visual examination (counting) I
spices)
Allspice, juniper berry and star anise Insect fragments (for AOAC 975.49 Flotation I
powdered/pieces)
Allspice, juniper berry and star anise Insect defiled ISO 927 Visual examination followed by gravimetry I
Allspice, juniper berry and star anise Rodent hair AOAC 965.40 Flotation I
Cloves Moisture ISO 939 Distillation I
Cloves Volatile oil on dry basis ISO 939 and ISO 6571 Calculation from moisture and volatile oils, I
distillation and distillation
Cloves Total ash on dry basis ISO 939 and ISO 928 Calculation from moisture and ash (at 550 I
0C), distillation and gravimetry

Cloves Acid-insoluble ash on dry basis ISO 939 and ISO 930 Calculation from moisture and ash (at 550 I
0C), distillation and gravimetry

Cloves Extraneous matter ISO 927 Visual examination followed by gravimetry I

Cloves Foreign matter ISO 927 Visual examination followed by gravimetry I
Cloves Insect damage ISO 927 Visual examination followed by gravimetry I
Cloves Insects/insect fragments ISO 927 Visual examination (counting) I
Cloves Crude fibre ISO 5498 Gravimetry I
Cloves Mould visible (for whole) Method V-8 Spices, Condiments, Visual examination followed by gravimetry IV
Flavors and Crude Drugs
(Macroanalytical Procedure
Manual, FDA Technical Bulletin
Number 5)
https://www.fda.gov/food/laborat
ory-methods-food/mpm-v-8-
spices-condiments-flavors-and-
crude-drugs#v32
CXS 234-1999 79
Spices and culinary herbs

Commodity Provisions Method Principle Type

Cloves Live insects ISO 927 Visual examination (counting) I
Cloves Mammalian or/and other excreta MPM V-8 Spices, Condiments, Visual examination followed by gravimetry IV
(for whole) Flavors and Crude Drugs A,
General methods for spices,
herbs and botanicals (v 32)
https://www.fda.gov/food/laborat
ory-methods-food/mpm-v-8-
spices-condiments-flavors-and-
crude-drugs#v32
Cumin Moisture ISO 939 Distillation I
Cumin Total ash ISO 928 Gravimetry I

Cumin Acid-insoluble ash ISO 930 Gravimetry I

Cumin Volatile oils ISO 6571 Distillation/Volumetric I

Cumin Extraneous vegetable matter ISO 927 Visual examination/Gravimetry I

Cumin Foreign matter ISO 927 Visual examination/Gravimetry I

CXS 234-1999 80

Spices and culinary herbs

Provisions Method Principle Type
Commodity

Cumin Insect damage Method V-8 Spices, Condiments, Visual examination IV

Flavors and Crude Drugs
(Macroanalytical Procedure
Manual, FDA)

http://www.fda.gov/Food/FoodSc
ienceResearch/LaboratoryMetho
ds/ucm084394.htm#v-32

Cumin Mammalian excreta Macroanalytical Procedure Visual examination IV

Manual USFDA Technical
Bulletin V.39 B (for whole)
Cumin Mammalian excreta AOAC 993.27 (for ground) Enzymatic detection method IV
Cumin Mould damage Method V-8 Spices, Condiments, Visual examination IV
Flavours and Crude Drugs
(Macroanalytical Procedure
Manual, FDA)
http://www.fda.gov/Food/FoodSc
ienceResearch/LaboratoryMetho
ds/ucm084394.htm#v-32

Dried basil Moisture ISO 939 Distillation I

Dried basil Total ash on dry basis ISO 939 and ISO 928 Calculation from moisture and ash at (at I
550 0C), distillation and gravimetry
Dried basil Acid-insoluble ash on dry basis ISO 928 and ISO 930 Calculation from moisture and ash at (at I
550 0C), distillation and gravimetry
Dried basil Volatile oil on dry basis ISO 939 and ISO 6571 Calculation from moisture and volatile oils, I
distillation and distillation
Dried basil Extraneous matter ISO 927 Visual examination followed by gravimetry I
Dried basil Foreign matter ISO 927 Visual examination followed by gravimetry I
CXS 234-1999 81

Spices and culinary herbs

Commodity Provisions Method Principle Type
Dried basil Insect damage (whole leaves) Method V-8 Spices, Condiments, Visual examination followed by gravimetry IV
Flavors and Crude Drugs
(Macroanalytical Procedure
Manual, FDA Technical Bulletin
Number 5)
https://www.fda.gov/food/laborat
ory-methods-food/mpm-v-8-
spices-condiments-flavors-and-
crude-drugs#v32
Dried basil Insects / insect fragments ISO 927 Visual examination (counting) I
Dried basil Mould damage (for whole Method V-8 Spices, Condiments, Visual examination followed by gravimetry IV
leaves) Flavors and Crude Drugs
(Macroanalytical Procedure
Manual, FDA Technical Bulletin
Number 5)
https://www.fda.gov/food/laborat
ory-methods-food/mpm-v-8-
spices-condiments-flavors-and-
crude-drugs#v32
Dried basil Mammalian excreta and other Method V-8 Spices, Condiments, Visual examination followed by gravimetry IV
excreta (for whole leaves) Flavors and Crude Drugs A.
General methods for spices,
herbs and botanicals (V 32)
(https://www.fda.gov/food/laborat
ory-methods-food/mpm-v-8-
spices-condiments-flavors-and-
crude-drugs#v32
Dried or dehydrated chilli pepper and Moisture ISO 939 Distillation I
paprika
Dried or dehydrated chilli pepper and Total ash on dry basis ISO 939 and ISO 928 Calculation from moisture and ash (at 550 0C), I
paprika distillation and gravimetry
Dried or dehydrated chilli pepper and Acid-insoluble ash on dry basis ISO 939 and ISO 930 Calculation from moisture and ash (at 550 0C), I
paprika distillation and gravimetry
CXS 234-1999 82

Spices and culinary herbs

Commodity Provisions Method Principle Type
Dried or dehydrated chilli pepper and Pungency, Scoville Heat Units ISO 3513 Sensory evaluation I
paprika
Dried or dehydrated chilli pepper and Colour value ISO 7541 Spectrophotometry I
paprika
Dried or dehydrated chilli pepper and Mammalian excreta (whole) ISO 927 Visual examination followed by gravimetry I
paprika
Dried or dehydrated chilli pepper and Mould damage (for whole chilli MPM V-8 Spices, Condiments, Visual examination followed by gravimetry IV
paprika peppers) Flavors and Crude Drugs A.
General methods for spices, herbs
and botanicals (V.32)
Dried or dehydrated chilli pepper and Mould damage (for ground) AOAC 945.94 Visual examination (Howard mould count) I
paprika
Dried or dehydrated chilli pepper and Insect damage (for whole chilli MPM V-8 Spices, Condiments, Visual examination followed by gravimetry IV
paprika peppers) Flavors and Crude Drugs A.
General methods for spices, herbs
and botanicals (V.32)
Dried or dehydrated chilli pepper and Extraneous matter ISO 927 Visual examination followed by gravimetry I
paprika
Dried or dehydrated chilli pepper and Foreign matter ISO 927 Visual examination followed by gravimetry I
paprika
Dried or dehydrated chilli pepper and Live insects ISO 927 Visual examination (counting) I
paprika
Dried or dehydrated chilli pepper and Insect fragments ISO 927 Visual examination (counting) I
paprika
Dried or dehydrated chilli pepper and Rodent hair (ground chilli) AOAC 978.22 Flotation I
paprika

Dried or dehydrated chilli pepper and Rodent hair (ground paprika) AOAC 977.25 B Microscopic examination I
paprika
Dried or dehydrated ginger Moisture ISO 939 Distillation I
CXS 234-1999 83

Spices and culinary herbs

Commodity Provisions Method Principle Type
Dried or dehydrated ginger Total ash on dry basis ISO 939 and ISO 928 Calculation from moisture and ash (at 600 0C), I
distillation and gravimetry
Dried or dehydrated ginger Acid-insoluble ash on dry basis ISO 939 and ISO 930 Calculation from moisture and ash (at 600 0C), I
distillation and gravimetry
Dried or dehydrated ginger Volatile oil on dry basis ISO 939 and ISO 6571 Calculation from moisture and ash (at 600 0C), I
distillation and distillation
Dried or dehydrated ginger Extraneous matter ISO 927 Visual examination followed by gravimetry I
Dried or dehydrated ginger Foreign matter ISO 927 Visual examination followed by gravimetry I
Dried or dehydrated ginger Insect damage Method V-8 Spices, Condiments, Visual examination followed by gravimetry IV
Flavors and Crude Drugs
(Macroanalytical Procedure
Manual) MPM: V-8 Spices
Dried or dehydrated ginger Whole dead insect ISO 927 Visual examination I
Dried or dehydrated ginger Mammalian/other excreta (for Macroanalytical Procedure Visual examination followed by gravimetry IV
whole) Manual, USFDA, Technical
Bulletin V.39 B
https://www.fda.gov/food/laborator
y-methods-food/mpm-v-8-spices-
condiments-flavors-and-crude-
drugs#v32
Dried or dehydrated ginger Mould visible ISO 927 Visual examination followed by gravimetry I
Dried or dehydrated ginger Live insects ISO 927 Visual examination I
Dried or dehydrated ginger Calcium (as oxide) on dry basis ISO 939 and ISO 928 and ISO Calculation from moisture and ash (at 600 0C), IV
1003-Annex A and titrimetry
Dried or dehydrated ginger Sulphur dioxide AOAC 990.28 Distillation followed by titrimetry IV
Dried oregano Moisture ISO 939 Distillation I
Dried oregano Total ash ISO 939 and ISO 928 Calculation from moisture and ash I
(dry weight basis) Distillation and gravimetry
CXS 234-1999 84

Spices and culinary herbs

Commodity Provisions Method Principle Type
Dried oregano Acid-insoluble ash ISO 939 and ISO 930 Calculation from moisture and ash I
(dry weight basis) Distillation and gravimetry
Dried oregano Volatile oils ISO 939 and ISO 6571 Calculation from moisture and volatile oils I
(dry weight basis) distillation and distillation
Dried oregano Extraneous matter ISO 927 Visual examination followed by gravimetry I
Dried oregano Foreign matter ISO 927 Visual examination followed by gravimetry I
CXS 234-1999 85

Spices and culinary herbs

Provisions Method Principle Type
Commodity

Dried oregano Mammalian excreta Macroanalytical Procedure Visual examination IV

other excreta Manual, USFDA, Technical
Bulletin V.39 B (for whole)
https://www.fda.gov/food/laborat
ory-methods-food/mpm-v-8-
spices-condiments-flavors-and-
crude-drugs#v32
Dried oregano Whole dead insect ISO 927 Visual examination IV
Dried oregano Whole dead insect MPM V-8 Spices, Condiments, Visual examination IV
Flavors and Crude Drugs A.
General methods for spices,
herbs and botanicals (V 32)
https://www.fda.gov/food/laborat
ory-methods-food/mpm-v-8-
spices-condiments-flavors-and-
crude-drugs#v32
Dried oregano Mould visible Method V-8 Spices, Condiments, Visual examination IV
Flavors and Crude Drugs
(Macroanalytical Procedure
Manual, FDA Technical Bulletin
Number 5)
https://www.fda.gov/food/laborat
ory-methods-food/mpm-v-8-
spices-condiments-flavors-and-
crude-drugs#v32
Dried oregano Insect damage ISO 927 Visual examination I
Nutmeg Piece of mace ISO 927 Visual examination followed by I
gravimetry
Nutmeg Moisture ISO 939 Distillation I
Nutmeg Total ash on dry basis ISO 939 and ISO 928 Calculation from moisture and ash I
(at 550 0C), distillation and
gravimetry
CXS 234-1999 86

Spices and culinary herbs

Provisions Method Principle Type
Commodity

Nutmeg Acid-insoluble ash on dry basis ISO 939 and ISO 930 Calculation from moisture and ash I
(at 550 0C), distillation and
gravimetry
Nutmeg Water-insoluble ash on dry basis ISO 939 and ISO 929 Calculation from moisture and ash I
(at 550 0C), distillation and
gravimetry
Nutmeg Volatile oil content on dry basis ISO 939 and ISO 6571 Calculation from moisture and I
volatile oils, distillation and
distillation
Nutmeg Extraneous matter ISO 927 Visual examination followed by I
gravimetry
Nutmeg Foreign matter ISO 927 Visual examination followed by I
gravimetry
Nutmeg Mould visible ISO 927 Visual examination followed by I
gravimetry
Nutmeg Insect defiled/infested ISO 927 Visual examination followed by I
gravimetry
Nutmeg Dead insect, insect fragments, ISO 927 Visual examination (counting) I
rodent contamination (hair)
Nutmeg Live insects ISO 927 Visual examination (counting) I
Nutmeg Mammalian and/or other excreta Microanalytical Procedure Visual examination followed by IV
(for whole and broken) Manual, USFDA, Technical gravimetry
Bulletin V.41
https://www.fda.gov/food/laborat
ory-methods-food/mpm-v-8-
spices-condiments-flavors-and-
crude-drugs#v32
Saffron Moisture ISO 363202 Gravimetry (drying at 103 oC) I
CXS 234-1999 87

Spices and culinary herbs

Provisions Method Principle Type
Commodity

Saffron Total ash on dry basis ISO 3632-2 and ISO 928 Calculation from moisture and ash I
(at 550 oC), gravimetry and
gravimetry
Saffron Acid-insoluble ash on dry basis ISO 3632-2 and ISO 928 and Calculation from moisture and ash I
ISO 930 (at 550 oC), gravimetry and
gravimetry
Saffron Soluble extract in cold water on ISO 3632-2 and ISO 941 Calculation from moisture and I
dry basis soluble extract, gravimetry and
extraction
Saffron Taste strength (expressed as ISO 3632-2 Absorbance I
picrocrocin) 𝐴1%
1𝑐𝑚 257 nm

Saffron Aroma strength (expressed as ISO 3632-2 Absorbance I

safranal) 𝐴 1%
1𝑐𝑚 330 nm

Saffron Colouring strength (expressed as ISO 3632-2 Absorbance I

crocin) 𝐴 1%
1𝑐𝑚 440 nm

Saffron Extraneous matter ISO 3632-2 Visual examination followed by I

gravimetry
Saffron Foreign matter ISO 3632-2 Visual examination followed by I
gravimetry
Saffron Insect damage ISO 927 Visual examination followed by I
gravimetry
Saffron Whole dead insects/insect ISO 927 Visual examination (counting) I
fragments
Saffron Mould visible ISO 927 Visual examination followed by I
gravimetry
Saffron Mammalian excreta (whole MPM V-8 Spices, Condiments, Visual examination followed by I
spice) Flavors and Crude Drugs MPM: gravimetry
V-8 Spices, Condiments, Flavors
and Crude Drugs FDA
CXS 234-1999 88

Spices and culinary herbs

Provisions Method Principle Type
Commodity

Saffron Rodent filth ISO 927 Visual examination followed by I

gravimetry
Thyme Moisture ISO 939 Distillation I
Thyme Total ash ISO 928 Gravimetry I
Thyme Acid-insoluble ash ISO 930 Gravimetry I
Thyme Volatile oils ISO 6571 Distillation/Volumetric I
Thyme Extraneous vegetable matter ISO 927 Visual examination/Gravimetry I
Thyme Foreign matter ISO 927 Visual examination/Gravimetry I
Thyme Insect damage Method V-8 Spices, Condiments, Visual examination IV
Flavors and Crude Drugs
(Macroanalytical Procedure
Manual, FDA)

http://www.fda.gov/Food/FoodSc
ienceResearch/LaboratoryMetho
ds/ucm084394.htm#v-32
Thyme Mammalian excreta Macroanalytical Procedure Visual examination IV
Manual USFDA Technical
Bulletin V.39 B (for whole)

Thyme Mammalian excreta AOAC 993.27 (for ground) Enzymatic detection method IV
Thyme Mould damage Method V-8 Spices, Condiments, Visual examination IV
Flavors and Crude Drugs
(Macroanalytical Procedure
Manual, FDA)
http://www.fda.gov/Food/FoodSc
ienceResearch/LaboratoryMetho
ds/ucm084394.htm#v-32
Turmeric Moisture ISO 939 Distillation I
CXS 234-1999 89

Spices and culinary herbs

Provisions Method Principle Type
Commodity

Turmeric Total ash on dry basis ISO 939 and ISO 928 Calculation from moisture and ash at (at I
550 oC), distillation and gravimetry
Turmeric Acid-insoluble ash on dry basis ISO 939 AND iso 930 Calculation from moisture and ash at (at I
550 oC), distillation and gravimetry
Turmeric Extraneous matter ISO 927 Visual examination followed by gravimetry I
Turmeric Foreign matter ISO 927 Visual examination followed by gravimetry I
Turmeric Insect defiled ISO 927 Visual examination followed by gravimetry I
Turmeric Whole insects live/dead (for ISO 927 Visual examination (counting) I
whole)
Turmeric Whole insects live/dead (for AOAC 975.49 Flotation I
powdered/pieces)
Turmeric Mammalian or/and other excreta Method V-8 Spices, Condiments, Visual examination followed by gravimetry IV
(whole) Flavors and Crude Drugs
(Macroanalytical Procedure
Manual) MPM: V-8 Spices
https://www.fda.gov/food/laborat
ory-methods-food/mpm-v-8-
spices-condiments-flavors-and-
crude-drugs
Turmeric Mould visible ISO 927 Visual examination followed by gravimetry I
Black and white pepper Bulk density ISO 959-1 Annex B (black) Gravimetry IV
ISO 959-2 Annex A (white)
Black pepper Light berries ISO 959-1 Annex A (black) Flotation IV

Black, white and green pepper Extraneous vegetable matter ISO 927 Visual examination/Gravimetry I

Black, white and green pepper Foreign matter ISO 927 Visual examination/Gravimetry I
CXS 234-1999 90

Spices and culinary herbs

Provisions Method Principle Type
Commodity

Black, white and green pepper Black berries Physical separation and Visual examination IV
weighing
ISO 959-2
Black, white and green pepper Broken berries Physical separation and Visual examination IV
weighing
ISO 959-2
Black, white and green pepper Mouldy berries Macroanalytical Procedure Visual examination IV
Manual USFDA Technical
Bulletin V.39 B
Black, white and green pepper Insect damage Macroanalytical Procedure Visual examination IV
Manual USFDA Technical
Bulletin V.39 B

Black, white and green pepper Pinheads or broken berries Physical separation and Visual examination IV
weighing ISO 959-1

Black, white and green pepper Mammalian excreta Macroanalytical Procedure Visual examination (for whole IV
Manual USFDA Technical pepper)
Bulletin V.39 B (for pepper
whole)

Black, white and green pepper Mammalian excreta AOAC 993.27 (for ground Enzymatic detection method (for I
pepper) ground pepper)

