Commission Regulation (EC) No 900/2008 of 16 September 2008 laying down the methods of analysis and other technical provisions necessary for the application of the arrangements for imports of certain goods resulting from the processing of agricultural products (Codified version)

Article 1

Scope

This Regulation lays down the following:

(a) the methodology and methods of analysis to be used for determining the content of the agricultural products within the meaning of Article 2(1)(a) of Council Regulation (EC) No 1216/2009 (⁵) or their specific components considered to have been incorporated in imported goods within the meaning of Article 2(1)(b) of Regulation (EC) No 1216/2009;

(b) the necessary methods of analysis to be used for the implementation of Regulation (EC) No 1216/2009 as far as imports of certain goods are concerned, of Annex I to Regulation (EEC) No 2658/1987 and of Commission Implementing Regulation (EU) No 514/2011 (⁶) or in the absence of a method of analysis, the nature of the analytical operations to be carried out or the principle of a method to be applied.

Article 2

Calculation of contents

In accordance with the definitions set out in footnotes 1, 2 and 3 of Annex III to Regulation (EU) No 514/2011 and in footnotes 1, 2 and 3 of Part Three, Section I, Annex 1, Table 1 of Annex I to Regulation (EEC) No 2658/87 concerning milk protein content, starch/glucose content and sucrose/invert sugar/isoglucose content, the following formulas, procedures and methods shall be used:

(a) for the application of Annexes II and III to Regulation (EU) No 514/2011;

(b) for the determination of milk fat content, milk protein content, starch/glucose content and sucrose/invert sugar/isoglucose content for the purpose of selecting the appropriate agricultural element, additional duties for sugar and additional duties for flour in the case of non-preferential imports as provided for in Part Two and in Part Three, Section I, Annex 1, of Annex I to Regulation (EEC) No 2658/87:

(expressed as 100 % anhydrous starch content of the goods as presented) (a) (Z – F) × 0,9, if the glucose content is not less than the fructose content; or (b) (Z – G) × 0,9, if the glucose content is less than the fructose content where: Z = is the glucose content determined by the method set out in Annex I to this Regulation; F = is the fructose content determined by HPLC (high performance liquid chromatography); G = is the glucose content determined by HPLC. In the case of point (a), where the presence of a lactose hydrolysate is declared and/or quantities of lactose and galactose are detected, a glucose content equivalent to the galactose content (determined by HPLC) shall be deducted from the glucose content (Z) before any other calculation is made.

(expressed as sucrose content of the goods as presented) (a) S + (2F) × 0,95, if the glucose content is not less than the fructose content; (b) S + (G + F) × 0,95, if the glucose content is less than the fructose content where: S = is the sucrose content determined by HPLC; F = is the fructose content determined by HPLC; G = is the glucose content determined by HPLC. Where the presence of a lactose hydrolysate is declared and/or quantities of lactose and galactose are detected, a glucose content equivalent to the galactose content (determined by HPLC) shall be deducted from the glucose (G) content before any other calculation is made.

(a) Save as otherwise provided in point (b) or (c), the milk fat content by weight of the goods as presented shall be determined by extraction with light petroleum after hydrolysis with hydrochloric acid; (b) Where fats other than milk fats are also declared in the composition of the goods, the following procedure shall be applied: — the percentage of weight of the total fats in the goods shall be determined as mentioned in point (a), — for the purposes of determining the milk fat content, a method based on extraction with light petroleum, preceded by hydrolysis with hydrochloric acid and followed by gas chromatography of the methyl esters of the fatty acids shall be used. If the presence of milk fats is detected, the percentage proportion thereof shall be calculated by multiplying the percentage concentration of methyl butyrate by 25, multiplying the product by the total percentage fat content by weight of the goods and dividing by 100. (c) Where fats other than milk fats are also declared in the composition of goods which are charged by an agricultural component as provided for in Part Two and in Annex 1 to Section I of Part Three of the Combined nomenclature set out in Annex I to Regulation (EEC) No 2658/87, and contain 30 % milk protein or more as determined in accordance with point 4 of this Article, and less than 6 % milk fat as declared by the declarant, the following procedure shall be applied instead of the procedure set out in point (b): — the percentage of weight of the total fats in the goods shall be determined as set out in point (a), — for the purposes of determining the milk fat content, a method based on extraction with light petroleum, preceded by hydrolysis with hydrochloric acid and followed by gas chromatography of the methyl esters of the fatty acids shall be used. If the presence of milk fats is detected, the percentage proportion thereof shall be calculated by multiplying the percentage concentration of methyl butyrate by 50, multiplying the product by the total percentage fat content by weight of the goods and dividing by 100.

