1、September 2012 Translation by DIN-Sprachendienst.English price group 11No part of this translation 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).
2、ICS 65.120!$UB“1915031www.din.deDDIN EN 16277Animal feeding stuffs Determination of mercury by cold-vapour atomic absorptionspectrometry (CVAAS) after microwave pressure digestion (extractionwith 65 % nitric acid and 30 % hydrogen peroxide);English version EN 16277:2012,English translation of DIN EN
3、 16277:2012-09Futtermittel Bestimmung von Quecksilber mit Kaltdampf-Atomabsorptionsspektrometrie (KD-AAS)nach Mikrowellen-Druckaufschluss (Extraktion mit 65 % Salpetersure und 30 %Wasserstoffperoxid);Englische Fassung EN 16277:2012,Englische bersetzung von DIN EN 16277:2012-09Aliments des animaux Do
4、sage du mercure par spectromtrie dabsorption atomique vapeur froide (SAVVF)aprs digestion sous pression par micro-ondes (extraction avec de lacide nitrique 65 %et du peroxyde dhydrogne 30 %);Version anglaise EN 16277:2012,Traduction anglaise de DIN EN 16277:2012-09www.beuth.deIn case of doubt, the G
5、erman-language original shall be considered authoritative.Document comprises 19 pages08.12 DIN EN 16277:2012-09 2 A comma is used as the decimal marker. National foreword This standard has been prepared by Technical Committee CEN/TC 327 “Animal feeding stuffs Methods of sampling and analysis” (Secre
6、tariat: NEN, Netherlands). The responsible German body involved in its preparation was the Normenausschuss Lebensmittel und landwirtschaftliche Produkte (Food and Agricultural Products Standards Committee), Working Committee NA 057-03-03 AA Futtermittel. DIN EN 16277 is based on a method of test dev
7、eloped by the Verband Deutscher Landwirtschaftlicher Untersuchungs- und Forschungsanstalten (VDLUFA) e.V. and verified by interlaboratory testing. The DIN Standards corresponding to the International Standards referred to in this document are as follows, whereby EN ISO Standards are only listed belo
8、w if these have not been published as DIN EN ISO Standards with the same number: EN ISO 3696 DIN ISO 3696 National Annex NA(informative) Bibliography DIN ISO 3696, Water for analytical laboratory use Specification and test methods EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 16277 July 2012
9、ICS 65.120 English Version Animal feeding stuffs - Determination of mercury by cold-vapour atomic absorption spectrometry (CVAAS) after microwave pressure digestion (extraction with 65 % nitric acid and 30 % hydrogen peroxide) Aliments des animaux - Dosage du mercure par spectromtrie dabsorption ato
10、mique vapeur froide (SAVVF) aprs digestion sous pression par micro-ondes (extraction avec de lacide nitrique 65 % et du peroxyde dhydrogne 30 %) Futtermittel - Bestimmung von Quecksilber mit Kaltdampf-Atomabsorptionsspektrometrie (KD-AAS) nach Mikrowellen-Druckaufschluss (Extraktion mit 65 % Salpete
11、rsure und 30 % Wasserstoffperoxid) This European Standard was approved by CEN on 17 May 2012. 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-dat
12、e lists and bibliographical references concerning such national standards may be obtained on application to the CEN-CENELEC 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
13、under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC 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, Form
14、er Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. Management Centre: Avenue Marnix 17, B-1000 B
15、russels 2012 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 16277:2012: EEUROPEAN COMMITTEE FOR STANDARDIZATIONCOMIT EUROPEN DE NORMALISATIONEUROPISCHES KOMITEE FR NORMUNGEN 16277:2012 (E) 2 Contents Page Foreword 31 Scope 42 Norm
16、ative references 43 Principle 44 Reagents .46 Procedure .77 Calculation 98 Precision .99 Test report . 10Annex A (informative) Results of the interlaboratory tests . 11Annex B (informative) Flowchart Determination of mercury by CVAAS after microwave digestion 12Annex C (informative) Alternative dige
17、stion procedure with the same extraction efficiency: Acid digestion with a mixture of 65 % nitric acid and 70 % perchloric acid (7:3 v/v) at atmospheric pressure 13Annex D (informative) Alternative digestion procedure with the same extraction efficiency: Acid digestion with a mixture of 65 % nitric
