# Commission Regulation (EU) No 519/2014Show full title

Commission Regulation (EU) No 519/2014 of 16 May 2014 amending Regulation (EC) No 401/2006 as regards methods of sampling            of large lots, spices and food supplements, performance criteria for T-2, HT-2 toxin and citrinin and screening methods of            analysis (Text with EEA relevance)         

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U.K.

# Commission Regulation (EU) No 519/2014                     

# of 16 May 2014

# amending Regulation (EC) No 401/2006 as regards methods of sampling of large lots, spices and food supplements, performance                        criteria for T-2, HT-2 toxin and citrinin and screening methods of analysis                     

# (Text with EEA relevance)

THE EUROPEAN COMMISSION,

Having regard to the Treaty on the Functioning of the European Union,

Having regard to Regulation (EC) No 882/2004 of the European Parliament and of the Council of 29 April 2004 on official controls                           performed to ensure the verification of compliance with feed and food law, animal health and animal welfare rules(1), in particular Article 11(4) thereof,                        

Whereas:

(1)Commission Regulation (EC) No 1881/2006(2) provides for maximum limits for certain mycotoxins in certain foodstuffs.

(2)Sampling plays a crucial part in the precision of the determination of the levels of mycotoxins, which are heterogeneously                              distributed in a lot. It is therefore necessary to set out criteria which the sampling methods should fulfil.

(3)Commission Regulation (EC) No 401/2006(3) establishes the criteria for the sampling for the control of the levels of mycotoxins.

(4)It is necessary to amend the rules concerning the sampling of spices in order to take into account the differences in particle                              size which leads to the heterogeneous distribution of mycotoxin contamination in spices. Furthermore it is appropriate to                              establish rules for the sampling of large lots in order to ensure a uniform enforcement approach across the Union. It is also                              appropriate to clarify which method of sampling has to be applied for the sampling of apple juice.

(5)The performance criteria for T-2 and HT-2 toxin need to be updated in order to take into account scientific and technological                              progress. Performance criteria for citrinin need to be established given the maximum level established for citrinin in food                              supplements based on rice fermented with the red yeast *Monascus purpureus.*

(6)For the analysis of mycotoxins, screening methodologies are used more and more. It is appropriate to establish criteria with                              which the screening methods have to comply with for use for regulatory purposes.

(7)The measures provided for in this Regulation are in accordance with the opinion of the Standing Committee on the Food Chain                              and Animal Health,

HAS ADOPTED THIS REGULATION:

# Article 1U.K.

Regulation (EC) No 401/2006 is amended as follows:

Annex I is amended as follows:

In part B, footnote (1) is replaced by the following:

## ‘(1)The sampling of such lots shall be performed in accordance with the rules set out in part L. Guidance for sampling large lots                                             shall be provided in a guidance document available on the following website: http://ec.europa.eu/food/food/chemicalsafety/contaminants/guidance-sampling-final.pdf

The application of sampling rules in accordance with EN ISO 24333:2009 or GAFTA Sampling Rules 124, applied by food business                                          operators to ensure compliance with provisions in legislation is equivalent to the sampling rules set out in part L.                                    

For the sampling of lots for *Fusarium*-toxins, the application of sampling rules in accordance with EN ISO 24333:2009 or GAFTA Sampling Rules 124, applied by food                                          business operators to ensure compliance with provisions in legislation is equivalent to the sampling rules set out in part                                          B;’                                    

In part B.2, Table 1 is replaced by the following table:

## *‘Table 1*

**Subdivision of lots into sublots depending on product and lot weight**

In part B.3, the following sentence is added at the end of the first indent:

‘For lots > 500 tonnes, the number of incremental samples is provided for in part L.2 of Annex I.’                                    

In part D.2 the following sentence is added after the first sentence:

‘This method of sampling is of also of application for the official control of the maximum levels established for ochratoxin                                          A, aflatoxin B1 and total aflatoxins in spices with a relatively large particle size (particle size comparable with peanuts                                          or larger e.g. nutmeg).’;                                    

In part E, the first sentence is replaced by the following:

‘This method of sampling is of application for the official control of the maximum levels established for ochratoxin A, aflatoxin                                          B1 and total aflatoxins in spices except in cases of spices with a relatively large particle size (heterogeneous distribution                                          of mycotoxin contamination).’;                                    

In part I, the heading and the first sentence are replaced by the following:

## ‘I.METHOD OF SAMPLING FOR SOLID APPLE PRODUCTS

This method of sampling is of application for the official control of the maximum levels established for patulin in solid                                          apple products, including solid apple products for infants and young children.’                                    

