1、BS EN1014-3:2010ICS 71.100.50NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBRITISH STANDARDWood preservatives Creosote and creosotedtimber Methods ofsampling and analysisPart 3: Determination of thebenzo(a)pyrene content of creosoteThis British Standardwas published under the
2、authority of the StandardsPolicy and StrategyCommittee on 31 July 2010 BSI 2010ISBN 978 0 580 64046 9Amendments/corrigenda issued since publicationDate CommentsBS EN 1014-3:2010National forewordThis British Standard is the UK implementation of EN 1014-3:2010. Itsupersedes BS EN 1014-3:1998 which is
3、withdrawn.The UK participation in its preparation was entrusted to TechnicalCommittee B/515, Wood preservation.A list of organizations represented on this committee can be obtained onrequest to its secretary.This publication does not purport to include all the necessary provisionsof a contract. User
4、s are responsible for its correct application.Compliance with a British Standard cannot confer immunityfrom legal obligations.BS EN 1014-3:2010EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 1014-3 June 2010 ICS 71.100.50 Supersedes EN 1014-3:1997English Version Wood preservatives - Creosote an
5、d creosoted timber - Methods of sampling and analysis - Part 3: Determination of the benzo(a)pyrene content of creosote Produits de prservation du bois - Crosote et bois crosot - Mthodes dchantillonnage et danalyse - Partie 3: Dtermination de la teneur en benzoapyrne dans la crosote Holzschutzmittel
6、 - Kreosot (Teerimprgnierl) und damit imprgniertes Holz - Probenahme und Analyse - Teil 3: Bestimmung des Gehaltes an Benzo(a)pyren im Kreosot This European Standard was approved by CEN on 12 May 2010. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the cond
7、itions for giving 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 thr
8、ee official 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 Management Centre has the same status as the official versions. CEN members are the national standards bodies of
9、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 United Kingdom.
10、EUROPEAN COMMITTEE 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 1014-3:2010: EBS EN 1014
11、-3:2010EN 1014-3:2010 (E) 2 Contents Page Foreword 31 Scope 42 Normative references 43 Principle 44 Reagents .45 Apparatus .46 Preparation of the calibration solutions and of the test samples 56.1 Preparation of the calibration solutions 56.2 Preparation of the test samples .57 Procedure .68 Calcula
12、tion 69 Expression of results 610 Precision .710.1 Repeatability .710.2 Reproducibility .711 Test report 7Annex A (informative) Examples of chromatograms .8Annex B (informative) Interpretation of the test results . 11Bibliography . 12BS EN 1014-3:2010EN 1014-3:2010 (E) 3 Foreword This document (EN 1
13、014-3:2010) has been prepared by Technical Committee CEN/TC 38 “Durability of wood and wood-based products”, the secretariat of which is held by AFNOR. 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
14、 by December 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 s
15、uch patent rights. This document supersedes EN 1014-3:1997. This standard forms part of a series of standards relating to the sampling and analysis of creosote and creosoted timber. The other standards of the series are: EN 1014-1, Wood preservatives Creosote and creosoted timber Methods of sampling
16、 and analysis Part 1: Procedure for sampling creosote EN 1014-2, Wood preservatives Creosote and creosoted timber Methods of sampling and analysis Part 2: Procedure for obtaining a sample of creosote from creosoted timber for subsequent analysis EN 1014-4, Wood preservatives Creosote and creosoted t
17、imber Methods of sampling and analysis Part 4: Determination of the water-extractable phenols content of creosote According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgar
