1、August 2007DEUTSCHE NORM English price group 9No 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 65.080!,zS“9875748www.di
2、n.deDDIN EN 15360Fertilizers Determination of dicyandiamide Method using high-performance liquid chromatography (HPLC)English version of DIN EN 15360:2007-08Dngemittel Bestimmung von Dicyandiamid Verfahren mit Hochleistungs-Flssigchromatographie (HPLC)Englische Fassung DIN EN 15360:2007-08Supersedes
3、DIN CEN/TS 15360:2006-06www.beuth.deDocument comprises 11 pages10.07DIN EN 15360:2007-08 2 National foreword This standard has been prepared by CEN/TC 260 “Fertilizers and liming materials” (Secretariat: DIN, Germany) under the mandate M/335. The responsible German body involved in its preparation w
4、as the Normenausschuss Lebensmittel und land-wirtschaftliche Produkte (Foodstuffs and Agricultural Products Standards Committee), Technical Committee Dngemittel. DIN EN 15360 is based on a method of test developed by the Verband Deutscher Landwirtschaftlicher Untersuchungs- und Forschungsanstalten (
5、VDLUFA) e.V. and verified by interlaboratory testing. The DIN Standards corresponding to the International Standards referred to in the EN are as follows: EN ISO 3696 DIN ISO 3696 ISO 5725-1 DIN ISO 5725-1 Amendments This standard differs from DIN CEN/TS 15360:2006-06 as follows: a) The standard has
6、 been editorially revised. b) Regulation (EC) No. 2003/2003 has been added to the bibliography. Previous editions CEN/TS 15360: 2006-06 National Annex NA (informative) Bibliography DIN ISO 3696, Water for analytical laboratory use Specification and test methods DIN ISO 5725-1, Accuracy (trueness and
7、 precision) of measurement methods and results Part 1: General principles and definitions EUROPEAN STANDARDNORME EUROPENNEEUROPISCHE NORMEN 15360May 2007ICS 65.080 Supersedes CEN/TS 15360:2006 English VersionFertilizers - Determination of dicyandiamide - Method using high-performance liquid chromato
8、graphy (HPLC)Engrais - Dtermination de la teneur en dicyandiamide -Mthode par chromatographie liquide haute performance(HPLC)Dngemittel - Bestimmung von Dicyandiamid - Verfahrenmit Hochleistungs-Flssigchromatographie (HPLC)This European Standard was approved by CEN on 13 April 2007.CEN members are b
9、ound 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 bibliographical references concerning such nationalstandards may be obtained on application to the C
10、EN 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 member into its own language and notified to the CEN Management Centre has the same statu
11、s 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, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal,Romania, Slo
12、vakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMIT EUROPEN DE NORMALISATIONEUROPISCHES KOMITEE FR NORMUNGManagement Centre: rue de Stassart, 36 B-1050 Brussels 2007 CEN All rights of exploitation in any form and by any means reservedworldwide f
13、or CEN national Members.Ref. No. EN 15360:2007: EEN 15360:2007 (E) 2 Contents Page Foreword3 1 Scope 4 2 Normative references 4 3 Principle4 4 Reagents.4 5 Apparatus .4 6 Sampling and sample preparation.5 7 Procedure .5 8 Calculation6 9 Precision.6 10 Notes on procedure.7 11 Test report 7 Annex A (i
14、nformative) Results of the inter-laboratory trial .8 Bibliography 9 EN 15360:2007 (E) 3 Foreword This document (EN 15360:2007) has been prepared by Technical Committee CEN/TC 260 “Fertilizers and liming materials”, the secretariat of which is held by DIN. This European Standard shall be given the st
15、atus of a national standard, either by publication of an identical text or by endorsement, at the latest by November 2007, and conflicting national standards shall be withdrawn at the latest by November 2007. This document supersedes CEN/TS 15360:2006. This document has been prepared under a mandate
16、 given to CEN by the European Commission and the European Free Trade 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, Cyprus, Czech Republic, Denm