In part I.1, second paragraph, the following sentences are deleted:

‘In case of liquid products the lot shall be thoroughly mixed insofar as possible by either manual or mechanical means immediately                                          prior to sampling. In this case, a homogenous distribution of patulin can be assumed within a given lot. It is therefore sufficient                                          to take three incremental samples from a lot to form the aggregate sample.’                                    

New Parts L and M as set out in Annex I to this Regulation, are added.

In Annex II, points 4.2 ‘General requirements’, 4.3 ‘Specific requirements’ and 4.4 ‘Estimation of measurement uncertainty,                              recovery calculation and reporting of results’ are replaced by the text set out in Annex II to this Regulation.                           

# Article 2U.K.

This Regulation shall enter into force on the twentieth day following that of its publication in the *Official Journal of the European Union*.                  

It shall apply from 1 July 2014.

This Regulation shall be binding in its entirety and directly applicable in all Member States.

Done at Brussels, 16 May 2014.

*For the Commission*                            

*The President*                            

José Manuel Barroso                            

## ANNEX IU.K.

### ‘L.METHOD OF SAMPLING FOR VERY LARGE LOTS OR LOTS STORED OR TRANSPORTED IN A WAY WHEREBY SAMPLING THROUGHOUT THE LOT IS NOT FEASIBLE

#### L.1.**General principles**

In case the way of transport or storage of a lot does not enable to take incremental samples throughout the whole lot, sampling                        of such lots should preferably be done when the lot is in flow (dynamic sampling).                  

In the case of large warehouses destined to store food, operators should be encouraged to install equipment in the warehouse                        enabling (automatic) sampling across the whole stored lot.                  

When the sampling procedures as provided for in this part L are applied, the food business operator or his representative                        should be informed of the sampling procedure. If the sampling procedure is questioned by the food business operator or his                        representative, the food business operator or his representative shall enable the competent authority to sample throughout                        the whole lot at his/her own cost.                  

Sampling of a part of the lot is allowed, on the condition that the quantity of the sampled part is at least 10 % of the lot                        to be sampled. If a part of a lot of food of the same class or description has been sampled and identified as not satisfying                        Union requirements, it shall be presumed that the entire lot is also affected, unless further detailed assessment shows no                        evidence that the rest of the lot is unsatisfactory.                  

The relevant provisions, such as weight of the incremental sample, provided for in the other parts of this Annex are applicable                        for the sampling for very large lots or lots stored or transported in a way whereby sampling throughout the lot is not feasible.                  

#### L.2.**Number of incremental samples to be taken in the case of very large lots**

In the case of large sampled portions (sampled portions > 500 tonnes), the number of incremental samples to be taken = 100                        incremental samples + √tonnes. However in case the lot is less than 1 500 tonnes and can be subdivided into sublots in accordance                        with the table 1 of part B and on the condition that the sublots can be separated physically, the number of incremental samples                        as provided for in part B have to be taken.                  

#### L.3.**Large lots transported by ship**

##### L.3.1.*Dynamic sampling of large lots transported by ship*

The sampling of large lots in ships is preferably carried out while the product is in flow (dynamic sampling).                  

The sampling is to be done per hold (entity that can physically be separated). Holds are however emptied partly one after                        the other so that the initial physical separation no longer exists after transfer into storage facilities. Sampling can therefore                        be performed based on initial physical separation or based on the separation after transfer into the storage facilities.                  

The unloading of a ship can last for several days. Normally, sampling has to be performed at regular intervals during the                        whole duration of unloading. It is however not always feasible or appropriate for an official inspector to be present for                        sampling during the whole operation of unloading. Therefore sampling of part of the lot is allowed to be undertaken (sampled                        portion). The number of incremental samples is determined by taking into account the size of the sampled portion.                  

