1、October 2007DEUTSCHE NORM English price group 15No part of this standard may be reproduced without prior permission ofDIN Deutsches Institut fr Normung e. V., Berlin. Beuth Verlag GmbH, 10772 Berlin, Germany,has the exclusive right of sale for German Standards (DIN-Normen).ICS 65.120!$I.“1386111www.
2、din.deDDIN EN 15510Animal feeding stuffs Determination of calcium, sodium, phosphorus, magnesium, potassium,iron, zinc, copper, manganese, cobalt, molybdenum, arsenic, lead andcadmium by ICP-AESEnglish version of DIN EN 15510:2007-10Futtermittel Bestimmung von Calcium, Natrium, Phosphor, Magnesium,
3、Kalium, Eisen, Zink, Kupfer,Mangan, Cobalt, Molybdn, Arsen, Blei und Cadmium mittels ICP-AESEnglische Fassung DIN EN 15510:2007-10www.beuth.deDocument comprises 31 pages DIN EN 15510:2007-10 2 National foreword This standard has been prepared by Technical Committee CEN/TC 327 “Animal feeding stuffs
4、Methods of sampling and analysis” (Secretariat: NEN, the Netherlands). The responsible German body involved in its preparation was the Normenausschuss Lebensmittel und landwirtschaftliche Produkte (Food and Agricultural Products Standards Committee), Technical Committee Futtermittel. The DIN Standar
5、d corresponding to the International Standard referred to in the EN is as follows: ISO 5725-1-1994 DIN ISO 5725-1:1997-11 National Annex NA (informative) Bibliography DIN ISO 5725-1:1997-11, Accuracy (trueness and precision) of measurement methods and results Part 1: General principles and definitio
6、ns EUROPEAN STANDARDNORME EUROPENNEEUROPISCHE NORMEN 15510August 2007ICS 65.120English VersionAnimal feeding stuffs - Determination of calcium, sodium,phosphorus, magnesium, potassium, iron, zinc, copper,manganese, cobalt, molybdenum, arsenic, lead and cadmium byICP-AESAliments des animaux - Dtermin
7、ation des teneurs encalcium, sodium, phosphore, magnsium, potassium, fer,zinc, cuivre, manganse, cobalt, molybdne, arsenic,plomb et cadmium par ICP-AESFuttermittel - Bestimmung von Calcium, Natrium,Phosphor, Magnesium, Kalium, Eisen, Zink, Kupfer,Mangan, Cobalt, Molybdn, Arsen, Blei und Cadmiummitte
8、ls ICP-AESThis European Standard was approved by CEN on 30 June 2007.CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this EuropeanStandard the status of a national standard without any alteration. Up-to-date lists and bibliographica
9、l references concerning such nationalstandards 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 translationunder the responsibility of a CEN mem
10、ber into its own language and notified to the CEN Management Centre has the same status as theofficial versions.CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland,France, Germany, Greece, Hungary, Iceland, Ireland, Italy, La
11、tvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal,Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMIT EUROPEN DE NORMALISATIONEUROPISCHES KOMITEE FR NORMUNGManagement Centre: rue de Stassart, 36 B-1050 Brussel
12、s 2007 CEN All rights of exploitation in any form and by any means reservedworldwide for CEN national Members.Ref. No. EN 15510:2007: EEN 15510:2007 (E) 2 Contents Page Foreword3 1 Scope 4 2 Normative references 4 3 Terms and definitions .4 4 Principle5 5 Reagents.5 6 Apparatus, usual laboratory app
13、aratus and, in particular, the following. .7 7 Sampling.7 8 Preparation of the test sample .8 9 Procedure .9 10 Calculation and expression of the result 13 11 Precision.15 12 Test report 20 Annex A (informative) Results of the interlaboratory test.21 Annex B (informative) Notes on the detection tech
14、nique, interferences and quantification 26 Bibliography 29 EN 15510:2007 (E) 3 Foreword This document (EN 15510:2007) has been prepared by Technical Committee CEN/TC 327 “Animal feeding stuffs Methods of sampling and analysis”, the secretariat of which is held by NEN. This European Standard shall be
15、 given the status of a national standard, either by publication of an identical text or by endorsement, at the latest by February 2008, and conflicting national standards shall be withdrawn at the latest by February 2008. According to the CEN/CENELEC Internal Regulations, the national standards orga
16、nizations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, R
17、omania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom. EN 15510:2007 (E) 4 1 Scope This European Standard specifies inductively coupled plasma atomic emission spectroscopy (ICP-AES) method for the determination of: minerals calcium, sodium, phosphorus, magnesium and potassium and
18、 the elements iron, zinc, copper, manganese, cobalt, molybdenum in animal feeding stuffs, elements arsenic, lead and cadmium in minerals on their own, in pre-mixtures or mixtures for use in animal nutrition. The method detection limit for each element is dependent on the sample matrix as well as of
