DIN EN 15705-2010 Fertilizers - Determination of urea condensates using high-performance liquid chromatography (HPLC) - Isobutylidenediurea and crotonylidenediurea (method A) and m.pdf

1、July 2010 Translation by DIN-Sprachendienst.English price group 12No 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).ICS 6

2、5.080!$iJH“1703937www.din.deDDIN EN 15705Fertilizers Determination of urea condensates using high-performance liquidchromatography (HPLC) Isobutylidenediurea and crotonylidenediurea (method A) andmethylen-urea oligomers (method B)English translation of DIN EN 15705:2010-07Dngemittel Bestimmung von H

3、arnstoffkondensaten mit Hochleistungs-Flssigchromatographie(HPLC) Isobutylidendiharnstoff und Crotonylidendiharnstoff (Verfahren A) undMethylenharnstoff-Oligomere (Verfahren B)Englische bersetzung von DIN EN 15705:2010-07Engrais Dosage des condensats dure par chromatographie liquide haute performanc

4、e (HPLC) Isobutylidne diure et crotonylidne diure (mthode A) et oligomres deTraduction anglaise de DIN EN 15705:2010-07SupersedesDIN CEN/TS 15705:2009-06www.beuth.deIn case of doubt, the German-language original shall be considered authoritative.Document comprises 24 pages(mthode B)mthylne-ure 07.10

5、 DIN EN 15705:2010-07 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/335. The responsible German body involved in its preparation was the N

6、ormenausschuss Lebensmittel 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: ISO 5725-1 DIN ISO 5725-1 ISO 5

7、725-2 DIN ISO 5725-2 Amendments This standard differs from DIN CEN/TS 15705:2009-06 as follows: a) Based on the results of the unique acceptance procedure, the document has been adopted as a European Standard without any modifications. b) An introduction has been included to clarify the correlation

8、with the requirements of Regulation (EC) 2003/2003 relating to fertilizers (see Bibliography). c) A footnote has been added concerning the preparation of the standard substances MDU and DMTU. Previous editions DIN CEN/TS 15705: 2009-06 National Annex NA (informative) Bibliography DIN ISO 5725-1, Acc

9、uracy (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

10、method EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 15705 March 2010 ICS 65.080 Supersedes CEN/TS 15705:2009English Version Fertilizers - Determination of urea condensates using high-performance liquid chromatography (HPLC) - Isobutylidenediurea and crotonylidenediurea (method A) and methyle

11、n-urea oligomers (method B) Engrais - Dosage des condensats dure par chromatographie liquide haute performance (HPLC) - Isobutylidne diure et crotonylidne diure (mthode A) et oligomres de mthylne-ure (mthode B) Dngemittel - Bestimmung von Harnstoffkondensaten mit Hochleistungs-Flssigchromatographie

12、(HPLC) - Isobutylidendiharnstoff und Crotonylidendiharnstoff (Verfahren A) und Methylenharnstoff-Oligomere (Verfahren B) This European Standard was approved by CEN on 21 February 2010. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving

13、 this European Standard the 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 versi

14、ons (English, French, German). 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,

15、 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 United Kingdom. EUROPEAN COMMITTE

16、E FOR STANDARDIZATION COMIT 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 15705:2010: EEN 15705:2010 (E) 2 Contents

17、 Page Foreword 3Introduction .41 Scope 52 Normative references 53 Terms and definitions .54 Sampling and sample preparation .55 Method A: Determination of CDU and IBDU .56 Method B: Determination of methylen-urea oligomers (MU) 87 Precision method A and method B . 138 Test report . 14Annex A (inform

18、ative) Results of the inter-laboratory tests 15Annex B (informative) Chromatogram and calibration curves method A 17Annex C (informative) Chromatogram and calibration curves method B 19Bibliography . 22DIN EN 15705:2010-07 EN 15705:2010 (E) 3 Foreword This document (EN 15705:2010) has been prepared

19、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 September 2010, and conflicting natio

20、nal standards shall be withdrawn at the latest by September 2010. This document supersedes CEN/TS 15705:2009. 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