17、ark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom. EN 15360:2007 (E) 4 1 Scope This document specifies a method for
18、the selective determination of dicyandiamide (DCD) in addition to all the other forms of nitrogen fixations, particularly in fertilizers to which DCD has been added as a nitrification inhibiting agent. 2 Normative references The following referenced documents are indispensable for the application of
19、 this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. EN 1482-2, Fertilizers and liming materials Sampling and sample preparation Part 2: Sample preparation EN ISO 3696, Water fo
20、r analytical laboratory use Specification and test methods (ISO 3696:1987) 3 Principle The sample is dissolved or suspended in water using an ultrasonic bath. Methyl dicyandiamide is added to the filtered solution as the internal standard. The solution is then transferred onto a C18 reversed-phase c
21、olumn using a bypass injector and then separated. For detection, a UV-detector is used at a wavelength of 220 nm. 4 Reagents Use only reagents of recognized analytical grade and water conforming to grade 2 of EN ISO 3696. 4.1 Dicyandiamide standard solution Weigh 50 mg of dicyandiamide of known puri
22、ty into a 1 000 ml volumetric flask and dissolve in water; make up the volume to the mark. Pipette 10 ml of this solution into a 100 ml volumetric flask and, after having added 10 ml of internal standard solution (4.2), make up the volume to the mark with water. 4.2 Internal standard solution Weigh
23、50 mg of methyl dicyandiamide of known purity into a 1 000 ml volumetric flask, dissolve in water and make up the volume to the mark. 4.3 Methanol HPLC-grade purity 5 Apparatus 5.1 Ultrasonic bath 5.2 Membrane filter 0,45 m, with the usual filtration equipment. EN 15360:2007 (E) 5 5.3 HPLC apparatus
24、 UV-detector for variable wavelengths and an electronic integrator, sample injection valve equipped with a 20 l bypass injector. 6 Sampling and sample preparation Sampling is not part of the method specified in this document; however, a recommended sampling method is given in EN 1482-1. Sample prepa
25、ration shall be carried out in accordance with EN 1482-2. 7 Procedure 7.1 Preparing the analytical solution Weigh to the nearest 0,001 g, between 0,8 g and 1,5 g of the ground and thoroughly homogenized test sample (corresponding to approximately 50 mg of DCD) and mix with 750 ml of water into a 1 0
26、00 ml volumetric flask and dissolve using an ultrasonic bath (5.1). Those portions that have not dissolved after 5 min are disregarded. Make up the volume to the mark with water. Filter one part of the homogenized sample solution (approximately 50 ml) through the membrane filter (5.2) into a dry ves
27、sel. Pipette 10 ml of this filtrate into a 100 ml volumetric flask and, after having added 10 ml of the internal standard solution (4.2), make up the volume to the mark with water. 7.2 HPLC conditions Eluent: Mixture of water and methanol (4.3), (99 + 1) parts by volume Separation column and packing
28、: 250 mm x 4,6 mm C18 reversed-phase column Column temperature: Room temperature Flow rate: 1,0 ml/min Wavelength: 220 nm7.3 HPLC determination Alternately, transfer the standard solution (4.1) and the test solution (7.1) onto the separation column three times, applying the standard solution before
29、the test solution. Measure the peak areas for DCD and methyl dicyandiamide. EN 15360:2007 (E) 6 8 Calculation Calculate the proportion in the standard solution (4.1), Pxaccording to the following equation: 21xAAP = (1) where A1is the peak area for DCD (4.1); A2is the peak area of the internal standa
30、rd. Calculate the proportion in the test solution (7.1), Pxaccording to the following equation: 23xAAP = (2) where A2is the peak area of the internal standard; A3is the peak area for DCD (of 7.1). Take from each of the groups three values for Pxand Px, the mean values PxMand PxM, and calculate the D
31、CD content of the sample, expressed in g/100 g using the following equation: mPxmPxw=MMDCD(3) where PxMis the mean value of the proportions for the sample; PxMis the mean value of the proportions for the standard; m is the mass of DCD in the standard solution (4.1) (50 mg), in milligrams; m is the m