Black, white and green pepper Moisture ISO 939 Distillation I

Black, white and green pepper Total ash ISO 928 Gravimetry I
Black, white and green pepper Non-volatile ether extract ISO 1108 Soxhlet extraction I
Black, white and green pepper Volatile oils ISO 6571 Distillation I
Black, white and green pepper Piperine content ISO 5564 Spectrophotometry I
CXS 234-1999 91

Spices and culinary herbs

Commodity Provisions Method Principle Type

Black, white and green pepper Acid-insoluble ash ISO 930 Gravimetry I
Black, white and green pepper Crude fibre ISO 5498 Gravimetry I
Small cardamom Moisture ISO 939 Distillation I
Small cardamom Total ash, on dry basis ISO 939 and ISO 928 Calculation from moisture and ash I
(at 550 oC), distillation and
gravimetry
Small cardamom Acid-insoluble ash, on dry basis ISO 939 and ISO 930 Calculation from moisture and ash I
(at 550 oC), distillation and
gravimetry
Small cardamom Volatile oil on dry basis ISO 939 and ISO 6571 Calculation from moisture and I
volatile oils, distillation and
distillation
Small cardamom Extraneous matter ISO 927 Visual examination followed I
gravimetry
Small cardamom Foreign matter ISO 927 Visual examination followed I
gravimetry
Small cardamom Insect defiled/infested ISO 927 Visual examination followed I
gravimetry
Small cardamom Immature and shrivelled ISO 882-1 and ISO 927 Visual examination followed I
capsules gravimetry
Small cardamom Mammalian or/and other excreta Method V-8 Spices, Condiments, Visual examination followed IV
(for whole) Flavors and Crude Drugs gravimetry
(Macroanalytical Procedure
Manual MPM: V-8 Spices
https://www.fda.gov/food/laborat
ory-methods-food/mpm-v-8-
spices-condiments-flavors-and-
crude-drugs
Small cardamom Mould visible ISO 927 Visual examination followed I
gravimetry
CXS 234-1999 92

Spices and culinary herbs

Provisions Method Principle Type
Commodity

Small cardamom Empty and malformed capsules ISO 882-1 Visual examination (counting) I
Small cardamom Whole insects live/dead (for ISO 927 Visual examination (counting) I
whole)
Small cardamom Whole insects live/dead (for AOAC 975.49 Flotation I
powdered/pieces)

Sugars and honey

Commodity Provisions Method Principle Type
Honey Acidity MAFF Validated Method V19 Titrimetry I
J. Assoc. Public Analysts (1992)
28 (4) 171-175
Honey Diastase activity IHC Method for determination of
diastase activity with Phadebas,
2009 except that the incubation IV
time should be increased from
15 to 30 minutes
Honey Moisture AOAC 969.38B Refractometry I
or
MAFF Validated Method V21
Honey Sample preparation AOAC 920.180 - -
Honey Solids, water-insoluble MAFF Validated Method V22 Gravimetry I
J. Assoc. Public Analysts (1992)
28(4) 189-193
Honey Sugars added (for sugar profile) AOAC 998.18 Carbon isotope ratio mass I
spectrometry
Honey Sugars added: detection of corn AOAC 978.17 Carbon isotope ratio mass I
and cane sugar products spectrometry
CXS 234-1999 93

Sugars and honey

Commodity Provisions Method Principle Type
Sugars (dextrose anhydrous and D-Glucose ISO 5377 Titrimetry I
dextrose monohydrate)
Sugars (dextrose anhydrous and Solids, total ISO 1741 Gravimetry (vacuum oven) I
dextrose monohydrate)
CXS 234-1999 94

Sugars and honey

Commodity Provisions Method Principle Type

Sugars (dextrose anhydrous and Sulphated ash ISO 5809 Single sulphonation I
dextrose monohydrate, dried glucose
syrup, glucose syrup, powdered
dextrose, lactose)
Sugars (dextrose anhydrous and Sulphur dioxide ISO 5379 Acidimetry and nephelometry IV
dextrose monohydrate)
Sugars (fructose) pH ICUMSA GS 1/2/3/4/7/8-23 Potentiometry I
Sugars (fructose) Conductivity ash ICUMSA GS 2/3-17 Conductimetry I
Sugars (fructose) D-Fructose ISO 10504 Liquid chromatography II
(refractive index detection)
Sugars (fructose) D-Glucose ISO 10504 Liquid chromatography II
(refractive index detection)
Sugars (fructose) Loss on drying ISO 1742 Gravimetry I
Sugars (fructose) Sulphur dioxide ISO 5379 Acidimetry and IV
nephelometry
Sugars (glucose syrup and dried Reducing sugar ISO 5377 Titrimetry I
glucose syrup)
Sugars (glucose syrup and dried Solids, total ISO 1742 Gravimetry (vacuum oven) I
glucose syrup)
Sugars (glucose syrup and dried Sulphur dioxide ISO 5379 Acidimetry and IV
glucose syrup) nephelometry
Sugars (lactose) Lactose, anhydrous ICUMSA GS 4/3-3 Titrimetry II
Sugars (lactose) Loss on drying USP General Chapter 731 Gravimetry (drying at 120 °C I
for 16 h)
Sugars (lactose) pH ICUMSA GS 1/2/3/4/7/8-23 Potentiometry I
Sugars (plantation and mill white Colour ICUMSA GS 9/1/2/3-8 Photometry I
sugar)
Sugars (plantation or mill white sugar) Conductivity ash ICUMSA GS 1/3/4/7/8-13 Conductimetry I
CXS 234-1999 95

Sugars and honey

Commodity Provisions Method Principle Type

Sugars (plantation or mill white sugar) Invert sugar ICUMSA GS 1/3/7-3 Titrimetry (Lane & Eynon) I
Sugars (plantation or mill white sugar) Loss on drying ICUMSA GS 2/1/3-15 Gravimetry I
Sugars (plantation or mill white sugar) Polarization ICUMSA GS 1/2/3-1 Polarimetry II
Sugars (plantation or mill white sugar) Sulphur dioxide ICUMSA GS 2/3-35 Enzymatic method II
NMKL 135
EN 1988-2
Sugars (powdered sugar and Sulphur dioxide ICUMSA GS 2/3-35 Enzymatic method II
powdered dextrose) NMKL 135
EN 1988-2
Sugars (powdered sugar) Colour ICUMSA GS 2/3-9 Photometry I
Sugars (powdered sugar) Conductivity ash ICUMSA GS 2/3-17 Conductimetry I
Sugars (powdered sugar) Invert sugar ICUMSA GS 2/3-5 after filtration if Titrimetry I
necessary to remove any anticaking
agents
Sugars (powdered sugar) Loss on drying ICUMSA GS 2/1/3-15 Gravimetry I
Sugars (powdered sugar) Polarization ICUMSA GS 2/3-1 after filtration if Polarimetry II
necessary to remove any anticaking
agents
Sugars (raw cane sugar) Sulphur dioxide ICUMSA GS 2/3-35 Enzymatic method II
NMKL 135
EN 1988-2
Sugars Conductivity ash ICUMSA GS 1/3/4/7/8-13 Conductimetry I
(soft white sugar and soft brown sugar)
Sugars Invert sugar ICUMSA GS 4/3-3 (applicable at levels Titrimetry (Lane and I
(soft white sugar and soft brown sugar) >10% m/m) Eynon)
Sugars Invert sugar ICUMSA GS 1/3/7-3 Titrimetry (Lane and I
(soft white sugar and soft brown sugar) (applicable at levels <10% m/m) Eynon)
Sugars Loss on drying ICUMSA GS 2/1/3-15 Gravimetry I
(soft white sugar and soft brown sugar)
CXS 234-1999 96

Sugars and honey

Commodity Provisions Method Principle Type

Sugars Sucrose plus invert sugar ICUMSA GS 4/3-7 Titrimetry I
(soft white sugar and soft brown sugar)
Sugars Sulphated ash ICUMSA GS 1/3/4/7/8-11 Gravimetry I
(soft brown sugar)
Sugars Sulphur dioxide ICUMSA GS 2/3-35 NMKL 135 Enzymatic method II
(soft white sugar and soft brown sugar) EN 1988-2
Sugars (soft white sugar) Colour ICUMSA GS 2/3-9 Photometry I
Sugars (white sugar) Conductivity ash ICUMSA GS 2/3-17 Conductimetry I
Sugars (white sugar) Invert sugar ICUMSA GS 2/3-5 Titrimetry I
Sugars (white sugar) Loss on drying ICUMSA GS 2/1/3-15 Gravimetry I
Sugars (white sugar) Polarization ICUMSA GS 2/3-1 Polarimetry II
Sugars (white sugar) Sulphur dioxide ICUMSA GS 2/3-35 NMKL 135 Enzymatic method II
EN 1988-2
Miscellaneous products

Commodity Provisions Method Principle Type

Chilli sauce pH NMKL 179 (Codex general Potentiometry II
method) / AOAC 981.12
Chilli sauce pH AOAC 981.12 Potentiometry III
(Codex general method)
Chilli sauce Fill of containers CAC/RM 46 (see Gravimetry I
Appendix X)
(Codex general method)
(for glass container)
Cooked rice wrapped in plant leaves Peroxide value ISO 3960 / AOCS Cd 8b-90 Titrimetry IV
Extraction of oils from
product (see Appendix XXII)
Date paste Moisture AOAC 934.06 Gravimetry I

Date paste Mineral impurities ISO 762 Gravimetry I

Date paste Ash AOAC 940.26 Gravimetry I
CXS 234-1999 97
Miscellaneous products

Commodity Provisions Method Principle Type

Date paste Acid soluble ash AOAC 900.02D Gravimetry, calculation I
Dried fruits Identification of defects Described in the standard Visual examination I
Dried fruits (except prunes and raisins) Moisture AOAC 934.06 Gravimetry (vacuum oven) I
Dried meat Chloride as sodium chloride ISO 1841-1 Titrimetry III
(≥ 1.0%) (Volhard method)
Dried meat Chloride as sodium chloride ISO 1841-2 Titrimetry II
(≥ 0.25%) (potentiometry)
Dried meat Ash ISO 936 Gravimetry I
Dried meat Water activity ISO 18787 Electrometry II
Dried meat Moisture content AOAC 950.46B Gravimetry I
Dried meat Protein* ISO 937 Calculation and titrimetry I
(*nitrogen-to-protein conversion
factor = 6.25)
Dried meat Total fat ISO 1443 Gravimetry I

Edible cassava flour Ash ISO 2171 and ISO 712 Calculation from moisture I
and gravimetry
(incineration at 550 oC)
Edible cassava flour Fibre, crude ISO 5498 Gravimetry I

Edible cassava flour Moisture ISO 712 Gravimetry (oven drying at I

98 – 100 oC)
Edible cassava flour Particle size ICC Recommendation 207 Sieving and gravimetry I
Fermented noni fruit juice Brix value (soluble solids) AOAC 983.17 / EN 12143 / Refractometry IV
IFUMA 8 / ISO 2173
Fermented noni fruit juice Ethanol AOAC 2017.07 Enzymatic determination IV
CXS 234-1999 98
Miscellaneous products

Commodity Provisions Method Principle Type

Fermented noni fruit juice Ethanol IFUMA 52 Enzymatic determination IV
Fermented noni fruit juice Ethanol AOAC 2016.12 Headspace GC-FID IV
Fermented noni fruit juice Identification of scopoletin Method described in Solid-phase extraction IV
Appendix XXIII, Part A and thin layer
chromatography
Fermented noni fruit juice Identification of Method described in Thin layer IV
deacetylasperulosidic acid Appendix XXIII, Part B chromatography
Fermented noni fruit juice pH value NMKL 179 / AOAC 981.12 Potentiometry II
Fermented soybean paste Total nitrogen AOAC 984.13 Kjeldahl I
Fermented soybean paste Amino nitrogen AOAC 920.154 on the Volumetry I
conditions specified in the
standardxliv
Fermented soybean paste Moisture AOAC 934.01 Gravimetry I
(≤ 70°C, ≤ 50 mm Hg)
Food-grade salt Arsenic EuSalt/AS 015 ICP-OES IV
Food-grade salt Cadmium EuSalt/AS 015 ICP-OES III
Food-grade salt Cadmium EuSalt/AS 014 Atomic absorption spectrophotometry IV
Food-grade salt Calcium and magnesium ISO 2482 Complexometric titrimetry II

xliv
Section 9.2 Determination of amino nitrogen
Preparation of test samples: Weigh 2 g of sample into a 250 ml beaker and mix the sample with 100 ml of cold (15 °C) NH3-free H2O and then stir the mixture for 60 min. Next, decant the mixture through a
quantitative filter and collect the filtrate in a 100 ml volumetric flask.
End-point – A pH metre shall be used to determine the end-point instead of optical verification of colours.
CXS 234-1999 99
Miscellaneous products

Food-grade salt Calcium and magnesium EuSalt/AS 009 Flame atomic absorption III
spectrometry
Food-grade salt Calcium and magnesium EuSalt/AS 015 ICP-OES III
Food-grade salt Copper EuSalt/AS 015 ICP-OES III
Food-grade salt Insoluble matter ISO 2479 Gravimetry II

Miscellaneous products

Commodity Provisions Method Principle Type

Food-grade salt Iodine EuSalt/AS 002 Titrimetry using sodium thiosulphate II
Food-grade salt Iodine EuSalt/AS 019 ICP-OES III
Food-grade salt Iodine WHO/UNICEF/ICCIDD Titrimetry using sodium thiosulphate IV
methodxlv
Only applicable to a product
which has been fortified with
iodate

Food-grade salt Loss on drying ISO 2483 Gravimetry (drying at 110 °C) I
Food-grade salt Mercury EuSalt/AS 012 Cold vapour atomic absorption IV
spectrophotometry
Food-grade salt Potassium EuSalt/AS 008 Flame atomic absorption II
spectrophotometry
Food-grade salt Potassium EuSalt/AS 015 ICP-OES III
Food-grade salt Sodium chloride Described in the standard Calculation I
Food-grade salt Sulphate ISO 2480 Gravimetry II
Food-grade salt Sulphate EuSalt/AS 015 ICP-OES III

xlv
Assessment of iodine deficiency disorders and monitoring their elimination. A guide for programme managers. Third edition, Annex 1: Titration method for determining salt iodate and salt iodine content.
World Health Organization, Geneva, 2007. The report is available from http://www.who.int/nutrition/publications/micronutrients/iodine_deficiency/WHO_NHD_01.1/en/index.html
CXS 234-1999 100
Miscellaneous products

Food-grade salt Sulphate EuSalt/AS 018 Ion chromatography III

Foul medames Sample preparation AOAC 945.68 _
Foul medames Salt content AOAC 971.27 Potentiometry II
NMKL 178
Foul medames Drained weight AOAC 968.30 Sieving I
CXS 234-1999 101
Miscellaneous products

Commodity Provisions Method Principle Type

Gari Ash ISO 2171 and ISO 712 Calculation from moisture and I
Gravimetry (incineration at 550 oC)
Gari Fibre, crude ISO 5498 and ISO 712 Gravimetry (separation) I
Gari Moisture ISO 712 Gravimetry (oven drying at 130 – I
133 oC)
Gari Particle size ICC Recommendation 207 Sieving and gravimetry I
Gari Total acidity ISO 7305 and ISO 712 Titrimetry (ethanol extraction) I
Ginseng products Moisture AOAC 925.45 B (dried Gravimetry I
ginseng)
Quantity of sample: 2 g
Ginseng products Moisture AOAC 925.45 D (ginseng Gravimetry I
extract)
Quantity of sample: 1.5 g
(mixing with 20 g of sea sand)
Ginseng products Solids AOAC 925.45 B (dried Calculation I
ginseng)
calculated by subtracting the
content of water from 100%
Quantity of sample: 2 g
Ginseng products Ash AOAC 923.03 Gravimetry I
AACC Intl 08-01.01
Ginseng products Water-insoluble solids Described in the standard Gravimetry I
(Annex I)
Ginseng products Water-saturated n-butanol Described in the standard Gravimetry I
extracts (Annex II)
Ginseng products Identification of ginsenosides Described in the standard TLC or HPLC IV
Rb1 and Rf (Annex III)
Gochujang Capsaicin Journal of AOAC HPLC-Fluorescence IV
International Vol. 91 No. 2,
2008, pp 387-391
CXS 234-1999 102
Miscellaneous products

Commodity Provisions Method Principle Type

Gochujang Capsaicin Journal of AOAC Gas chromatography-FID IV
International Vol. 91 No. 2,
2008, pp 387-391
Gochujang Crude protein AOAC 984.13 Titrimetry, Kjeldahl I
(Nitrogen conversion factor:
6.25)
Gochujang Moisture AOAC 945.43 Gravimetry I
Guideline level for acrylonitrile Acrylonitrile AOAC 985.13 Gas chromatography II
Guideline levels for vinyl chloride Vinyl chloride monomer ISO 6401 Gas chromatography II
monomer
Guideline levels for vinyl chloride Vinyl chloride monomer Commission Directive Gas chromatography ("headspace") III
monomer 81/432/EEC O.J. No. L.167,
p. 6, 24.6.81

Guidelines for nutrition labelling Polyunsaturated fatty acids AOCS Ce 1h-05xlvi Gas-liquid chromatography II

Guidelines for nutrition labelling AOAC 996.06; or AOCS Ce II

Saturated fat Gas-liquid chromatography
1h-05

Guidelines for nutrition labelling Saturated fatty acids AOCS Ce 1h-05 Gas-liquid chromatography II

Harissa Acidity ISO 750 Titrimetry I

Harissa Acid-insoluble ash ISO 763 Gravimetry I

Harissa Dry extract – soluble solids ISO 2173 Refractometry I

Halwa tehenia Acidity AOAC 924.53, AOAC 942.15 Titrimetry IV

xlvi
Can also be used to measure trans unsaturated fatty acids.
CXS 234-1999 103
Miscellaneous products

Commodity Provisions Method Principle Type

Halwa tehenia Ash AOAC 900.02 | AACC Intl Gravimetry I
8.14.01

Halwa tehenia Fat AOAC 963.15 Gravimetry I

Halwa tehenia Moisture AOAC 925.45 | AACC Intl Gravimetry I

44.60.01

Halwa tehenia IV
Sugars ISI 28-1exlvii Titrimetry

Humus with tehenia Salt content AOAC 971.27 Potentiometry II

NMKL 178

Humus with tehenia Total acidity AOAC 925.53 Titrimetry I

xlvii
http://www.starch.dk/isi/methods/28luff.htm
CXS 234-1999 104
Miscellaneous products

Commodity Provisions Method Principle Type

Kava products for use as a beverage Moisture AOAC 925.45 Gravimetry I

when mixed with water

Mixed zaatar Sodium chloride ISO 939 and Calculation by moisture and ash I
(dry weight basis) Distillation and
AOAC 971.27 titrimetry