18、ia, 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 Kingdom. BS EN 1014-3:2010EN
19、1014-3:2010 (E) 4 1 Scope This European Standard specifies a method for the determination of benzo(a)pyrene in creosote using high performance liquid chromatography (HPLC). This standard is only applicable to creosotes containing more than 30 mg/kg benzo(a)pyrene. 2 Normative references The followin
20、g 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 ISO 3696:1995, Water for analytical laboratory use Specifi
21、cation and test methods (ISO 3696:1987) 3 Principle The creosote sample is diluted to an appropriate concentration with an acetonitrile/water mixture. The diluted sample is analyzed using high performance liquid chromatography (HPLC) at constant temperature with a reverse phase packed column and iso
22、cratic elution. The result is compared with that from a known reference standard. 4 Reagents 4.1 Acetonitrile water mixture. Add 220 ml of water (according to grade 1 of EN ISO 3696:1995) to 780 ml of acetonitrile (CH3CN, HPLC grade) and mix thoroughly. The mixture shall be at room temperature befor
23、e use. 4.2 Benzo(a)pyrene solution, containing approximately 0,1 mg/ml of benzo(a)pyrene. Weigh to the nearest 0,1 mg, approximately 0,01 g of benzo(a)pyrene with a purity of min. 98 % and transfer it quantitatively to a 100 ml one-mark volumetric flask. Add 50 ml of the acetonitrile (CH3CN, HPLC gr
24、ade). Dissolve the benzo(a)pyrene and make up to the mark with the acetonitrile-water mixture (4.1). Store the prepared solution in the stoppered brown glass storage flasks (5.6) in the dark, at a temperature below 10 C. CAUTION Care should be taken to prevent benzo(a)pyrene contacting the skin. NOT
25、E 1 Under normal conditions, this solution is stable for six months, although frequent use may result in faster ageing. NOTE 2 Commercially available certified standard solutions, including those containing additional components, may be used. 5 Apparatus Usual laboratory apparatus and glassware toge
26、ther with the following: 5.1 Volumetric glassware, which shall have an accuracy of at least 0,5 %. 5.2 Single mark pipettes class A of 1 ml, 2 ml, 5 ml and 20 ml capacity. BS EN 1014-3:2010EN 1014-3:2010 (E) 5 5.3 High performance liquid chromatograph (HPLC) which shall consist of a solvent delivery
27、 pump with constant flow regulation; on-line degasser; 20 l loop injector; reverse phase stainless steel column, 250 mm in length with an internal diameter of 4 mm, packed with C18 bonded silica stationary phase, having a particle size of 5 m; the use of a column specially designed for polyaromatic
28、hydrocarbons analysis is recommended; fluorescence detector capable of being set at (380 3) nm excitation wavelength and (403 5) nm emission wavelength; a potentiometric recorder. NOTE As an alternative, any other HPLC configuration giving at least the same resolution (see Figures A.1 and A.2 or A.3
29、) could be used. 5.4 Analytical balance, capable of weighing to 0,1 mg. 5.5 Ultrasonic bath capable of containing a 100 ml volumetric flask. 5.6 Brown glass storage flasks of 100 ml capacity, fitted with ground glass stoppers. 5.7 Glass or metal receptacles fitted with stoppers. 5.8 Glass syringe fo
30、r HPLC, of 50 I capacity. 6 Preparation of the calibration solutions and of the test samples 6.1 Preparation of the calibration solutions Transfer by pipette (5.2) 1 ml of the benzo(a)pyrene solution (4.2) to a 100 ml one-mark volumetric flask and dilute to the mark with the acetonitrile-water mixtu
31、re (4.1). To a series of 100 ml one-mark volumetric flasks, transfer by pipette (5.2) 1 ml, 2 ml, 5 ml and 20 ml of this solution and dilute to the mark with acetonitrile-water mixture (4.1). This provides calibration solutions containing 0,01 mg/I, 0,02 mg/I, 0,05 mg/I and 0,20 mg/I benzo(a)pyrene.