19、the instrument. The method is not applicable for determination of low concentrations of elements. The limit of quantification should be 3 mg/kg or lower. NOTE This method can also be used for the determination of minerals in products with high mineral content ( 5%), yet for this purpose, other more
20、precise analytical techniques are available. 2 Normative references The following referenced documents are indispensable for the application of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any am
21、endments) applies. EN ISO 3696, Water for analytical laboratory use Specification and test methods (ISO 3696:1987) ISO 6498, Animal feeding stuffs Preparation of test samples 3 Terms and definitions For the purposes of this document, the following terms and definitions apply. 3.1 limit of detection
22、(LOD) smallest measured content from which it is possible to deduce the presence of the analyte with reasonable statistical certainty NOTE The limit of detection is numerically equal to three times the standard deviation of the mean of blank determinations (n 10, were n = number of measures) perform
23、ed under reproducibility conditions. 3.2 limit of quantification (LOQ) lowest content of the analyte that can be measured with reasonable statistical certainty NOTE If both trueness and precision are constant over a concentration range around the limit of detection, then the limit of quantification
24、is numerically equal to ten times the standard deviation of the mean of blank determinations (n 10, were n = number of measures) performed under reproducibility conditions. 3.3 feed additives substances are feed additives when they comply with the definition of feed additives given in regulation EU
25、1831/2003 3.4 animal feeding stuffs substances that comply with the definition of animal feeding stuffs given in regulation EU 178/2002 EN 15510:2007 (E) 5 4 Principle For the determination of the minerals calcium, sodium, phosphorus, magnesium and potassium and the elements iron, zinc, copper, mang
26、anese, cobalt, molybdenum, a test portion of the sample is ashed and dissolved in hydrochloric acid (in the case of organic feeding stuffs) or wet digested with hydrochloric acid (in the case of mineral compounds). For the determination of the elements arsenic, cadmium and lead, a test portion of th
27、e sample is wet digested with nitric acid. The concentration of the elements calcium, sodium, phosphorus, magnesium, potassium, iron, zinc, copper, manganese, cobalt, molybdenum, arsenic, cadmium and lead is determined by inductively coupled plasma atomic emission spectrometry (ICP-AES) using extern
28、al calibration or standard addition technique. WARNING Use of this European 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 European Standard to
29、 establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use. 5 Reagents Use only reagents of recognized analytical grade, unless otherwise specified. 5.1 Water, complying with grade 2 as defined in EN ISO 3696. 5.2 Nitric acid, concentrat
30、ed, not less than 65 % (mass fraction), having a density of approximately (HNO3) 1,42 g/ml. 5.3 Dilute nitric acid, to be prepared by mixing 1 volume of nitric acid (5.2) with 1 volume of water. 5.4 Nitric acid solution of 5 % (m/v), to be prepared: pipette 160 ml of dilute nitric acid (5.3) into a
31、1 000 ml volumetric flask (6.7) and fill to the mark with water. 5.5 Nitric acid solution of 2 % (v/v), to be prepared: pipette 20 ml of nitric acid (5.2) into a 1 000 ml volumetric flask (6.7) and fill to the mark with water. 5.6 Hydrochloric acid, concentrated, not less than 30 % (mass fraction),
32、having a density of approximately (HCl) 1,15 g/ml. 5.7 Dilute hydrochloric acid, to be prepared by mixing 1 volume of hydrochloric acid (5.6) with 1 volume of water. 5.8 Hydrochloric acid solution of 1 % (m/v), to be prepared: pipette 60 ml of dilute hydrochloric acid (5.7) into a 1 000 ml volumetri
33、c flask (6.7) and fill to the mark with water. 5.9 Element stock solutions Ca, Na, P, Mg, K, Fe, Zn, Cu, Mn, Co, Mo, Cd, Pb, As c = 1 000 mg/l. The user should choose a suitable stock solution. Both single-element stock solutions and multi-element stock solutions with adequate specification stating
34、the acid used and the preparation technique are commercially available. It is advisable to use certified stock solutions. EN 15510:2007 (E) 6 Stock solutions are not to be used after the expiry date. NOTE Element stock solutions with concentrations different from 1 000 mg/l may also be used. 5.10 St