32、ass of the sample in the aliquot part of the test solution (7.1) used, in milligrams. If necessary, correct the final result depending on the purity of the DCD used for the standard solution. 9 Precision 9.1 General The precision of the method has been determined in an inter-laboratory trial, carrie
33、d out and evaluated in 1997 according to ISO 5725-1. A summary of the results is given in Annex A. The values derived from this inter-laboratory trial might not be applicable to concentration ranges and matrices other than those given. EN 15360:2007 (E) 7 9.2 Repeatability The absolute difference be
34、tween two independent single test results obtained using the same method on identical test material in the same laboratory by the same operator using the same equipment within short time intervals, will exceed the values for the repeatability limit r, given in Table 1, on average in no more than 5 %
35、 of cases. 9.3 Reproducibility The absolute difference between two independent single test results obtained using the same method on identical test material in different laboratories by different operators using different equipment will exceed the values for the reproducibility limit R, given in Tab
36、le 1, on average in no more than 5 % of cases. Table 1 Precision data Sample Level g/100 g r g/100 g R g/100 g No 1 (Alzon 27/1) 2,41 0,089 98 0,466 7 No 2 (Alzon 27/2) 2,18 0,073 7 0,329 10 Notes on procedure 10.1 As described, the method covers a range between 2 g and 10 g of DCD per 100 g. By var
37、iation of the weighed portion and the extent of dilution, DCD contents of between (0,1 g and 100 g) per 100 g can be determined. 10.2 The method may also be carried out using an external standard. In this case the equation for the calculation has to be modified accordingly. 11 Test report The test r
38、eport shall include at least the following information: a) information necessary for complete identification of the sample; b) test method used, making reference to this document, i.e. EN 15360; c) test results together with the units used to express them; d) date the test was finished; e) statement
39、 as to whether the requirement for the repeatability limit has been fulfilled; f) procedural steps not specified in this document or carried out optionally, as well as details of any circumstances that occurred while carrying out the method that might have influenced the result(s). EN 15360:2007 (E)
40、 8 Annex A (informative) Results of the inter-laboratory trial The precision of the method was determined in 1997 during an inter-laboratory trial with respectively 13 and 12 laboratories participating and carried out on 2 samples of fertilizer. The statistical results are given in Table A.1. Table
41、A.1 Statistical results of the inter-laboratory trial Parameter Sample No 1 Sample No 2 Year of the test 1997 Number of participating laboratories 13 12 Number of laboratories after eliminating outliers 13 12 Level mean value, (g/100 g) 2,41 2,18 Repeatability standard deviation sr, (g/100 g) 0,03 0
42、,03 Coefficient of variation CVr(%) 1,348 1,218 Repeatability limit r (2,83 sr) (g/100 g) 0,089 98 0,073 7 Reproducibility standard deviation, sR(g/100 g) 0,168 4 0,118 7 Coefficient of variation CVR(%) 6,99 5,44 Reproducibility limit R (2,83 sR) (g/100 g) 0,466 7 0,329 EN 15360:2007 (E) 9 Bibliogra
43、phy 1 Vilsmeier, K., 1984: Bestimmung von Dicyandiamid, Nitrit und Nitrat in Bodenextrakten mit Hochdruckflssigkeitschromatographie. Zeitschrift fr Pflanzenernhrung, Dngung und Bodenkunde 147, page 264 to 268 2 ISO 5725-1, Accuracy (trueness and precision) of measurement methods and results Part 1:
44、General principles and definitions 3 EN 1482-1, Fertilizers and liming materials Sampling and sample preparation Part 1: Sampling 4 Regulation (EC) No 2003/2003 of the European Parliament and of the Council of 13 October 2003 relating to fertilisers, Official Journal L 304, 21/11/2003, P. 0001-0194