Even if the official sample is taken automatically, the presence of an inspector is necessary. However if the automatic sampling                        is done with pre-set parameters which cannot be changed during the sampling and the incremental samples are collected in a                        sealed receptacle, preventing any possible fraud, then the presence of an inspector is only required at the beginning of the                        sampling, every time the receptacle of the samples needs to be changed and at the end of the sampling.                  

##### L.3.2.*Sampling of lots transported by ship by static sampling*

In cases where the sampling is done in a static way the same procedure as foreseen for storage facilities (silos) accessible                        from above has to be applied (see point L.5.1).                  

The sampling has to be performed on the accessible part (from above) of the lot/hold. The number of incremental samples is                        determined by taking into account the size of the sampled portion.                  

#### L.4.**Sampling of large lots stored in warehouses**

The sampling has to be performed on the accessible part of the lot. The number of incremental samples is determined by taking                        into account the size of the sampled portion.                  

#### L.5.**Sampling of storage facilities (silos)**

##### L.5.1.*Sampling of silos (easily) accessible from above*

The sampling has to be performed on the accessible part of the lot. The number of incremental samples is determined by taking                        into account the size of the sampled portion.                  

##### L.5.2.*Sampling of silos not accessible from above (closed silos)*

###### L.5.2.1.*Silos not accessible from above (closed silos) with individual sizes > 100 tonnes*

Food stored in such silos cannot be sampled in a static way. Therefore when the food in the silo has to be sampled and there                        is no possibility to move the consignment, the agreement has to be made with the operator that he or she has to inform the                        inspector about when the silo will be unloaded, partially or completely, in order to enable sampling when the food is in flow.                  

###### L.5.2.2.*Silos not accessible from above (closed silos) with individual sizes < 100 tonnes*

Contrary to the provision in part point L.1 (sampled part at least 10 %), the sampling procedure involves the release into                        a receptacle of a quantity of 50 to 100 kg and taking the sample from it. The size of the aggregate sample corresponds to                        the whole lot and the number of incremental samples relate to the quantity of the food from the silo released into the receptacle                        for sampling.                  

#### L.6.**Sampling of loose food in large closed containers**

Such lots can often only be sampled when unloaded. In certain cases it is not possible to unload at the point of import or                        control and therefore the sampling should take place when such containers are unloaded. The operator has to inform the inspector                        about the place and time of unloading the containers.                  

### M.METHOD OF SAMPLING OF FOOD SUPPLEMENTS BASED ON RICE FERMENTED WITH RED YEAST *MONASCUS PURPUREUS*

This method of sampling is applicable to the official control of the maximum level established for citrinin in food supplements                        based on rice fermented with red yeast *Monascus purpureus.*                  

#### **Sampling procedure and sample size**

The sampling procedure is on the supposition that the food supplements based on rice fermented with red yeast *Monascus purpureus* are marketed in retail packages containing usually 30 to 120 capsules per retail package.                  

## ANNEX IIU.K.

### ‘4.2.**General requirements**

Confirmatory methods of analysis used for food control purposes shall comply with the provisions of items 1 and 2 of Annex                        III to Regulation (EC) No 882/2004.                  

### 4.3.**Specific requirements**

#### 4.3.1.*Specific requirements for confirmatory methods*

##### 4.3.1.1.*Performance criteria*

It is recommended that fully validated confirmatory methods (i.e. methods validated by collaborative trials for relevant matrices)                        are used where appropriate and available. Other suitable validated confirmatory methods (e.g. methods validated in-house on                        relevant matrices belonging to the commodity group of interest) may also be used provided they fulfil the performance criteria                        set out in the following tables.                  

Where possible, the validation of in-house validated methods shall include a certified reference material.                  

Performance criteria for aflatoxins                           

###### *Note:*

                -  Values to apply to both B1 and sum of B1 + B2 + G1 + G2                                  
                -  If sum of individual aflatoxins B1 + B2 + G1 + G2 are to be reported, then response of each to the analytical system must be either known or equivalent.                                  