35、andard solutions 5.10.1 General Depending on the scope, different multi-element standard solutions may be necessary. In general, when combining multi-element standard solutions, their chemical compatibility and the possible hydrolysis of the components shall be regarded. Spectral interferences from
36、other elements in multi-element standard solutions also need to be considered (Annex B.2.2). The examples given below also consider the measuring range of various inductively coupled plasma atomic emission spectrometers and the expected concentration of the element in animal feeding stuffs. The mult
37、i-element standard solutions are considered to be stable for several months, if stored in the dark. Other combinations of elements at different concentrations can be used, provided that the element stock solutions (5.9) are diluted with the same acid and equal concentration as the acid in the test s
38、olution to a range of standards that covers the concentrations of the elements to be determined. 5.10.2 Multi-element standard solution Minerals in 1 % HCl c (Ca, Na, P, Mg, K) = 40 mg/l Pipette 40,0 ml of each element stock solution (Ca, Na, P, Mg, K) (5.9) into a 1 000 ml volumetric flask (6.7). A
39、dd 60 ml of dilute hydrochloric acid (5.7). Fill to the mark with water and transfer to a suitable storage bottle. 5.10.3 Multi-element standard solution - Fe, Zn, Cu, Mn, Co, Mo in 1 % HCl c (Fe, Zn, Cu, Mn, Co, Mo) = 50 mg/l Pipette 50,0 ml of each element stock solution (Fe, Zn, Cu, Mn, Co, Mo) (
40、5.9) into a 1 000 ml volumetric flask (6.7). Add 60 ml of dilute hydrochloric acid (5.7). Fill to the mark with water and transfer to a suitable storage bottle. 5.10.4 Multi-element standard solution Cd, Pb, As in 5 % HNO3c (Cd, Pb, As) = 100 mg/l Pipette 100,0 ml of each element stock solution (Cd,
41、 Pb, As) (5.9) into a 1000 ml volumetric flask (6.7). Add 160 ml dilute nitric acid (5.3). Fill to the mark with water and transfer to a suitable storage bottle. EN 15510:2007 (E) 7 6 Apparatus, usual laboratory apparatus and, in particular, the following. 6.1 Laboratory grinder 6.1.1 Use laboratory
42、 grinders that are equipped such that they do not lead to contamination of the samples. 6.1.2 Laboratory grinder capable of grinding to a particle size of less than or equal to 1 mm, e.g. a knife mill or equivalent. 6.1.3 Laboratory grinder capable of grinding to a particle size of less than or equa
43、l to 0,1 mm, e.g. a ball mill or equivalent. 6.1.4 Mortar with pestle, free of contamination. 6.2 Analytical balance, capable of weighing to an accuracy of 1 mg. 6.3 Electric hot plate, with temperature control. 6.4 Ashing crucibles, of platinum, quartz or porcelain. 6.5 Electric muffle-furnace, cap
44、able of being maintained at a temperature of 450 C 20 C. The real temperature in the furnace has to be checked, because this temperature may be substantially different from the adjust temperature. 6.6 Beaker, of capacities 100 ml, 250 ml. 6.7 One-mark volumetric flasks, of capacities 100 ml, 500 ml,
45、 1 000 ml. 6.8 Inductively coupled plasma Atomic Emission Spectrometer The instrument shall be equipped with a radial plasma as a minimum requirement; an axial plasma is equally acceptable. Background correction shall also be performed when necessary. Settings of the working conditions (e.g. viewing
46、 height, gas flows, RF or plasma power, sample uptake rate, integration time, number of replicates, ) shall be optimised according the manufacturers instructions. 6.9 Freeze drying equipment, capable of freeze-drying liquid animal feeding stuffs. 7 Sampling Sampling is not part of the method specifi
47、ed in this Standard. A recommended sampling method is given in EN ISO 6497. It is important that the laboratory receives a sample that is truly representative and has not been damaged or changed during transport or storage. EN 15510:2007 (E) 8 8 Preparation of the test sample 8.1 General Prepare the
48、 test sample in accordance with ISO 6498. Grinding must be carried out in conditions such that the substance is not appreciably heated. Operation is to be repeated as many times as is necessary and it must be affected as quickly as possible in order to prevent any gain or loss of constituents (water
49、). Whole ground product is placed in a flask made of e.g. polypropylene, which can be stoppered and stored in such way to prevent any change in composition. Before any weighing is carried out for the analysis, the whole test sample must be thoroughly mixed for reasons of homogeneity. 8.2 Animal feeding stuffs which can be ground as such Grind the laboratory sample (usually 500 g), using a grinder (6.1.2) or mortar (6.1.4), until a particle size of 1 mm or less has been reached. 8.3 Liquid animal feeding stuffs 8.3.1 General Li
