70/373/EC - Council Directive 70/373/EEC of 20 July 1970 on the introduction of Community methods of sampling and analysis for the official control of feeding-stuffs

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.

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.

This European Standard specifies a method for the determination of decoquinate. This high-performance liquid chromatographic (HPLC) method with a fluorescence detection is applicable to the quantification of decoquinate content in complete and complementary compound feeds, medicated feeds, semi-liquid feeds, premixtures and feed additives.
The method was fully validated from LOQ to 60 000 mg/kg on different matrices during an international collaborative study [11], especially on complete compound feeds for poultry, at trace contamination level of 3 mg/kg and at European authorized level of 20 mg/kg to 40 mg/kg [12].
The limit of detection is between 0,1 mg/kg and 0,3 mg/kg and the limit of quantification is around 0,5 mg/kg. These limits were validated during the collaborative study [11], from results on the blank feed. Lower limits of detection or quantification could be reached but a single laboratory validation is then requested.

This European Standard specifies an Ion-Selective Electrode method (ISE) after hydrochloric acid treatment for the determination of fluoride from animal feeding stuffs. The content of fluoride (F-) corresponds to that of fluorine (F) specified in Commission Regulation (EU) 574/2011[3].
This European Standard is strictly based on several conventions such as those contained in the following example:
EXAMPLE   0,5 g test portion for extraction of fluoride from animal feeds by means of an acid treatment with 20 ml of 1 mol/l hydrochloric acid solution at ambient temperature (20 °C to 25 ºC) for 20 min. The pH is controlled and adjusted to 5,5 in the buffered test solution before determination of fluoride by ISE using standard addition technique.
The method was successfully tested in an interlaboratory study in concentrations between 100 mg/kg up to 500 mg/kg. If this method is followed strictly, then theoretically all concentrations from 40 mg/kg up to 4 000 mg/kg can be analysed within the linear calibration function.
Only for concentrations lower than 40 mg/kg is the use of an interpolation technique required instead of standard addition Annex C.
The quantification limit for fluoride using the conventions of the method including the standard addition technique is 40 mg/kg or lower than 2,5 mg/kg when using interpolation Annex C.

This European Standard is applicable to the quantitative analysis of (bound and free) hydrocyanic acid (HCN) in feed materials of plant origin and compound feed by High Performance Liquid Chromatography (HPLC).
The method is validated from 10 mg HCN/kg to 350 mg HCN/kg. When the method is used outside this range it should be validated at least within the laboratory. A limit of quantification of 2 mg HCN/kg should normally be obtained.

This European Standard specifies a method for the determination of mercury in animal feeding stuffs by Cold-Vapour Atomic Absorption Spectrometry (CVAAS) after microwave pressure digestion. The limit of quantification in the test solution should be 0,25 µg/l or lower. Using a test portion of 0,5 g and a volume of the test solution of 25 ml a limit of quantification of 0,0125 mg/kg or lower should be obtained.

This European Standard describes a procedure for the determination of inorganic arsenic in animal feeding stuffs of marine origin by Solid Phase Extraction (SPE) and Hydride Generation Atomic Absorption Spectrometry (HG-AAS). The method has been successfully tested in a collaborative trial with a working range from 0,19 mg/kg to 2,7 mg/kg (HORRATvalues < 2). The LOQ of the method is usually approximately 0,1 mg/kg or lower.

This European Standard specifies an Ion-Selective Electrode method (ISE) after hydrochloric acid treatment for the determination of fluoride from animal feeding stuffs. The content of fluoride (F-) corresponds to that of fluorine (F) specified in Commission Regulation (EU) 574/2011[3].
This European Standard is strictly based on several conventions such as those contained in the following example:
EXAMPLE   0,5 g test portion for extraction of fluoride from animal feeds by means of an acid treatment with 20 ml of 1 mol/l hydrochloric acid solution at ambient temperature (20 °C to 25 ºC) for 20 min. The pH is controlled and adjusted to 5,5 in the buffered test solution before determination of fluoride by ISE using standard addition technique.
The method was successfully tested in an interlaboratory study in concentrations between 100 mg/kg up to 500 mg/kg. If this method is followed strictly, then theoretically all concentrations from 40 mg/kg up to 4 000 mg/kg can be analysed within the linear calibration function.
Only for concentrations lower than 40 mg/kg is the use of an interpolation technique required instead of standard addition Annex C.
The quantification limit for fluoride using the conventions of the method including the standard addition technique is 40 mg/kg or lower than 2,5 mg/kg when using interpolation Annex C.

