1、BRITISH STANDARD BS EN 13273:2001 Surface active agents Determination of the content of non-ionic substances in anionic surface active agents by high performance liquid chromatography (HPLC) The European Standard EN 13273:2001 has the status of a British Standard ICS 71.100.40 NO COPYING WITHOUT BSI
2、 PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBS EN 13273:2001 This British Standard, having been prepared under the direction of the Sector Committee for Materials and Chemicals, was published under the authority of the Standards Committee and comes into effect on 15 June 2001 BSI 06-2001 ISBN 0
3、580 37402 5 National foreword This British Standard is the official English language version of EN 13273:2001. The UK participation in its preparation was entrusted to Technical Committee CII/34, Methods of test for surface active agents, which has the responsibility to: A list of organizations repr
4、esented on this committee can be obtained on request to its secretary. Additional information Some textual errors were discovered when the English language version of EN 13273:2001 was adopted as the national standard. Some of the chemical names are not consistently presented and do not conform to I
5、UPAC recommendations. These textual errors have been reported to CEN in a proposal to amend the text of the European Standard. Cross-references The British Standards which implement international or European publications may be found in the BSI Standards Catalogue under the section entitled “Interna
6、tional Standards Correspondence Index”, or by using the “Find” facility of the BSI Standards Electronic Catalogue. A British Standard does not purport to include all the necessary provisions of a contract. Users of British Standards are responsible for their correct application. Compliance with a Br
7、itish Standard does not of itself confer immunity from legal obligations. aid enquirers to understand the text; present to the responsible European committee any enquiries on the interpretation, or proposals for change, and keep the UK interests informed; monitor related international and European d
8、evelopments and promulgate them in the UK. Summary of pages This document comprises a front cover, an inside front cover, the EN title page, pages 2 to 14, an inside back cover and a back cover. The BSI copyright date displayed in this document indicates when the document was last issued. Amendments
9、 issued since publication Amd. No. Date CommentsEUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 13273 April 2001 ICS 71.100.40 English version Surface active agents Determination of the content of non-ionic substances in anionic surface active agents by high performance liquid chromatography (H
10、PLC) Agents de surface Dtermination de la teneur en substances non-ioniques dans les agents de surfaces anioniques par chromatographie liquide haute performance (CLHP) Grenzflchenaktive Stoffe Bestimmung des Gehaltes von nichtionischen Substanzen in anionischen grenzflchenaktiven Stoffen mittels Hoc
11、hleistungs- Flssigchromatographie (HPLC) This European Standard was approved by CEN on 20 January 2001. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European Standard the status of a national standard without any alteration.
12、 Up-to-date lists and bibliographical references concerning such national standards may be obtained on application to the 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 translation un
13、der the responsibility of a CEN member into its own language and notified to the Management Centre has the same status as the official versions. CEN members are the national standards bodies of Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece, Iceland, Ireland, Italy, Luxe
14、mbourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: rue de Stassart, 36 B-1050 Brussels 2001 CEN All rights of exploitation in any form and by any
15、 means reserved worldwide for CEN national Members. Ref. No. EN 13273:2001 EPage 2 EN 13273:2001 BSI 06-2001 Contents Page Foreword.3 1 Scope .4 2 Normative references .4 3 Principle .4 4 Reagents 4 5 Apparatus 4 6 Calibration .5 7 Sampling and preparation of the test sample7 8 Procedure 7 9 Calcula
16、tion and expression of results7 10 Precision 8 11 Test report .8 Annex A (informative) Flow diagram for the backflush valve in position 1 and position 29 Annex B (informative) Example chromatograms 10 Annex C (informative) Ring test results .14Page 3 EN 13273:2001 BSI 06-2001 Foreword This European
17、Standard has been prepared by Technical Committee CEN/TC 276, Surface active agents, 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 by October 2001, and co
18、nflicting national standards shall be withdrawn at the latest by October 2001. Annexes A, B and C are informative. According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Czech
19、 Republic, Denmark, Finland, France, Germany, Greece, Iceland, Ireland, Italy, Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and the United Kingdom.Page 4 EN 13273:2001 BSI 06-2001 1 Scope This European Standard specifies a method for the determination of the contents of non-
20、ionic substances in anionic surface active agents (sulfates, sulfonates and ethoxysulfates) by high performance liquid chromatography (HPLC). 2 Normative references This European Standard incorporates by dated or undated reference, provisions from other publications. These normative references are c