Mixed zaatar Moisture ISO 939 Distillation I

Mixed zaatar Acid-insoluble ash ISO 939 and Calculation by moisture and ash I
(dry weight basis) AOAC 941.12 (corrected for Distillation and
moisture by ISO 930) gravimetry, Furnace, 550 0C

Mixed zaatar Extraneous matter ISO 927 Visual examination I

Gravimetry

Mixed zaatar Foreign matter ISO 927 Visual examination I

Gravimetry

Mixed zaatar Insects/Insect fragments ISO 927 Visual examination IV

Mixed zaatar Insects/Insect fragments AOAC 969.44 Visual examination IV

Mixed zaatar Insects/Insect fragments AOAC 975.49 Visual examination IV

Mixed zaatar Mould damage Method V-8 Spices, Visual examination IV

Condiments, Flavors and
Crude Drugs (Macroanalytical
Procedure Manual, FDA,
Technical Bulletin Number 5)
CXS 234-1999 105

Miscellaneous products

Commodity Provisions Method Principle Type

Mixed zaatar Mammalian excreta Macroanalytical Procedure Visual examination IV

Manual, USFDA, Technical
Bulletin V.39 B (for whole)

Mixed zaatar Mammalian excreta AOAC 993.27 (for ground) Enzymatic detection method IV

Non-fermented soybean products Moisture content AOAC 925.09 | AACCI 44- Gravimetry (vacuum oven) I
40.01

Non-fermented soybean products Protein content NMKL 6 or AACCI 46-16.01 Titrimetry, Kjeldahl digestion I
or AOAC 988.05 or AOCS Bc
4-91 or AOCS Ba 4d-90
(Nitrogen factor 5.71)

Sago flour Moisture content ISO 712 Gravimetry I

Sago flour Ash (inorganic extraneous matter) ISO 2171 Gravimetry I

Sago flour Acidity AOAC 939.05 Titrimetry I

Sago flour Crude fibre ISO 6541 Gravimetry I

Sago flour Starch AOAC 920.44 Gravimetry I
CXS 234-1999 106

Miscellaneous products

Commodity Provisions Method Principle Type

Soybean products fermented with Bacillus species

Natto Lipid content 4 g quantity of AOAC 963.15 Gravimetry (Soxhlet) I

samples

Natto Moisture content AOAC 925.09 Gravimetry I

Natto Protein content AOAC 988.05 Titrimetry I

(*nitrogen-to-protein conversion (Kjeldahl)
factor = 5.71)

Cheonggukjang Moisture content AOAC 934.01 Gravimetry I

Cheonggukjang Protein content AOAC 988.05 Titrimetry (Kjeldahl) I

(*nitrogen-to-protein conversion
factor = 5.71)

Cheonggukjang Lipid content 5 g quantity of AOAC 963.15 Gravimetry (Soxhlet) I

samples
Thua Nao Moisture content AOAC 925.09 Gravimetry I

Thua Nao Protein content AOAC 988.05 Titrimetry (Kjeldahl) I

(*nitrogen-to-protein conversion
factor = 5.71)

Tehena Moisture content ISO 934 Gravimetry I

Tehena Protein content ISO 1871 Titrimetry, Kjeldahl I

Tehena Total ash ISO 6884 Gravimetry I

Tehena Acid-insoluble ash ISO 735 Gravimetry I

Tehena Total acidity ISO 729 Titrimetry I

CXS 234-1999 107

Miscellaneous products

Commodity Provisions Method Principle Type

Tehena Sesame oil AOCS Cb 2-40 Colour reaction I

(Baudouin test)

Tempe Moisture content AOAC 925.09 | AACCI 44- Gravimetry (vacuum oven) I
40.01

Tempe Protein content NMKL 6 or AOAC 988.05 or Titrimetry, Kjeldahl digestion I

AACCI 46-16.01
(Nitrogen factor 5.71)

Tempe Lipid content AOAC 963.15 Gravimetry (Soxhlet extraction) I

Tempe Crude fibre ISO 5498 or AOAC 962.09 or Gravimetry I
AACCI 32-10.01

Laver products Moisture content AOAC 925.45B Gravimetry, drying at atmospheric IV

pressure

Laver products Acidity: acid value for the See Appendix XXIV Extraction of oil I
extracted oil and
ISO 660 | AOCS Cd 3d-63 Titrimetry

Laver products Moisture content AOAC 925.45 Gravimetry, drying at atmospheric I

pressure

Unrefined shea butter Moisture content ISO 662 Gravimetry I

Unrefined shea butter Free fatty acid content acid value ISO 660 Titrimetry I
and acidity
AOCS Cd 3d-63

Unrefined shea butter Relative density AOCS Cc 10c-95/ Pycnometry I

ISO 6883
CXS 234-1999 108

Miscellaneous products

Commodity Provisions Method Principle Type

Unrefined shea butter Saponification value ISO 3657 Titrimetry I

AOCS Cd 3d-25

Unrefined shea butter Iodine value AOAC 993.20/ Wijs-titrimetry I

ISO 3961/
AOCS Cd 1d-92/
NMKL 39

Unrefined shea butter Peroxide value AOCS Cd 8b-90/ Titrimetry I

ISO 3960/
NMKL 158
Unrefined shea butter Unsaponifiable matter ISO 3596/ Gravimetry I
AOCS Ca 6a-40

Unrefined shea butter Insoluble impurities content ISO 663/ Gravimetry I

AOCS Ca 3a-46

Unrefined shea butter Melting point ISO 6321 Open ended capillary tube I
AOCS Cc 3b-92
CXS 234-1999 109
IRRADIATED FOODS

Commodity Provision Method Principle Type

Food containing fat (e.g. raw meat Detection of irradiated food – Detection of radiation-induced EN 1784 Gas chromatographic analysis of II
and chicken, cheese, fruits) hydrocarbons hydrocarbons

Food containing fat (e.g. raw meat Detection of irradiated food – Detection of radiation-induced 2- EN 1785 Gas chromatographic mass III
and chicken, liquid whole egg) alkylcyclobutanones spectrometric analysis of 2-
alkylcyclobutanones

Food containing bone Detection of irradiated food – Radiation-induced Electron Spin EN 1786 ESR spectroscopy II
Resonance (ESR) signal attributed to hydroxyapatite (principal
component of bones)

Food containing cellulose (e.g. nuts Detection of irradiated food – Radiation-induced Electron Spin EN 1787 ESR spectroscopy II
and spices) Resonance (ESR) signal attributed to crystalline cellulose

Food containing silicate minerals Detection of irradiated foods – Thermoluminescence glow ratio EN 1788 Thermoluminescence II
(e.g. herbs, spices, their mixtures used to indicate the irradiation treatment of the food
and shrimps)

Food containing silicate minerals Detection of irradiated foods – Measurement of photo- EN Photo-stimulated luminescence III
(e.g. shellfish, herbs, spices, stimulated luminescence intensity 13751
seasonings)

Food containing crystalline sugar Detection of irradiated food – Radiation-induced Electron Spin EN ESR spectroscopy II
(e.g. dried fruits and raisins) Resonance (ESR) signal attributed to crystalline sugar 13708
CXS 234-1999 110
Table 11. Numeric performance criteria for lead and cadmium in foods

Method performance criteria

ML Minimum Limit of Limit of Precision Example of

Commodity Provision applicable detection quantification (RSDR) applicable
Recovery
(mg/kg) range (LOD) (LOQ) (%) no methods that Principle
(%)
more meet the
(mg/kg) (mg/kg) (mg/kg) than criteria

FDA Method ICP-MS

Infant formula, formula 4.7 Ver.1.2, ICP-MS
for special medical AOAC ICP-MS
lead 0.01 0.006–0.014 0.002 0.004 44 60–115
purposes intended for 2013.06, ICP-MS
infants and follow-up EN 15763,
formula EN 17851

FDA Method
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
2013.06, ICP-MS
Milk lead 0.02 0.011–0.029 0.004 0.008 44 60–115
EN 15763, ICP-MS
EN 17851, GF-AAS
EN 14083, ICP-MS
NMKL 186

FDA Method
4.7 Ver.1.2,
AOAC ICP-MS
Secondary milk 2013:06, ICP-MS
products (including EN 15763, ICP-MS
butter, edible casein lead 0.02 0.011–0.029 0.004 0.008 44 60–115 EN 17851, ICP-MS
products and whey NMKL 186 ICP-MS
powders) EN 14083, GF-AAS
AOAC 999.11 AAS or GF-AAS
(edible
casein)
CXS 234-1999 111

Method performance criteria

ML Minimum Limit of Limit of Precision Example of

Commodity Provision applicable detection quantification (RSDR) applicable
Recovery
(mg/kg) range (LOD) (LOQ) (%) no methods that Principle
(%)
more meet the
(mg/kg) (mg/kg) (mg/kg) than criteria

FDA Method
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
Fruit juices, except
2013:06, ICP-MS
juices exclusively from
lead 0.03 0.017–0.043 0.006 0.012 44 60–115 EN 15763, ICP-MS
berries and other small
EN 17851, GF-AAS
fruits
EN 14083, ICP-MS
NMKL 186, GF-AAS
NMKL 161

EN 15763, ICP-MS
Fat spreads and EN 17851. ICP-MS
lead 0.04 0.022–0.058 0.008 0.016 44 60–115
blended spreads NMKL 186 ICP- MS
NMKL 161 GF-AAS

EN 15763, ICP-MS
EN 17851, ICP-MS
Grape juice lead 0.04 0.022–0.058 0.008 0.016 44 60–115 NMKL 186 ICP-MS
EN 14083, GF-AAS
NMKL 161 GF-AAS
EN 15763, ICP-MS
Canned chestnuts and EN 17851, ICP-MS
canned chestnuts lead 0.05 0.028–0.072 0.010 0.020 44 60–115 NMKL 186, ICP-MS
puree EN 14083, GF-AAS
NMKL 161 GF-AAS
Fruit juices obtained FDA Method ICP-MS
exclusively from berries 4.7 Ver.1.2, ICP-MS
lead 0.05 0.028–0.072 0.010 0.020 44 60–115
and other small fruits, AOAC ICP-MS
except grape juice 2013:06, ICP-MS
CXS 234-1999 112

Method performance criteria

ML Minimum Limit of Limit of Precision Example of

Commodity Provision applicable detection quantification (RSDR) applicable
Recovery
(mg/kg) range (LOD) (LOQ) (%) no methods that Principle
(%)
more meet the
(mg/kg) (mg/kg) (mg/kg) than criteria
EN 15763, ICP-MS
EN 17851, GF-AAS
NMKL 186, GF-AAS
EN 14083.
NMKL 161
FDA Method
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
Fruiting vegetables, 2013:06, ICP-MS
except fungi and lead 0.05 0.028–0.072 0.010 0.020 44 60–115 EN 15763, ICP-MS
mushrooms EN 17851, ICP-MS
NMKL 186 GF-AAS
EN 14083. GF-AAS
NMKL 161
FDA Method
4.7 Ver.1.2,
ICP-MS
AOAC
ICP-MS
2013.06,
ICP-MS
EN 15763,
Preserved tomatoes lead 0.05 0.028–0.072 0.010 0.020 44 60–115 ICP-MS
EN 17851,
ICP-MS
NMKL 186
GF-AAS
EN 14083.
GF-AAS
NMKL 161

Edible fats and oils GF-AAS

AOAC
(including fats and oils ICP-OES
994.02,
(all); named vegetable lead 0.08 0.045–0.115 0.016 0.032 44 60–115 GF-AAS
AOCS Ca
oils; olive oils and olive ICP-MS
17a-18,
pomace oils) ICP-MS
CXS 234-1999 113

Method performance criteria

ML Minimum Limit of Limit of Precision Example of

Commodity Provision applicable detection quantification (RSDR) applicable
Recovery
(mg/kg) range (LOD) (LOQ) (%) no methods that Principle
(%)
more meet the
(mg/kg) (mg/kg) (mg/kg) than criteria
ISO 12193, ICP-OES
EN 17851,
NMKL 186
ISO 21033
FDA Method
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
Berries and other small 2013:06, ICP-MS
fruits, except cranberry, lead 0.1 0.03–0.17 0.01 0.02 44 80–110 EN 15763, ICP-MS
currant and elderberry EN 17851,
ICP-MS
NMKL 161,
GF-AAS
EN 14083
FDA Method
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
Brassica vegetables, 2013:06, ICP-MS
except kale and leafy lead 0.1 0.03–0.17 0.01 0.02 44 80–110 EN 15763, ICP-MS
Brassica vegetables EN 17851,
ICP-MS
NMKL 161 GF-AAS
EN 14083
FDA Method ICP-MS
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
Bulb vegetables lead 0.1 0.03–0.17 0.01 0.02 44 80–110
2013:06, ICP-MS
EN 15763, ICP-MS
EN 17851 GF-AAS
CXS 234-1999 114

Method performance criteria

ML Minimum Limit of Limit of Precision Example of

Commodity Provision applicable detection quantification (RSDR) applicable
Recovery
(mg/kg) range (LOD) (LOQ) (%) no methods that Principle
(%)
more meet the
(mg/kg) (mg/kg) (mg/kg) than criteria
NMKL 186
GF-AAS
EN 14083.
NMKL 161
FDA Method
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
2013:06, ICP-MS
Canned fruits lead 0.1 0.03–0.17 0.01 0.02 44 80–110 EN 15763, ICP-MS
EN 17851, ICP-MS
NMKL 186 GF-AAS
EN 14083 GF-AAS
NMKL 161
FDA Method
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
2013:06, ICP-MS
Canned vegetables lead 0.1 0.03–0.17 0.01 0.02 44 80–110 EN 15763, ICP-MS
EN 17851, ICP-MS
NMKL 186 GF-AAS
EN 14083 GF-AAS
NMKL 161
ICP-MS
FDA Method
ICP-MS
4.7 Ver.1.2,
Fruits, except ICP-MS
AOAC
cranberry, currants and lead 0.1 0.03–0.17 0.01 0.02 44 80–110 ICP-MS
2013:06,
elderberry ICP-MS
EN 15763,
GF-AAS
EN 17851,
GF-AAS
CXS 234-1999 115

Method performance criteria

ML Minimum Limit of Limit of Precision Example of

Commodity Provision applicable detection quantification (RSDR) applicable
Recovery
(mg/kg) range (LOD) (LOQ) (%) no methods that Principle
(%)
more meet the
(mg/kg) (mg/kg) (mg/kg) than criteria
NMKL 186
EN 14083
NMKL 161
FDA Method
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
2013.06, ICP-MS
Legume vegetables lead 0.1 0.03–0.17 0.01 0.02 44 80–110 EN 15763, ICP-MS
EN 17851, ICP-MS
NMKL 186 GF-AAS
EN 14083 GF-AAS
NMKL 161
FDA Method
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
2013.06, ICP-MS
Meat and fat of poultry lead 0.1 0.03–0.17 0.01 0.02 44 80–110 EN 15763, ICP-MS
EN 17851, ICP-MS
NMKL 186 GF-AAS
EN 14083 GF-AAS
NMKL 161
ICP-MS
FDA Method
ICP-MS
4.7 Ver.1.2
ICP-MS
Meat of cattle, pigs and AOAC
lead 0.1 0.03–0.17 0.01 0.02 44 80–110 ICP-MS
sheep 2013.06,
ICP-MS
EN 15763,
GF-AAS
EN 17851,
GF-AAS
CXS 234-1999 116

Method performance criteria

ML Minimum Limit of Limit of Precision Example of

Commodity Provision applicable detection quantification (RSDR) applicable
Recovery
(mg/kg) range (LOD) (LOQ) (%) no methods that Principle
(%)
more meet the
(mg/kg) (mg/kg) (mg/kg) than criteria
NMKL 186
EN 14083
NMKL 161
EN 15763, ICP-MS
EN 17851, ICP-MS
Pickled cucumbers
lead 0.1 0.03–0.17 0.01 0.02 44 80–110 NMKL 186 ICP-MS
(cucumber pickles)
EN 14083 GF-AAS
NMKL 161 GF-AAS
FDA Method
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
2013.06, ICP-MS
Poultry, edible offal of lead 0.1 0.03–0.17 0.01 0.02 44 80–110 EN 15763, ICP-MS
EN 17851, ICP-MS
NMKL 186 GF-AAS
EN 14083 GF-AAS
NMKL 161
FDA Method
ICP-MS
4.7 Ver.1.2,
ICP-MS
AOAC
ICP-MS
2013.06,
ICP-MS
Pulses lead 0.1 0.03–0.17 0.01 0.02 44 80–110 EN 15763,
ICP-MS
EN 17851,
GF-AAS
NMKL 186
EN 14083 GF-AAS
NMKL 161
CXS 234-1999 117

Method performance criteria

ML Minimum Limit of Limit of Precision Example of

Commodity Provision applicable detection quantification (RSDR) applicable
Recovery
(mg/kg) range (LOD) (LOQ) (%) no methods that Principle
(%)
more meet the
(mg/kg) (mg/kg) (mg/kg) than criteria
FDA Method
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
2013:06, ICP-MS
Root and tuber
lead 0.1 0.03–0.17 0.01 0.02 44 80–110 EN 15763, ICP-MS
vegetables
EN 17851, ICP-MS
NMKL 186 GF-AAS
EN 14083 GF-AAS
NMKL 161
Wine from grapes EN 15763,
ICP-MS
harvested after July lead 0.1 0.03–0.17 0.01 0.02 44 80–110 OIV-MA-
ICP-MS
2019 AS323-07.

