Commission Regulation (EC) No 333/2007Show full title

PART C U.K. SAMPLE PREPARATION AND ANALYSIS

C.1. LABORATORY QUALITY STANDARDS U.K.

Laboratories shall comply with the provisions of Article 12 of Regulation (EC) No 882/2004 .

Laboratories shall participate in appropriate proficiency testing schemes which comply with the ‘International Harmonised Protocol for the Proficiency Testing of (Chemical) Analytical Laboratories’(1) developed under the auspices of IUPAC/ISO/AOAC.

Laboratories shall be able to demonstrate that they have internal quality control procedures in place. Examples of these are the ‘ISO/AOAC/IUPAC Guidelines on Internal Quality Control in Analytical Chemistry Laboratories’(2).

Wherever possible the trueness of analysis shall be estimated by including suitable certified reference materials in the analysis.

C.2. SAMPLE PREPARATION U.K.

C.2.1. Precautions and general considerations U.K.

The basic requirement is to obtain a representative and homogeneous laboratory sample without introducing secondary contamination.

All of the sample material received by the laboratory shall be used for the preparation of the laboratory sample.

Compliance with maximum levels laid down in Regulation (EC) No 1881/2006 shall be established on the basis of the levels determined in the laboratory samples.

C.2.2. Specific sample preparation procedures U.K.

[F1 C.2.2.1. Specific procedures for lead, cadmium, mercury, inorganic tin and inorganic arsenic U.K.

The analyst shall ensure that samples do not become contaminated during sample preparation. Wherever possible, apparatus and equipment coming into contact with the sample shall not contain those metals to be determined and be made of inert materials, e.g. plastics such as polypropylene, polytetrafluoroethylene (PTFE) etc. These should be acid cleaned to minimise the risk of contamination. High quality stainless steel may be used for cutting edges.

There are many satisfactory specific sample preparation procedures which may be used for the products under consideration. For those aspects not specifically covered by this Regulation, the CEN Standard ‘Foodstuffs. Determination of elements and their chemical species. General considerations and specific requirements’(3) has been found to be satisfactory but other sample preparation methods may be equally valid.

In the case of inorganic tin, care shall be taken to ensure that all the material is taken into solution as losses are known to occur readily, particularly because of hydrolysis to insoluble hydrated Sn(IV) oxide species.]

Textual Amendments

F1 Substituted by Commission Regulation (EU) 2016/582 of 15 April 2016 amending Regulation (EC) No 333/2007 as regards the analysis of inorganic arsenic, lead and polycyclic aromatic hydrocarbons and certain performance criteria for analysis (Text with EEA relevance).

[F2 C.2.2.2. Specific procedures for polycyclic aromatic hydrocarbons U.K.

The analyst shall ensure that samples do not become contaminated during sample preparation. Containers shall be rinsed with high purity acetone or hexane before use to minimise the risk of contamination. Wherever possible, apparatus and equipment coming into contact with the sample shall be made of inert materials such as aluminium, glass or polished stainless steel. Plastics such as polypropylene or PTFE shall be avoided because the analytes can adsorb onto these materials.

[F3 For the analysis of PAH in cocoa and cocoa derived products, the determination of the fat content is performed in accordance with AOAC Official method 963.15 for the determination of the fat content of cocoa beans and derived products. Equivalent fat determination procedures can be applied for which it can be demonstrated that the used fat determination procedure provides an equal (equivalent) fat content value.]]

Textual Amendments

F3 Inserted by Commission Regulation (EU) 2016/582 of 15 April 2016 amending Regulation (EC) No 333/2007 as regards the analysis of inorganic arsenic, lead and polycyclic aromatic hydrocarbons and certain performance criteria for analysis (Text with EEA relevance).

Textual Amendments

F2 Substituted by Commission Regulation (EU) No 836/2011 of 19 August 2011 amending Regulation (EC) No 333/2007 laying down the methods of sampling and analysis for the official control of the levels of lead, cadmium, mercury, inorganic tin, 3-MCPD and benzo(a)pyrene in foodstuffs (Text with EEA relevance).

C.2.3. Treatment of the sample as received in the laboratory U.K.

The complete aggregate sample shall be finely ground (where relevant) and thoroughly mixed using a process that has been demonstrated to achieve complete homogenisation.

C.2.4. Samples for enforcement, defence and referee purposes U.K.