Performance criteria for ochratoxin A                           

Performance criteria for patulin                           

Performance criteria for deoxynivalenol                           

Performance criteria for zearalenone                           

Performance criteria for Fumonisin B1 and B2 individually                           

Performance criteria for T-2 and HT-2 toxin individually                           

Performance criteria for citrinin                           

Notes to the performance criteria for the mycotoxins:                           
                -  The detection limits of the methods used are not stated as the precision values given at the concentrations of interest.                                  
                -  The precision values are calculated from the Horwitz equation, in particular the original Horwitz equation (for concentrations                                       1,2 × 10–7 ≤ C ≤ 0,138) (*) and the modified Horwitz equation (for concentrations C < 1,2 × 10–7) (**).                                   (*)
Horwitz equation for concentrations 1,2 × 10–7 ≤ C ≤ 0,138:                                          

RSDR = 2(1-0.5logC)                                          

(ref: *W. Horwitz, L.R. Kamps, K.W. Boyer, J.Assoc.Off.Analy.Chem.,1980, 63, 1344)*                                          
(**)
Modified Horwitz equation (*) for concentrations C < 1,2 × 10–7:                                          

RSDR = 22 %                                          

(ref: *M. Thompson, Analyst, 2000, 125, p. 385-386)*                                          

Where:                                          
-  RSDR is the relative standard deviation calculated from results generated under reproducibility conditions [(sR/) × 100]                                                 
-  C is the concentration ratio (i.e. 1 = 100g/100g, 0,001 = 1 000 mg/kg)                                                 

This is a generalised precision equation which has been found to be independent of analyte and matrix but solely dependent                                                on concentration for most routine methods of analysis.                                          
 

##### 4.3.1.2.*“Fitness-for-purpose” approach*

For in-house validated methods, as an alternative, a “fitness-for-purpose” approach (***) may be used to assess their suitability                        for official control. Methods suitable for official control must produce results with a standard measurement uncertainty (u)                        less than the maximum standard measurement uncertainty calculated using the formula below:                  

where:                  
          -  *Uf* is the maximum standard measurement uncertainty (μg/kg)                         
          -  LOD is the limit of detection of the method (μg/kg)                         
          -  α is a constant, numeric factor to be used depending on the value of C. The values to be used are set out in Table hereafter.                         
          -  C is the concentration of interest (μg/kg)                         

If the analytical method provides results with uncertainty measurements less than the maximum standard uncertainty the method                        shall be considered being equally suitable to one which meets the performance criteria given in point 4.3.1.1.                  

###### *Table*

**Numeric values to be used for α as constant in formula set out in this point, depending on the concentration of interest**

###### (***)Ref: *M. Thompson and R. Wood, Accred. Qual. Assur., 2006, 10, p. 471-478*.

#### 4.3.2.*Specific requirements for semi-quantitative screening methods*

##### 4.3.2.1.*Scope*

The scope applies to bioanalytical methods based on immuno-recognition or receptor binding (such as ELISA, dip-sticks, lateral                        flow devices, immuno-sensors) and physicochemical methods based on chromatography or direct detection by mass spectrometry                        (e.g. ambient MS). Other methods (e.g. thin layer chromatography) are not excluded provided the signals generated relate directly                        to the mycotoxins of interest and allow that the principle described hereunder is applicable.                  

The specific requirements apply to methods of which the result of the measurement is a numerical value, for example a (relative)                        response from a dip-stick reader, a signal from LC-MS, etc., and that normal statistics apply.                  

The requirements do not apply to methods that do not give numerical values (e.g. only a line that is present or absent), which                        require different validation approaches. Specific requirements for these methods are provided in point 4.3.3.                  

This document describes procedures for the validation of screening methods by means of an inter-laboratory validation, the                        verification of the performance of a method validated by means of an inter-laboratory exercise and the single-laboratory validation                        of a screening method.                  

##### 4.3.2.2.*Terminology*

Screening target concentration (STC): the concentration of interest for detection of the mycotoxin in a sample. When the aim                        is to test compliance with regulatory limits, the STC is equal to the applicable maximum level. For other purposes or in case                        no maximum level has been established, the STC is predefined by the laboratory.                  

Screening method: means method used for selection of those samples with levels of mycotoxins that exceed the screening target                        concentration (STC), with a given certainty. For the purpose of mycotoxin screening, a certainty of 95 % is considered fit-for-purpose.                        The result of the screening analysis is either “negative” or “suspect”. Screening methods shall allow a cost-effective high                        sample-throughput, thus increasing the chance to discover new incidents with high exposure and health risks to consumers.                        These methods shall be based on bio-analytical, LC-MS or HPLC methods. Results from samples exceeding the cut-off value shall                        be verified by a full re-analysis from the original sample by a confirmatory method.                  

