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This document defines terms relating to animal feeding stuffs. NOTE It includes the most common and frequently used terms in the field of animal feeding stuffs.

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This document specifies minimum specifications for maize (Zea mays L.) intended for human consumption and which is the subject of international trade.

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ISO 12099:2017 gives guidelines for the determination by near infrared spectroscopy of constituents such as moisture, fat, protein, starch and crude fibre and parameters such as digestibility in animal feeding stuffs, cereals and milled cereal products. The determinations are based on spectrometric measurement in the near infrared spectral region.

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ISO 13904:2016 specifies a method for determination of the total and free tryptophan (Trp) content in feeding stuffs (e.g. complete and complementary feeds, supplementary feeds, raw materials, ingredients, and concentrates) and determination of free tryptophan in commercial pure substances and premixtures containing more than 2 % of tryptophan. It does not distinguish between D- and L-forms.

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ISO 6495-1:2015 specifies a method for the determination of water-soluble chloride content, expressed as sodium chloride, of animal feeding stuffs. This method is applicable to animal feeding stuffs containing water-soluble chloride content, expressed as sodium chloride, ≥0,05 %.

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ISO 17180:2013 specifies a method for the quantitative determination of free (non-protein-bound) lysine, methionine, and threonine in commercial products and premixtures containing more than about 10 % mass fraction of the respective amino acid. It does not distinguish between d- and l-forms.

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ISO 6498:2012 specifies guidelines for the preparation of test samples from laboratory samples of animal feeding stuffs, including pet foods. The guidelines are overruled by special instructions and regulations for sample preparation demanded by specific analysis methods.

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ISO 30024:2009 specifies the determination of phytase activity in feed samples. The method does not distinguish between phytase added as a feed additive and endogenous phytase already present in the feed materials. The method cannot be used to evaluate or compare the in vivo efficacy of the phytase product. It is not a predictive method of the in vivo efficacy of phytases present on the market as they can develop different in vivo efficacy per unit of activity. The method is suitable and validated exclusively for the determination of phytase activity and exclusively in complete feeds.

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ISO 5983-2:2009 specifies a method for the determination of nitrogen content of animal feeding stuffs according to the Kjeldahl method, and a method for the calculation of the crude protein content. It is suitable for use as a semi-micro rapid routine method using block digestion, copper catalyst, and steam distillation into boric acid. The method is applicable to the determination of greater than 0,5 % mass fraction Kjeldahl nitrogen in animal feeding stuffs, pet foods, and their raw materials. The method does not measure oxidized forms of nitrogen nor heterocyclic nitrogen compounds. The method does not distinguish between protein nitrogen and non-protein nitrogen.

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ISO 13906:2008 specifies a method for the determination of acid detergent fibre (ADF) insoluble residue and acid detergent lignin (ADL) in all types of animal feeding stuffs. The limit of determination is 1 % mass fraction for ADF and 1,5 % mass fraction for ADL. A gravimetric routine and reference method is used.

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ISO 17372:2007 is applicable to the analysis of zearalenone in animal feed and feed ingredients, including barley, corn, oats, rye, wheat, soybean meal, canola (rapeseed) meal, corn gluten, dried distillers' grains, lentils, and sugar beet pulp. The limit of quantification is 0,05 mg/kg (50 µg/kg). A lower limit of quantification may be achievable subject to appropriate validation being conducted by the user laboratory.

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ISO 17375:2006 specifies a method for the determination of aflatoxin B1 in animal feeding stuffs using high-performance liquid chromatography with post-column derivatization. It is applicable to animal feeding stuffs with a fat content of up to 50 %. The limit of quantification of this method has been demonstrated to be better than 0,5 g/kg for aflatoxin B1 for a signal-to-noise ratio of 6.

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ISO 16472:2006 specifies methods for the determination of amylase-treated neutral detergent insoluble fibrous residue content in all types of animal feed. It includes a gravimetric routine method and a reference method.

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ISO 14183:2005 specifies a high-performance liquid chromatographic (HPLC) method for the determination of the monensin, narasin and salinomycin contents of animal feeding stuffs, supplements (dry and liquid) and mineral premixtures. The method is not applicable to drug premixes (pharmaceutical products). Lasalocid and semduramicin cannot be determined by this method. The limit of quantitation is approximately 1 mg/kg, 2 mg/kg and 2 mg/kg for monensin, salinomycin and narasin, respectively. A lower limit of quantitation can be achievable but this is to be validated by the user.