(a) Save as otherwise provided in point (b), the milk protein content of the goods shall be calculated by multiplying the nitrogen content (determined by the Kjeldahl method) by the factor 6,38; (b) Where components containing proteins other than milk proteins are also declared in the composition of the goods: — the total percentage nitrogen content by weight shall be determined by the Kjeldahl method, — the milk protein content shall be calculated as indicated in point (a) by subtracting from the total percentage nitrogen content the nitrogen content corresponding to the non-milk proteins.

Article 3

Classification of Goods

For the purpose of applying Annex I to Regulation (EU) No 514/2011 and Annex I to Regulation (EEC) No 2658/87, the following methods and procedures shall be used for the classification of the following goods:

Article 4

Test report

The test report shall include the following particulars:

— all the information necessary for identifying the sample,

— the Community method used and precise reference to the legal instrument in which it is laid down, or, where appropriate, detailed reference to a method, specifying the nature of the analytical operations to be carried out, or the principle of the method to be applied, as indicated in this Regulation,

— any factors liable to have influenced the results,

— the results of the analysis, with due regard to the way in which they are expressed in the method used and the means of expression dictated by the needs of the customs or administrative departments that requested the analysis.

Article 4a

The information referred to in paragraph 1 shall be communicated in an electronic form and consist of:

(a) the date of acceptance of the customs declaration;

(b) the quantity of the goods concerned;

(c) tariff classification of the goods concerned;

(d) the additional code declared and, for goods to which the method of analysis set out in point (c) of Article 2(3) has been applied, the additional code applicable on the basis of the results of that method;

(e) the milk protein content as determined by the method of analysis referred to in Article 2(4);

(f) indication of the method of analysis applied for the determination of milk fat content in accordance with Article 2(3);

(g) the total fat content as determined by the method of analysis referred to in point (a) of Article 2(3);

(h) the milk fat content as determined by one of the methods of analysis referred to in Article 2(3);

(i) where relevant, the type of fat other than milk fat contained in the processed agricultural product.

Article 5

Final provision

Regulation (EEC) No 4154/87 is repealed.

References to the repealed Regulation shall be construed as references to this Regulation and shall be read in accordance with the correlation table in Annex V.

Article 6

This Regulation shall enter into force on the 20th day following its publication in the Official Journal of the European Union.

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

ANNEX I

Enzymatic determination of starch and its degradation products including glucose in food products using high performance liquid chromatography (HPLC)

1. Scope

This method describes the determination of the content of starch and its degradation products including glucose in food products for human consumption hereafter referred to as ‘starch’. The starch content is determined from the quantitative analysis of glucose by high-performance liquid chromatography (HPLC) after enzymatic conversion of starch and its degradation products into glucose.

2. Definition of the total glucose content and of the total glucose content expressed as starch

The total glucose content means the value Z as calculated in point 7.2.1 of this Annex. It represents the content of starch and all its degradation products, glucose included.

The starch/glucose content as defined in Annex III to Regulation (EC) No 1460/96 shall be calculated on the basis of the total glucose content Z and as set out in Article 2 point 1 of this Regulation.

The starch (or dextrin) content as referred to in column 3 of Annex IV to Commission Regulation (EC) No 1043/2005 (⁷) shall be calculated on the basis of the total glucose content Z as set out in Article 2 point 2.1 of Commission Regulation (EC) No 904/2008 (⁸).

The Starch content referred to in point 1 of this Annex means the value E, as calculated in point 7.2.2 of this Annex. It is expressed in % (m/m). It is equivalent to the total glucose content Z, expressed as starch. This value E does not interfere in the above mentioned calculations.

3. Principle

The samples are homogenised and suspended in water. The starch and its degradation products, present in the samples, are enzymatically converted into glucose in two steps:

Products containing a high content of proteins or fat are clarified and filtrated.

The determination of sugars is performed by HPLC analysis.

Because a partial inversion of sucrose may occur during the enzymatic treatment, the determination of free sugars is also performed by HPLC analysis to calculate the corrected glucose content.

4. Reagents and other materials

Use reagents of recognised analytical grade and demineralised water.

