BS EN 15850-2010 Foodstuffs - Determination of zearalenone in maize based baby food, barley flour, maize flour, polenta.pdf
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1、 May EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 15850 April 2010 ICS 67.060 English Version Foodstuffs - Determination of zearalenone in maize based baby food, barley flour, maize flour, polenta, wheat flour and cereal based foods for infants and young children - HPLC method with immunoaff
2、inity column cleanup and fluorescence detection Produits alimentaires - Dosage de la zaralnone dans la farine dorge, de mas et de bl, la polenta et les produits pour nourrissons et jeunes enfants base de crales - Mthode par chromatographie liquide haute performance avec purification sur colonne dimm
3、unoaffinit et dtection par fluorescence Lebensmittel - Bestimmung von Zearalenon in Suglingsnahrung auf Maisbasis, Gerstenmehl, Maismehl, Maisgrie, Weizenmehl und Lebensmittel auf Getreidebasis fr Suglinge und Kleinkinder - HPLC- Verfahren mit Reinigung an einer Immunoaffinittssule und Fluoreszenzde
4、tektion This European Standard was approved by CEN on 27 February 2010. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European Standard the status of a national standard without any alteration. Up-to-date lists and bibliograp
5、hical references concerning such national standards may be obtained on application to the CEN Management Centre or to any CEN member. This European Standard exists in three official versions (English, French, German). A version in any other language made by translation under the responsibility of a
6、CEN member into its own language and notified to the CEN Management Centre has the same status as the official versions. CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland,
7、 Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Mar
8、nix 17, B-1000 Brussels 2010 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 15850:2010: E EN 15850:2010 (E) 2 Contents Page Foreword 3 1Scope 4 2Normative references 4 3Principle 4 4Reagents .4 5Apparatus .7 6Procedure .9 7HPLC an
9、alysis . 10 8Calculation . 12 9Precision 12 10Test report . 13 Annex A (informative) Typical chromatograms 15 Annex B (informative) Precision data . 16 Bibliography . 18 EN 15850:2010 (E) 3 Foreword This document (EN 15850:2010) has been prepared by Technical Committee CEN/TC 275 “Food analysis - Ho
10、rizontal methods”, the secretariat of which is held by DIN. This European Standard shall be given the status of a national standard, either by publication of an identical text or by endorsement, at the latest by October 2010, and conflicting national standards shall be withdrawn at the latest by Oct
11、ober 2010. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. CEN and/or CENELEC shall not be held responsible for identifying any or all such patent rights. This document has been prepared under a mandate given to CEN by the Europea
12、n Commission and the European Free Trade Association. WARNING The use of this standard can involve hazardous materials, operations and equipment. This standard does not purport to address all the safety problems associated with its use. It is the responsibility of the user of this standard to establ
13、ish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use. According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulg
14、aria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and the United Kingdom. EN 15850:2010 (E)
15、4 1 Scope This European Standard specifies a method for the determination of zearalenone in maize based baby food, barley flour, maize flour, polenta, wheat flour and cereal based foods for infants and young children by high performance liquid chromatography (HPLC) with immunoaffinity cleanup and fl
16、uorescence detection. This method has been validated in two interlaboratory studies. The first study was for the analysis of samples of maize based baby food, barley flour, maize flour, polenta and wheat flour ranging from 10 g/kg to 335 g/kg, and the second study was for samples of cereal based foo
17、ds for infants and young children ranging from 9 g/kg to 44 g/kg. Further information on validation, see Clause 9 and Annex B. 2 Normative references The following referenced documents are indispensable for the application of this document. For dated references, only the edition cited applies. For u
18、ndated references, the latest edition of the referenced document (including any amendments) applies. EN ISO 3696:1995, Water for analytical laboratory use Specification and test methods (ISO 3696: 987). 3 Principle A test portion is extracted with aqueous acetonitrile or methanol according to the pr
19、oducts analyzed. The extract is then diluted with phosphate buffered saline (PBS) to give an aqueous extract that is applied to an immunoaffinity column containing antibodies specific for zearalenone. Zearalenone is purified and concentrated on the column and removed from the antibodies using aceton
20、itrile or methanol as eluent. Zearalenone is quantified by reverse-phase high performance liquid chromatography (RP-HPLC) with fluorescence detection. 4 Reagents 4.1 General Use only reagents of recognised analytical grade and water complying with grade 1 of EN ISO 3696:1995, unless otherwise specif
21、ied. Solvents shall be of quality for HPLC analysis, unless otherwise specified. Commercially available solutions with equivalent properties to those listed may be used. 4.2 Disodium hydrogen phosphate, Na2HPO4 anhydrous or Na2HPO412 H2O. 4.3 Potassium chloride (KCl). 4.4 Potassium dihydrogen phosph
22、ate, KH2PO4. 4.5 Sodium chloride (NaCl). 4.6 Sodium hydroxide (NaOH). 4.7 Hydrochloric acid solution, mass fraction w(HCl) = 37 % in water. 4.8 Hydrochloric acid solution, substance concentration c(HCl) = 0,1 mol/l. Dilute 8,28 ml of hydrochloric acid solution (4.7) to 1 l with water. EN 15850:2010
23、(E) 5 4.9 Sodium hydroxide solution, c(NaOH) = 0,1 mol/l. Dissolve 4 g of sodium hydroxide (4.6) in 1 l of water. 4.10 Phosphate buffered saline (PBS) solution, c(NaCl) = 120 mmol/l, c(KCl) = 2,7 mmol/l, c(phosphate buffer) = 10 mmol/l, pH = 7,4. Dissolve 8,0 g of sodium chloride (4.5), 1,2 g of anh
24、ydrous disodium hydrogen phosphate or 2,9 g of Na2HPO412 H2O (4.2), 0,2 g of potassium dihydrogen phosphate (4.4) and 0,2 g of potassium chloride (4.3) in 900 ml of water. After dissolution, adjust the pH to 7,4 with hydrochloric acid solution (4.8) or sodium hydroxide solution (4.9) as appropriate,
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