Fortified/liqueur wine EN 15763,

ICP-MS
from grapes harvested lead 0.15 0.05–0.25 0.015 0.03 43 80–110 OIV-MA-
ICP-MS
after 2019 AS323-07.
FDA Method
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
2013.06, ICP-MS
Pig, edible offal of lead 0.15 0.05–0.25 0.015 0.03 43 80–110 EN 15763, ICP-MS
EN 17851, ICP-MS
NMKL 186 GF-AAS
EN 14083 GF-AAS
NMKL 161
FDA Method
ICP-MS
Cattle, edible offal of lead 0.2 0.08–0.32 0.02 0.04 41 80–110 4.7 Ver.1.2,
ICP-MS
AOAC
CXS 234-1999 118

Method performance criteria

ML Minimum Limit of Limit of Precision Example of

Commodity Provision applicable detection quantification (RSDR) applicable
Recovery
(mg/kg) range (LOD) (LOQ) (%) no methods that Principle
(%)
more meet the
(mg/kg) (mg/kg) (mg/kg) than criteria
2013:06, ICP-MS
EN 15763, ICP-MS
EN 17851, ICP-MS
NMKL 186 GF-AAS
EN 14083 GF-AAS
NMKL 161
FDA Method
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
Cereal grains, except 2013:06, ICP-MS
buckwheat, cañihua lead 0.2 0.08–0.32 0.02 0.04 41 80–110 EN 15763, ICP-MS
and quinoa EN 17851, ICP-MS
NMKL 186 GF-AAS
EN 14083 GF-AAS
NMKL 161
FDA Method
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
2013:06, ICP-MS
Cranberry lead 0.2 0.08–0.32 0.02 0.04 41 80–110 EN 15763, ICP-MS.
EN 17851, ICP-MS
NMKL 186 GF-AAS
EN 14083 GF-AAS
NMKL 161
FDA Method ICP-MS
4.7 Ver.1.2, ICP-MS
Currants lead 0.2 0.08–0.32 0.02 0.04 41 80–110
AOAC ICP-MS
2013:06, ICP-MS
CXS 234-1999 119

Method performance criteria

ML Minimum Limit of Limit of Precision Example of

Commodity Provision applicable detection quantification (RSDR) applicable
Recovery
(mg/kg) range (LOD) (LOQ) (%) no methods that Principle
(%)
more meet the
(mg/kg) (mg/kg) (mg/kg) than criteria
EN 15763, ICP-MS
EN 17851, GF-AAS
NMKL 186 GF-AAS
EN 14083.
NMKL 161
FDA Method
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
2013:06, ICP-MS
Elderberry lead 0.2 0.08–0.32 0.02 0.04 41 80–110 EN 15763, ICP-MS
EN 17851, ICP-MS
NMKL 186 GF-AAS
EN 14083 GF-AAS
NMKL 161
Wine (wine and ICP-MS
fortified/ liqueur wine) EN 15763,
ICP-MS.
made from grapes lead 0.2 0.08–0.32 0.02 0.04 41 80–110 OIV-MA-
harvested before July AS323-07
2019
GF-AAS
AOAC 999.11,
ICP-MS
FDA Method
ICP-MS
4.7 Ver.1.2,
ICP-MS
Fish lead 0.3 0.13–0.47 0.03 0.06 38 80–110 AOAC
ICP-MS
2013:06,
ICP-MS
EN 15763,
GF-AAS
EN 17851
GF-AAS
CXS 234-1999 120

Method performance criteria

ML Minimum Limit of Limit of Precision Example of

Commodity Provision applicable detection quantification (RSDR) applicable
Recovery
(mg/kg) range (LOD) (LOQ) (%) no methods that Principle
(%)
more meet the
(mg/kg) (mg/kg) (mg/kg) than criteria
NMKL 186
EN 14083
NMKL 161
Fresh farmed
mushrooms (common EN 15763, ICP-MS
mushrooms (Agaricus EN 17851, ICP-MS
bisporous), shiitake
lead 0.3 0.13–0.47 0.03 0.06 38 80–110 NMKL 186 ICP-MS
mushrooms (Lentinula
EN 14083 GF-AAS
edodes), and oyster
mushrooms (Pleurotus NMKL 161 GF-AAS
ostreatus))
FDA Method
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
2013:06, ICP-MS
Leafy vegetables,
lead 0.3 0.13–0.47 0.03 0.06 38 80–110 EN 15763, ICP-MS
except spinach
EN 17851, ICP-MS
NMKL 186 GF-AAS
EN 14083 GF-AAS
NMKL 161
FDA Method
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
2013:06, ICP-MS
Jams, jellies and
lead 0.4 0.18–0.62 0.04 0.08 37 80–110 EN 15763, ICP-MS
marmalades
EN 17851, ICP-MS
NMKL 186 GF-AAS
EN 14083 GF-AAS
NMKL 161
CXS 234-1999 121

Method performance criteria

ML Minimum Limit of Limit of Precision Example of

Commodity Provision applicable detection quantification (RSDR) applicable
Recovery
(mg/kg) range (LOD) (LOQ) (%) no methods that Principle
(%)
more meet the
(mg/kg) (mg/kg) (mg/kg) than criteria
FDA Method
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
2013.06, ICP-MS
Mango chutney lead 0.4 0.18–0.62 0.04 0.08 37 80–110 EN 15763, ICP-MS
EN 17851, ICP-MS
NMKL 186 GF-AAS
EN 14083 GF-AAS
NMKL 161
FDA Method
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
2013.06, ICP-MS
EN 15763, ICP-MS
Table olives lead 0.4 0.18–0.62 0.04 0.08 37 80–110 EN 17851, ICP-MS
NMKL 186 GF-AAS
EN 14083, AAS (Flame
AOAC 999.11 Absorption)
NMKL 139 GF-AAS
NMKL 161
EUsalt/AS
ICP-OES
015,
Salt, food grade lead 1 0.5–1.5 0.1 0.2 32 80–110 ICP-MS
EN 17851,
GF-AAS
EN 14083
FDA Method ICP-MS
Brassica vegetables, 4.7 Ver.1.2, ICP-MS
except Brassica leafy cadmium 0.05 0.03–0.07 0.01 0.02 44 60–115
AOAC ICP-MS
vegetables
2013:06, ICP-MS
CXS 234-1999 122

Method performance criteria

ML Minimum Limit of Limit of Precision Example of

Commodity Provision applicable detection quantification (RSDR) applicable
Recovery
(mg/kg) range (LOD) (LOQ) (%) no methods that Principle
(%)
more meet the
(mg/kg) (mg/kg) (mg/kg) than criteria
EN 15763, ICP-MS
EN 17851, GF-AAS
NMKL 186 GF-AAS
EN 14083
NMKL 161
FDA Method
ICP-MS
4.7 Ver.1.2,
ICP-MS
AOAC
ICP-MS
2013:06,
ICP-MS
EN 15763,
Bulb vegetables cadmium 0.05 0.03–0.07 0.01 0.02 44 60–115 ICP-MS
EN 17851,
GF-AAS
NMKL 186
GF-AAS
EN 14083.
NMKL 161
FDA Method ICP-MS
4.7 Ver.1.2 ICP-MS
AOAC ICP-MS
Fruiting vegetables, 2013:06, ICP-MS
except tomatoes and cadmium 0.05 0.03–0.07 0.01 0.02 44 60–115 EN 15763, ICP-MS
edible fungi EN 17851, GF-AAS
NMKL 186 GF-AAS
EN 14083
NMKL 161
ISO 23637, GF-AAS
Cereal grains, except EN 17851, ICP-MS
buckwheat, cañihua, cadmium 0.1 0.03–0.17 0.01 0.02 44 80–110 NMKL 186 ICP-MS
quinoa, wheat and rice EN 14083 GF-AAS
NMKL 161 GF-AAS
CXS 234-1999 123

Method performance criteria

ML Minimum Limit of Limit of Precision Example of

Commodity Provision applicable detection quantification (RSDR) applicable
Recovery
(mg/kg) range (LOD) (LOQ) (%) no methods that Principle
(%)
more meet the
(mg/kg) (mg/kg) (mg/kg) than criteria
FDA Method ICP-MS
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
2013:06, ICP-MS
Legume vegetables cadmium 0.1 0.03–0.17 0.01 0.02 44 80–110 EN 15763, ICP-MS
EN 17851, GF-AAS
NMKL 186 GF-AAS
EN 14083
NMKL 161
EN 15763, ICP-MS
EN 17851, ICP-MS
Pulses, except soya
cadmium 0.1 0.03–0.17 0.01 0.02 44 80–110 NMKL 186 ICP-MS
bean (dry)
EN 14083 GF-AAS
NMKL 161 GF-AAS
FDA Method ICP-MS
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
Root and tuber 2013:06, ICP-MS
vegetables, except cadmium 0.1 0.03–0.17 0.01 0.02 44 80–110 EN 15763, ICP-MS
celeriac EN 17851, GF-AAS
NMKL 186 GF-AAS
EN 14083
NMKL 161
FDA Method ICP-MS
4.7 Ver.1.2, ICP-MS
Stalk and stem AOAC ICP-MS
cadmium 0.1 0.03–0.17 0.01 0.02 44 80–110
vegetables 2013:06, ICP-MS
EN 15763, ICP-MS
EN 17851, GF-AAS
CXS 234-1999 124

Method performance criteria

ML Minimum Limit of Limit of Precision Example of

Commodity Provision applicable detection quantification (RSDR) applicable
Recovery
(mg/kg) range (LOD) (LOQ) (%) no methods that Principle
(%)
more meet the
(mg/kg) (mg/kg) (mg/kg) than criteria
NMKL 186 GF-AAS
EN 14083
NMKL 161
FDA Method
4.7 Ver.1.2, ICP-MS
AOAC ICP-MS
2013:06, ICP-MS
Leafy vegetables cadmium 0.2 0.08–0.32 0.02 0.04 41 80–110 EN 15763, ICP-MS
EN 17851, ICP-MS
NMKL 186 GF-AAS
EN 14083 GF-AAS
NMKL 161
ISO 23637, GF-AAS
Wheat (common wheat, EN 17851, ICP-MS
durum wheat, spelt and cadmium 0.2 0.08–0.32 0.02 0.04 41 80–110 NMKL 186 ICP-MS
emmer) EN 14083 GF-AAS
NMKL 161 GF-AAS
EN 15763,
ICP-MS
EN 17851,
Chocolate containing or ICP-MS
NMKL 186
declaring < 30% total ICP-MS
cadmium 0.3 0.13–0.47 0.03 0.06 38 80–110 EN 14083
cocoa solids on a dry GF-AAS
matter basis NMKL 161
GF-AAS

ISO 23637 GF-AAS

Rice, polished cadmium 0.4 0.18–0.62 0.04 0.08 37 80–110
EN 17851, ICP-MS
CXS 234-1999 125

Method performance criteria

ML Minimum Limit of Limit of Precision Example of

Commodity Provision applicable detection quantification (RSDR) applicable
Recovery
(mg/kg) range (LOD) (LOQ) (%) no methods that Principle
(%)
more meet the
(mg/kg) (mg/kg) (mg/kg) than criteria
NMKL 186 ICP-MS
EN 14083 GF-AAS
NMKL 161 GF-AAS
EUsalt/AS
ICP-OES
015,
Salt, food grade cadmium 0.5 0.23–0.77 0.05 0.10 36 80–110 ICP-MS
EN 17851,
GF-AAS
EN 14083.

Chocolate containing or EN 15763, ICP-MS

declaring > 30% to EN 17851 ICP-MS
< 50% total cocoa cadmium 0.7 0.35–1.05 0.07 0.14 34 80–110
NMKL 186 ICP-MS
solids on a dry matter
basis EN 14083 GF-AAS

Chocolate containing or
declaring ≥ 50% to
< 70% total cocoa ICP-MS
solids on a dry matter EN 15763, ICP-MS
basis, including sweet EN 17851, ICP-MS
chocolate, gianduja cadmium 0.8 0.40–1.20 0.08 0.16 33 80–110
NMKL 186 GF-AAS
chocolate, semi – bitter
table chocolate, EN 14083
vermicelli chocolate/
chocolate flakes, and
bitter table chocolate
CXS 234-1999 126

Method performance criteria

ML Minimum Limit of Limit of Precision Example of

Commodity Provision applicable detection quantification (RSDR) applicable
Recovery
(mg/kg) range (LOD) (LOQ) (%) no methods that Principle
(%)
more meet the
(mg/kg) (mg/kg) (mg/kg) than criteria

Chocolate containing or
declaring ≥ 70% total
cocoa solids on a dry
matter basis, including EN 15763 ICP-MS
sweet chocolate, EN 17851 ICP-MS
cadmium 0.9 0.46–1.34 0.09 0.18 33 80–110
gianduja chocolate, NMKL 186 ICP-MS
semi – bitter table EN 14083 GF-AAS
chocolate, vermicelli
chocolate/ chocolate
flakes, and bitter table
EN 15763, ICP-MS
EN 17851, ICP-MS
Cephalopods cadmium 2 1.1–2.9 0.2 0.4 29 80–110 NMKL 186 ICP-MS
EN 14083 GF-AAS
NMKL 161 GF-AAS
ICP-MS
EN 15763
Marine bivalve ICP-MS
EN 17851
molluscs (clams, ICP-MS
NMKL 186
cockles and mussels), cadmium 2 1.1–2.9 0.2 0.4 29 80–110 GF-AAS
except oysters and GF-AAS
EN 14083
scallops
NMKL 161
CXS 234-1999 127

PART B – METHODS OF SAMPLING BY COMMODITY CATEGORIES AND NAMES

Commodity categories Method of sampling Notes

Cereals, pulses and legumes and derived products
Wheat protein products including wheat gluten ISO 13690
Fats and oils
Olive oils and olive pomace oils ISO 661 and ISO 5555
Fish oils ISO 5555
Milk and milk products
Milk products ISO 707 | IDF 50 General instructions for obtaining a sample from a bulk
Milk products ISO 5538 | IDF 113 Inspection by attributes
Milk products ISO 3951-1 Inspection by variables
Processed fruits and vegetables
Desiccated coconut Described in the standard
Certain canned vegetables, jams and jellies Described in the standard
Chilli sauce Described in the standard
Table olives Described in the standard
 CXS 234-1999 128

APPENDIX I
PART A – EXTRACTION OF OIL FROM INSTANT NOODLES

1. Extraction of oil from instant noodles

1.1 Apparatus
(a) Rotary evaporator
(b) Water bath
1.2 Preparation of test sample
Remove instant noodles from package and leave garnishing and seasoning in package. Transfer the noodles
to plastic bag to prevent moisture change and then break these into small fragments with hands or wooden
hammer. Select broken noodles in the size range of 2.36 mm to 1.7 mm by using two sieves with 2.36 mm and
1.7 mm openings and mix well. Use these noodles for the test sample. If the noodles are too thin to screen
with sieves, cut them into 1 cm to 2 cm lengths, mix well and use these cut noodles for the test sample.
1.3 Extraction
Weigh 25 g test portion into 200 Ml Erlenmeyer flask. Add 100 Ml petroleum ether to the flask after replacing
air in flask by N2 gas. Stopper flask and leave for 2 hours. Decant supernatant through filter paper into
separating funnel. Add 50 Ml petroleum ether to residue and filtrate supernatant through filter paper into the
separating funnel. Add 75 Ml water to the separating funnel and shake well. Allow layers to separate and drain
the lower aqueous layer. Add water, shake and remove aqueous layer again as done previously. Decant the
petroleum ether layer after dehydration with Na2SO4 into pear-shaped flask. Evaporate petroleum ether in the
flask on rotary evaporator at not over 40 °C. Spray N2 gas on extract in the flask to remove all petroleum ether.

PART B – DETERMINATION OF ACID VALUE

2. Determination of acid value
2.1 Definition and principle
Acid value of oil from fried instant noodles = mg KOH required to neutralize 1 g oil. Oil extracted from noodle
is dissolved in alcohol-ether mixture and titrated with alcoholic KOH standard solution.
2.2 Apparatus
Air-tight desiccator: silica gel heated at 150 °C is satisfactory drying agent.
2.3 Reagents
(a) Alcoholic potassium hydroxide standard solution: 0.05 mol/L. Dissolve 3.5 g potassium hydroxide in
equal volume of water (CO2-free) and add ethanol (95%) to 1 L. After mixing, let solution stand for
several days keeping the solution CO2-free. Use supernatant after standardization.
Standardization:
Weigh required quantity of amidosulfuric acid (certified reference material for volumetric analysis)
and place it into desiccator (< 2.0 kPa) for 48 hours. Next, accurately weigh 1 g to 1.25 g (recording
the weight to 0.1 mg), dissolve in water (CO2-free), and dilute to 250 mL. Put 25 mL solution into
Erlenmeyer flask, add 2 to 3 drops of bromothymol blue indicator and titrate with 0.05 mol/L
alcoholic potassium hydroxide solution until colour of solution change to faint blue.
Calculation:
Factor of molarity = (g amidosulfuric acid × purity × 25) / 1.2136 / mL KOH
(b) Alcohol-ether mixture: equal volumes ethanol (99.5%) and ether.
(c) Phenolphthalein solution: 1% in alcohol.
 CXS 234-1999 129

2.4 Titration
Before sampling, liquefy extracted oil using water bath. Weigh 1 g to 2 g liquefied test portion into Erlenmeyer
flask. Add 80 mL alcohol-ether mixture and a few drops of phenolphthalein solution. Titrate with 0.05 mol/L
alcoholic KOH until faint pink colour appears and retain for more than 30 s. Perform blank test using only
alcohol-ether mixture and phenolphthalein solution.
2.5 Calculation
Calculate using following equation:
Acid value [mg/g] = (mL test portion – mL blank) × factor of molarity × 2.806 / g test portion
2.5.1 Definition and principle
Acid value of oil from fried instant noodles = mg KOH required to neutralize 1 g oil. Oil extracted from noodle
is dissolved in alcohol-ether mixture and titrated with alcoholic KOH standard solution.
2.5.2 Apparatus
Air-tight desiccator: silica gel heated at 150 °C is satisfactory drying agent.
2.5.3 Reagents
(d) Alcoholic potassium hydroxide standard solution: 0.05 mol/L. Dissolve 3.5 g potassium hydroxide in
equal volume of water (CO2-free) and add ethanol (95%) to 1 L. After mixing, let solution stand for
several days keeping the solution CO2-free. Use supernatant after standardization.
Standardization:
Weigh required quantity of amidosulfuric acid (certified reference material for volumetric analysis)
and place it into desiccator (< 2.0 kPa) for 48 hour. Next, accurately weigh 1 g to 1.25 g (recording
the weight to 0.1 mg), dissolve in water (CO2-free), and dilute to 250 mL. Put 25 mL solution into
Erlenmeyer flask, add 2 to 3 drops of bromothymol blue indicator and titrate with 0.05 mol/L
alcoholic potassium hydroxide solution until colour of solution change to faint blue.
Calculation:
Factor of molarity = (g amidosulfuric acid × purity × 25) / 1.2136 / mL KOH
(e) Alcohol-ether mixture: equal volumes ethanol (99.5%) and ether.
(f) Phenolphthalein solution: 1% in alcohol.
2.5.4 Titration
Before sampling, liquefy extracted oil using water bath. Weigh 1 g to 2 g liquefied test portion into Erlenmeyer
flask. Add 80 mL alcohol-ether mixture and a few drops of phenolphthalein solution. Titrate with 0.05 mol/L
alcoholic KOH until faint pink colour appears and retain for more than 30 s. Perform blank test using only
alcohol-ether mixture and phenolphthalein solution.
2.5.5 Calculation
Calculate using following equation:
Acid value [mg/g] = (mL test portion – mL blank) × factor of molarity × 2.806 / g test portion

PART C– DETERMINATION OF MOISTURE

3. Determination of moisture
3.1 Apparatus
(a) Aluminium dish: diameter ≥ 55 mm, height ≥ 15 mm, and with inverted tight-fitting lid.
(b) Air oven: with control accuracy ±1 °C.
(c) Air-tight desiccator: silica gel heated at 150 °C is satisfactory drying agent.
 CXS 234-1999 130

3.2 Preparation of test sample

Remove instant noodles from package and leave garnishing and seasoning in package. Transfer the noodles
to plastic bag to prevent moisture change and then break these into small fragments with hands or wooden
hammer. Select broken noodles in the size range of 2.36 mm to 1.7 mm by using two sieves with 2.36 mm and
1.7 mm openings (mesh size 12–8) and mix well. Use these noodles for test sample. If noodles are too thin to
screen with sieves, cut them into 1 cm to 2 cm lengths, mix well and use these cut noodles for test sample.
3.3 Determination
3.3.1 Fried noodles
In cooled and weighed dish (with lid), previously heated to 105 °C, weigh ca 2 g well-mixed test portion to
1 mg. Uncover test portion and dry dish, lid, and contents 2 h in oven provided with opening for ventilation and
maintained at 105 °C. (The 2 h drying period begins when oven temperature is actually 105 °C.) After drying
period, cover dish while still in oven, transfer to desiccator, and weigh to 1 mg soon after reaching room
temperature. Report loss in weight as moisture (indirect method).
3.3.2 Non-fried noodles
For non-fried noodles, follow the directions for fried noodles, but dry test portion for 4 h.
3.4 Calculation
Calculate using the following equation:
Moisture (%) = {(g test portion before drying – g test portion after drying) / g test portion before drying}×100
CXS 234-1999 131

APPENDIX II

DETERMINATION OF PRESENTATION IN CANNED TUNA AND BONITO

The presentation of all sample units shall be determined by the following procedure.
(i) Open the can and drain the contents, following the procedures outlined in Appendix III
determination of drained weight.
(ii) Remove and place the contents onto a tared 1.2 cm mesh screen equipped with a collecting pan.
(iii) Separate the fish with a spatula being careful not to break the configuration of the pieces. Ensure
that the smaller pieces of fish are moved to the top of a mesh opening to allow them to fall through
the screen onto the collecting pan.
(iv) Segregate the material on the pan according to flaked, grated (shredded) or paste and weigh the
individual portions to establish the weight of each component.
(v) If declared as a "chunk" pack weigh the screen with the fish retained and record the weight.
Subtract the weight of the sieve from this weight to establish the weight of solid and chunk fish.
(vi) If declared as "solid" pack remove any small pieces (chunks) from the screen and reweigh.
Subtract the weight of the sieve from this weight to establish the weight of "solid" fish.