“Negative sample” means the mycotoxin content in the sample is < STC with a certainty of 95 % (i.e. there is a 5 % chance                        that samples will be incorrectly reported as negative).                  

“False negative sample” means the mycotoxin content in the sample is > STC but it has been identified as negative.                  

“Suspect sample” (screen positive) means the sample exceeds the cut-off level (see below) and may contain the mycotoxin at                        a level higher than the STC. Any suspect result triggers a confirmatory analysis for unambiguous identification and quantification                        of the mycotoxin.                  

“False suspect sample” is a negative sample that has been identified as suspect.                  

“Confirmatory methods” means methods that provide full or complementary information enabling the mycotoxin to be identified                        and quantified unequivocally at the level of interest.                  

Cut-off level: the response, signal, or concentration, obtained with the screening method, above which the sample is classified                        as “suspect”. The cut-off is determined during the validation and takes the variability of the measurement into account.                  

Negative control (blank matrix) sample: a sample known to be free(4) of the mycotoxin to be screened for, e.g. by previous determination using a confirmatory method of sufficient sensitivity.                        If no blank samples can be obtained, then material with the lowest obtainable level might be used as long as the level allows                        the conclusion that the screening method is fit for purpose.                  

Positive control sample: sample containing the mycotoxin at the screening target concentration, e.g. a certified reference                        material, a material of known content (e.g. test material of proficiency tests) or otherwise sufficiently characterised by                        a confirmatory method. In the absence of any of the above, a blend of samples with different levels of contamination or a                        spiked sample prepared within laboratory and sufficiently characterised can be used, provided it can be proven that the contamination                        level has been verified.                  

##### 4.3.2.3.*Validation procedure*

The aim of the validation is to demonstrate the fitness of purpose of the screening method. This is done by determination                        of the cut-off value and determination of the false negative and false suspect rate. In these two parameters performance characteristics                        such as sensitivity, selectivity, and precision are embedded.                  

Screening methods can be validated by inter-laboratory or by single laboratory validation. If inter-laboratory validation                        data is already available for a certain mycotoxin/matrix/STC combination, a verification of method performance is sufficient                        in a laboratory implementing the method.                  

###### 4.3.2.3.1.Initial validation by single laboratory validation

Mycotoxins:                  

The validation shall be performed for every individual mycotoxin in the scope. In case of bio-analytical methods that give                        a combined response for a certain mycotoxin group (e.g. aflatoxins B1, B2, G1 & G2; fumonisins B1 & B2), applicability must be demonstrated and limitations of the test mentioned in the scope of the method. Undesired cross-reactivity                        (e.g. DON-3-glycoside, 3- or 15-acetyl-DON for immuno-based methods for DON) is not considered to increase the false negative                        rate of the target mycotoxins, but may increase the false suspect rate. This unwanted increasing will be diminished by confirmatory                        analysis for unambiguous identification and quantification of the mycotoxins.                  

Matrices:                  

An initial validation should be performed for each commodity, or, when the method is known to be applicable to multiple commodities,                        for each commodity group. In the latter case, one representative and relevant commodity is selected from that group (see table                        A).                  

Sample set:                  

The minimum number of different samples required for validation is 20 homogeneous negative control samples and 20 homogeneous                        positive control samples that contain the mycotoxin at the STC, analysed under intermediate precision (RSDRi) conditions spread over 5 different days. Optionally, additional sets of 20 samples containing the mycotoxin at other levels                        can be added to the validation set to gain insight to what extent the method can distinguish between different mycotoxin concentrations.                  

Concentration:                  

For each STC to be used in routine application, a validation has to be performed.                  

###### 4.3.2.3.2.Initial validation through collaborative trials

Validation through collaborative trials shall be done in accordance with an internationally recognised protocol on collaborative                        trials (e.g. ISO 5725:1994 or the IUPAC International Harmonised Protocol) which requires inclusion of valid data from at                        least eight different laboratories. Other than that, the only difference compared to single laboratory validations is that                        the ≥ 20 samples per commodity/level can be evenly divided over the participating laboratories, with a minimum of two samples                        per laboratory.                  