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ISO 5983-1:2005 specifies a method for the determination of the nitrogen content of animal feeding stuffs by the Kjeldahl process, and a method for the calculation of the crude protein content. The method does not measure oxidized forms of nitrogen or heterocyclic nitrogen compounds. This method does not distinguish between protein nitrogen and non-protein nitrogen. If it is important to determine the content of non-protein nitrogen, an appropriate method should be used.

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ISO 13903:2005 describes the determination of free (synthetic and natural) and total (peptide-bound and free) amino acids in feeding stuffs, using an amino acid analyser or HPLC equipment. It is applicable to the following amino acids: sum of cystine and cysteine; methionine; lysine; threonine; alanine; arginine; aspartic acid; glutamic acid; glycine; histidine; isoleucine; leucine; phenylalanine; proline; serine; tyrosine; valine. The method does not distinguish between the salts of amino acids, nor does it differentiate between D and L forms of amino acids. It is not valid for the determination of tryptophan or hydroxy analogues of amino acids. Limits of quantification depend on the chromatographic equipment, but levels as low as: 0,3 g/kg total lysine; 0,25 g/kg total methionine; 0,35 g/kg total cystine plus cysteine; 0,2 g/kg total threonine; 0,035g/kg free lysine; 0,035g/kg free methionine; and 0,03g/kg free threonine can typically be analysed.

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ISO 15914:2004 specifies a method for the enzymatic determination of the total starch content of animal feeding stuffs and raw materials for animal feeding stuffs. The method is also applicable to the determination of starch in starch. It is important that in the sample matrix no components are present which contribute to the measured extinction at 340 nm. The analytical range of the method is 40 g/kg to 1 000 g/kg starch. The standard procedure is applicable to the range 200 g/kg to 1000 g/kg. For the lower range, 40 g/kg to 200 g/kg, another dilution procedure for the standard glucose solution and samples can be used.

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ISO 6497:2002 specifies methods of sampling animal feeding stuffs, including fish feed, for quality control for commercial, technical and legal purposes. It is not applicable to pet foods. Nor are the methods intended for sampling for the purpose of microbiological examination. Conditions of, and requirements for, sampling are specified separately for feeding stuffs of different physical natures. For certain categories of animal feeding stuff, specific methods of sampling are specified in other International Standards. A list of these can be found in the bibliography. When sampling the products specified, it is these methods which shall be used. Methods of sampling for the determination of substances likely to be non-uniformly distributed are described in an annex.

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ISO/TS 17764-2:2002 specifies the application of gas chromatography with capillary columns and flame ionization detection for the determination of the quantitative content of fatty acids in a fat by making use of the methyl esters of the fatty acids obtained in accordance with the method specified in ISO/TS 17764-1:2002. ISO/TS 17764-2:2002 is applicable to the investigation of animal and vegetable fats, oils and fatty acid mixtures for incorporation in animal feeding stuffs and fat extracts of animal feeding stuffs and raw materials for compound animal feeds, including fats and fatty acid mixtures containing butyric acid. This method is not applicable to polymerized fatty acids.

  • Technical specification
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ISO 5985:2002 specifies two procedures for animal feeding stuffs for the determination of the ash which is insoluble in hydrochloric acid. The applicable procedure depends on the nature of the sample. Procedure A is applicable to simple organic animal feeding stuffs and to compound feeding stuffs, except those mentioned under procedure B. Procedure B is applicable to minerals, mineral mixtures and compound feeding stuffs of which the ash insoluble in hydrochloric acid exceeds 1 % (mass fraction), as determined by procedure A.

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ISO 5984:2002 specifies a method for the determination of crude ash of animal feeding stuffs.

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ISO/TS 17764-1:2002 specifies two methods for preparing the methyl esters of fatty acids of animal and vegetable fats, oils and fatty acid mixtures for raw materials for compound animal feeds, and fatty acids originating from fat extracts of animal feeding stuffs, including fats and fatty acid mixtures containing butyric acid. The general method, the boron trifluoride (BF3) method is concerned with the preparation of methyl esters of fatty acids with six or more C atoms, originating from fats, oils and free fatty acids. The KOH/HCl method is concerned with the preparation of methyl esters of fatty acids with four or more C atoms. This method can also be used for the quantitative determination of fatty acids with a chain length shorter than 10 C atoms in free fatty acid mixtures. The methyl esters produced can be used for gas-liquid chromatography (GLC). Unsaponifiable materials are not removed and can, when present in considerable amounts, interfere with the chromatographic analysis. ISO/TS 17764-2:2002 describes the application of gas chromatography with capillary columns and flame ionization detection for the determination of the content of fatty acids in a fat by making use of the methyl esters of the fatty acids obtained in accordance with the methods specified in ISO/TS 17764-1:2002.