The samples for enforcement, defence and referee purposes shall be taken from the homogenised material unless this conflicts with [F4 the rules applicable in each constituent territory of Great Britain'] on sampling as regards the rights of the food business operator.

Textual Amendments

F4 Words in Annex para. C.2.4 substituted (31.12.2020) by S.I. 2019/639, reg. 30(d) (as substituted by The Food and Feed Hygiene and Safety (Miscellaneous Amendments etc.) (EU Exit) Regulations 2020 (S.I. 2020/1504), regs. 1(2), 8(10))

C.3. METHODS OF ANALYSIS U.K.

C.3.1. Definitions U.K.

The following definitions shall apply:

‘r’

=

Repeatability the value below which the absolute difference between single test results obtained under repeatability conditions (i.e., same sample, same operator, same apparatus, same laboratory, and short interval of time) may be expected to lie within a specific probability (typically 95 %) and hence r = 2,8 × s_r.

‘s_r’

=

Standard deviation calculated from results generated under repeatability conditions.

‘RSD_r’

=

Relative standard deviation calculated from results generated under repeatability conditions [(s_r/) × 100].

‘R’

=

Reproducibility the value below which the absolute difference between single test results obtained under reproducibility conditions (i.e., on identical material obtained by operators in different laboratories, using the standardised test method), may be expected to lie within a certain probability (typically 95 %); R = 2,8 × s_R.

‘s_R’

=

Standard deviation, calculated from results under reproducibility conditions.

‘RSD_R’

=

Relative standard deviation calculated from results generated under reproducibility conditions [(s_R/) × 100].

‘[F5 LOD’

=

Limit of detection, smallest measured content, from which it is possible to deduce the presence of the analyte with reasonable statistical certainty.

‘LOQ’

=

Limit of quantification, lowest content of the analyte which can be measured with reasonable statistical certainty.]

‘[F2 HORRAT (4) _r ’

=

The observed RSD _r divided by the RSD _r value estimated from the (modified) Horwitz equation(5) (cf. point C.3.3.1 (‘Notes to the performance criteria’)) using the assumption r = 0,66 R.]

‘[F2 HORRAT (6) _R ’

=

The observed RSD _R divided by the RSD _R value estimated from the (modified) Horwitz equation(7) (cf. point C.3.3.1 (‘Notes to the performance criteria’)).]

‘[F2 u’

=

Combined standard measurement uncertainty obtained using the individual standard measurement uncertainties associated with the input quantities in a measurement model](8)

‘U’

=

The expanded measurement uncertainty, using a coverage factor of 2 which gives a level of confidence of approximately 95 % (U = 2u).

‘Uf’

=

Maximum standard measurement uncertainty.

Textual Amendments

F5 Substituted by Commission Implementing Regulation (EU) 2019/2093 of 29 November 2019 amending Regulation (EC) No 333/2007 as regards the analysis of 3-monochloropropane-1,2-diol (3-MCPD) fatty acid esters, glycidyl fatty acid esters, perchlorate and acrylamide (Text with EEA relevance).

[F1 C.3.2. General requirements U.K.

Methods of analysis used for food control purposes shall comply with the provisions of Annex III to Regulation (EC) No 882/2004.

Methods for analysis for total tin are appropriate for control on inorganic tin levels.

For the analysis of lead in wine, the methods and rules established by the OIV(9) apply in accordance with Article 80(5) of Regulation (EU) No 1308/2013(10).

Methods for analysis for total arsenic are appropriate for screening purpose for control on inorganic arsenic levels. If the total arsenic concentration is below the maximum level for inorganic arsenic, no further testing is required and the sample is considered to be compliant with the maximum level for inorganic arsenic. If the total arsenic concentration is at or above the maximum level for inorganic arsenic, follow-up testing shall be conducted to determine if the inorganic arsenic concentration is above the maximum level for inorganic arsenic.]

C.3.3. Specific requirements U.K.

[F2 C.3.3.1. Performance criteria U.K.

Where no specific methods for the determination of contaminants in foodstuffs are prescribed F6 ..., laboratories may select any validated method of analysis for the respective matrix provided that the selected method meets the specific performance criteria set out in Tables 5, 6 and 7.

