M/382 - Animal Feedingstuffs

This document is applicable to the determination of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), (together termed ‘dioxins’ (PCDD/Fs)) and dioxin-like PCBs and non-dioxin-like PCBs (dl-PCBs and ndl-PCBs) in animal feeding stuffs. Collaborative studies have been carried out. The method is suitable for the determination of dioxins, dl-PCBs and ndl-PCBs at the appropriate MRL in compound feed and ingredients e.g. oil, mineral clay. The method is applicable to samples containing trace level amounts of one or more dioxins, dioxin-like PCBs and non-dioxin-likePCBs. The limit of quantification (LOQ) is
-   0,05 pg/g (OCDD/F = 0,1 pg/g) for the relevant individual congeners of dioxins/furans,
-   0,05 pg/g for non-ortho PCBs,
-   10 pg/g for mono-ortho PCBs, and
-   100 pg/g for non-dioxin-like-PCBs.
For determination of dioxins and dioxin-like PCBs, the procedure can be used as confirmatory method as defined by Commission Regulation (EC) No 152/2009 for dioxins and dl-PCB in feed [1]. Confirmatory methods as described in this standard are high-resolution gas chromatography/high resolution mass spectrometry (HRGC/HRMS) methods. If only the analysis of non-dioxin-like PCBs is required, a GC-LRMS method can be used (e.g. EN 15741 [2]) provided that appropriate analytical performance criteria are met in the relevant range for the matrix of interest.
This document is split into four modules. Each module describes a part of the whole procedure (see Figure 1 and Figure 2) to be followed:
a)   Module A:   Description of standards which might be used;
b)   Module B:   Description of extraction procedures;
c)   Module C:   Description of clean-up procedures;
d)   Module D:    GC/HRMS determination.
Each module describes a part of the whole method as well as, when applicable, alternatives which should be equivalent. Each module has to be regarded as an example. Combining modules and/or alternatives gives a highly flexible, "performance based" procedure. It is permitted to modify the method if all performance criteria laid down in Commission Regulation (EC) No 152/2009 [1] are met.
Any deviation of the described method, combination of modules needs to be recorded as part of the QA/QC procedures of accredited laboratories and should be available on request.
Figure 1 - Flow scheme for the determination of dioxins, dl-PCBs and non-dioxin-like-PCBs in feed
Figure 2 - Flow scheme for the determination of dioxins, dl-PCBs and non-dioxin-like-PCBs in oil and fat

This document is applicable to the determination of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), (together termed ‘dioxins’ (PCDD/Fs)) and dioxin-like PCBs and non-dioxin-like PCBs (dl-PCBs and ndl-PCBs) in animal feeding stuffs. Collaborative studies have been carried out. The method is suitable for the determination of dioxins, dl-PCBs and ndl-PCBs at the appropriate MRL in compound feed and ingredients e.g. oil, mineral clay. The method is applicable to samples containing trace level amounts of one or more dioxins, dioxin-like PCBs and non-dioxin-likePCBs. The limit of quantification (LOQ) is
-   0,05 pg/g (OCDD/F = 0,1 pg/g) for the relevant individual congeners of dioxins/furans,
-   0,05 pg/g for non-ortho PCBs,
-   10 pg/g for mono-ortho PCBs, and
-   100 pg/g for non-dioxin-like-PCBs.
For determination of dioxins and dioxin-like PCBs, the procedure can be used as confirmatory method as defined by Commission Regulation (EC) No 152/2009 for dioxins and dl-PCB in feed [1]. Confirmatory methods as described in this standard are high-resolution gas chromatography/high resolution mass spectrometry (HRGC/HRMS) methods. If only the analysis of non-dioxin-like PCBs is required, a GC-LRMS method can be used (e.g. EN 15741 [2]) provided that appropriate analytical performance criteria are met in the relevant range for the matrix of interest.
This document is split into four modules. Each module describes a part of the whole procedure (see Figure 1 and Figure 2) to be followed:
a)   Module A:   Description of standards which might be used;
b)   Module B:   Description of extraction procedures;
c)   Module C:   Description of clean-up procedures;
d)   Module D:    GC/HRMS determination.
Each module describes a part of the whole method as well as, when applicable, alternatives which should be equivalent. Each module has to be regarded as an example. Combining modules and/or alternatives gives a highly flexible, "performance based" procedure. It is permitted to modify the method if all performance criteria laid down in Commission Regulation (EC) No 152/2009 [1] are met.
Any deviation of the described method, combination of modules needs to be recorded as part of the QA/QC procedures of accredited laboratories and should be available on request.
Figure 1 - Flow scheme for the determination of dioxins, dl-PCBs and non-dioxin-like-PCBs in feed
Figure 2 - Flow scheme for the determination of dioxins, dl-PCBs and non-dioxin-like-PCBs in oil and fat

This International Standard 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 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 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 , 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. 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 , 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. This European Standard is strictly based on several conventions such as those contained in the following 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 ?C) 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 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; HORRAT value is Horwitz -Ratio value). The LOQ of the method is usually approximately 0,1 mg/kg or lower.

