1、November 2010 Translation by DIN-Sprachendienst.English price group 8No 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).IC
2、S 65.080!$l1“1731456www.din.deDDIN EN 15905Fertilizers Determination of 3-methylpyrazole (MP) usinghigh-performance liquid chromatography (HPLC)English translation of DIN EN 15905:2010-11Dngemittel Bestimmung von 3-Methylpyrazol (MP) mit Hochleistungs-Flssigchromatographie (HPLC)Englische bersetzung
3、 von DIN EN 15905:2010-11Engrais Dosage du 3-mthylpyrazole (MP) parchromatographie liquide haute performance (CLHP)Traduction anglaise de DIN EN 15905:2010-11www.beuth.deDocument comprises pagesIn case of doubt, the German-language original shall be considered authoritative.1110.10 DIN EN 15905:2010
4、-11 2 A comma is used as the decimal marker. National foreword This standard has been prepared by Technical Committee CEN/TC 260 “Fertilizers and liming materials” (Secretariat: DIN, Germany) under the mandate M/418. The responsible German body involved in its preparation was the Normenausschuss Leb
5、ensmittel und landwirtschaftliche Produkte (Food and Agricultural Products Standards Committee), Working Committee NA 057-03-02 AA Dngemittel. The DIN Standards corresponding to the International Standards referred to in this document are as follows, whereby EN ISO Standards are only listed below if
6、 these have not been published as DIN EN ISO Standards with the same number: EN ISO 3696 DIN ISO 3696 ISO 5725-1 DIN ISO 5725-1 ISO 5725-2 DIN ISO 5725-2 National Annex NA (informative) Bibliography DIN ISO 3696, Water for analytical laboratory use Specification and test methods DIN ISO 5725-1, Accu
7、racy (trueness and precision) of measurement methods and results Part 1: General principles and definitions DIN ISO 5725-2, Accuracy (trueness and precision) of measurement methods and results Part 2: Basic method for the determination of repeatability and reproducibility of a standard measurement m
8、ethod EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 15905 June 2010 ICS 65.080 English Version Fertilizers - Determination of 3-methylpyrazole (MP) using high-performance liquid chromatography (HPLC) Engrais - Dosage du 3-mthylpyrazole (MP) par chromatographie liquide haute performance ( HP)
9、Dngemittel - Bestimmung von 3-Methylpyrazol (MP) mit Hochleistungs-Flssigchromatographie (HPLC) This European Standard was approved by CEN on 29 April 2010. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European Standard the
10、status of a national standard without any alteration. Up-to-date lists and bibliographical 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
11、). A version in any other language made by translation under the responsibility of a 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,
12、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 United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT
13、EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 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 15905:2010: ECLEN 15905:2010 (E) 2 Contents Page Foreword 31 Scope 42
14、 Normative references 43 Terms and definitions .44 Principle 45 Reagents .46 Apparatus .47 Sampling and sample preparation .58 Procedure .58.1 Preparation of the test solution 58.2 Preparation of the calibration solutions 58.3 HPLC conditions 58.4 HPLC determination 69 Calculation and expression of
15、the results 69.1 Calculation 69.2 Expression of results 610 Precision .710.1 Inter-laboratory test .710.2 Repeatability .710.3 Reproducibility .711 Test report 7Annex A (informative) Results of the inter-laboratory test .8Bibliography 9DIN EN 15905:2010-11 EN 15905:2010 (E) 3 Foreword This document
16、(EN 15905:2010) 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 status of a national standard, either by publication of an identical text or by endorsement, at the latest by Dec
17、ember 2010, and conflicting national standards shall be withdrawn at the latest by December 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 pat
18、ent rights. This document has been prepared under a mandate 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 Standar
19、d: Austria, Belgium, Bulgaria, 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 Ki
20、ngdom. DIN EN 15905:2010-11 EN 15905:2010 (E) 4 1 Scope This European Standard specifies a method for the determination of the 3-methylpyrazole (MP) content in fertilizers, in particular in urea and materials containing urea using high-performance liquid chromatography (HPLC). 2 Normative references
21、 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 amendments) applies. EN 1482-2, Fertilizers and liming materials Samp
22、ling and sample preparation Part 2: Sample preparation EN 12944-1:1999, Fertilizers and liming materials and soil improvers Vocabulary Part 1: General terms EN 12944-2:1999, Fertilizers and liming materials and soil improvers Vocabulary Part 2: Terms relating to fertilizers1)(including corrigendum A
23、C:2000) EN ISO 3696:1995, Water for analytical laboratory use Specification and test methods (ISO 3696:1987) 3 Terms and definitions For the purposes of this document, the terms and definitions given in EN 12944-1:1999 and EN 12944-2:1999 apply. 4 Principle The sample of fertilizer is dissolved in w
24、ater or extracted with water. The 3-methlypyrazole is determined in the solution using reversed phase high-performance liquid chromatography with an UV detector. 5 Reagents Use only reagents of recognized analytical grade and distilled or demineralized water (grade 3 according to EN ISO 3696:1995).