Textual Amendments

F6 Words in Annex para. C.3.3.1 omitted (31.12.2020) by virtue of S.I. 2019/639, reg. 30(e) (as substituted by The Food and Feed Hygiene and Safety (Miscellaneous Amendments etc.) (EU Exit) Regulations 2020 (S.I. 2020/1504), regs. 1(2), 8(10))

It is recommended that fully validated methods (i.e. methods validated by collaborative trial for the respective matrix) are used where appropriate and available. Other suitable validated methods (e.g. in-house validated methods for the respective matrix) may also be used provided that they fulfil the performance criteria set out in Tables 5, 6 and 7.

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

(a)

[F1 Performance criteria for methods of analysis for lead, cadmium, mercury, inorganic tin and inorganic arsenic

Table 5

Parameter	Criterion
Applicability	Foods specified in Regulation (EC) No 1881/2006
Specificity	Free from matrix or spectral interferences
Repeatability (RSD r )	HORRAT r less than 2
Reproducibility (RSD R )	HORRAT R less than 2
Recovery	The provisions of point D.1.2 apply
LOD	= three tenths of LOQ
LOQ	Inorganic tin	≤ 10 mg/kg
	Lead	ML ≤ 0,01 mg/kg	0,01 < ML ≤ 0,02 mg/kg	0,02 < ML < 0,1 mg/kg	ML ≥ 0,1 mg/kg
		≤ ML	≤ two thirds of the ML	≤ two fifths of the ML	≤ one fifth of the ML
	Cadmium, mercury, inorganic arsenic	ML is < 0,100 mg/kg		ML is ≥ 0,100 mg/kg
		≤ two fifths of the ML		≤ one fifth of the ML]

(b)

[F5 Performance criteria for methods of analysis for 3-monochloropropane-1,2-diol (3-MCPD), 3-MCPD fatty acid esters and glycidyl fatty acid esters:

• Performance criteria for methods of analysis for 3-MCPD in foods specified in point 4.1 of the Annex to Regulation (EC) No 1881/2006

  Table 6a

  Parameter	Criterion
  Applicability	Foods specified in point 4.1 of the Annex to Regulation (EC) No 1881/2006
  Specificity	Free from matrix or spectral interferences
  Field blanks	Less than LOD
  Repeatability (RSD r )	0,66 times RSD R as derived from (modified) Horwitz equation
  Reproducibility (RSD R )	as derived from (modified) Horwitz equation
  Recovery	75-110 %
  Limit of Detection (LOD)	≤ 5 μg/kg (on dry matter basis)
  Limit of Quantification (LOQ)	≤ 10 μg/kg (on dry matter basis)

• Performance criteria for methods of analysis for 3-MCPD in foods specified in point 4.3 of the Annex to Regulation (EC) No 1881/2006

  Table 6b

  Parameter	Criterion
  Applicability	Foods specified in point 4.3 of the Annex to Regulation (EC) No 1881/2006
  Specificity	Free from matrix or spectral interferences
  Field blanks	Less than LOD
  Repeatability (RSD r )	0,66 times RSD R as derived from (modified) Horwitz equation
  Reproducibility (RSD R )	as derived from (modified) Horwitz equation
  Recovery	75-110 %
  Limit of Detection (LOD)	≤ 7 μg/kg
  Limit of Quantification (LOQ)	≤ 14 μg/kg

• Performance criteria for methods of analysis for 3-MCPD fatty acid esters, expressed as 3-MCPD , in foods specified in point 4.3 of the Annex to Regulation (EC) No 1881/2006

  Table 6c

  Parameter	Criterion
  Applicability	Foods specified in point 4.3 of the Annex to Regulation (EC) No 1881/2006
  Specificity	Free from matrix or spectral interferences
  Repeatability (RSD r )	0,66 times RSD R as derived from (modified) Horwitz equation
  Reproducibility (RSD R )	as derived from (modified) Horwitz equation
  Recovery	70-125 %
  Limit of Detection (LOD)	Three tenths of LOQ
  Limit of Quantification (LOQ) for foods specified in 4.3.1 and 4.3.2	≤ 100 μg/kg in oils and fats
  Limit of Quantification (LOQ) for foods specified in 4.3.3 and in 4.3.4 with a fat content < 40 %	≤ two fifths of the ML
  Limit of Quantification (LOQ) for foods specified in 4.3.4 with a fat content ≥ 40 %	≤ 15 μg/kg fat