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.

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 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 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.
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 extraction 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.

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.

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.

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 describes the determination of phytase activity in complete feedingstuff 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.

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.

This European Standard defines general rules for the enumeration of probiotic lactobacilli in feed samples (additives, premixtures and feeding stuffs) that contain lactobacilli as a single bacterial component or in a mixture with other microorganisms. This standard is not applicable to 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 defines general rules for the enumeration of probiotic bacilli in feeds containing bacilli (Bacillus species) as a single microorganism, component or mixed with other microorganisms. This method is not applicable to 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 limits are 500 (5 x 102) colony forming units per gram (CFU/g). The limits of determination are 2 x 104 CFU/g.

This International 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 g/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 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 defines general rules for the enumeration of probiotic yeasts in feed samples (additives, premixtures and feeding stuffs) that contain yeast as a single microorganism component or in a mixture with other microorganisms. The standard is not applicable to mineral feeds which are defined as complementary feedingstuffs composed mainly of minerals and containing at least 40% crude ash (Council Directive 79/373/EEC) [4].
There are different categories of feed samples:
a)   Additives which contain about 109 CFU/g to 1010 CFU/g (CFU = colony forming units).
b)   Premixtures which contain about 108 CFU/g
c)   Feeds, meal or pellets, which contain about 106 CFU/g and include complete feedingstuffs, and milk replacers.
The detection limit is as defined in ISO 7218.

This European Standard defines general rules for the enumeration of enterococci in feed samples (additives, premixtures and feeding stuffs) that contain enterococci (E. faecium) as a single microorganism component or in a mixture with other microorganisms. This standard is not applicable to mineral feeds which are defined as complementary feedingstuffs 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 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 defines general rules for the enumeration of probiotic pediococci in feed samples (additives, premixtures and feeding stuffs) that contain pediococci 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 defines general rules for the enumeration of probiotic bacilli in feeds containing bacilli (Bacillus species) as a single microorganism, component or mixed with other microorganisms. This method is not applicable to 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 limits are 500 (5 x 102) colony forming units per gram (CFU/g). The limits of determination are 2 x 104 CFU/g.

This European Standard defines general rules for the enumeration of probiotic yeasts in feed samples (additives, premixtures and feeding stuffs) that contain yeast as a single microorganism component or in a mixture with other microorganisms. The standard is not applicable to mineral feeds which are defined as complementary feedingstuffs composed mainly of minerals and containing at least 40% crude ash (Council Directive 79/373/EEC) [4].
There are different categories of feed samples:
a)   Additives which contain about 109 CFU/g to 1010 CFU/g (CFU = colony forming units).
b)   Premixtures which contain about 108 CFU/g
c)   Feeds, meal or pellets, which contain about 106 CFU/g and include complete feedingstuffs, and milk replacers.
The detection limit is as defined in ISO 7218.

This European Standard defines general rules for the enumeration of enterococci in feed samples (additives, premixtures and feeding stuffs) that contain enterococci (E. faecium) as a single microorganism component or in a mixture with other microorganisms. This standard is not applicable to mineral feeds which are defined as complementary feedingstuffs 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 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 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 defines general rules for the enumeration of probiotic pediococci in feed samples (additives, premixtures and feeding stuffs) that contain pediococci 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 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 European Standard defines general rules for the enumeration of probiotic lactobacilli in feed samples (additives, premixtures and feeding stuffs) that contain lactobacilli as a single bacterial component or in a mixture with other microorganisms. This standard is not applicable to 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.

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.

This document applies to medical compression hosiery and thrombosis prophylaxis hosiery.
An important property of hosiery is the compression it exerts on the limb. This document is intended to provide a reference for testing the compressive properties in medical hosiery.

This document applies to medical compression hosiery and thrombosis prophylaxis hosiery.
An important property of hosiery is the compression it exerts on the limb. This document is intended to provide a reference for testing the compressive properties in medical hosiery.