25、5.1 Acetonitrile, HPLC purity. 5.2 De-ionized water, conductivity less than 0,5 mS/m according to ISO 3696, grade 3. 5.3 3-methylpyrazole. 6 Apparatus 6.1 Analytical balance, measuring accuracy 0,1 mg or better. 1) See corrigendum EN 12944-2:1999/AC:2000 which corrects EN 12944-2:1999. DIN EN 15905:
26、2010-11 EN 15905:2010 (E) 5 6.2 Ultrasonic bath. 6.3 Filtration equipment, (manual or pressure filtration). 6.4 Membrane filter, pore size 0,45 m or smaller. NOTE For samples which are difficult to filter, to the purpose a pressure filtration can be used. 6.5 Volumetric flasks,100 ml, 1 000 ml. 6.6
27、HPLC device, for isocratic working procedure. 6.7 Sample issuing system. 6.8 UV detector. 6.9 Evaluation unit, e.g. electronic integrator. 6.10 Reversed phase HPLC separator column, e.g. C18 10 m, 250 mm 4 mm. NOTE With suitable separation performance, it is also possible to use a shorter separation
28、 column. Instead of a C18 column, a separation column with hydroxyl phase can be used. 7 Sampling and sample preparation Sampling is not part of the method specified in this document. A recommended sampling method is given in EN 1482-1. Sample preparation shall be carried out in accordance with EN 1
29、482-2. 8 Procedure 8.1 Preparation of the test solution Weigh a test portion corresponding to approximately 8 mg of 3-methlypyrazole into a 1 000 ml volumetric flask. Add approximately 750 ml of water (5.2) and dissolve in the ultrasonic bath (6.2). Make up to the mark with water (5.2) and filter a
30、part of the sample solution through a membrane filter (6.3, 6.4) in order to remove any un-dissolved substances which may still be present. This solution is used for the determination. NOTE The amount of ammonium nitrate-urea solution weighed in is 10 g, precise to 10 mg. For solid fertilizers, for
31、reasons of homogeneity, an amount weighed in should be used of 100 g, precise to 0,1 g. In this case, the dilution of the filtrate by 1+9 is necessary; this is to be taken into account during the calculation. 8.2 Preparation of the calibration solutions Weigh 1 g of 3-methylpyrazole (5.3) to the nea
32、rest of 0,1 mg into a 1 000 ml volumetric flask. Dissolve with water (5.2), make up to the mark with water (5.2) and mix well. Pipette 10 ml of this solution into a 100 ml volumetric flask, make up to the mark with water (5.2) and mix well. Pipette 5 ml, 10 ml, and 15 ml of this solution into three
33、100 ml volumetric flasks. Make up each flask to the mark with water (5.2) and mix well. Take the solutions to create the calibration curve. 8.3 HPLC conditions Separation column: filling for reversed phase HPLC DIN EN 15905:2010-11 EN 15905:2010 (E) 6 Column temperature: room temperature Elution age
34、nt: water/acetonitrile (5.1) mixture 80+20 (volume fraction) Flow rate: 0,8 ml/min to 1,2 ml/min Injection volume: 20 l Wavelength: 214 nm The elution agent shall be degassed, e.g. in the ultrasonic bath (6.2). 8.4 HPLC determination To determine the calibration curve, inject an amount of 20 l of ea
35、ch calibration solution (see 8.2) three times. The calibration curve can be used to for the content determination if the correlation coefficient 0,99. The correlation coefficient is calculated in accordance with the method of smallest squares. Inject the test solution two times in succession. 9 Calc
36、ulation and expression of the results 9.1 Calculation Carry out the evaluation on the basis of the calibration curve over the peak heights or the peak areas. Calculate the mass fraction of 3-methylpyrazole, wMP, in percent according to the following equation: maFbAw=100)(dpkMP(1) where Apkis the pea