##### 4.3.2.4.*Determination of cut-off level and rate of false suspected results of blank samples*

The (relative) responses for the negative control and positive control samples are taken as basis for the calculation of the                        required parameters.                  

Screening methods with a response proportional with the mycotoxin concentration                  

For screening methods with a response proportional with the mycotoxin concentration the following applies:                  

*Cut-off = RSTC – t-value0,05 * SDSTC*                  

R =

mean response of the positive control samples (at STC)                     

t-value:

one tailed t-value for a rate of false negative results of 5 % (see table B)                     

SD =

standard deviationScreening methods with a response inversely proportional with the mycotoxin concentration                     

Similarly, for screening methods with a response inversely proportional with the mycotoxin concentration, the cut-off is determined                        as:                  

*Cut-off = RSTC + t-value0,05 * SDSTC*                  

By using this specific t-value for establishing the cut-off value, the rate of false negative results is by default set at                        5 %.                  

Fitness for purpose assessment                  

Results from the negative control samples are used to estimate the corresponding rate of false suspect results. The t-value                        is calculated corresponding to the event that a result of a negative control sample is above the cut off value, thus erroneously                        classified as suspect.                  

t-value

=

(cut off – meanblank)/SDblankfor screening methods with a response proportional with the mycotoxin concentration                     

or                  

t-value

=

(*mean*blank – cut off)/SDblankfor screening methods with a response inversely proportional with the mycotoxin concentration                     

From the obtained t-value, based on the degrees of freedom calculated from the number of experiments, the probability of false                        suspect samples for a one tailed distribution can either be calculated (e.g.. spread sheet function “TDIST”) or taken from                        a table for t-distribution.                  

The corresponding value of the one tailed t-distribution specifies the rate of false suspect results.                  

This concept is described in detail with an example in Analytical and Bioanalytical Chemistry DOI 10.1007/s00216 -013-6922-1.                  

##### 4.3.2.5.*Extension of the scope of the method*

###### 4.3.2.5.1.Extension of scope to other mycotoxins:

When new mycotoxins are added to the scope of an existing screening method, a full validation is required to demonstrate the                        suitability of the method.                  

###### 4.3.2.5.2.Extension to other commodities:

If the screening method is known or expected to be applicable to other commodities, the validity to these other commodities                        shall be verified. As long as the new commodity belongs to a commodity group (see Table A) for which an initial validation                        has already been performed, a limited additional validation is sufficient. For this, a minimum of 10 homogeneous negative                        control and 10 homogeneous positive control (at STC) samples shall be analysed under intermediate precision conditions. The                        positive control samples shall all be above the cut-off value. In case this criterion is not met, a full validation is required.                  

##### 4.3.2.6.*Verification of methods already validated through collaborative trials*

For screening methods that have already been successfully validated through a collaborative laboratory trial, the method performance                        shall be verified. For this a minimum of 6 negative control and 6 positive control (at STC) samples shall be analysed. The                        positive control samples shall all be above the cut-off value. In case this criterion is not met, the laboratory has to perform                        a root-cause analysis to identify why it cannot meet the specification as obtained in the collaborative trial. Only after                        taking corrective action it shall re-verify the method performance in its laboratory. In case the laboratory is not capable                        to verify the results from the collaborative trial, it will need to establish its own cut-off in a complete single laboratory                        validation.                  

##### 4.3.2.7.*Continuous method verification/on-going method validation*

After initial validation, additional validation data are acquired by including at least two positive control samples in each                        batch of samples screened. One positive control sample is a known sample (e.g. one used during initial validation), the other                        is a different commodity from the same commodity group (in case only one commodity is analysed, a different sample of that                        commodity is used instead). Inclusion of a negative control sample is optional. The results obtained for the two positive                        control samples are added to the existing validation set.                  

At least once a year the cut-off value is re-established and the validity of the method is re-assessed. The continuous method                        verification serves several purposes:                  
          -  quality control for the batch of samples screened                         
          -  providing information on robustness of the method at conditions in the laboratory that applies the method                         
          -  justification of applicability of the method to different commodities                         
          -  allowing to adjust cut-off values in case of gradual drifts over time.                         

##### 4.3.2.8.*Validation report*

The validation report shall contain:                  
          -  A statement on the STC                         
          -  A statement on the obtained cut-off.                         