  • Technical specification
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This International Standard specifies a qualitative method for the determination of zearalenone in animal feeding stuffs and, in particular, in maize. This method is for screening purposes only. The limit of determination of zearalenone is approximately 50 μg/kg . NOTE Although sorghum gives interfering fluorescent spots identical to those of zearalenone, the method is still applicable to this feed because Rf the values are different after development of the chromatogram in the second direction. These spots are not developed by the specified confirmation technique.

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This International Standard specifies a method for the determination of castor oil seed (Ricinus communis) husks in single and compound animal feeding stuffs and, in particular, in oilseed residues. The limits of detection is 5 mg/kg. The method requires final microscopical identification of the isolated husks. This last phase therefore requires a specialist who has had sufficient experience in this type of identification and who is experienced in microscopic techniques.

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This International Standard specifies a method for the determination of the trypsin inhibitor activity (TIA) of soya products. This trypsin inhibitor activity is indicative of the degree of toasting of these products. The detection limit of the method is 0,5 mg/g.

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This International Standard specifies a high-performance liquid chromatographic (HPLC) method for the determination of the carbadox content in premixtures and animal feeding stuffs. The method is applicable to animal feeding stuffs with a mass fraction of carbadox of 0,5 mg/kg (limit of quantification) to 100 mg/kg, and to premixtures with a mass fraction of carbadox up to 10 %. The lower limit of detection is 0,1 mg/kg. NOTE 1 For animal feeding stuffs the mass fraction of carbadox is expressed in milligrams per kilogram, and for premixtures as a percentage by mass. NOTE 2 Carbadox is a chemotherapeuticum belonging to the quinoxaline group. Carbadox is used as a growth-promoting feed additive for piglets.

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This International Standard specifies a method for the determination of the vitamin E (DL-_-tocopherol) content of animal feeding stuffs and pet foods using high performance liquid chromatography.

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Cette norme internationale prescrit une méthode de détermination de la teneur en cellulose brute par filtration intermédiaire. Elle décrit un mode opératoire manuel et un mode opératoire semi-automatique. Cette méthode est applicable aux aliments des animaux ayant une teneur en cellulose brute supérieure à 10 g/kg. NOTE Pour des aliments pour animaux contenant de la cellulose brute inférieure ou égale à 10 g/kg, la méthode décrite dans l'ISO 6541 doit être utilisée. Cette Norme internationale est également applicable aux céréales et aux légumes.»

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This International Standard specifies a gas chromatographic method for the determination of residues of organochlorine pesticides in animal feeding stuffs. The method is applicable to animal feeding stuffs containing residues of one or more of the following organochlorine pesticides and some of their isomers and degradation products: aldrin, op'-DDE, pp'-DDE, op'-DDT, pp'-DDT, dieldrin, endosulfan, endrin, HCB, _-HCH (_-BHC), _-HCH (_-BHC), _-HCH (_-BHC), _-HCH (_-BHC), heptachlor, heptachlor epoxide, op'-TDE (op'-DDD), pp'-TDE (pp'-DDD) and methoxychlor. The lower determination limit for these organochlorine pesticides is 0,005 _g/g. However, the lower determination limit is 0,01 _g/g for op'-DDT and pp'-DDT, and 0,05 _g/g for methoxychlor.

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This International Standard specifies a calibration method and a standard addition method for the determination of potassium and sodium contents of animal feeding stuffs by flame-emission spectrometry. The standard addition method applies if it is not known whether the emissions measured for potassium or sodium are subject to matrix effects. The limit of determination for potassium and sodium is 0,04 g/kg. The calibration method applies if such matrix effects have been shown to be absent.

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This International Standard specifies a method for the determination of the fat content of animal feeding stuffs. The method is applicable to animal feeding stuffs except oilseeds and oilseed residues. For the purposes of this method, the following two categories of animal feeding stuffs are distinguished. Samples of products in category B need a hydrolysis step prior to extraction. Category B: straight feeds of animal origin including milk products; straight feeds of vegetable origin from which fats cannot be extracted without prior hydrolysis; in particular: gluten, yeast, soya and potato proteins, and heat-treated feeds; compound feeds containing the preceding products in such quantities that at least 20 % of the fat content stems from these products. Category A: animal feeding stuffs not mentioned under category B. NOTE For oilseed residues, a method for the determination of the "oil content" by hexane extraction is specified in ISO 734-1 [2], whereas a method for the determination of the "oil content" by diethyl ether extraction is specified in ISO 736 [3]. For oilseeds, a method for the determination of the "oil content" by hexane extraction is specified in ISO 659 [1].