Calculations

(i) Express the weight of flaked, grated (shredded and paste) as a percentage of the total drained
weight of fish.

Weight of flakes
% flakes  x 100
Total weight of drained fish

(ii) Calculate the weight of solid and chunk fish retained on the screen by difference and express as
a % of the total drained weight of fish.

Weight of solid & chunk fish

% solid chunk fish  x 100
Total weight of drained fish

(iii) Calculate the weight of solid fish retained on the screen by difference and express as a % of the
total drained weight of the fish.

Weight of solid fish

% of solid fish = x 100
Total weight of drainedfish
CXS 234-1999 132

APPENDIX III

DETERMINATION OF NET WEIGHT, DRAINED WEIGHT, AND WASHED DRAINED WEIGHT IN FISH AND FISHERY PRODUCTS

Commodity Provision Method

Net weight Net contents of all sample units shall be determined by the following procedure:
(i) Weigh the unopened container.
(ii) Open the container and remove the contents.
(iii) Weigh the empty container, (including any ends) after removing excess liquid and
adhering meat.
(iv) Subtract the weight of the empty container from the weight of the unopened container.
(v) The resultant figure will be the net content.
 Canned salmon
Drained weight The drained weight of all sample units shall be determined by the following procedure:
 Canned shrimps or prawns
(i) Maintain the container at a temperature between 20 C and 30 C for a minimum of
 Canned tuna and bonito 12 hours prior to examination.
 Canned crab meat (ii) Open and tilt the container to distribute the contents on a pre-weighed circular sieve
 Canned sardines and sardine-type which consists of wire mesh with square openings of 2.8 mm x 2.8 mm.
products (iii) Remove any wrapping material and incline the sieve at an angle of approximately
 Canned finfish 17– 20° and allow sample to drain for two minutes, measured from the time the
product is poured into the sieve.
(iv) Weigh the sieve containing the drained sample.
(v) The weight of drained sample is obtained by subtracting the weight of the sieve from
the weight of the sieve and drained product.
 Canned sardines and sardine-type Washed drained (i) Maintain the container at a temperature between 20 C and 30 C for a minimum of
products weight (for packs 12 hours prior to examination.
with sauces)
 Canned finfish (ii) Open and tilt the container and wash the covering sauce and then the full contents
with hot tap water (approx. 40 C), using a wash bottle (e.g. plastic) on the tared
circular sieve.
(iii) Wash the contents of the sieve with hot water until free of adhering sauce; where
necessary separate optional ingredients (spices, vegetables, fruits) with pincers.
CXS 234-1999 133

Commodity Provision Method

Incline the sieve at an angle of approximately 17–20 and allow the fish to drain two
minutes, measured from the time the washing procedure has finished.
(iv) Remove adhering water from the bottom of the sieve by use of paper towel. Weigh the
sieve containing the washed drained fish.
(v) The washed drained weight is obtained by subtracting the weight of the sieve from the
weight of the sieve and drained product
Quick-frozen fish sticks (fish fingers), Net weight The net weight (exclusive of packaging material) is determined on each whole primary container of
fish portions and fish fillets – breaded or each sample representing a lot and shall be determined in the frozen state.
in batter

Salted fish and dried salted fish of the Net weight The net weight (excluding packaging material and excess salt) of each sample unit in the sample
Gadidae family of fishes lot shall be determined.
Dried shark fins Net weight The net weight (exclusive of packaging material) of each sample unit in the sample lot shall be
determined.

Salted Atlantic herring and salted sprat Net weight The net weight (excluding packaging material) of each sample unit in the sample lot shall be
determined. Remove the herring from the container (barrel) and put it on an appropriate sieve.
Allow to drain for 5 min and remove adhering salt crystals. Weigh the herring and calculate net
weight.
Sturgeon caviar Net weight The net weight (excluding packaging material) of each sample unit in the sample lot shall be
determined by deducting the weight of the empty container from the total weight.

Smoked fish, smoke-flavoured fish and Net weight The net weight is determined as the weight of the product, exclusive of packaging material,
smoke-dried fish interleaving material, etc.

Raw bivalve molluscs Net weight (i) Weigh the unopened container.
(ii) Open the container and remove the contents.
(iii) Dry the empty container and weigh.
(iv) Subtract the weight of the empty container from the weight of the unopened container.
The resultant figure will be the total net weight.
CXS 234-1999 134

Commodity Provision Method

Net weight of The net weight (exclusive of packaging material) of each sample unit representing a lot shall be
frozen products determined in the frozen state.
not covered by
glaze
Net weight of Additionally, AOAC 963.26– Net weight of products with water added that is inside a "block-frozen"
products covered product.
by glaze
(Live abalone and) raw fresh chilled or Net weight (i) Remove any frost and ice from outside of package.
frozen abalone for direct consumption or
(ii) Weigh the unopened container.
for further processing
(iii) Open the container and remove the contents.
(iv) Dry the empty container and weigh.
(v) Subtract the weight of the empty container from the weight of the unopened container.
The resultant figure will be the total net weight.
Net weight of The net weight (exclusive of packaging material) of each sample unit representing a lot shall be
frozen products determined in the frozen state.
not covered by
glaze
 Quick-frozen finfish, uneviscerated Net weight of The net weight (exclusive of packaging material) of each sample unit representing a lot shall be
and eviscerated frozen products determined in the frozen state.
not covered by
 Quick-frozen shrimps or prawns glaze
 Quick-frozen lobsters
 Quick-frozen blocks of fish fillets,
minced fish flesh and mixtures of
fillets and minced fish flesh
 Quick-frozen fish fillets
 Quick-frozen raw squid
CXS 234-1999 135

Commodity Provision Method

Quick-frozen shrimps or prawns Net weight of (1) Open the package with quick-frozen shrimps or prawns immediately after removal from low
products covered temperature storage.
by glaze
(i) For the raw product, place the contents in a container into which fresh water at room
temperature is introduced from the bottom at a flow of approximately 25 litres per minute.
(ii) For the cooked product place the product in a container containing an amount of fresh
potable water of 27 ºC (80 º F) equal to 8 times the declared weight of the product. Leave
the product in the water until all ice is melted. If the product is block frozen, turn block over
several times during thawing. The point at which thawing is complete can be determined
by gently probing the block apart.
(2) Weigh a dry clean sieve with woven wire cloth with nominal size of the square aperture 2.8 mm
(ISO Recommendation R565) or alternatively 2.38 mm (US No. 8 Standard Screen).
(i) If the quantity of the total contents of the package is 500 g (1.1 lbs) or less, use a sieve
with a diameter of 20 cm (8 inches).
(ii) If the quantity of the total contents of the package is more than 500 g (1.1 lbs) use a sieve
with a diameter of 30 cm (12 inches).
(3) After all glaze that can be seen or felt has been removed and the shrimps or prawns separate
easily, empty the contents of the container on the previously weighed sieve. Incline the sieve at an
angle of about 20º and drain for two minutes.
(4) Weigh the sieve containing the drained product. Subtract the mass of the sieve; the resultant
figure shall be considered to be the net content of the package.
Quick-frozen lobsters Net weight of (1) As soon as the package is removed from frozen temperature storage, open immediately and
products covered place the contents under a gentle spray of cold water until all ice glaze that can be seen or felt is
by glaze removed. Remove adhering water by the use of paper towel and weigh the product.
(alternate
(2) The pre-weighed glazed sample is immersed into a water bath by hand, until all glaze is
methods)
removed, which preferably can be felt by the fingers. As soon as the surface becomes rough, the
still frozen sample is removed from the water bath and dried by use of a paper towel before
estimating the net product content by second weighing. By this procedure thaw drip losses and/or
re-freezing of adhering moisture can be avoided.
(i) As soon as the package is removed from frozen temperature storage, place the product in
a container containing an amount of fresh potable water of 27 C (80 F) equal to 8 times
the declared weight of the product. Leave the product in the water until all ice is melted. If
CXS 234-1999 136

Commodity Provision Method

the product is block frozen, turn block over several time during thawing. The point at which
thawing is complete can be determined by gently probing the block.
(ii) Weigh a dry clean sieve with woven wire cloth with nominal size of the square aperture
2.8 mm (ISO Recommendation R565) or alternatively 2.38 mm (U.S. No. 8 Standard
Screen.) (a) If the quantity of the total contents of the package is 500 g (1.1 lbs) or less,
use a sieve with a diameter of 20 cm (8 inches). (b) If the quantity of the total contents of
the package is more than 500 g (1.1 lbs) use a sieve with a diameter of 30 cm (12 inches).
(iii) After all glaze that can be seen or felt has been removed and the lobsters separate easily,
empty the contents of the container on the previously weighed sieve. Incline the sieve at
an angle of about 20 and drain for two minutes.
(iv) Weigh the sieve containing the drained product. Subtract the mass of the sieve; the
resultant figure shall be considered to be part of the net content of the package.
Quick-frozen blocks of fish fillets, Net weight of As soon as the package is removed from frozen temperature storage, open immediately and place
minced fish flesh and mixtures of fillets products covered the contents under a gentle spray of cold water until all ice glaze that can be seen or felt is removed.
and minced fish flesh by glaze Remove adhering water by the use of paper towel and weigh the product.
An alternate method is outlined in the annex.
Quick-frozen fish fillets Determination of As soon as the package is removed from low temperature storage, open immediately and place
net weight of the contents under a gentle spray of cold water.
products covered
Agitate carefully so that the product is not broken.
by glaze
Spray until all ice glaze that can be seen or felt is removed.
Remove adhering water by the use of paper towel and weight the product in a tared pan
Quick-frozen raw scallop products Net weight (i) AOAC 963.18.
(ii) Block frozen products: AOAC 967.13. The block-frozen scallops shall be thawed
inside waterproof bags to prevent contact with, and absorption of, the water used to
thaw the product
 CXS 234-1999 137

ANNEX TO APPENDIX III

METHOD FOR THE DETERMINATION OF NET CONTENT OF FROZEN FISH BLOCKS
COVERED BY GLAZE
Glazing is not used for quick-frozen blocks of white fish. Only quick-frozen blocks of herring, mackerel and other
brown (fat) fish are glazed, which are destined for further processing (canning, smoking). For such blocks the
following procedure may be applicable (tested with block-frozen shrimps).
1. PRINCIPLE
The pre-weighed glazed sample is immersed into a water bath by hand till all glaze is removed (as felt by fingers).
As soon as the surface becomes rough, the still frozen sample is removed from the water bath and dried by use
of a paper towel before estimating the net product content by repeated weighing. By this procedure thaw drip
losses and/or re-freezing of adhering moisture can be avoided.
2. EQUIPMENT
Balance – sensitive to 1 g
Water bath, preferably with adjustable temperature
Circular sieve with a diameter of 20 cm and 1–3 mm mesh apertures (ISO R 565)
Paper or cloth towels with smooth surface
A freezer box should be available at the working place
3. PREPARATION OF SAMPLES AND WATER BATH

The product temperature should be adjusted to -18/-20 C to achieve standard deglazing conditions (especially
necessary if a standard deglazing period shall be defined in case of regular shaped products).
After sampling from the low temperature store remove, if present, external ice crystals or snow from the
package with the frozen product.
The water bath shall contain an amount of fresh potable water equal to about 10 times of the declared weight
of the product; the temperature should be adjusted on about 15 C to 35 C.
4. DETERMINATION OF GROSS-WEIGHT "A"
After removal of the package, the weight of the glazed product is determined: In case of single fish fillets, single
weights are recorded (A 1-A n). The weighed samples are placed intermediately into the freezer box.
5. REMOVAL OF GLAZE
The pre-weighed samples/sub-samples are transferred into the water bath and kept immersed by hand. The
product may be carefully agitated, till no more glaze can be felt by the fingertips on the surface of the product:
change from slippery to rough. Needed time, depending on size/shape and glaze content of the product, 10 to
60 sec (and more in case of higher glaze contents or if frozen together).
For block-frozen products in consumer packs (also for single glaze products, which are frozen together during
storage) the following (preliminary) procedure may be applicable: The pre-weighed block or portion is
transferred onto a suitable sized sieve and immersed into the water bath. By slight pressure of the fingers
separating deglazed portions are removed fractionally. Short immersing is repeated, if glaze residues are still
present.
6. DETERMINATION OF NET WEIGHT "B"
The deglazed sample/subsample, after removal of adhering water by use of a towel (without pressure) is
immediately weighed. Single net-weights of sub-samples are summed up: B1-n.
 CXS 234-1999 138

7. DETERMINATION OF GLAZE-WEIGHT "C"

Grossweight " A"  Net weight " B "  Glaze weight " C"
8. CALCULATION OF PERCENTAGE PROPORTIONS

" B"
% net content of the product " F"  x 100
" A"
" C"
% glaze  related to the gross weight of the product " G"  x 100
" A"
" C"
% glaze  related to the net weight of the product " H"  x 100
" B"
 CXS 234-1999 139

APPENDIX IV

SENSORY AND PHYSICAL EXAMINATION OF FISH AND FISHERY PRODUCTS

Canned salmon, canned crab meat, canned shrimps or prawns, canned tuna and bonito, canned finfish,
canned sardine and sardine-type products
1. Complete external can examination for the presence of container integrity defects or can ends which may be
distorted outward.
2. Open can and complete weight determination according to defined procedures in Appendix III.
3. For canned salmon and canned crab meat
 Examine product for discolouration, foreign and objectionable matter.
4. For canned shrimps or prawns
 Carefully remove the product and examine for size designation in accordance with the procedure in
Section 7.2.2 of CXS 37-1991.2
 Examine product for discolouration, foreign and objectionable matter.
5. For canned tuna and bonito
 Examine the product for discolouration.
 Carefully remove the product and determine the presentation according to the defined procedures in
Appendix III.
 Examine product for discolouration, foreign matter and struvite crystals.
6. For canned finfish
 Examine the product for the form of presentation.
 Examine product for discolouration, foreign and objectionable matter.
7. For canned sardines and sardine-type products
 Carefully remove product and examine for discolouration, foreign matter and struvite crystals.
8. The presence of hard bone in canned salmon, canned tuna and bonito, canned finfish, canned sardines
and sardine-type products is an indicator of underprocessing and will require an evaluation for sterility.
9. Assess odour, flavour and texture in accordance with the Guidelines for the sensory evaluation of fish and shellfish
in laboratories (CXG 31-1999).3
Quick-frozen finfish, uneviscerated and eviscerated, quick-frozen shrimps or prawns, quick-frozen lobsters,
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), fish portions and fish fillets – breaded or in batter, quick-frozen raw
squid, fresh and quick-frozen raw scallop products
1. For quick-frozen fish sticks (fish fingers), fish portions and fish fillets– breaded or in batter, the sample
used for sensory evaluation should not be the same as that used for other examinations.
2. Complete net weight determination, according to defined procedures in Appendix III (de-glaze as required).
3. For quick-frozen finfish, uneviscerated and eviscerated
 Examine the frozen sample unit for the presence of deep dehydration by measuring those areas or
counting instances which can only be removed with a knife or other sharp instrument. Measure the total
surface area of the sample unit, and calculate the percentage affected.
 Thaw and individually examine each fish in the sample unit for the presence of foreign matter.
 Examine each fish using the criteria outlined in the respective standard. Flesh odours are examined by
tearing or making a cut across the back of the neck such that the exposed surface of the flesh can be
evaluated.
 CXS 234-1999 140