32、 Store the calibration solutions in stoppered brown glass storage flasks (5.6.) in the dark at a temperature below 10 C. NOTE Under normal conditions, the calibration solutions are stable for three months, although frequent use may result in faster ageing. 6.2 Preparation of the test samples Prepare
33、 duplicate test samples. The laboratory sample shall be in a closed receptacle (5.7). Heat the closed receptacle to 70 C for (30 2) min to ensure that the sample is completely liquid. Weigh to the nearest 0,1 mg, 30,0 mg of the laboratory sample into a 100 ml one-mark volumetric flask. NOTE Attentio
34、n should be paid that no re-crystallization occurs during this transfer. Record the masses taken as m1and m2. Add approximately 80 ml of the acetonitrile-water mixture (4.1). Place the volumetric flask in the ultrasonic bath (5.5) for 5 min to dissolve the sample. BS EN 1014-3:2010EN 1014-3:2010 (E)
35、 6 When the sample has dissolved completely, make up to the mark with the acetonitrile-water mixture (4.1). 7 Procedure 7.1 Set-up the HPLC (5.3) in accordance with the manufacturers instructions. Adjust the fluorescence detector to the following wavelengths: excitation: (380 3) nm; emission: (403 5
36、) nm. The fluorescence detector shall be fine-tuned to maximize the signal for benzo(a)pyrene. Under isocratic conditions set the flow rate through the column to 1,2 ml/min using the acetonitrile-water mixture (4.1) as the eluent. 7.2 Analyze the test samples and the calibration solutions at the sam
37、e temperature ( 0,5 C). At the same temperature (i.e. within 0,5 C) and using the syringe (5.8) fill up the loop injector and inject successively the series of calibration solutions (6.1) and then the two test samples into the HPLC apparatus (5.3). 7.3 Repeat 7.2 in reverse order by successively inj
38、ecting portions of the two test samples followed by the calibration solutions. 7.4 Measure the peaks heights for benzo(a)pyrene produced by the HPLC recorder for all test samples and calibration solutions. 8 Calculation Calculate the benzo(a)pyrene content of the two samples, Bs1and Bs2, expressed i
39、n milligrams benzo(a)pyrene per kilogram creosote, using the equation: 6ccscs10=HCHBB where Bcis the concentration of the benzo(a)pyrene calibration solution nearest to the test sample in milligrams per litre (mg/l); Hcis the mean of the duplicated benzo(a)pyrene peak heights obtained with the calib
40、ration solution in millimetres (mm); Ccis the concentration of creosote in the test sample (6.2) in milligrams per litre (mg/l); Hsis the peak height obtained for the test sample (6.2) in millimetres (mm). 9 Expression of results Report the benzo(a)pyrene content B, of the laboratory sample as the a
41、verage of Bs1 and Bs2in milligrams per kilogram benzo(a)pyrene in creosote, rounded to the nearest 1 mg/kg (see Annex B). BS EN 1014-3:2010EN 1014-3:2010 (E) 7 10 Precision 10.1 Repeatability Results obtained by the same operator shall be considered suspect if the duplicates differ by more than 10 %
42、 of the lower in the concentration range up to 100 mg/kg benzo(a)pyrene in creosote and 5 % of the lower above 100 mg/kg benzo(a)pyrene in creosote. 10.2 Reproducibility Single results obtained by two laboratories shall be considered suspect if they differ by more than 30 % of the lower. 11 Test rep
43、ort The test report shall include at least the following information: a) the number and date of this European Standard; b) full identification of the sample tested and details of its preparation for analysis; c) the date of the test; d) the results of the analysis, expressed as milligrams per kilogr
44、am benzo(a)pyrene in creosote (see Clause 9); e) whether the repeatability has been verified; f) any particular points observed in the course of the test; g) any operations not specified in the method or regarded as optional which might have affected the result. BS EN 1014-3:2010EN 1014-3:2010 (E) 8
45、 Annex A (informative) Examples of chromatograms Column: Packing C 18-particle size 5 m; Length 250 mm; Internal diameter 4,6 mm. Operating conditions: Flow: 1,2 ml/min; Temperature: 25 C. NOTE Retention times depend on the column used and on the operational conditions. Figure A.1 Standard solution
46、BS EN 1014-3:2010EN 1014-3:2010 (E) 9 Column: Packing C 18-particle size 5 m; Length 250 mm; Internal diameter 4,6 mm. Operating conditions: Flow: 1,2 ml/min; Temperature: 25 C. NOTE Retention times depend on the column used and on the operational conditions. Figure A.2 Creosote oil with approximate
47、ly 500 mg/kg benzo(a)pyrene in creosote BS EN 1014-3:2010EN 1014-3:2010 (E) 10 Column: Packing C 18-particle size 5 m; Length 250 mm; Internal diameter 4,6 mm. Operating conditions: Flow: 1,2 ml/min; Temperature: 25 C. NOTE Retention times depend on the column used and on the operational conditions.
48、 Figure A.3 Detail of chromatogram BS EN 1014-3:2010EN 1014-3:2010 (E) 11 Annex B (informative) Interpretation of the test results This method can e.g. be used to determine the benzo(a)pyrene content of unused creosote (for instance sampled according to EN 1014-1) or of creosote extracted from creos
49、oted timber (for example as obtained using the procedure defined in EN 1014-2). In the latter case, however, care should be taken when drawing conclusions as to the benzo(a)pyrene content of the creosote with which the timber was originally treated. In the course of the service-life the chemical composition of the creosote in treated timber changes as a result of the preferential evaporation or leaching of certain compone