This European Standard specifies a method for the determination of mercury in animal feeding stuffs by Cold-Vapour Atomic Absorption Spectrometry (CVAAS) after microwave pressure digestion. The limit of quantification in the test solution should be 0,25 µg/l or lower. Using a test portion of 0,5 g and a volume of the test solution of 25 ml a limit of quantification of 0,0125 mg/kg or lower should be obtained.

This European Standard describes a procedure for the determination of inorganic arsenic in animal feeding stuffs of marine origin by Solid Phase Extraction (SPE) and Hydride Generation Atomic Absorption Spectrometry (HG-AAS). The method has been successfully tested in a collaborative trial with a working range from 0,19 mg/kg to 2,7 mg/kg (HORRATvalues < 2). The LOQ of the method is usually approximately 0,1 mg/kg or lower.

This European Standard 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.

This European Standard specifies a method for the determination of decoquinate. This high-performance liquid chromatographic (HPLC) method with a fluorescence detection is applicable to the quantification of decoquinate content in complete and complementary compound feeds, medicated feeds, semi-liquid feeds, premixtures and feed additives.
The method was fully validated from LOQ to 60 000 mg/kg on different matrices during an international collaborative study [11], especially on complete compound feeds for poultry, at trace contamination level of 3 mg/kg and at European authorized level of 20 mg/kg to 40 mg/kg [12].
The limit of detection is between 0,1 mg/kg and 0,3 mg/kg and the limit of quantification is around 0,5 mg/kg. These limits were validated during the collaborative study [11], from results on the blank feed. Lower limits of detection or quantification could be reached but a single laboratory validation is then requested.

This European Standard is applicable to the quantitative analysis of (bound and free) hydrocyanic acid (HCN) in feed materials of plant origin and compound feed by High Performance Liquid Chromatography (HPLC).
The method is validated from 10 mg HCN/kg to 350 mg HCN/kg. When the method is used outside this range it should be validated at least within the laboratory. A limit of quantification of 2 mg HCN/kg should normally be obtained.

This European standard specifies a high-performance liquid chromatographic (HPLC) method for the determination of the semduramicin content at authorized level in animal feeding stuffs [2], using mass spectrometry detection or post-column derivatization and (UV)-VIS detection (hereinafter UV detection). This method is applicable to poultry feed. The limit of quantitation is 1,0 mg/kg when mass spectrometry is used for detection and 3,0 mg/kg when the detection is performed by UV with post-column derivatization. Lower limits of quantitation are achievable but this is to be validated by the user.
The method allows the discrimination of semduramicin from monensin, salinomycin, narasin, maduramicin and lasalocid.

This European Standard specifies a method for the determination of selenium in animal feeding stuffs by hydride generation atomic absorption spectrometry (HGAAS) after microwave pressure digestion.
The method was successfully tested by an inter-laboratory study of CEN/TC 327/WG 4 in the range of 0,25 mg/kg to 74 mg/kg.
The limit of quantification is 0,5 µg/l of the test solution which corresponds to the calibration standard 2. Using a test portion of 0,5 g and a volume of the test solution of 25 ml after pressure digestion the limit of quantification is calculated as 0,125 mg/kg in the feed material.
NOTE   A lower limit of quantification could be achieved – each laboratory has to prove it.

This European Standard specifies a method for the determination of total arsenic in animal feeding stuffs by hydride generation atomic absorption spectrometry (HGAAS) after microwave pressure digestion. The limit of quantification is 0,5 µg/l of the test solution. Using a test portion of 0,5 g, a volume of the test solution of 25 ml and an aliquot of 5 ml for pre-reduction the limit of quantification is 0,125 mg/kg in the feed material.
NOTE   For feed materials containing organic arsenic species from compounds of marine origin (i.e. arsenobetaine and tetramethylarsine oxide) a higher digestion temperature of the microwave system up to 300 °C may be necessary in order to enable the hydridisation of these arsenic compounds and in order to determine all different kinds of arsenic species in the corresponding feeding stuffs. Alternatively, the digestion procedure of Annex D can be used if the microwave system does not reach higher temperatures up to 300 °C to ensure complete mineralization for HGAAS determination.

2010-03-19 EMA: Corrigendum on the French version only.