4. For quick-frozen blocks of fish fillet, minced fish flesh and mixtures of fillets and minced fish flesh
 Examine the frozen block or fillets for the presence of dehydration by measuring those areas which
can only be removed with a knife or other sharp instrument. Measure the total surface area of the
sample unit, and calculate the percentage affected.
 Thaw and individually examine each block in the sample unit for the presence of foreign matter, bone
where applicable, odour and textural defects.
5. For quick-frozen fish fillets
 Examine the frozen block or fillets for the presence of dehydration by measuring those areas which can
only be removed with a knife or other sharp instrument. Measure the total surface area of the sample
unit, and calculate the percentage affected.
 Thaw and individually examine each block in the sample unit for the presence of foreign matter,
parasites, bone where applicable, odour and flesh abnormality defects.
6. For quick-frozen shrimps and prawns
 Examine the frozen shrimp in the sample unit or the surface of the block for the presence of dehydration.
Determine the percentage of shrimp or surface area affected.
 Thaw using the procedure described in Appendix V and individually examine each shrimp in the sample
unit for the presence of foreign matter and presentation defects. Determine the weight of shrimp affected
by presentation defects.
 Examine product for count declarations in accordance with procedures in CXS 92-1981.4
 Assess the shrimp for odour and discolouration as required.
7. For quick-frozen lobsters
 Examine the frozen lobster for the presence of deep dehydration. Determine the percentage of lobster
affected.
 Thaw using the procedure described in Appendix V and individually examine each sample unit for the
presence of foreign and objectionable matter.
 Examine product count and weight declarations in accordance with procedures in CXS 95-1981.5
 Assess the lobster for odour and discolouration as required.
8. For quick-frozen fish sticks (fish fingers), fish portions and fish fillets– breaded or in batter
 Complete fish core determination on one set of the sample units according to defined procedures in
Appendix VI.
 Complete the estimation of the proportion of fillets and minced flesh, if required.
 Cook the other set of sample units and examine for odour, flavour, texture, foreign matter and bones.
9. For quick-frozen raw squid
 Examine the frozen squid for the presence of deep dehydration by measuring those areas which can
only be removed with a knife or other sharp instrument. Measure the total surface area of the sample
unit, and determine the percentage affected using the following formula;
area affected
×100% = % affected by deep dehydration
total surface area
 Thaw and individually examine each squid in the sample unit for the presence of foreign matter and
colour.
 Examine each squid using the criteria outlined in CXS 191-1995.6 Flesh odours are examined by
making a cut parallel to the surface of the flesh so that the exposed surface can be evaluated.
10. For fresh and quick-frozen raw scallop products
 Examine the frozen scallop product in the sample unit or the surface of the block for the presence of
dehydration. Determine the percentage of scallop meat or surface area affected.
 CXS 234-1999 141

 Thaw using the procedure described in Appendix III and individually examine each scallop product in
the sample unit for the presence of foreign matter, objectionable matter and presentation defects.
 Determine the weight of scallop product affected by presentation defects.
 Examine product for pieces and count declarations in accordance with procedures in CXS 315-2014.7
 Assess the scallop product for odour and parasites as required.
 A small portion of the sample unit (100 g to 200 g) is cooked without delay and the odour/flavour/texture
and presence of sand is determined. If necessary, additional portions may be cooked and examined for
confirmation.
11. In cases where a final decision regarding the odour or texture cannot be made in the thawed uncooked state:
 In quick-frozen finfish, uneviscerated and eviscerated, a small portion of the flesh (approximately
200 g) is sectioned from the product and the odour, flavour or texture confirmed without delay by using
one of the cooking methods defined as follows: AOAC Methods – "Moisture in Meat and Meat Products,
Preparation of Sample Procedure"; 883.18 and "Moisture in Meat" (Method A); 950.46; AOAC 1990.
 In quick-frozen blocks of fish fillet, minced fish flesh and mixtures of fillets and minced fish flesh,
a small portion of the disputed material (approximately 200 g) is sectioned from the block and the odour
and flavour confirmed without delay by using one of the cooking methods defined in Appendix V.
 In quick-frozen raw squid, a portion of the sample unit is sectioned off and the odour, flavour and
texture confirmed without delay by using one of the cooking methods defined in Appendix V.
12. In cases where a final decision regarding the odour cannot be made in the thawed uncooked state:
 In quick-frozen fish fillets, a small portion of the disputed material (approximately 200 g) is sectioned
from the sample unit and the odour and flavour confirmed without delay by using one of the cooking
methods defined in Appendix V.
13. In cases where a final decision on gelatinous condition cannot be made in the thawed uncooked state:
 In quick-frozen finfish, uneviscerated and eviscerated, the disputed material is sectioned from the
product and gelatinous condition confirmed by cooking as defined in Appendix IV or by using one of the
cooking methods defined as follows to determine if greater than 86 percent moisture is present in any
fish: AOAC Methods – "Moisture in Meat and Meat Products, Preparation of Sample Procedure"; 883.18
and "Moisture in Meat" (Method A); 950.46; AOAC 1990. If a cooking evaluation is inconclusive, then
one of the cooking methods defined as follows would be used to make the exact determination of
moisture content: AOAC Methods – "Moisture in Meat and Meat Products, Preparation of Sample
Procedure"; 883.18 and "Moisture in Meat" (Method A); 950.46; AOAC 1990.
 In quick-frozen blocks of fish fillet, minced fish flesh and mixtures of fillets and minced fish flesh,
the disputed material is sectioned from the block and the gelatinous condition confirmed by cooking as
defined in Appendix V or by using procedure in Appendix VII to determine if greater than 86 percent
moisture is present in any fillet. If cooking evaluation is inconclusive, then procedure in Appendix VI
would be used to make the exact determination of moisture content.
 In quick-frozen fish fillets and quick-frozen fish sticks (fish fingers), fish portions and fish fillets
– breaded or in batter, the disputed material is sectioned from the product and gelatinous condition
confirmed by cooking as defined in Appendix V or AOAC Methods– "Moisture in Meat and Meat
Products, Preparation of Sample Procedure"; 983.18 and "Moisture in Meat" (Method A); 950.46 to
determine if greater than 86 percent moisture is present in any fillet or product unit. If a cooking
evaluation is inconclusive, then AOAC Methods– "Moisture in Meat and Meat Products, Preparation of
Sample Procedure"; 983.18 and "Moisture in Meat" (Method A); 950.46 would be used to make the
exact determination of moisture content.
14. In cases where a final decision regarding the odour/flavour cannot be made in the thawed state in quick-
frozen shrimps or prawns, quick-frozen lobsters, a small portion of the sample unit (100 g to 200 g) is
prepared without delay for cooking and the odour/flavour confirmed by using one of the cooking methods
defined in Appendix V.
Crackers from marine and freshwater fish, crustacean and molluscan shellfish and boiled dried salted
anchovies
The sample used for sensory evaluation should not be same as that used for other examination.
 CXS 234-1999 142

1. For crackers from marine and freshwater fish, crustacean and molluscan shellfish, examine the sample
unit for foreign matter, bones and discolouration.
2. For boiled dried salted anchovies, examine every fish in the sample unit for foreign matter, breakage, pink
condition and mould growth.
3. For all the mentioned commodities above, assess the odour in the uncooked sample in accordance with the
Guidelines for the sensory evaluation of fish and shellfish in laboratories (CXG 31-1999).3
4. For all the mentioned commodities above, assess the flavour in cooked sample in accordance with CXG 31-
1999.
5. For crackers from marine and freshwater fish, crustacean and molluscan shellfish, the sample shall be
deep fried in fresh cooking oil at 190 oC for 20–60 seconds as appropriate to the thickness of the crackers.
6. For boiled dried salted anchovies, the sample shall be cooked prior to assessment according to the cooking
instructions on the package. When such instructions are not given, the sample shall be deep fried in fresh
cooking oil at 190 ºC for 1–2 minutes as appropriate to the size.
Fish sauce
1. Complete external packaging unit examination for the presence of any integrity defects, particularly cracks or
leakage or loose pieces of the packaging units.
2. Examination of the product for translucence and foreign matter.
3. Evaluation of odour and taste.
Salted fish and dried salted fish of the Gadidae family of fishes
1. Examine every fish in the sample in its entirety.
2. Examine the product for the form of presentation.
3. Examine the fish for foreign matter, pink conditions, halophilic mould, liver stains, intense bruising, severe
burning and texture.
4. Assess odour in accordance with CXG 31-1999.3
Salted Atlantic herring and salted sprat
See CXS 244-2004.8
Sturgeon caviar
See CXS 291-2010.9
Live and raw bivalve molluscs
See Appendices III, V and IX, and CXS 292-2008.10
Smoked fish, smoke-flavoured fish and smoke-dried fish
See Appendices III, V and IX.
Live abalone and for raw fresh chilled or frozen abalone for direct consumption or for further
processing
See CXS 312-2013.11
CXS 234-1999 143

APPENDIX V

PART 1: THAWING PROCEDURES IN FISH AND FISHERY PRODUCTS

Quick-frozen shrimps or prawns, quick-frozen lobsters and quick-frozen raw squid

The sample unit is thawed by enclosing it in a film type bag and immersing in water at room temperature (not
greater than 35 ºC). The complete thawing of the product is determined by gently squeezing the bag
occasionally so as not to damage the texture of the sample, until no hard core or ice crystals are left.
Raw bivalve molluscs
For frozen product, the sample unit is thawed by enclosing it in a film type bag and immersing in water at room
temperature (not greater than 35 ºC). The complete thawing of the product is determined by gently squeezing
the bag occasionally so as not to damage the texture of the bivalve molluscs, until no hard core or ice crystals
are left.
Smoked fish, smoke-flavoured fish and smoke-dried fish
Temperatures for thawing
Frozen samples of final products shall be thawed at refrigeration temperatures to maintain quality and safety.
Raw fresh chilled or frozen abalone
For frozen product, the sample is thawed by enclosing it in a film type bag allowing it to thaw at room
temperature or in a refrigerator (at 2–6 °C). The complete thawing of the product is determined by gently
squeezing the bag occasionally so as not to damage the texture of the abalone, until no hard core or ice
crystals are left.
Quick-frozen blocks of fish fillet, minced fish flesh and mixtures of fillets and minced fish flesh
Air thaw method:
Frozen fish blocks are removed from the packaging. The frozen fish blocks are individually placed into snug
fitting impermeable plastic bags or a humidity controlled environment with a relative humidity of at least
80 percent. Remove as much air as possible from the bags and seal. The frozen fish blocks sealed in plastic
bags are placed on individual trays and thawed at air temperature of 25 C (77 F) or lower. Thawing is
completed when the product can be readily separated without tearing. Internal block temperature should not
exceed 7 C (44.6 F).
Water immersion method:
Frozen fish blocks are removed from the packaging. The frozen fish blocks are sealed in plastic bags. Remove
as much air as possible from the bags and seal. The frozen fish blocks are placed into a circulating water bath
with temperatures maintained at 21 C + 1.5 C (70 F + 3 F). Thawing is completed when the product can
be easily separated without tearing. Internal block temperature should not exceed 7 C (44.6 F).
PART 2: COOKING PROCEDURES IN FISH AND FISHERY PRODUCTS
Quick-frozen finfish, uneviscerated and eviscerated, quick-frozen shrimps or prawns, quick-frozen
lobsters, quick-frozen blocks of fish fillet, minced fish flesh and mixtures of fillets and minced fish
flesh, quick-frozen fish fillets, quick-frozen raw squid.
The following procedures are based on heating the product to an internal temperature of 65–70 C. The
product must not be overcooked. Cooking times vary according to the size of the product and the temperatures
used. The exact times and conditions of cooking for the product should be determined by prior experimentation.
Baking procedure: Wrap the product in aluminium foil and place it evenly on a flat cookie sheet or shallow
flat pan.
Steaming procedure: Wrap the product in aluminium foil and place it on a wire rack suspended over boiling
water in a covered container.
Boil-in-bag procedure: Place the product into a boilable film-type pouch and seal. Immerse the pouch into
boiling water and cook.
Microwave procedure: Enclose the product in a container suitable for microwave cooking. If plastic bags
are used, check to ensure that no odour is imparted from the plastic bags. Cook according to equipment
specifications.
CXS 234-1999 144

Quick-frozen fish sticks (fish fingers), fish portions and fish fillets – breaded or in batter
The frozen sample shall be cooked prior to sensory assessment according to the cooking instructions on the
package. When such instructions are not given, or equipment to cook the sample according to the instructions
is not obtainable, the frozen sample shall be cooked according to the applicable method(s) given below:
Use procedure 976.16 of the AOAC. It is based on heating product to an internal temperature of 65–70 C.
Cooking times vary according to size of product and equipment used. If determining cooking time, cook extra
samples, using a temperature measuring device to determine internal temperature.
CXS 234-1999 145

APPENDIX VI

DETERMINATION OF FISH CONTENT (DECLARATION) IN QUICK-FROZEN FISH STICKS (FISH

FINGERS), FISH PORTIONS AND FISH FILLETS – BREADED OR IN BATTER
Estimation of fish content
AOAC Method 996.15. (End-product method)
Calculation:
% fish content = (Wd/Wb) X 100 + adjustment factor*
Wd = weight of debattered and/or debreaded test unit
Wb = weight of battered and/or breaded test unit
* Raw breaded frozen coated fish and fishery products: 2.0%
* Batter-dipped frozen coated fish and fishery products: 2.0%
* Precooked frozen coated fish and fishery products: 4.0%
Reference: J. AOAC Int. 80, 1235 (1997)
Other methods
(1) Chemical analysis method (nitrogen factor end-product method)
Appropriate in cases where there is reason to doubt the composition of the fish core (i.e. appears to contain
non-fish ingredients). Except for fully cooked products, this method requires confirmation with the
AOAC Method 996.15, or with Method #2 (Determination of fish content) in conjunction with investigation at
the processing plant when determining product compliance with the labelling provisions in CXS 166-1989.12
This method should trigger in-factory investigation (e.g. raw ingredient recipe checks) when suspect products
are identified.
The percentage fish content, corrected for the non-fish flesh nitrogen contributed by the carbohydrate coating,
is calculated as follows.
(% total nitrogen - % nonfish flesh nitrogen)
% Fish  x100
N factor*
*appropriate N (nitrogen) factor
The non-fish flesh nitrogen is calculated as follows:
% non-fish flesh nitrogen = % carbohydrate X 0.02
Where the carbohydrate is calculated by difference:
% carbohydrate = 100 – (%water + % fat + % protein + % ash)
References
Determination of nitrogen: ISO 937
Determination of moisture: ISO 1442
Determination of total fat: ISO 1443
Determination of ash: ISO 936
Average nitrogen factors to be used for fish flesh for specific fish species used as raw material for the product
can be found at the following website:
http://www.fao.org/in-action/globefish/fishery-information/resource-detail/en/c/338604/
http://www.fao.org/fishery/topic/1514/en
The uncertainty of each nitrogen factor should be taken into account from the statistical data presented with
the published nitrogen factor (e.g. 2 standard errors about the mean).
CXS 234-1999 146

(2) Rapid method used during production

The fish content of a fish finger (fish stick) is calculated by using the following equation:

Weight of ingoing fish

% Fish Content  x100
Weight of final product

For most products, therefore, the fish ingredient weight is that of the raw ingredient. Any figure placed or
declared on a product label would be a typical quantity reflecting the producer’s normal manufacturing
variations, in accordance with good manufacturing practice.
 CXS 234-1999 147

APPENDIX VII

ESTIMATION OF PROPORTION OF FISH FILLETS AND MINCED FISH FLESH

(West European Fish Technologists Association– WEFTA Method)
a) Equipment
Balance, sensitive to 0.1 g
Circular sieve – 200 mm diameter, 2.5 or 2.8 mesh opening (ISO) soft rubber edge (or blunt) spatula, forks,
suitable sized plates, water tight plastic bags.
b) Preparation of samples
Fish portions/sticks: Take as many portions as needed to provide a fish core sample of about 200 g (2 kg). If
breaded and/or battered first strip coating according to the method described in Appendix VI.
c) Detemination of weights "A" of the frozen fish samples
Weight the single fish portions/decoated fish cores while they are still frozen. Smaller portions are combined
to a sample subunits of about 200 g (e.g. 10 fish sticks of about 20 g each). Record the weight "A" n of the
subunits. Place the pre-weighed sample subunits into water tight bags.
d) Thawing
Thaw the samples by immersing the bags into a gently agitated water bath of about 20 ºC, but not more than
35 ºC.
e) Draining
After thawing has been completed (duration about 20-30 minutes) take each sample unit, one at a time, and
drain the exuded fluid (thaw drip) for 2 minutes on a pre-weighed circular sieve incluned at an angle of
17– 20 degrees. Remove adhering drip from the bottom of the sieve by use of a paper towel when draining is
completed.
f) Determination of weight "B" of the drained fish sample and weight ”C” of the thaw drip
Determine the weight of the drained fish sample "B" - sieve plus fish minus sieve weight. The difference of "A"
- "B" is the weight of exuded fluid - thaw drip.
g) Separation
Place the drained fish core on a plate and separate the minced flesh from the fillet using a fork to hold the fillet
flesh and a soft, rubber edge spatula to scrape off the minced flesh.
 CXS 234-1999 148

APPENDIX VIII

PREPARATION OF FISH SAMPLES AND DETERMINATION OF SALT AND WATER CONTENT IN FISH
AND FISHERY PRODUCTS

PART 1: PREPARATION OF FISH SAMPLES

Salted fish and dried salted fish of the Gadidae family of fishes
1. Before preparing of a subsample adhering salt crystals should be removed by brushing from the surface of the
sample without using water.
2. The preparation of fish samples for the determination of salt content, and water content in order to calculate
the % salt saturation of the fish should be carried out according to AOAC 937.07. The analysis should be on
the edible portion of the fish.
3. Determination should be performed at least in duplicate.
PART 2: DETERMINATION OF SALT CONTENT
Salted fish and dried salted fish of the Gadidae family of fishes, salted Atlantic herring and salted sprat,
and sturgeon caviar
1. Principle
The salt is extracted by water from the pre-weighed sample. After the precipitation of the proteins, the chloride
concentration is determined by titration of an aliquot of the solution with a standardized silver nitrate solution
(Mohr method) and calculated as sodium chloride.
2. Equipment and chemicals
- Brush
- Sharp knife or saw
- Balance, accurate to  0.01 g
- Calibrated volumetric flasks, 250 ml
- Erlenmeyer flasks
- Electric homogenizer
- Magnetic stirrer
- Folded paper filter, quick running
- Pipettes
- Funnel
- Burette
- Potassium hexacyano ferrate (II), K4Fe(CN)6·3H2O, 15% w/v (aq)
- Zinc sulphate, ZnSO4·6H2O, 30% w/v (aq)
- Sodium hydroxide, NaOH, 0.1 N, 0.41% w/v (aq)
- Silver nitrate, AgNO3, 0.1 N, 1.6987% w/v (aq), standardized
- Potassium chromate, K2CrO4 5% w/v (aq)
- Phenolphthalein, 1% in ethanol
- Distilled or deionized water
3. Procedure
(i) Five grams of homogenized subsample is weighted into a 250 ml volumetric flask and vigorously
shaken with approximately 100 ml water.
(ii) Five millilitres of potassium hexacyano-ferrate solution and 5 ml of zinc sulphate solution are added,
the flask is shaken.
(iii) Water is added to the graduation mark.
 CXS 234-1999 149

(iv) After shaking again and allowing to stand for precipitation, the flask content is filtered through a folded
paper filter.
(v) An aliquot of the clear filtrate is transferred into an Erlenmeyer flask and two drops of phenolphthalein
are added. Sodium hydroxide is added dropwise until the aliquot takes on a faint red colour. The aliquot
then diluted with water to approximately 100 ml.
(vi) After addition of approximately 1 ml potassium chromate solution, the diluted aliquot is titrated under
constant stirring, with silver nitrate solution. End-point is indicated by a faint, but distinct, change in
colour. This faint reddish-brown colour should persist after brisk shaking.
To recognize the colour change, it is advisable to carry out the titration against a white background.
(vii) Blank titration of reagents used should be done.
(viii) End-point determination can also be made by using instruments like potentiometer or colorimeter.
4. Calculation of results
In the equation of the calculation of results the following symbols are used:
A= volume of aliquot (ml)
C= concentration of silver nitrate solution in N
V= volume of silver nitrate solution in ml used to reach end-point and corrected for blank value
W= sample weight (g)
The salt content in the sample is calculated by using the equation:
Salt concentration (%) = (V x C x 58.45 x 250 x 100) / (A x W x 1000)
Results should be reported with one figure after the decimal point.
5. Reference method
As reference method a method should be used which includes the complete ashing of the sample in a muffle
furnace at 550 °C before chloride determination according to the method described above (leaving out steps (ii)
and (iv)).
6. Comments
By using the given equation all chloride determined is calculated as sodium chloride. However it is impossible
to estimate sodium by this methodology, because other chlorides of the alkali and earth alkali elements are
present which form the counterparts of chlorides.
The presence of natural halogens other than chloride in fish and salt is negligible.
A step, in which proteins are precipitated (ii), is essential to avoid misleading results.
PART 3: DETERMINATION OF WATER CONTENT
Salted fish and dried salted fish of the Gadidae family of fishes
i) Determination of % salt saturation as required by the standard, should be in accordance to AOAC 950.46.B
(air-drying (a)).
ii) Determination of water content in the whole fish, when needed in the commercial trade of klippfish and wet
salted fish, the method of sampling the fish should be carried out according to the “Determination of water
content in whole fish by cross section method” defined in the annex to this appendix.
Salted Atlantic herring and salted sprat
Determination of water content is performed according to AOAC 950.46B (air-drying).
 CXS 234-1999 150

ANNEX TO APPENDIX VIII

DETERMINATION OF WATER CONTENT IN WHOLE FISH BY CROSS SECTION METHOD
1 PRINCIPLE
The fish is cut in sections as described in method. The sections are cut in smaller bits to a collected sample.
The water content of the collected sample is determined by drying. Examinations and experience have shown
that the water content of this collected sample is closed to the “true” water content of the fish.
2 EQUIPMENT
- Soft brush
- Basins (steel, glass, porcelain)
- Scissors
- Band saw
- Knife
- Weight, 1 g precision
- Oven, 103–105 °C
- Desiccator

3 PREPARATION OF SAMPLE
Salt particles on the surface of the fish are brushed away.
The weight of the fish is determined to 1 g accuracy.
The length of the fish is measured as the distance between the cleft in the tail and a line drawn between the
tips of the ear bones.
4 PROCEDURE
(i) The sampling of the fish is described in the enclosed figure.