• Performance criteria for methods of analysis for glycidyl fatty acid esters, expressed as glycidol, in foods specified in point 4.2 of the Annex to Regulation (EC) No 1881/2006

  Table 6d

  Parameter	Criterion
  Applicability	Foods specified in point 4.2 of the Annex to Regulation (EC) No 1881/2006
  Specificity	Free from matrix or spectral interferences
  Repeatability (RSD r )	0,66 times RSD R as derived from (modified) Horwitz equation
  Reproducibility (RSD R )	as derived from (modified) Horwitz equation
  Recovery	70-125 %
  Limit of Detection (LOD)	Three tenths of LOQ
  Limit of Quantification (LOQ) for foods specified in 4.2.1 and 4.2.2	≤ 100 μg/kg in oils and fats
  Limit of Quantification (LOQ) for foods specified in 4.2.3 with a fat content < 65 % and in 4.2.4 with a fat content < 8 %	≤ two fifths of the ML
  Limit of Quantification (LOQ) for foods specified in 4.2.3 with a fat content ≥ 65 % and in 4.2.4 with a fat content ≥ 8 %	≤ 31 μg/kg fat]

(c)

Performance criteria for methods of analysis for polycyclic aromatic hydrocarbons:

The four polycyclic aromatic hydrocarbons to which these criteria apply are benzo(a)pyrene, benz(a)anthracene, benzo(b)fluoranthene and chrysene.

Table 7

Parameter	Criterion
Applicability	Foods specified in Regulation (EC) No 1881/2006
Specificity	Free from matrix or spectral interferences, verification of positive detection
Repeatability (RSD r )	HORRAT r less than 2
Reproducibility (RSD R )	HORRAT R less than 2
Recovery	50-120 %
LOD	≤ 0,30 μg/kg for each of the four substances
LOQ	≤ 0,90 μg/kg for each of the four substances

(d)

[F5 Performance criteria for methods of analysis for acrylamide:

Table 8

Parameter	Criterion
Applicability	All foods
Specificity	Free from matrix or spectral interferences
Field blanks	Less than Limit of Detection (LOD)
Repeatability (RSD r )	0,66 times RSD R as derived from (modified) Horwitz equation
Reproducibility (RSD R )	as derived from (modified) Horwitz equation
Recovery	75-110 %
Limit of Detection (LOD)	Three tenths of LOQ
Limit of Quantification (LOQ)	For foods with benchmark levels < 125 μg/kg: ≤ two fifths of the benchmark level, however not required to be lower than 20 μg/kg For foods with benchmark level ≥ 125 μg/kg: ≤ 50 μg/kg]

(e)

[F7 Performance criteria for methods of analysis for perchlorate:

Table 9

Parameter	Criterion
Applicability	All foods
Specificity	Free from matrix or spectral interferences
Repeatability (RSD r )	0,66 times RSD R as derived from (modified) Horwitz equation
Reproducibility (RSD R )	as derived from (modified) Horwitz equation
Recovery	70-110 %
Limit of Detection (LOD)	Three tenths of LOQ
Limit of Quantification (LOQ)	≤ two fifths of the ML

(f)

Notes to the performance criteria:

The Horwitz equation(11) (for concentrations 1,2 x 10 ^-7 ≤ C ≤ 0,138) and the modified Horwitz equation(12) (for concentrations C < 1,2 × 10 ^-7 ) are generalised precision equations which are independent of analyte and matrix but solely dependent on concentration for most routine methods of analysis.

Modified Horwitz equation for concentrations C < 1,2 × 10 ^-7 :

RSD _R = 22 %

where:

• RSD _R is the relative standard deviation calculated from results generated under reproducibility conditions

• C is the concentration ratio (i.e. 1 = 100g/100g, 0,001 = 1 000 mg/kg). The modified Horwitz equation applies to concentrations C < 1,2 × 10 ^-7 .

Horwitz equation for concentrations 1,2 x 10 ^-7 ≤ C ≤ 0,138:

RSD _R = 2C ^{(- 0,15 )}

where:

• RSD _R is the relative standard deviation calculated from results generated under reproducibility conditions

• C is the concentration ratio (i.e. 1 = 100g/100g, 0,001 = 1 000 mg/kg). The Horwitz equation applies to concentrations 1,2 x 10 ^-7 ≤ C ≤ 0,138.]]