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This International Standard specifies a method for the determination of the moisture and other volatile matter content of animal feeding stuffs. The method is applicable to animal feeding stuffs with the exception of: a) milk products; b) mineral substances; c) mixtures containing a considerable amount of milk products or mineral substances, for example milk replacers; d) animal feeding stuffs containing humectants (e.g. propylene glycol); e) the following simple animal feeding stuffs: animal and vegetable fats and oils (for which method A specified in ISO 662 [1] is applicable); oilseeds (for which a method is specified in ISO 665 [2]); oilseed residues (for which a method is specified in ISO 771 [3]); cereals, except maize, and cereal products (for which a method is specified in ISO 712 [4]); maize (for which the reference method specified in ISO 6540 [5] is applicable)

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This International Standard specifies a high-performance liquid chromatographic (HPLC) method for the determination of the furazolidone content of premixtures and animal feeding stuffs. The method is applicable to animal feeding stuffs with a furazolidone content of 25 mg/kg to 5 000 mg/kg and to premixtures with a mass fraction of furazolidone of up to 20 % [formerly written as 20 % (m/m)]. NOTE 1 For animal feeding stuffs, the furazolidone content is expressed in milligrams per kilogram; for premixtures, as a mass fraction in percent [% (m/m)]. NOTE 2 Furazolidone is a chemotherapeuticum belonging to the group of nitrofuranes. Nitrofuranes are bacteriostatic or bactericidal against Gram-positive and Gram-negative microorganisms and against some moulds and protozoa.

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This International Standard specifies a high-performance liquid chromatographic (HPLC) method for the determination of aflatoxin B1 content of animal feeding stuffs including those containing citrus pulp. The lower limit of determination is 1 mg/kg. NOTE 1 This International Standard may be applicable for the determination of the aflatoxin B1 content of a number of raw materials and straight feeding stuffs such as corn gluten, groundnut, palm kernel, copra, citrus pulp, tapioca, soya bean, rice bran, pollard, rape seed, niger seed and cotton seed (see references [1] and [2]). These materials were, however, not included in the collaborative testing of the method. NOTE 2 This International Standard may also be applicable for the determination of the content of the sum of the aflatoxins B1, B2, G1 and G2. However, the method has not been validated for this parameter by collaborative testing.

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This International Standard specifies a spectrometric method for the determination of the phosphorus content of animal feeding stuffs. The method is applicable to animal feeding stuffs with a phosphorus content less than 50 g/kg. It is particularly appropriate for the analysis of products with low phosphorus content. For products with higher phosphorus content, application of a gravimetric method is advised, using for instance quinoline phosphomolybdate.

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Specifies a spectrometric method for the determination of the urea content. Gives a normative reference to ISO 6498:1983. Defines the term urea content. Describes principle, reagents, apparatus, sampling, preparation of the test sample, procedure, expression of results and test report.

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The method is applicable to cottonseed and cottonseed meals and cakes. The method consists in extracting of the compound in the presence of 3-aminopropane-1-ol, either with a mexture of 2-propanol and hexane for the determination of free gossypol or with dimethylformamide for the determination of total gossypol. Conversion to gossypol-dianiline using aniline, and measurement of the absorbance at the wavelength of maximum absorbance (between 435 and 445 nm).

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The method is applicable to products having contents greater than 1 g/kg. It consists in ashing of a test portion, treatment of the ash with hydrochloric acid and precipitation of the calcium as calcium oxalate, dissolution of the precipitation in sulfuric acid and titration with standard volumetric potassium permanganate solution of the oxalic acid formed.

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The method consists in hydrolysing with hydrochloric acid of a ground test portion, dollowed by separation of the total lysine by column chromatography and determination by spectrometry at 570 nm. Reaction of a second ground test portion with an ethanolic solution of 2,4-dinitrofluorobenzene in an alkaline medium, purification, then hydrolysis with hydrochloric acid, separation of the non-available lysine by column chromatography, and determination by spectrometry at 570 nm.

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ISO 12099:2010 gives guidelines for the determination by near infrared spectroscopy of constituents such as moisture, fat, protein, starch, and crude fibre as well as parameters such as digestibility in animal feeding stuffs, cereals and milled cereal products. The determinations are based on spectrometric measurement in the near infrared spectral region.

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