*Note:* The cut-off must have the same number of significant figures as the STC. Numerical values used to calculate the cut-off need                        at least one more significant figure than the STC.                  
          -  A statement on calculated false suspected rate                         
          -  A statement on how the false suspected rate was generated.                         

*Note:* The statement on the calculated false suspected rate indicates if the method is fit-for-purpose as it indicates the number                        of blank (or low level contamination) samples that will be subject to verification.                  

###### *Table A*

**Commodity groups for the validation of screening methods**

###### *Table B*

**One tailed t-value for a false negative rate of 5 %**

#### 4.3.3.*Requirements for qualitative screening methods (methods that do not give numerical values)*

The development of validation guidelines for binary test methods is currently subject of various standardization bodies (e.g.                        AOAC, ISO). Very recently AOAC has drafted a guideline on this matter. This document can be regarded as the current state                        of the art in its field. Therefore methods that give binary results (e.g. visual inspection of dip-stick tests) should be                        validated according to this guideline                  

http://www.aoac.org/imis15_prod/AOAC_Docs/ISPAM/Qual_Chem_Guideline_Final_Approved_031412.pdf                  

### 4.4.**Estimation of measurement uncertainty, recovery calculation and reporting of results(5)**

#### 4.4.1.*Confirmatory methods*

The analytical result must be reported as follows:                  

Corrected for recovery, the level of recovery being indicated. The correction for recovery is not necessary in case the recovery                                 rate is between 90-110 %.                           

As x +/– U whereby x is the analytical result and U is the expanded measurement uncertainty, using a coverage factor of 2                                 which gives a level of confidence of approximately 95 %.                           

For food of animal origin, the taking into account of the measurement uncertainty can also be done by establishing the decision                        limit (CCα) in accordance with Commission Decision 2002/657/EC(6) (point 3.1.2.5 of Annex I — the case of substances with established permitted limit).                  

However if the result of the analysis is significantly (> 50 %) lower than the maximum level or much higher than the maximum                        level (i.e. more than 5 times the maximum level), and on the condition that the appropriate quality procedures are applied                        and the analysis serves only the purpose of checking compliance with legal provisions, the analytical result might be reported                        without correction for recovery and the reporting of the recovery rate and measurement uncertainty might be omitted in these                        cases.                  

The present interpretation rules of the analytical result in view of acceptance or rejection of the lot apply to the analytical                        result obtained on the sample for official control. In case of analysis for defence or referee purposes, the national rules                        apply.                  

#### 4.4.2.*Screening methods*

The result of the screening shall be expressed as compliant or suspected to be non-compliant.                  

“Suspected to be non-compliant” means the sample exceeds the cut-off level and may contain the mycotoxin at a level higher                        than the STC. Any suspect result triggers a confirmatory analysis for unambiguous identification and quantification of the                        mycotoxin.                  

“Compliant” means that the mycotoxin content in the sample is < STC with a certainty of 95 % (i.e. there is a 5 % chance that                        samples will be incorrectly reported as negative). The analytical result is reported as “< level of STC” with the level of                        STC specified.’                  

1

OJ L 165, 30.4.2004, p. 1.                           

2

Commission Regulation (EC) No 1881/2006 of 19 December 2006 setting maximum levels for certain contaminants in foodstuffs                              (OJ L 364, 20.12.2006, p. 5).                           

3

Commission Regulation (EC) No 401/2006 of 23 February 2006 laying down the methods of sampling and analysis for the official                              control of the levels of mycotoxins in foodstuffs (OJ L 70, 9.3.2006, p. 12).                           

4

Samples are considered free of analyte if the amount present in the sample does not exceed more than 1/5th of the STC. If the level can be quantified with a confirmatory method, the level must be taken into consideration for the                              validation assessment.                           

5

More details on procedures for the estimation of measurement uncertainty and on procedures for assessing recovery can be found                              in the report “Report on the relationship between analytical results, measurement uncertainty, recovery factors and the provisions                              of EU food and feed legislation” — http://ec.europa.eu/food/food/chemicalsafety/contaminants/report-sampling_analysis_2004_en.pdf                           

6

Commission Decision 2002/657/EC of 14 August 2002 implementing Council Directive 96/23/EC concerning the performance of analytical                              methods and the interpretation of results (OJ L 221, 17.8.2002, p. 8).