Textual Amendments

F7 Inserted by Commission Implementing Regulation (EU) 2019/2093 of 29 November 2019 amending Regulation (EC) No 333/2007 as regards the analysis of 3-monochloropropane-1,2-diol (3-MCPD) fatty acid esters, glycidyl fatty acid esters, perchlorate and acrylamide (Text with EEA relevance).

[F2 C.3.3.2. ‘Fitness-for-purpose’ approach U.K.

For in-house validated methods, as an alternative a ‘fitness-for-purpose’ approach(13) may be used to assess their suitability for official control. Methods suitable for official control must produce results with a combined 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). The LOD must meet the performance criteria set in point C.3.3.1 for the concentration of interest.

• C is the concentration of interest (μg/kg);

• α is a numeric factor to be used depending on the value of C. The values to be used are given in [F5 Table 10].

[F5 Table 10]

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

C (μg/kg)	α
≤ 50	0,2
51-500	0,18
501- 1 000	0,15
1 001 - 10 000	0,12
> 10 000	0,1

The analyst shall note the ‘Report on the relationship between analytical results, measurement uncertainty, recovery factors and the provisions of EU food and feed legislation’(14).]

(1)

‘The international harmonized protocol for the proficiency testing of analytical chemistry laboratories’ by M. Thompson, S.L.R. Ellison and R. Wood, Pure Appl. Chem., 2006, 78, 145-96.

(2)

Edited by M. Thompson and R. Wood, Pure Appl. Chem., 1995, 67, 649-666.

(3)

[F1 Standard EN 13804:2013, ‘ Foodstuffs. Determination of elements and their chemical species. General considerations and specific requirements ’ , CEN, Rue de Stassart 36, B-1050 Brussels .]

(4)

[F2 Horwitz W. and Albert, R., 2006, The Horwitz Ratio (HorRat): A useful Index of Method Performance with respect to Precision, Journal of AOAC International, Vol. 89, 1095-1109.]

(5)

[F2 M. Thompson, Analyst, 2000, p. 125 and 385-386.]

(6)

[F2 Horwitz W. and Albert, R., 2006, The Horwitz Ratio (HorRat): A useful Index of Method Performance with respect to Precision, Journal of AOAC International, Vol. 89, 1095-1109.]

(7)

[F2 M. Thompson, Analyst, 2000, p. 125 and 385-386.]

(8)

[F2 International vocabulary of metrology – Basic and general concepts and associated terms (VIM), JCGM 200:2008.]

(9)

[F1 Organisation internationale de la vigne et du vin.]

(10)

[F1 Regulation (EU) No 1308/2013 of the European Parliament and of the Council of 17 December 2013 establishing a common organisation of the markets in agricultural products and repealing Council Regulations (EEC) No 922/72, (EEC) No 234/79, (EC) No 1037/2001 and (EC) No 1234/2007 ( OJ L 347, 20.12.2013, p. 671 ).]

(11)

[F2 [F7 W. Horwitz, L.R. Kamps, K.W. Boyer, J.Assoc.Off.Analy.Chem.,63, 1980, 1344-1354.] ]

(12)

[F2 [F7 M. Thompson, Analyst, 125, 2000, 385-386.] ]

(13)

[F2 M. Thompson and R. Wood, Accred. Qual. Assur., 2006, p. 10 and 471-478.]

(14)

[F2 http://ec.europa.eu/food/food/chemicalsafety/contaminants/report-sampling_analysis_2004_en.pdf]

Textual Amendments

F1 Substituted by Commission Regulation (EU) 2016/582 of 15 April 2016 amending Regulation (EC) No 333/2007 as regards the analysis of inorganic arsenic, lead and polycyclic aromatic hydrocarbons and certain performance criteria for analysis (Text with EEA relevance).

F2 Substituted by Commission Regulation (EU) No 836/2011 of 19 August 2011 amending Regulation (EC) No 333/2007 laying down the methods of sampling and analysis for the official control of the levels of lead, cadmium, mercury, inorganic tin, 3-MCPD and benzo(a)pyrene in foodstuffs (Text with EEA relevance).

F7 Inserted by Commission Implementing Regulation (EU) 2019/2093 of 29 November 2019 amending Regulation (EC) No 333/2007 as regards the analysis of 3-monochloropropane-1,2-diol (3-MCPD) fatty acid esters, glycidyl fatty acid esters, perchlorate and acrylamide (Text with EEA relevance).