1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards PublicationFertilizers Determination of 3,4-dimethyl-1H-pyrazole phosphate (DMPP) Methodusing high-performance liquidchromatography (HPLC)BS EN 16328:2012Copyright European Committee for St
2、andardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-National forewordThis British Standard is the UK implementation of EN 16328:2012.The UK participation in its preparation was entrusted to Technical CommitteeCII/37,
3、Fertilisers and related chemicals.A list of organizations represented on this committee can be obtained on request to its secretary.This publication does not purport to include all the necessary provisions of a contract. Users are responsible for its correct application. The British Standards Instit
4、ution 2013.Published by BSI Standards Limited 2013 ISBN 978 0 580 76198 0 ICS 65.080Compliance with a British Standard cannot confer immunity from legal obligations.This British Standard was published under the authority of the Standards Policy and Strategy Committee on 31 January 2013.Amendments is
5、sued since publicationDate Text affectedBRITISH STANDARDBS EN 16328:2012Copyright European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 16328
6、 December 2012 ICS 65.080 English Version Fertilizers - Determination of 3,4-dimethyl-1H-pyrazole phosphate (DMPP) - Method using high-performance liquid chromatography (HPLC) Engrais - Dosage du 3,4-dimthyl-1H-pyrazole phosphate (DMPP) - Mthode par chromatographie liquide haute performance (HPLC) D
7、ngemittel - Bestimmung von 3,4-Dimethyl-1H-pyrazolphosphat (DMPP) - Verfahren mit Hochleistungs-Flssigchromatographie (HPLC) This European Standard was approved by CEN on 27 October 2012. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giv
8、ing 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-CENELEC Management Centre or to any CEN member. This European Standard exists in three off
9、icial versions (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-CENELEC Management Centre has the same status as the official versions. CEN members are the national standards bodies o
10、f Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain,
11、 Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 17, B-1000 Brussels 2012 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN natio
12、nal Members. Ref. No. EN 16328:2012: EBS EN 16328:2012Copyright European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-EN 16328:2012 (E) 2 Contents Page Foreword 31 Scope 42 Normative references
13、43 Terms and definitions .44 Principle 45 Reagents .46 Apparatus .47 Sampling .58 Procedure .58.1 Preparation of the test solution 58.2 Preparation of calibration solution 58.3 HPLC conditions 58.4 HPLC determination 69 Calculation and expression of the results 610 Precision .610.1 Inter-laboratory
14、test .610.2 Repeatability .610.3 Reproducibility .611 Test report 7Annex A (informative) Results of the inter-laboratory test .8A.1 Inter-laboratory tests .8A.2 Statistical results .8Bibliography 9BS EN 16328:2012Copyright European Committee for Standardization Provided by IHS under license with CEN
15、Not for ResaleNo reproduction or networking permitted without license from IHS-,-,-EN 16328:2012 (E) 3 Foreword This document (EN 16328:2012) 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
16、be given the status of a national standard, either by publication of an identical text or by endorsement, at the latest by June 2013, and conflicting national standards shall be withdrawn at the latest by June 2013. Attention is drawn to the possibility that some of the elements of this document may
17、 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 Trade Association. According to the CEN/CENELEC Internal Reg
18、ulations, the national standards organisations 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 Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland,
19、Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. BS EN 16328:2012Copyright European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduc
20、tion or networking permitted without license from IHS-,-,-EN 16328:2012 (E) 4 1 Scope This European Standard specifies a method for the determination of 3,4-dimethyl-1H-pyrazole phosphate (DMPP, CAS-No: 202842-98-6) in mineral N containing fertilizers using high-performance liquid chromatography (HP
21、LC). 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. For undated references, the latest edition of the referenced document (including any
22、amendments) applies. EN 1482-2, Fertilizers and liming materials Sampling 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 Vocab
23、ulary Part 2: Terms relating to fertilizers 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 fertilizer sample is dissolved in or extracted with water. The concentration of 3,4-dimethyl-1H-pyrazol
24、e phosphate is determined in diluted aqueous solution by high performance liquid chromatography (HPLC) at a reversed phase using a UV detector. 5 Reagents WARNING The described method includes the handling with hazardous materials. Attention must be paid to technical, organizational and personal saf
25、ety measures. Use only reagents of recognized analytical grade. 5.1 Water, de-ionized. 5.2 Acetonitrile, HPLC grade. 5.3 Sodiumdihydrogenphosphate-monohydrate, NaH2PO4H2O. 5.4 3,4-dimethyl-1H-pyrazole, 99 % purity. 6 Apparatus 6.1 Ultrasonic bath and magnetic stirrer, respectively. BS EN 16328:2012C
26、opyright European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-EN 16328:2012 (E) 5 6.2 Filtration unit, manual or high-pressure filtration with membrane filter 0,45 m or comparable filter. NOTE
27、High pressure filtration is advantageous for difficult samples. 6.3 HPLC unit, for isocratic operation with UV detector. 6.4 Evaluation unit. 6.5 Reverted phase-HPLC separation column, e.g. C18, 5 m, 150 mm 4,6 mm. NOTE With satisfactory separation performance, a shorter separation column can be use