21、all such patent rights. This document has been prepared under a mandate given to CEN by the European Commission and the European Free Trade Association. The following has been added to the former edition of the European Standard: a) introduction; b) information concerning the preparation of the stan

22、dard substances MDU and DMTU. 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, France, Germany, Gre

23、ece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom. DIN EN 15705:2010-07 EN 15705:2010 (E) 4 Introduction Fertilizers containing the condensates of urea and spe

24、cified aldehydes (with crotonaldehyde called crotonyliden diurea or CDU, with isobutyraldehyde called isobutylidene diurea or IBDU, with formaldehyde called urea formaldehyde or methylene urea or MU) are covered by Annex I of the Regulation (EC) 2003/2003 1 as nitrogenous fertilizers. The methods de

25、scribed in this European Standard enable the quantitative determination of these condensates and the determination of the solubility of the MU-oligomers according to the Regulation. DIN EN 15705:2010-07 EN 15705:2010 (E) 5 1 Scope This European Standard specifies methods for the determination of iso

26、butylidenediurea (IBDU), crotonylidenediurea (CDU) (method A) and methylene-urea oligomers (MU) (method B) in fertilizers using high-performance liquid chromatography (HPLC). The method is applicable to all fertilizers which do not contain interfering organic compounds. 2 Normative references The fo

27、llowing 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 Sampling an

28、d 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 fertilizers EN ISO 3696:1995, Water for anal

29、ytical 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 Sampling and sample preparation Sampling is not part of the method specified in this Europea

30、n Standard. A recommended sampling method is given in EN 1482-1. Sample preparation shall be carried out in accordance with EN 1482-2. 5 Method A: Determination of CDU and IBDU 5.1 Principle The sample is extracted with water and, after appropriate dilution, analyzed using a suitable HPLC system. 5.

31、2 Reagents 5.2.1 Reagents of recognized analytical grade and distilled or demineralized water (grade 3 according to EN ISO 3696:1995). 5.2.2 Acetonitrile, p.a., HPLC-grade. DIN EN 15705:2010-07 EN 15705:2010 (E) 6 5.2.3 Isobutylidenediurea and crotonylidenediurea, in their pure form. 5.3 Apparatus 5

32、.3.1 Laboratory equipment and glassware, for preparation of solutions and dilutions. 5.3.2 Analytical balance, capable of weighing to an accuracy of 0,1 mg. 5.3.3 HPLC-system, with UV-detector. 5.3.4 Ultrasonic bath 5.3.5 Magnetic stirrer 5.3.6 Disposable filter, 0,45 m. 5.4 Procedure 5.4.1 System p

33、arameters of HPLC Analytical/separating column: silica column with C18 reverse phase 1)Detection wavelength: 200 nm Eluent: acetonitrile/water: 10/90 (volume fraction) Flow rate: 1 ml/min Temperature: ambient temperature Injection volume: 20 l 5.4.2 Calibration 5.4.2.1 Stock solution IBDU (IBDU) = 1

34、00 mg/l Weigh 100/R mg of IBDU (5.2.3), where R is the purity of IBDU, into a 1 000 ml flask and add about 900 ml of water (5.2.1). Dissolve in an ultrasonic bath (5.3.4) for about 10 min, followed by stirring on a magnetic stirrer (5.3.5) for about 1 h. Make up to volume. Filtration is not necessar

35、y. 5.4.2.2 Stock solution CDU (CDU) = 100 mg/l Weigh 100/R mg of CDU (5.2.3) , where R is the purity of CDU, into a 1 000 ml flask and add about 900 ml of water (5.2.1). Dissolve in an ultrasonic bath (5.3.4) for about 10 min, followed by stirring on a magnetic stirrer (5.3.5) for about 1 h. Make up

36、 to volume. Filtration is not necessary. 5.4.2.3 Calibration solution For calibration, prepare three solutions according to Table 1 using one-mark (bulb) pipettes and dilute to the mark with water (5.2.1). 1)LiChrosorb RP-18 7 m 250/4 mm is an example of a suitable product available commercially. Th

37、is information is given for the convenience of users of this European Standard and does not constitute an endorsement by CEN or CENELEC of this product. DIN EN 15705:2010-07 EN 15705:2010 (E) 7 For the determination of the retention time, dilute 10 ml of the stock solution 5.4.2.1 or respectively 5.