A) Wet salted fish is sliced in sections by knife

B) Salted and dried salted fish is sliced in sections by band saw.

1) A section of 20 mm measured from a line drawn between the ear bones, dotted line on figure, is
cut.

2) The next cut is a 40 mm section.

3) A 2 mm section is cut from the front part of the 40 mm section and collected (see Section 7
[Comments]).

4) The next cut is a new cut of a 40 mm section.

5) A 2 mm section is cut from the front part of the 40 mm section and collected.

6) The entire fish is cut in 40 mm sections from which are cut 2 mm sections (see enclosed figure).

7) All sections of 2 mm, marked II, IV, VI, VIII in the figure, even numbers, are collected toa collected
sample.
(ii) The 2 mm sections in the collected sample are cut with scissors in smaller pieces directly in tared basins
just after the fish is cut.
(iii) The basins containing the sample are weighted.
(iv) The basins containing the samples are put in the oven at 103 °C –105 °C for drying to constant weight
(18 hours over night).
(v) The basins are taken from the oven to a desiccator and cooled.
(vi) The basins are weighted.
 CXS 234-1999 151

5. CALCULATION OF RESULTS
In the equation of the calculation of results the following symbols are used:
W1 = Weight of fish and basins before drying, g.
W2 = Weight of fish and basins after drying, g.
Ws = Weight of tared basins, g
The water content in the fish is calculated by using the equation:
Water content, g/100 g = 100*(W1-W2)/ (W1 – Ws)
The result is reported to the nearest gram, together with the length and the weight of the analysed fish.
6. CONTROL ANALYSIS OF WHOLE FISH.
The determination of water content in whole fish by cross section method appears to give the closest result
compared to water content determined by the drying of the whole fish
7. COMMENTS
Each sampled fish should be packed and sealed in a plastic bag before analysis. The samples should be
stored under chilled or refrigerated conditions from the time of sampling to the time of analysis.
The analysis must be performed as soon as possible after the fish has been sampled.
It might be difficult to cut sections of 2 mm when the fish has a water content above 50 percent, but the section
must be close to 2 mm.
To minimize the loss of water from the 2 mm sections it is important to weight the collected sample immediately
after the fish is cut in sections.
Determination should be performed at least in duplicate.
CXS 234-1999 152

FIGURE
CXS 234-1999 153

APPENDIX IX

METHOD OF ANALYSIS OF PATHOGENS IN FISH AND FISHERY PRODUCTS

PART 1: ESCHERICHIA COLI IN LIVE AND RAW BIVALVE MOLLUSCS

The ISO/TS 16649-3 – Horizontal method for the enumeration of beta-glucuronidase-positive Escherichia coli
– Part 3: Most probable number technique using 5-bromo-4-chloro-3-indolyl beta-D-glucuronide or other
validated methods in accordance with the protocol set out in the ISO 16140 or other internationally accepted
similar protocol.

PART 2: DETERMINATION OF LISTERIA MONOCYTOGENES IN SMOKED FISH, SMOKE-FLAVOURED

FISH AND SMOKE-DRIED FISH
The microbiological criteria for products in which growth of L. monocytogenes will not occur are based on the
use of the ISO 11290-2 method. Other methods that provide equivalent sensitivity, reproducibility, and
reliability can be employed if they have been appropriately validated (e.g. based on ISO 16140). The
microbiological criteria for products in which growth of L. monocytogenes can occur are based on the use of
ISO 11290-1 method. Other methods that provide equivalent sensitivity, reproducibility, and reliability can be
employed if they have been appropriately validated (e.g. based on ISO 16140).

PART 3: DETERMINATION OF CLOSTRIDIUM BOTULINUM IN SMOKED FISH, SMOKE-FLAVOURED

FISH AND SMOKE-DRIED FISH
AOAC 977.26 for the detection of C. botulinum and its toxins in foods or other scientifically equivalent validated
method. This method is not routinely performed on the product but may be used when there is a suspicion of
the presence of toxins.
 CXS 234-1999 154

APPENDIX X
DETERMINATION OF WATER CAPACITY OF CONTAINERS (CAC/RM 46)

1. SCOPE
This method applies to glass containers.
2. DEFINITION
The water capacity of a container is the volume of distilled water at 20 °C which the sealed container will hold
when completely filled.
3. PROCEDURE
3.1 Select a container which is undamaged in all respects.
3.2 Wash, dry and weigh the empty container.
3.3 Fill the container with distilled water at 20 °C to the level of the top thereof, and weigh the container thus filled.
4. CALCULATION AND EXPRESSION OF RESULTS
Subtract the weight found in Section 3.2 from the weight found in Section 3.3. The difference shall be
considered to be the weight of water required to fill the container. Results are expressed as ml of water.
 CXS 234-1999 155

APPENDIX XI

DETERMINATION OF MOISTURE IN POWDERED MILK, POWDERED CREAM AND BLEND OF

SKIMMED MILK POWDER WITH VEGETABLE FAT

TEST MOISTURE METHOD AT NORMAL PRESSURE (102 ± 2)°C IN POWDERED MILK, POWDERED
CREAM, AND BLEND OF SKIMMED MILK POWDER WITH VEGETABLE FAT

DESCRIPTION OF THE METHOD: DETERMINATION OF MOISTURE

1. SCOPE
This standard specifies a method for the determination of moisture content for all types of powdered milk,
powdered cream and mixtures of powdered skimmed milk with vegetable fat.
2. DEFINITION
The content is the mass loss determined by the procedure specified in this standard. It is expressed in
percentage by mass g/100 g.
3. PRINCIPLE
A portion of the sample is dried in an oven set at (102 ± 2) °C until constant weight and weighed to determine
the loss of mass.
4. EQUIPMENT
Common laboratory equipment and, in particular, the following.
4.1 Analytical balance, capable of weighing with a precision of 1 mg, with a minimum resolution of 0.1 mg.
4.2 Drying oven, with good ventilation, as far as possible with forced ventilation, capable of being thermostatically
maintained at (102 ± 2) °C throughout the workspace, with a temperature controller.
4.3 Desiccator, with freshly dried silica gel with hygrometric indicator or another effective desiccant.
4.4 Flat-bottomed dishes, approximately 25 mm deep, approximately 50 mm in diameter, and made of an
appropriate material (for example, glass, stainless steel, nickel, or aluminium), fitted with tight-fitting, removable
lids easily.
5. SAMPLING
It is important that the laboratory receive a truly representative sample and that it has not been damaged or
changed during transport or storage.
Sampling is not part of the method specified in this standard. A recommended sampling method is provided in
ISO 707 | IDF 50.
6. TEST SAMPLE PREPARATION
Transfer the entire sample to a dry, tightly closed container with a capacity of approximately twice the volume
of the sample. Mix thoroughly by turning and shaking the container.
7. PROCEDURE
7.1 Preparation of the dish
7.1.1 Heat the uncovered capsule and its lid (4.4) in the oven (4.2) controlled at (102 ± 2) °C, for 1 h.
7.1.2 Transfer the capped dish to the desiccator (4.3), allow it to cool to room temperature in the balance room, and
weigh (4.1) to the nearest 0.1 mg.
7.2 Test sample
7.2.1 Place 1 g-1.5 g of the prepared test sample (6) in the dish, cover with the lid and weigh to the nearest 0.1 mg.
7.3 Determination
7.3.1 Uncover the capsule and place it together with the lid in the oven (4.2), controlled at (102 ± 2) °C for 2 hrs.
7.3.2 Replace the cap, transfer the capped dish to the desiccator, allow to cool to balance room temperature, and
weigh to the nearest 0.1 mg.
 CXS 234-1999 156

7.3.3 Uncover the capsule and heat again, along with its lid, on the oven for 1 h. Then repeat operation 7.3.2.
7.3.4 Repeat this process until the difference in mass between two successive weightings does not exceed 0.5 mg.
Record the lowest mass.
8. CALCULATION AND EXPRESSION OF RESULTS
8.1 Calculation
The moisture content in the sample, expressed in g/100 g, is equal to:
moisture = (m 1 – m 2 ) x 100
(m 1 – m 0 )
where,
m 0 is the mass, in grams, of the dish and lid (Section 7.1.2)
m 1 is the mass, in grams, of the dish, lid and test sample before drying (Section 7.2.1)
m 2 is the mass, in grams, of the dish, lid and test sample after drying (Section 7.3.4)
8.2 Expression of test results
Express the sample results to two decimal places.
 CXS 234-1999 157

APPENDIX XII

STANDARD PROCEDURE FOR TOUGH STRING TEST OF CANNED AND QUICK-FROZEN GREEN
AND WAX BEANS
(CAC/RM 39-1970)
1. DEFINITION
A tough string is a string that will support the weight of 250 g for 5 seconds or longer when tested in accordance
with the procedure described below.
2. PRINCIPLE
Strings are removed from individual pods, fastened through a clamp assembly weighing 250 g, and hung so
that the string supports the entire weight. If the string supports the weight for 5 seconds or more, it is considered
a tough string.
3. APPARATUS
3.1 Weighted clamp
Use battery clamp (with teeth filed off or turned back), spring operated clothes pin or binder clip which presents
a flat clamping surface. Attach weight so that entire assembly of weight and clamp weighs 250 g. See Figure 1.
A bag containing lead pellets is convenient as a weight.
4. PROCEDURE
4.1 From the drained product select a representative sample of not less than 285 g. Record the weight of this test
sample.
4.2 Break the individual bean units and set aside those that show evidence of tough strings. Remove the strings
from the pods and retain the pod material for weighing.
4.3 Fasten the clamp assembly to one end of the string. Grasp the other end of the string with the fingers (a cloth
may be used to aid in holding the string) and lift gently.
4.4 If the string supports the 250 g assembly for at least five seconds consider the bean unit as containing tough
string. If the string breaks in less than five seconds, retest the broken parts that are 13 mm or longer to
determine if such portions are tough.
4.5 Weigh the bean units which contain tough strings.
5. CALCULATION AND EXPRESSION OF RESULTS

pods containing tough strings (g)

% m/m pods containing tough strings  x 100
test sample (g)

Figure 1: Tough string tester for green or wax beans

 CXS 234-1999 158

APPENDIX XIII

METHOD FOR DISTINGUISHING TYPE OF PEAS (CAC/RM 48-1972)

1. DEFINITION
This method is based on differentiation between starch granules of the wrinkled-seeded types and starch
granules of the smooth-seeded types.
2. REAGENTS AND MATERIALS
2.1 Compound microscope:
- 100 to 250 magnification
- phase contrast
2.2 Microscope slide and cover glass.
2.3 Spatula.
2.4 Ethanol – 95% (v/v).
2.5 Glycerine.
3. PROCEDURE
3.1 Preparing mount
3.1.1 Remove a small portion of the endosperm and place on glass slide;
3.1.2 Using a spatula grind the material with 95% (v/v) ethanol;
3.1.3 Add a drop of glycerine, place cover glass on material and examine under microscope.
3.2 Identification
3.2.1 Starch granules of the wrinkled-seeded types (garden peas, sweet) show up as clear cut, well defined,
generally spherical particles.
3.2.2 Starch granules of the smooth-seeded types (round, early, Continental) show up as an amorphous mass with
no well-defined geometric shape.
CXS 234-1999 159

APPENDIX XIV

DETERMINATION OF INTERNAL DEFECTS: DATES

Examine each date carefully for internal defects using a strong light. If the dates are pitted, open up the flesh
so that the internal cavity can be viewed. If the dates are unpitted, slit the date open so as to expose the pit,
remove the pit and examine the pit cavity.
CXS 234-1999 160

APPENDIX XV

EXTRANEOUS VEGETABLE MATTER IN DESICCATED COCONUT

The determination is carried out by spreading 100 g of the sample in a thin layer against a white background and
counting the extraneous material with the naked eye.
CXS 234-1999 161

APPENDIX XVI

DETERMINATION OF BROKEN, SLABS, DIRTY, MOULDY, DAMAGED AND IMMATURE FRUITS:

DRIED APRICOTS

Examine the fruits (sample size: 1 kg) visually and weigh the defective items. Calculate the percentage of
defects:

Weight of defective unites

x100  % defective
Total weight of sample
CXS 234-1999 162

APPENDIX XVII

DETERMINATION OF VOLUME OF FILL (BY DISPLACEMENT): PICKLED CUCUMBERS

METHOD 1
(1) This method may be used for all sub-types of pickles. Use a 4 to 8 litre size can with an overflow spout
constructed from 0.5 cm to 1 cm inside diameter metal tubing (see Figure 1). The tubing is soldered
to an opening in the side of the can 2 cm to 3 cm from the bottom and is bent upward parallel to sides.
The tube is bent over and slightly downward from the can at the upper end to form a spout about 4 cm
below the top of the can. The lower tip end of the spout is lower than the inside lower curve of the
spout (point A). The upper tip end of the spout is higher than the inside lower curve of the spout (point
A). The upper tip end of the spout is slightly shorter than the lower tip end of the spout. A brace near
the top of the can holds the tubing firmly in place. A woven wire basket made from screen wire with
about eight meshes to the inch with a handle is used for lowering the pickle ingredient into the overflow
can.
(2) Place overflow can on a level table so that overflow will discharge into a sink. Fill the overflow can with
water at room temperature (approximately 20 °C or 68 °F). Place the empty basket into the filled
overflow can.
(3) When overflow ceases, place a beaker or graduated cylinder under the spout.
(4) Remove basket and place drained pickle ingredient (at room temperature) in basket and lower slowly
into the overflow can. When overflow ceases, measure the volume of the fluid overflow. The percent
volume of pickle ingredient (volume occupied) is calculated as follows:

Overflow Volume
x100  percent volume of pickle ingredient
Total capacity (volume) of container (see Method E)

Figure 1

4 cm
Point "A"

brace
0.5 to 1 cm
inside diameter
tubing
2.5 cm

Overflow can

METHOD 2
(1) Use water to partially fill a graduated cylinder (or other technical measuring device) large enough so
that the pickle ingredient from one container may be completely submerged.
(2) Prior to adding the pickle ingredient, record the volume of water in the partially filled cylinder.
(3) Add all the drained pickle ingredient from one container so that it is entirely submerged.
(4) Measure the volume of liquid and pickle ingredient after submersion of pickle ingredient.
(5) Subtract the value in (2) from the value in (4) to obtain the pickle ingredient displacement.
(6) To determine Volume fill, calculate:

Pickle Ingredient Displacement

x100  percent volume of pickle ingredient
Total Capacity (volume) of Container (see Method E)
CXS 234-1999 163

METHOD 3
(1) Remove and collect the packing medium from the container for other quality determinations.
(2) With the pickle ingredient in the container fill it to capacity with water.
(3) Drain, collect and measure the water.