18、acid, 37 % hydrochloric acid and 30 % hydrogen peroxide under reflux . 15Bibliography . 16DIN EN 16277:2012-09 EN 16277:2012 (E) 3 Foreword This document (EN 16277:2012) has been prepared by Technical Committee CEN/TC 327 “Animal feeding stuffs Methods of sampling and analysis”, the secretariat of w
19、hich is held by NEN. 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 January 2013, and conflicting national standards shall be withdrawn at the latest by January 2013. Attention is drawn to the po
20、ssibility 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 European Commission and the European Free Trad
21、e Association. According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedon
22、ia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. DIN EN 16277:2012-09 EN 16277:2012 (E) 4 1 Scope This European Standard
23、specifies a method for the determination of mercury in animal feeding stuffs by Cold-Vapour Atomic Absorption Spectrometry (CVAAS) after microwave pressure digestion. The limit of quantification in the test solution should be 0,25 g/l or lower. Using a test portion of 0,5 g and a volume of the test
24、solution of 25 ml a limit of quantification of 0,0125 mg/kg or lower should be obtained. 2 Normative references The following documents, in whole or in part, are normatively referenced in this document and are indispensable for its application. For dated references, only the edition cited applies. F
25、or undated references, the latest edition of the referenced document (including any amendments) applies. EN ISO 3696, Water for analytical laboratory use Specification and test methods (ISO 3696) EN ISO 6497, Animal feeding stuffs Sampling (ISO 6497) EN ISO 6498, Animal feeding stuffs Guidelines for
26、 sample preparation (ISO/FDIS 6498) 3 Principle Mercury is determined in the test solution by cold-vapour atomic absorption spectrometry (CVAAS) after microwave pressure digestion. The homogenised feeding stuff test sample is digested with nitric acid and hydrogen peroxide under pressure and high te
27、mperatures in a microwave-heated pressure digestion system. The test solution is transferred to the reaction vessel of the mercury analysis unit. The mercury is reduced with sodium borohydride or tin(II) chloride to elemental volatile mercury and flushed into the cell of the AAS instrument using a c
28、arrier gas stream. As an option with an additional amalgamation step, sensitivity could be increased and matrix effects could be decreased. The absorption at 253,7 nm (mercury line) is used as a measure of the mercury concentration in the cell. Other digestion procedures with the same extraction eff
29、iciency (see Annex D and Annex E) or other measurement systems like Fl-CVAAS (flow injection cold-vapour atomic absorption spectroscopy) or CV-ICP-AES (cold-vapour inductively coupled plasma atomic emission spectroscopy) are possible. WARNING The use of this standard can involve hazardous materials,
30、 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 establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use. 4 R
31、eagents The concentration of the trace elements in the reagents and water used shall be low enough not to affect the results of the determination. A blank should be measured simultaneously with the test samples on each day of analysis to control contamination and carry over with mercury in the reage
32、nts and apparatus used. Use water conforming to grade 2 of EN ISO 3696. 4.1 Nitric acid (HNO3), not less than 65 % (mass fraction), of approximately (HNO3) = 1,4 g/ml. NOTE Use nitric acid available with high purity or perform a sub-boiling distillation. 4.2 Hydrogen peroxide (H2O2), not less than 3
33、0 % (mass fraction). DIN EN 16277:2012-09 EN 16277:2012 (E) 5 4.3 Hydrochloric acid (HCl), not less than or equal to 30 % (mass fraction), of approximately (HCl) 1,15 g/ml. 4.4 Diluted hydrochloric acid, e.g. about 3 % (mass fraction), as carrier solution for the use in the flow-injection-system and
34、 for dilution of the mercury stock solution to the standard solution and furthermore to the calibration solutions. EXAMPLE Dilute approximately 90 ml of hydrochloric acid (4.3) to 1 l with water. 4.5 Reducing agents. Tin(II) chloride or sodium borohydride may be used as the reducing agent, but it is