38、4.2.2 into two 100 ml flasks and make up to volume with water (5.2.1). The evaluation of calibration is carried out manually or by means of a suitable PC-aided (computerized) calculation method. Table 1 Preparation of calibration solutions Parameter Amount of stock solution IBDU/CDU ml (to be added

39、to the 100 ml flask) Content of IBDU mg/l Content of CDU mg/l Standard 1 10 10,0 10,0 Standard 2 25 25,0 25,0Standard 3 50 50,0 50,0 5.4.3 Preparation of the test portion Weigh 1 g of the sample grounded to 0,2 mm to the nearest 0,1 mg and flush into a 1 000 ml volumetric flask with water (5.2.1). F

40、ill the flask to an amount of approximately 900 ml and treat it for 10 min in the ultrasonic bath (5.3.4). Then make up to the mark and stir for 1 h at room temperature on a magnetic stirrer (5.3.5). Dilute 10 ml of the solution in a 100 ml volumetric flask and filter into the HPLC injection vial th

41、rough a disposable filter (5.3.6). 5.4.4 Measurement Measurement is performed manually or by means of an automatic sample loading system (autosampler). 5.4.5 Important annotations IBDU is able to form urea in aqueous solution. Therefore, the measurement of the calibration and sample solutions shall

42、be completed within one working day. The concentrations of CDU and IBDU in the sample solutions shall be kept within the calibration limits (5.4.2) to ensure sufficient reproducibility. 5.5 Calculation The calculation can be performed manually or by means of a PC using the calibration parameters in

43、respect to the amount used. In the case of PC-aided (computerized) calculation and application of Table 1 regarding the amounts of stock solution, the content of IBDU/CDU in milligrams per litre will be calculated by the system. The calculated values are equal to the percentage mass concentration of

44、 IBDU/CDU in the analysed sample of fertilizer. Following general rules for declaration in regulations to declare the content of the compounds as percentage mass fraction of nitrogen, calculate the contents, wN(IBDU)/wN(CDU)in percent (g/100 g), according to the following equations: 322,0IBDU(IBDU)N

45、= ww (1) DIN EN 15705:2010-07 EN 15705:2010 (E) 8 0,326CDUN(CDU)= ww (2) where 0,322 is the conversion factor for the content of IBDU in the fertilizer into nitrogen content; 0,326 is the conversion factor for the content of CDU in the fertilizer into nitrogen content. 6 Method B: Determination of m

46、ethylen-urea oligomers (MU) NOTE By the condensation of urea and formaldehyde several oligomers like methylendiurea (MDU), dimethylen-triurea (DMTU), trimethylen-tetraurea (TMTU) and higher oligomers are formed. These three molecules are the most soluble in water, the higher compounds are insoluble

47、in hot water, but their nitrogen is available for plants by microbiological decomposition. Also urea is always a companion of MU oligomers. 6.1 Principle The sample is extracted with boiling water and analyzed using a suitable HPLC system. The methylen-urea soluble oligomers are measured and detecte

48、d by the HPLC-method. In the HPLC-diagram methylen-urea oligomers are represented by different peaks: urea, methylen-diurea, dimethylen-triurea; trimethylen-tetraurea are, in the mean time, the most soluble and important. 6.2 Reagents 6.2.1 Reagents of recognized analytical grade and distilled or de

49、mineralized water (grade 3 according to EN ISO 3696:1995). 6.2.2 Acetonitrile, p.a., HPLC-grade. 6.2.3 Urea, p.a., 46,6 % of total nitrogen. 6.2.4 Methylen-diurea (MDU), synthesized and purified by a special laboratory, 42,4 % of total nitrogen.2)6.2.5 Dimethylen-triurea (DMTU), synthesized and purified by a special laboratory, 41,2 % of total nitrogen. 2)6.2.6 Trimethylen-tetraurea (TMTU), synthesized and purified by a special laboratory, 40,6 % of total nitrogen. 6.3