To determine ‘volume fill’, calculate:

V1  V2
percent volume of pickle ingredient 
V1
Where,
V1=Total capacity (volume) of container; and
V2=Volume of drained water from (3) above
 CXS 234-1999 164

APPENDIX XVIII
STANDARD PROCEDURE FOR THAWING OF QUICK-FROZEN FRUITS AND VEGETABLES

1. SCOPE
This thawing procedure is for the purposes of analysis and assessing the organoleptic the characteristics and
is generally applicable to all quick-frozen fruits and vegetables.
2. FIELD OF APPLICATION
2.1 Most on quick-frozen fruits and many vegetables can be examined on the basis of their organoleptic
characteristics in a thawed condition. Where a vegetable requires cooking prior to organoleptic testing the
prescribed procedure for the cooking of quick-frozen vegetables is to be followed (Appendix XIX, CAC/RM 33-
1970).
2.2 Where a particular quick-frozen fruit or vegetable requires special treatment not fully covered by this general
procedure for examination, the treatment outlined in the appropriate Codex commodity standard should be
followed.
3. DEFINITIONS
3.1 Thawing of quick-frozen fruits and vegetables for the purpose of this examination procedure, means subjecting
the product to controlled conditions of temperature until the product is sufficiently free from ice crystals so that
the individual units can be readily separated and handled.
3.2 Air thawing means thawing of the product in unopened container by exposure to air of an ambient temperature
in free or forced ventilation.
3.3 Water thawing by indirect contact, means thawing of the product in a tightly sealed container by immersion in
water, stationary or flowing, at a temperature not exceeding 30 oC.
3.4 Water thawing by direct contact, means thawing of the unpacked product by immersion in water at a
temperature not exceeding 30 oC. (This method is applicable only to some vegetables).
4. PRINCIPLE OF METHODS
By rapidly thawing quick-frozen products under controlled conditions, the quality factors of the original product
retained by the quick-freezing process are preserved to a high degree.
For the purpose of this examination procedure there are two general methods for thawing quick-frozen fruits
and vegetables: air thawing and water thawing, Water thawing is faster and in some instances more desirable
than air thawing, some quick-frozen commodities, especially those where the product consists of small
individual units surrounded, by air, thaw much faster than others, Through experience the analyst will learn to
judge the best procedure and time requirement for adequate thawing for each commodity.
5. APPARATUS
5.1 Electric fan (optional), for forced ventilation air thawing.
5.2 Water bath with thermostat and circulation pump, for indirect or direct water thawing.
5.3 Plastic bags or other suitable watertight and closable container, for samples to be subjected to water thawing.
5.4 Clamps or weights, to prevent agitation of package in water bath during thawing.
5.5 Screen, to remove excess water after water thawing by direct contact.
5.6 Tray, on which the product is placed after removal of excess water when thawed by direct contact with water.
6. SAMPLES
The entire package or sample unit is used intact, except that in the case of bulk or industrial size containers a
representative sample of 1–2 kg is adequate for testing and organoleptic examination.
7. PROCEDURE
For the rapid thawing of quick-frozen products contained in consumer-size packages, bulk or industrial
packages and subsamples of these in suitable containers, one of the following methods should be used:
 CXS 234-1999 165

7.1 Air thawing

Thaw in unopened containers at ambient temperature. To hasten the thawing process forced air ventilation
may be applied and the packages may be separated from each other.
7.2 Water thawing by indirect contact
Products packed in tightly sealed containers may be thawed by immersion of the container in water at a
temperature not exceeding 30 °C, e.g. a water bath with thermostat and circulation pump.
7.3 Water thawing b direct contact (applicable only to some vegetables)
The vegetable is removed from the pack and thawed by immersion in water at a temperature not exceeding
30 °C. As soon as the product is thawed sufficiently to permit easy separation of the individual units, it is
drained on a suitable screen to remove excess water and placed on a tray for final air thawing and examination.
8. NOTES ON PROCEDURE
8.1 Selection of thawing method
8.1.1 Certain quick-frozen vegetables should not be subjected to water thawing by direct contact in order to prevent
leaching of soluble solids or product material.
8.1.2 If there is an indication of off flavours or off odours in the quick-frozen product when the packages are opened,
water thawing by direct contact is not to be used as a preparatory step to cooking, as the off flavour or off
odour may be partially removed during such thawing. Such suspect samples are to be placed in a cooking
receptacle while still frozen.
8.2 Prevention of damage
Extreme care should be taken during the thawing process in order that the product is not damaged or exposed
to abuse that will alter or degrade the true characteristics of the product. Quick-frozen fruits are more
susceptible to abuse during thawing than quick-frozen vegetables. Some fruits, especially light-coloured fruits,
oxidize quite readily and should be examined for colour before thawing is completed. Also, some fruits show
a breakdown in texture or “bleed” when thawed more than necessary. Consequently, rapid thawing under
controlled conditions is most desirable in preparing the product for laboratory examination.
9. TEST REPORT
The identity of the sample and the thawing procedure used should be recorded.
10. ADDITIONAL NOTES
10.1 Quick-frozen corn (maize) or products containing corn should always be air thawed or water thawed by indirect
contact to avoid leaching of soluble solids or product material.
10.2 Quick-frozen peaches and apricots (light-coloured fruits) and red cherries oxidize quite readily and should be
examined while some ice crystals remain in the product.
 CXS 234-1999 166

APPENDIX XIX

STANDARD PROCEDURE FOR COOKING OF QUICK-FROZEN VEGETABLES

(CAC/RM 33-1970)
1. SCOPE
This cooking procedure is for the purposes of analysis and assessing the organoleptic characteristics and is
generally applicable to all quick-frozen vegetables.
2. FIELD OF APPLICATION
2.1 The cooking procedure described below applies to those quick-frozen vegetables which are normally cooked
prior to consumption for the proper evaluation of such organoleptic quality factors as texture, tenderness,
maturity or flavour.
2.2 Where a particular quick-frozen vegetable requires a special cooking procedure not fully covered by this
general procedure for examination, the method outlined in the appropriate Codex commodity standard shall
be followed.
3. DEFINITION
Cooking of vegetables, for the purpose of this examination procedure, means to prepare, food for the table by
subjecting quick-frozen vegetables to an appropriate standard (cooking) procedure by partial or whole
immersion of the product in boiling water for a specified time.
4. PRINCIPLE OF METHOD
By heating the quick-frozen vegetable, through partial or whole immersion in water at boiling temperature for
such a period of time as to undergo specific changes of conditions.
5. APPARATUS
5.1 Two-litre saucepan with cover.
5.2 Hot plate or gas fire.
5.3 Tray on which product is placed after cooking for cooling and presentation.
5.4 Graduated cylinder or similar measuring device for water.
6. SAMPLES
Generally, a separate set of samples for cooking purposes only need not be taken. Ordinarily part of the
contents of a larger retail size package or part of a sample of a bulk container, used for testing other product
characteristics can be used for the cooking procedure. Care should be taken, however, that the portion used
for cooking is not treated differently from the normal procedure, e.g. thawed prior to cooking whereas the
product would usually be put in boiling water while still in the frozen state.
 CXS 234-1999 167

APPENDIX XX

DETERMINATION OF THE ALCOHOL-INSOLUBLE SOLIDS CONTENT OF QUICK-FROZEN PEAS

1. PRINCIPLE OF THE METHOD

The alcohol-insoluble solids in peas consist mainly of insoluble carbohydrates (starch) and protein. A weighed
quantity of the sample is boiled with slightly diluted alcohol. The solids are washed with alcohol until the filtrate
is clear. The alcohol-insoluble solids are dried and weighed. The percentage by mass present is used as a
guide to maturity.
2. REAGENTS
2.1 Ethanol (95 percent) or denaturated ethanol.
Ethanol denaturated with 5 percent v/v methanol.
2.2 Diluted ethanol or diluted denaturated ethanol 80 percent v/v.
Dilute 8 parts by volume of reagent under Section 2.1 to 9.5 parts by volume with H2O.
3. APPARATUS
3.1 Analytical balance.
3.2 Beaker, 600 ml, if sample is boiled or 250 ml (standard taper ground-glass joint) flask with reflux condenser if
refluxed.
3.3 Buchner funnel.
3.4 Drying dish with lid, flat bottomed.
3.5 Hot plates or boiling water bath for refluxing or boiling.
3.6 Clamps or weights to prevent agitation of package in water bath during thawing.
3.7 Desiccator with active desiccant.
3.8 Drying oven, well-ventilated and thermostatically controlled and adjusted to operate at 100 ± 2 C.
3.9 Filter paper, Whatman No. 1 or equivalent.
3.10 Macerator or blender.
3.11 Plastics bag of sufficient capacity to hold the entire sample for thawing.
3.12 “Policemen” on glass rods, bent so as to facilitate cleaning flask or beaker.
3.13 Water bath, with continuous flow at room temperature or regulated at room temperature for thawing.
4. PREPARATION OF TEST SAMPLE
Place frozen peas or frozen peas with sauce in plastic bag and tie off. Immerse sample in water bath with
continuous flow at room temperature or regulated at room temperature. Avoid agitation of package during
thawing by using clamps or weights if necessary. When completely thawed, remove package from bath. Blot
off adhering water from the plastic bag. Transfer the peas from container to a sieve, the meshes of which are
made by so weaving wire as to from square openings of 2.8 mm by 2.8 mm. If sauce is present, wash with
gentle spray of water at room temperature until the sauce is removed. Without shifting the peas, incline the
sieve as to facilitate drainage, and drain for two minutes. Wipe the bottom of the sieve. Weight 250 g peas into
blender, add 250 ml distilled water and macerate to a smooth paste. If there is less than 250 g sample, use
the entire sample of peas with an equivalent quantity by mass of distilled water and macerate to a smooth
paste.
 CXS 234-1999 168

5. PROCEDURE
5.1 Dry a filter paper in flat bottomed dish, lid off, for two hours at 100 ± 2 C. Cover dish, cool in a desiccator and
weigh accurately. (The filter paper should be larger than the base of the funnel and folded at the circumference
to facilitate subsequent removal without loss of solids).
5.2 Weight 20 g ± 0.01 g paste into a 250 ml ground-joint flask, add 120 ml denaturated ethanol or ethanol, and
swirl to mix. Reflux on a steam or water bath for 30 minutes.
If boiling rather than refluxing is preferred, weight 40 g ± 0.01 g paste into a 600 ml beaker. Add 240 ml
denaturated ethanol or ethanol, stir and cover beaker. Bring solution in the beaker to a boil and simmer slowly
for 30 minutes on a hot plate.
Immediately filter with suction on a Buchner funnel through the dried and weighed filter paper. Decant most of
the supernatant liquid through the filter paper. Wash the solids in the flask or beaker without delay, with small
portions of 80 percent denaturated ethanol or 80 percent ethanol until the washings are colourless, allow solids
to become dry during the washing. Transfer solids to the filter paper, spreading the solids evenly.
5.3 Remove the filter paper containing the residue from the funnel, transfer to the dish used in preparing the filter
paper and dry uncovered in an air over for two hours at 100 ± 2 C. Cover the dish, cool in a desiccator and
weigh accurately. The weight of the dry residue is the difference between the weight under Section 5.1 and
this final weight.
6. CALCULATION AND EXPRESSION OF RESULTS
Calculate the alcohol-insoluble solids content of the sample by means of the following formula:
6.1 If 20 g sample is refluxed:
Alcohol-insoluble solids content (% m/m) = 10 M
Where:
M = the mass in g of dry residue (see Section 5.3)
6.2 If 40 g sample is refluxed:
Alcohol-insoluble solids content (% m/m) = 5 M
Where:
M = the mass in g of dry residue (see Section 5.3)
7. REPEATABILITY OF RESULTS
The difference between results of duplicate determination (results obtained simultaneously or in rapid
succession by the same analyst) should not exceed 0.6 g alcohol-insoluble solids for 100 g of the product.
8. EXPRESSION OF RESULTS
Results are expressed as g alcohol-insoluble solids per 100 g of the product (% m/m).
 CXS 234-1999 169

APPENDIX XXI

DETERMINATION OF SALT-FREE DRY MATTER (QUICK-FROZEN SPINACH)

PROCEDURE

1. Determine the total dry matter of the product by drying over sand for 4 hours at 105 °C.

2. From the value obtained in (1) deduct the amount of salt (NaCl) determine by either (a) electrometric titration
using a pH metre with a silver electrode; or (b) direct titration with AgNO 3. Express the result, after deducting
salt from total dry matter, as salt-free dry matter.
CXS 234-1999 170

APPENDIX XXII

DETERMINATION OF PEROXIDE VALUE IN COOKED RICE WRAPPED IN PLANT LEAVES:

EXTRACTION OF OILS FROM THE PRODUCT

Apparatus
(a) Rotary evaporator
(b) Water bath
Extraction
Remove the product package and plant leaves etc. take out the edible part of the representative sample; crush
it and put it in a homogenizer or glass mortar and grind it continuously to make the sample fully mashed and
mixed well and then put it in the wide-mouth bottle and add 2 to 3 times the sample volume of petroleum ether
(boiling range: 30 °C–60 °C). After fully mixing, stopper the bottle and leave for more than 12 hours. Filter all
the solution with a funnel filled with anhydrous sodium sulphate into a round-bottom flask. Rinse the residue
in the wide-mouth bottle with petroleum ether. Repeat the filtration once with a new anhydrous sodium sulphate
funnel, if the filtrate is not clear enough. Evaporate the petroleum ether in the round-bottom flask under reduced
pressure on a rotary evaporator at below 40 °C, and the residue is the test sample. A sufficient number of
representative samples should be selected to ensure that not less than 8 grams of the test sample can be
obtained. The test sample should be tested as soon as possible.
 CXS 234-1999 171

APPENDIX XXIII

PART A – IDENTIFICATION OF SCOPOLETIN IN FERMENTED NONI FRUIT JUICE

1. PREPARATION OF SAMPLES
Noni fruit juice is filtered through a 0.45 μm membrane filter and then purified by solid-phase extraction
(SPE) with Waters OASIS® HLB 6cc 200 mg extraction cartridges (or similar solid-phase extraction
cartridge), after first equilibrating with methanol (5 mL) followed by deionized water (5 mL). The filtered
juice samples (3 mL) are then loaded onto the equilibrated cartridge and washed with 5 percent methanol
(MeOH) in deionized water (5 mL). The cartridges are allowed to dry under flow of air for 5 mins and then,
eluted with MeOH (3 mL). The MeOH eluate is retained for thin layer chromatography (TLC) analysis. The
SPE flow rates of equilibration, wash and elution solvents through the cartridge is approximately 1 drop per
second.
2. PREPARATION OF REFERENCE STANDARD
2.1 A reference standard is prepared by dissolving 0.1 mg scopoletin in 1 millilitre of methanol.
2.2 Alternately, certified Morinda citrifolia reference plant material may be prepared in the same manner as the
samples to be analysed. The certified Morinda citrifolia reference material should be from the same part of
the plant as the samples to be analysed.
3. IDENTIFICATION
3.1 Thin layer chromatography
Spot 5 microlitres of sample solutions and reference standard solution on a silica gel 60 F254 TLC plate.
After spotting the plates are dried at 110 °C for 15 minutes in a drying oven. Develop the plate with a mobile
phase of dichloromethane: methanol (19:1, v/v). View bright fluorescent blue colours on developed plate
under UV lamp, 365 nm. Identify scopoletin in samples by comparing Rf values and colours to the standard.

PART B– IDENTIFICATION OF DEACETYLASPERULOSIDIC ACID IN FERMENTED NONI FRUIT JUICE

1. PREPARATION OF SAMPLES
Noni fruit juice is filtered through a 0.45 μm membrane filter and diluted 1:1 with MeOH.
2. PREPARATION OF REFERENCE STANDARD
2.1 A reference standard is prepared by dissolving 1 mg deacetylasperulosidic acid in 1 millilitre of methanol.
2.2 Alternately, certified Morinda citrifolia reference plant material may be prepared in the same manner as the
samples to be analysed. The certified Morinda citrifolia reference material should be from the same part of the
plant as the samples to be analysed.
3. PREPARATION OF p-ANISALDEHYDE SOLUTION
Anisaldehyde solution was prepared by dissolving 2 g of p-anisaldehyde in 96 mL of ethanol with stirring. The
solution was then acidified through dropwise addition of concentrated sulfuric acid (4 mL).
4. IDENTIFICATION
4.1 THIN LAYER CHROMATOGRAPHY
Spot 5 microlitres of sample solutions and reference standard solution on a silica gel 60 F254 TLC plate,
previously dried at 110 °C for 15 minutes in a drying oven. After spotting samples are again dried at 110 oC or
through application of heat via a heat gun for a period of 8-10 seconds. The TLC plates are developed with a
mobile phase of dichloromethane: methanol: water (13:6:1, v/v/v). Upon completion of elution, the plate is air
dried and developed by spraying with 2% anisaldehyde/4% sulfuric acid in ethanol (EtOH) solution and then
heat in oven at 110 °C for 1–5 minutes to reveal and maximize the blue colour. Identify deacetylasperulosidic
in samples by comparing spot Rf values and colour with reference standard solution on same TLC plate.
CXS 234-1999 172

APPENDIX XXIV

METHOD FOR THE EXTRACTION OF OIL (LAVER PRODUCTS)

Weigh 50 g of test sample into a 1 000 ml Erlenmeyer flask.

Add 500 ml of petroleum ether to the flask followed by replacing air in the flask by N2 gas

Put a stopper on the flask and let it stand for 2 hours.

Decant the extracted solution (A) through a filter paper, on which Na 2SO4 is mounted to remove moisture, on
a funnel into a 1 000 ml round flask-flat bottom.

Add an additional 250 ml of petroleum ether to the residue in the Erlenmeyer flask and decant the extracted
solution (B) into the round flask-flat bottom again as done previously.

Evaporate the whole extracted solution (mixture of solutions A and B) on the rotary evaporator in vacuum less
than 40 oC.
CXS 234-1999 173

NOTES
1 FAO and WHO. 1981. Standard for natural mineral waters. Codex Alimentarius Standard, No. CXS 108-1981. Codex
Alimentarius Commission. Rome.
2 FAO and WHO. 1991. Standard for Canned Shrimps or Prawns. Codex Alimentarius Standard, No. CXS 37-1991.

Codex Alimentarius Commission. Rome.

3 FAO and WHO. 1999. Guidelines for the sensory evaluation of fish and shellfish in laboratories. Codex Alimentarius

Guideline, No. CXG 31-1999. Codex Alimentarius Commission. Rome.

4 FAO and WHO. 1981. Standard for quick-frozen shrimps or prawns. Codex Alimentarius Standard, No. CXS 92-1981.

Codex Alimentarius Commission. Rome.

5 FAO and WHO. 1981. Standard for quick-frozen lobsters. Codex Alimentarius Standard, No. CXS 95-1981. Codex

Alimentarius Commission. Rome.

6 FAO and WHO. 1995. Standard for quick-frozen raw squid. Codex Alimentarius Standard, No. CXS 191-1995. Codex

Alimentarius Commission. Rome.

7 FAO and WHO. 2014. Standard for fresh and quick-frozen raw scallop products. Codex Alimentarius Standard, No.

CXS 315-2014. Codex Alimentarius Commission. Rome.

8 FAO and WHO. 2024. Standard for salted Atlantic herring and salted sprat. Codex Alimentarius Standard, No.

CXS 244-2004. Codex Alimentarius Commission. Rome.

9 FAO and WHO. 2010. Standard for sturgeon caviar. Codex Alimentarius Standard, No. CXS 291-2010. Codex

Alimentarius Commission. Rome.

10 FAO and WHO. 2008. Standard for live and raw bivalve molluscs. Codex Alimentarius Standard, No. CXS 292-2008.

Codex Alimentarius Commission. Rome.

11 FAO and WHO. 2013. Standard for live abalone and for raw fresh chilled or frozen abalone for direct consumption or

for further processing. Codex Alimentarius Standard, No. CXS 312-2013. Codex Alimentarius Commission. Rome.
12 FAO and WHO. 1989. Standard for quick-frozen fish sticks (fish fingers), fish portions and fish fillets - breaded or in

batter. Codex Alimentarius Standard, No. CXS 166-1989. Codex Alimentarius Commission. Rome.