28、d. 6.6 Analytical balance, capable of weighing to an accuracy of 0,001 g. 7 Sampling 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 1482-2. 8 Procedure 8.1 Preparation o
29、f the test solution Weigh (6.6) (30 0,001) g of sample (m) into a 1 000 ml volumetric flask. Add approximately 900 ml water (5.1) and dissolve the sample in the ultrasonic bath (6.1) for about 15 min. Fill up the flask with water (5.1) to capacity. Filter part of the sample solution through a membra
30、ne filter (6.2) to remove possible conditioners. If necessary, dilute the sample for the calibration range. 8.2 Preparation of calibration solution Prepare from at least two weighted samples (stock solutions) made with the present 3,4-dimethyl-1H-pyrazole (5.4) at least four concentrations for calib
31、ration. The concentration range should be between 3 mg/l and 100 mg/l. 8.3 HPLC conditions Separation column: filled for reversed phase-HPLC (6.5); column temperature: room temperature; elution agent: dissolve 1,38 g NaH2PO4H2O (5.3) in 1 l water and add 175 ml acetonitrile (5.2); the elution agent
32、shall be de-gassed, e.g. in an ultrasonic bath (6.1); flow rate: 1,5 ml/min; injection volume: 10 l; wave length: 224 nm; retention time DMPP (at 1,5 ml/min flow rate): approximately 5,3 min. BS EN 16328:2012Copyright European Committee for Standardization Provided by IHS under license with CENNot f
33、or ResaleNo reproduction or networking permitted without license from IHS-,-,-EN 16328:2012 (E) 6 8.4 HPLC determination To prepare a calibration curve, inject 10 I of each calibration solution three times consecutively. Calculate the compensation curve and correlation coefficient p by least-square
34、method. The calibration curve can be used for concentration determination when p 0,99. Inject the test solution two times consecutively. 9 Calculation and expression of the results Carry out the evaluation on the basis of the calibration line over the peak areas. Calculate the content of 3,4-dimethy
35、l-1H-pyrazole phosphate, wDMPP, in mg/kg according to formula (1): 000101,962,194100)(dpkDMPP=maFbAw (1) where Apkis the peak height or peak area; b is the y-axis section of straight calibration line; a is the slope of straight calibration line; Fdis the dilution factor; m is the sample weight, in m
36、illigrams. Calculate the arithmetic mean from both values obtained. Report the result to the nearest 0,001 mg/kg. 10 Precision 10.1 Inter-laboratory test Details of inter laboratory tests performed in 2010 on the precision of the method are summarized in Annex A. The values derived from this test ma
37、y not be applicable to concentration ranges and matrices other than those given. 10.2 Repeatability The absolute difference between 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 with
38、in a short interval of time, will in no more than 5 % of the cases be greater than the repeatability limit r given in Table 1. 10.3 Reproducibility The absolute difference between two single test results, obtained using the same method on identical test material in different laboratories with differ
39、ent operators using different equipment, will in no more than 5 % of the cases be greater than the reproducibility limit R given in Table 1. BS EN 16328:2012Copyright European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted wi
40、thout license from IHS-,-,-EN 16328:2012 (E) 7 Table 1 Mean values, repeatability and reproducibility limits for DMPP Sample x r R mg/kg mg/kg mg/kgSample 1 Ammonium sulfate nitrate (ASN) 1 700 90 170 Sample 2 NPK 24+8+7 1 270 70 410 Sample 3 NPK 13+10+20 706 56 210 11 Test report The test report sh
41、all contain at least the following information: a) all information necessary for the complete identification of the sample; b) test method used with reference to this document; c) test results obtained; d) date of sampling and sampling procedure (if known); e) date when the analysis was finished; f)
42、 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 incidents occurred when performing the method, which might have influenced the test result(s). BS EN 16328:2012Copyri
43、ght European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-EN 16328:2012 (E) 8 Annex A (informative) Results of the inter-laboratory test A.1 Inter-laboratory tests An inter-laboratory test has b
44、een carried out in 2010 with nine participating laboratories and three different samples of fertilizers. Repeatability and reproducibility were calculated according to ISO 5725-1 and ISO 5725-2. A.2 Statistical results The statistical results are given in Table A.1. Table A.1 Statistical results for
45、 the determination of DMPP using HPLC Parameter Sample 1 Sample 2 Sample 3 Ammonium sulfate nitrate (ASN) NPK 24+8+7 NPK 13+10+20Year of the test 2010 2010 2010 Number of participating laboratories 9 9 9 Number of laboratories after eliminating outliers 7 8 8 Mean value, x , (mg/kg) 1 700 1 270 706
46、Repeatability standard deviation sr, (mg/kg) 30 20 20Relative standard deviation of repeatability, RSDr2 1,9 3 Repeatability limit, r, (mg/kg) 90 70 56 Reproducibility standard deviation, sR,(mg/kg) 60 150 75Relative standard deviation of reproducibility, RSDR4 11,6 11 Reproducibility limit, R, (mg/
47、kg) 170 410 210BS EN 16328:2012Copyright European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-EN 16328:2012 (E) 9 Bibliography 1 EN 1482-1, Fertilizers and liming materials Sampling and sample
48、preparation Part 1: Sampling 2 EN ISO 3696:1995, Water for analytical laboratory use Specification and test methods (ISO 3696:1987) 3 ISO 5725-1, Accuracy (trueness and precision) of measurement methods and results Part 1: General principles and definitions 4 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 5 Reg