This European standard specifies a high-performance liquid chromatographic (HPLC) method for the determination of the semduramicin content at authorized level in animal feeding stuffs [2], using mass spectrometry detection or post-column derivatization and (UV)-VIS detection (hereinafter UV detection). This method is applicable to poultry feed. The limit of quantitation is 1,0 mg/kg when mass spectrometry is used for detection and 3,0 mg/kg when the detection is performed by UV with post-column derivatization. Lower limits of quantitation are achievable but this is to be validated by the user.
The method allows the discrimination of semduramicin from monensin, salinomycin, narasin, maduramicin and lasalocid.

This European Standard specifies a method for the determination of total arsenic in animal feeding stuffs by hydride generation atomic absorption spectrometry (HGAAS) after microwave pressure digestion. The limit of quantification is 0,5 µg/l of the test solution. Using a test portion of 0,5 g, a volume of the test solution of 25 ml and an aliquot of 5 ml for pre-reduction the limit of quantification is 0,125 mg/kg in the feed material.
NOTE   For feed materials containing organic arsenic species from compounds of marine origin (i.e. arsenobetaine and tetramethylarsine oxide) a higher digestion temperature of the microwave system up to 300 °C may be necessary in order to enable the hydridisation of these arsenic compounds and in order to determine all different kinds of arsenic species in the corresponding feeding stuffs. Alternatively, the digestion procedure of Annex D can be used if the microwave system does not reach higher temperatures up to 300 °C to ensure complete mineralization for HGAAS determination.

This European Standard specifies a method for the determination of selenium in animal feeding stuffs by hydride generation atomic absorption spectrometry (HGAAS) after microwave pressure digestion.
The method was successfully tested by an inter-laboratory study of CEN/TC 327/WG 4 in the range of 0,25 mg/kg to 74 mg/kg.
The limit of quantification is 0,5 µg/l of the test solution which corresponds to the calibration standard 2. Using a test portion of 0,5 g and a volume of the test solution of 25 ml after pressure digestion the limit of quantification is calculated as 0,125 mg/kg in the feed material.
NOTE   A lower limit of quantification could be achieved – each laboratory has to prove it.

This European Standard is applicable to the determination of Fumonisin B1 & B2 (FB1 & FB2) in compound animal feed at levels starting from 3 mg/kg up to 16 mg/kg.

This European Standard specifies a method for the determination of Ochratoxin A (OTA) in cereal based animal feed using immunoaffinity for clean-up followed by liquid-chromatography with fluorescence detection.
NOTE   The validated mass fraction range was 39 µg/kg to 338 µg/kg OTA.

This European Standard is applicable to the determination of Fumonisin B1 & B2 (FB1 & FB2) in compound animal feed at levels starting from 3 mg/kg up to 16 mg/kg.

This European Standard specifies a method for the determination of Ochratoxin A (OTA) in cereal based animal feed using immunoaffinity for clean-up followed by liquid-chromatography with fluorescence detection.
NOTE   The validated mass fraction range was 39 µg/kg to 338 µg/kg OTA.

This European Standard defines general rules for the enumeration of probiotic bifidobacteria in feed samples (additives, premixtures and feeding stuffs) that contain bifidobacteria as a single bacterial component or in a mixture with other microorganisms. This standard is not applicable for mineral feeds which are defined as complementary feeding stuffs composed mainly of minerals and containing at least 40% crude ash (Council Directive 79/373/EEC) [3].
There are different categories of feed samples:
a)   Additives containing about 1010 colony forming units (CFU)/g
b)   Premixtures containing  about 108 CFU/g
c)   Feeds, meal or pellets, which contain about 106 CFU/g and include complete feeding stuffs, and milk replacers.
The detection limit is as defined in ISO 7218.

This European Standard specifies a method for the determination of additive use of nicarbazin in animal feeding stuffs and premixtures (maximum concentration 2,5% nicarbazin) using high performance liquid chromatography. Nicarbazin is a 1:1 equimolar mixture of 4,4’-dinitrocarbanilide (DNC) and 4,6-dimethyl-2-pyriminol (HDP). Nicarbazin is generally determined by using DNC as the target compound. In this method the DNC moiety of nicarbazin is detected.
The limit of quantitation is 20 mg/kg. The limit of detection is 0,5 mg/kg
NOTE   A lower limit of quantitation may be achievable but should be validated by the user.