35、 not advisable to use the two reagents alternately. The concentration by mass of the reducing agent solutions may be varied to suit the system and the relevant information provided by the manufacturer of the apparatus shall be observed. 4.5.1 Tin(II) chloride solution, c(SnCl22H2O) = 100 g/l. Dissol
36、ve 50 g of tin(II) chloride in approximately 100 ml of hydrochloric acid (4.3) in a 500 ml volumetric flask (5.2) and dilute to the mark with water. Prepare a fresh solution daily. 4.5.2 Sodium borohydride solution, e.g. c(NaBH4) = 2 g/l. Dissolve 2 g of sodium hydroxide pellets in water using a 1 0
37、00 ml flask (5.2), add 2 g of sodium borohydride and dilute to the mark with water. Prepare a fresh solution daily and, when necessary, filter before use. When the analysis procedure requires a longer time it is recommended to cool the sodium borohydride solution, i.e. with ice around the flask, dur
38、ing its use in the CVAAS measurement. NOTE Sodium borohydride, stable aq. solution, 4,4 mol/l in 14 mol/l NaOH, is also commercially available. WARNING It is essential to observe the safety instructions for working with sodium borohydride. Sodium borohydride forms hydrogen with acids and this can re
39、sult in an explosive air/hydrogen mixture. A permanent extraction system shall be provided at the point where measurements are carried out. 4.6 Mercury stock solution, c(Hg) = 1 000 mg/l. The stock solution is commercially available. It is advisable to use certified stock solutions. Otherwise, disso
40、lve 1,080 g of mercury(II) oxide in 10 ml of potassium dichromate solution and dilute to 1 l with water. Use a potassium dichromate solution with a concentration of 5 g/l. Dissolve 5 g of potassium dichromate with 500 ml nitric acid (4.1) and dilute to 1l with water. 4.7 Mercury standard solution, c
41、(Hg) = 1 mg/l. Dilute 100 l the stock solution (4.6) with diluted hydrochloric acid (4.4) in a 100 ml flask (5.2) to a concentration of 1 mg/l. The standard solution is stable for at least three months. 4.8 Mercury calibration solutions. Dilute the standard solution (4.7) to the concentrations neede
42、d for calibration with diluted hydrochloric acid (4.4). The following calibration solutions are recommended (see Table 1). Take aliquots of 0 l, 25 l, 50 l, 250 l, 500 l of the mercury standard solution (4.7) into flasks of 50 ml (5.2) and fill up with diluted hydrochloric acid (4.4) to concentratio
43、ns of 0 g/l, 0,5 g/l, 1 g/l, 5 g/l and 10 g/l. DIN EN 16277:2012-09 EN 16277:2012 (E) 6 Table 1 Recommended calibration solutions (4.8) for the determination of mercury Mercury (Hg) Concentration of calibration solution (4.8) g/l Aliquots of standard solution (4.7), transferred in 50 ml flasks l Cal
44、ibration standard 1 0 0 Calibration standard 2 0,5 25 Calibration standard 3 1 50 Calibration standard 4 5 250 Calibration standard 5 10 500 Choose the concentrations of the calibration solutions so as not to exceed the linear range of the calibration curve. It is recommended to use a minimum of fiv
45、e calibration solutions with different concentrations. In general, the calibration curve should be linear. Using a non-linear calibration curve is possible if it is well-described. 5 Apparatus and equipment To minimise the contamination, all apparatus which come into direct contact with the sample a
46、nd the solutions should be carefully pre-treated. NOTE Recommendations are given in EN 13804. 5.1 Microwave-heated pressure digestion apparatus with inert reaction vessels, made of materials such as Polytetrafluorethen (PTFE), Polyfluoralkan (PFA), Perfluorethylenpropylen (FEP) or quartz and shich a
47、re suitable for digestion temperatures exceeding 200 C. The microwave oven should be generally resistant to corrosion. In particular, the whole electronic area of the microwave oven should be protected against corrosion to ensure safe operation. The ventilation should transfer the acid vapours to an
48、 extractor hood. The reaction vessels should have a safety valve designed for a pressure of 10 000 kPa. 5.2 Pipettes, Flasks, of the following capacities: 25 ml, 50 ml, 100 ml, 500 ml and 1 000 ml. 5.3 Flow injection cold-vapour system with sample loop, i.e. 500 l. 5.4 Atomic absorption spectrometer (AAS), with a heated quartz cell and optionally with an amalgamation system. 5.5 Element-specific lamp for mercury. NOTE An electrodeless discharge la