4.1. Glucose, min 99 %.

4.2. Fructose, min 99 %.

4.3. Sucrose, min 99 %.

4.4. Maltose-monohydrate, min 99 %.

4.5. Lactose-monohydrate, min 99 %.

4.6. Solution of thermostable alpha-amylase (1,4-alpha-D-Glucan-glucanohydrolase), with activity about 31 000 U/ml (1U will liberate 1,0 mg of maltose from starch in 3 minutes at pH 6,9 and 20 °C). This enzyme can contain a low amount of impurities (e.g. glucose or sucrose) and other interfering enzymes. Storage at ca. 4 °C. Alternatively, other sources of alpha-amylase may be used yielding a final solution with comparable enzyme activity.

4.7. Amyloglucosidase (1,4-alpha-D-Glucan glucohydrolase) from Aspergillus niger, powder with activity about 120 U/mg or about 70 U/mg (1U will liberate 1 micromol glucose from starch per minute at pH 4,8 and 60 °C). This enzyme can contain a low amount of impurities (e.g. glucose or sucrose) and other interfering enzymes (e.g., Invertase). Storage at ca. 4 °C. Alternatively, other sources of amyloglucosidase may be used yielding a final solution with comparable enzyme activity.

4.8. Zinc acetate dihydrate, p.a..

4.9. Potassium hexacyanoferrate (II) (K4[Fe(CN)]6.3H2O), extra pure.

4.10. Sodium acetate anhydrous, p.a..

4.11. Glacial acetic acid, 96 % (v/v) (minimum).

4.12. Sodium acetate buffer (0,2 mol/l). Weigh 16,4 gram sodium acetate (point 4.10) into a beaker glass. Dissolve in water and rinse into a volumetric flask of 1 000 ml. Dilute to the mark with water and adjust the pH to 4,7 with acetic acid (by use of a pH-meter (point 5.7). This solution may be used for max 6 months with storage at 4 °C.

4.13. Amyloglucosidase solution. Prepare a solution of amyloglucosidase powder (point 4.7) by using sodium acetate buffer (point 4.12). The enzyme activity must be sufficient and in accordance with the starch content in the amount of sample (for example, activity about 600 U/ml is obtained from 0,5 g amyloglucosidase powder 120 U/mg (point 4.7) in a final volume of 100 ml for 1 g starch in the amount of sample). Prepare immediately before use.

4.14. Reference solutions. Prepare solutions of glucose, fructose, sucrose, maltose and lactose in water, as conventionally used in the HPLC analysis of sugars.

4.15. Reagent for clarification (Carrez I). Dissolve 219,5 gram zinc acetate (point 4.8) in water in a beaker glass. Rinse into a volumetric flask of 1 000 ml and add 30 ml acetic acid (point 4.11). Mix thoroughly and dilute to the mark with water. This solution may be used for max 6 months while stored at ambient temperature. Other clarification reagents, equivalent to Carrez solution, may be used.

4.16. Reagent for clarification (Carrez II). Dissolve 106,0 gram potassium hexacyanoferrate (II) (point 4.9) in water in a beaker glass. Rinse into a volumetric flask of 1 000 ml. Mix thoroughly and dilute to the mark with water. This solution may be used for max. 6 months while stored at ambient temperature. Other clarification reagents, equivalent to Carrez solution, may be used.

4.17. HPLC Mobile phase. Prepare a mobile phase which is conventionally used in the HPLC analysis of sugars. In case of using an aminopropyl silicagel column, e.g., a common mobile phase is a mixture of HPLC grade water and acetonitrile.

5. Apparatus

5.1. Standard laboratory glass ware.

5.2. Fluted filters, e.g., 185 mm.

5.3. Syringe filters, 0,45 μm, suitable for aqueous solutions.

5.4. Sample vials suitable for the HPLC autosampler.

5.5. 100 ml volumetric flasks.

5.6. Plastic syringes, 10 ml.

5.7. pH-meter.

5.8. Analytical balance.

5.9. Water bath with thermostat, adjustable to 60 °C and 90 °C.

5.10. HPLC Apparatus suitable for analysis of sugars.

6. Procedure

The product is homogenised.

The amount of sample is estimated from the ingredient declaration and the conditions of the HPLC analysis (concentration of the glucose reference solution), and shall not exceed:

Weigh the sample to 0,1 mg accuracy.

The blank is determined by performing a complete analysis (as described in point 6.4), without adding sample. The result of the blank determination is used in the calculation of the starch content (point 7.2).

Homogenise the sample by shaking or stirring. The chosen test portion (point 6.2) is weighed into a volumetric flask (point 5.5) and about 70 ml warm water is added.