37、k height or peak area; b is the ordinate section of calibration curve; a is the rise of the calibration curve; Fdis the dilution factor; m is the mass of the test portion, in milligrams. 9.2 Expression of results Calculate the arithmetic mean from both values obtained. Indicate the result to the nea
38、rest 0,001 %. DIN EN 15905:2010-11 EN 15905:2010 (E) 7 10 Precision 10.1 Inter-laboratory test An inter-laboratory test has been carried out in 2008 with 9 participating laboratories and 2 different samples of fertilizers. This test yielded the data given in Annex A. Repeatability and reproducibilit
39、y were calculated according to ISO 5725-1 and ISO 5725-2. The values derived from this inter-laboratory test may not be applicable to concentration ranges and matrices other than those given in Annex A. 10.2 Repeatability The absolute difference between two independent single test results, obtained
40、with the same method on identical test material in the same laboratory by the same operator using the same equipment within a short interval of time, will in not more than 5 % of the cases exceed the values of r given in Table 1. 10.3 Reproducibility The absolute difference between two single test r
41、esults, obtained with the same method on identical test material in different laboratories by different operators using different equipment, will in not more than 5 % of the cases exceed the values of R given in Table 1. Table 1 Mean values, repeatability and reproducibility limits Sample x r R Liqu
42、id sample 1 0,030 4 0,000 3 0,003 9 Liquid sample 2 0,034 5 0,000 9 0,003 9 11 Test report The test report shall contain at least the following information: a) all information necessary for the complete identification of the sample; b) the test method used with reference to this document; c) the tes
43、t results obtained; d) date of sampling and sampling procedure (if known); e) date when the analysis was finished; f) whether the requirement of the repeatability limit has been fulfilled; g) all operating details not specified in this document, or regarded as optional, together with details of any
44、incidents occurred when performing the method, which may have influenced the test result(s). DIN EN 15905:2010-11 EN 15905:2010 (E) 8 Annex A (informative) Results of the inter-laboratory test The precision of the method has been determined in the year 2008 in an inter-laboratory trial with 9 labora
45、tories participating and carried out on 2 different liquid samples of fertilizer. The statistical results are given in Table A.1. Table A.1 Statistical results of the inter-laboratory test Parameter Liquid sample 1 Liquid sample 2 Year of the test 2008 2008 Number of participating laboratories 9 9 N
46、umber of laboratories after eliminating outliers 8 9 mean value, x , (%) (mass fraction) 0,030 4 0,034 5 Repeatability standard deviation sr, (%) (mass fraction) 0,000 1 0,000 3 RSDr(%) (mass fraction) 0,30 0,93 Repeatability limit r (2,83 sr) (%) (mass fraction) 0,000 3 0,000 9 Reproducibility stan
47、dard deviation, sR(%) (mass fraction) 0,001 4 0,001 4 RSDR(%) (mass fraction) 4,63 4,03 Reproducibility limit R (2,83 sR) (%) (mass fraction) 0,003 9 0,003 9 DIN EN 15905:2010-11 EN 15905:2010 (E) 9 Bibliography 1 EN 1482-1, Fertilizers and liming materials Sampling and sample preparation Part 1: Sa
48、mpling 2 ISO 5725-1, Accuracy (trueness and precision) of measurement methods and results Part 1: General principles and definitions 3 ISO 5725-2, Accuracy (trueness and precision) of measurement methods and results Part 2: Basic method for the determination of repeatability and reproducibility of a standard measurement method 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. 1-194 DIN EN 15905:2010-11