This European Standard specifies a high performance liquid chromatography (HPLC) method for the
determination of the content of maduramicin in feeding stuffs and premixtures.
The usual concentration of maduramicin in feedstuffs is 5 mg/kg, in premixtures 500 mg/kg. The limit of
quantification is 2 mg/kg. The limit of detection is 0,5 mg/kg.
NOTE A lower limit of quantification may be achievable but shall be validated by the user.

This International Standard is applicable to the determination of zearalenone in animal feed at concentrations from 30 µg/kg to 3 000 µg/kg.

This Standard is applicable to the determination of deoxynivalenol (DON) in animal compound feed at concentrations of 150 μg/kg up to at least 4 000 μg/kg.

This European Standard defines general rules for the enumeration of probiotic bifidobacteria in feed samples (additives, premixtures and feeding stuffs) that contain bifidobacteria as a single bacterial component or in a mixture with other microorganisms. This standard is not applicable for mineral feeds which are defined as complementary feeding stuffs composed mainly of minerals and containing at least 40% crude ash (Council Directive 79/373/EEC) [3].
There are different categories of feed samples:
a)   Additives containing about 1010 colony forming units (CFU)/g
b)   Premixtures containing  about 108 CFU/g
c)   Feeds, meal or pellets, which contain about 106 CFU/g and include complete feeding stuffs, and milk replacers.
The detection limit is as defined in ISO 7218.

This Standard is applicable to the determination of deoxynivalenol (DON) in animal compound feed at concentrations of 150 μg/kg up to at least 4 000 μg/kg.

This International Standard is applicable to the determination of zearalenone in animal feed at concentrations from 30 µg/kg to 3 000 µg/kg.

This European Standard specifies a high performance liquid chromatography (HPLC) method for the
determination of the content of maduramicin in feeding stuffs and premixtures.
The usual concentration of maduramicin in feedstuffs is 5 mg/kg, in premixtures 500 mg/kg. The limit of
quantification is 2 mg/kg. The limit of detection is 0,5 mg/kg.
NOTE A lower limit of quantification may be achievable but shall be validated by the user.

This European Standard specifies a method for the determination of additive use of nicarbazin in animal feeding stuffs and premixtures (maximum concentration 2,5% nicarbazin) using high performance liquid chromatography. Nicarbazin is a 1:1 equimolar mixture of 4,4’-dinitrocarbanilide (DNC) and 4,6-dimethyl-2-pyriminol (HDP). Nicarbazin is generally determined by using DNC as the target compound. In this method the DNC moiety of nicarbazin is detected.
The limit of quantitation is 20 mg/kg. The limit of detection is 0,5 mg/kg
NOTE   A lower limit of quantitation may be achievable but should be validated by the user.

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.

ISO 16634-1:2008 specifies a method for the determination of the total nitrogen content and the calculation of crude protein content of oilseeds and animal feeding stuffs.
This method, like the Kjeldahl method, does not distinguish between protein nitrogen and non-protein nitrogen. For the calculation of protein content, various conversion factors are used.
This method is not applicable to milk and milk products.

This Technical Specification defines a polymerase chain reaction (PCR) methodology for the identification of S. cerevisiae probiotic yeast strains. Additionally, a method for the extraction of high quality DNA from yeast is suggested.

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.

This Technical Specification defines a polymerase chain reaction (PCR) methodology for the identification of S. cerevisiae probiotic yeast strains. Additionally, a method for the extraction of high quality DNA from yeast is suggested.

ISO 16634-1:2008 specifies a method for the determination of the total nitrogen content and the calculation of crude protein content of oilseeds and animal feeding stuffs.
This method, like the Kjeldahl method, does not distinguish between protein nitrogen and non-protein nitrogen. For the calculation of protein content, various conversion factors are used.
This method is not applicable to milk and milk products.

This Technical Specification describes the quantitative determination of specific sugars (glucose, fructose, galactose, sucrose, maltose, and lactose) in dry animal feeding stuffs at the g/kg level by a sophisticated high performance anion exchange chromatography in combination with pulsed amperometric detection (HPAEC-PAD).

This Technical Specification describes the quantitative determination of specific sugars (glucose, fructose, galactose, sucrose, maltose, and lactose) in dry animal feeding stuffs at the g/kg level by a sophisticated high performance anion exchange chromatography in combination with pulsed amperometric detection (HPAEC-PAD).

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.

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.

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.

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.

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.

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.

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.

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.