21、ited at the appropriate places in the text and the publications are listed hereafter. For dated references, subsequent amendments to or revisions of any of these publications apply to this European Standard only when incorporated in it by amendment or revision. For undated references the latest edit
22、ion of the publication referred to applies (including amendments). EN ISO 3696, Water for analytical laboratory use Specification and test methods (ISO 3696:1987). ISO 607, Surface active agents and detergents Methods of sample division. 3 Principle A specified mass of sample is dissolved and then m
23、ade up to standard volume with methanol. For sulfonic acids only, the sample is neutralized with aqueous sodium hydroxide solution prior to the dissolution. The solution is then analysed by reverse phase high performance liquid chromatography (RP-HPLC). An octadecylsilane bonded stationary phase, me
24、thanol/water mobile phase, differential refractometer detector and external means of quantification are employed. A back flush technique is used to “focus” the non-ionic components after all the active matter components and water have eluted from the column. 4 Reagents During the analysis, only use
25、reagents of recognized analytical grade and water complying with grade 3 as defined in EN ISO 3696. 4.1 Methanol, CH 4 O HPLC grade. 4.2 Sodium hydroxide NaOH c(NaOH) = 1 mol/l solution. 4.3 Phenolphthalein indicator, ethanolic solution, C 20 H 14 O 4 , w(C 20 H 14 O 4 ) = 0,1 g/100 g. 4.4 Calibrati
26、on substances, based on the feedstock from which the surface active agent/sulfonic acid has been produced, e.g.: parent linear alkyl benzene (LAB) for alkyl benzene sulfonates; parent alcohols for alcohol sulfates; parent alcohol ethoxylates for alcohol ethoxy sulfates. 5 Apparatus Ordinary laborato
27、ry apparatus and the following. 5.1 High performance liquid chromatograph, suitable for analysis according to the conditions given in 6.3.Page 5 EN 13273:2001 BSI 06-2001 5.2 Data logger/plotter giving the chromatographic peak areas. 5.3 Burette, capacity 10 ml with polytetrafluorethylene (PTFE) key
28、, never lubricated with stopcock grease. 5.4 Syringe, capacity 1 ml with “Luer” fitting. 5.5 Needle with “Luer” fitting to suit sample injection valve, or use an automated injector. 5.6 Syringe filter or membrane suitable for use with methanol, 0,45 m, with “Luer” fittings. 6 Calibration 6.1 General
29、 Always wash glassware thoroughly with water and then acetone (propan-2-one). Dry thoroughly. 6.2 Determination of the optimum backflush time Prepare and analyse the appropriate calibration substance (4.4) according to clause 7, but keep the backflush valve in position 1 (see Figure A.1) throughout
30、the complete analysis. The chromatogram of the calibration substance alkyl ether sulfate, type 2, used in the ring test is shown in Figure B.1. Note the elution time just before the first peak of the calibration substance appears. This is the backflush valve switching time. The above valve switching
31、 time needs only be determined if the method is being set up or if it becomes apparent that the retention times have changed during routine use. 6.3 Determination of response factors Determine the response factors for each type of the surface active agents as follows. Prepare a calibration solution
32、by weighing (0,15 0,02) g of the calibration substance (4.4) to the nearest 0,1 mg into a 100 ml volumetric flask. Dissolve with methanol (4.1) and make up to volume. Load the syringe (5.4) with 1 ml of the calibration solution and mount a new syringe filter fitted with the needle (5.5). Use the fol
33、lowing conditions: Column: Type HPLC column with appropriate fittings. A radial compression type is suitable if used with appropriate conditions. Length internal diameter Dimensions which enable results similar to the chromatograms in annex B. For a radial compression column, 100 mm 8 mm is suitable
34、. For conventional columns, 250 mm 4,6 mm is suitable. Stationary phase Octadecylsilyl bonded micro-particulate silica of mean particle size not more than 5 m. This should be of the completely-end-capped or deactivated type.Page 6 EN 13273:2001 BSI 06-2001 Mobile Phase: Composition A mixture of meth
35、anol and water capable of use with the chosen HPLC column so as to give results similar to the examples in annex B. A mixture of methanol : water of 85:15 parts by volume can be suitable Flow rate 1 ml/min to 2 ml/min to the nearest 0,01 ml/min. Detector: Type Differential refractometer Sensitivity
36、A setting which gives a display of peak sizes such that deviations from normal appearance can be readily seen. Temperatures: Column 30,0 C NOTE Thermostatting of the column gives improved stability, but the method can be used with the column at ambient temperatures. Detector 40,0 C Injection system:
37、 High pressure syringe-loading injection valve or a suitable automated injector, fitted with an injection loop of appropriate volume, for example 50 l. Backflush valve: A 6 port high pressure switching valve for HPLC, fitted with a very low volume by-pass loop. Valve positioning: Position 1 at the b
38、eginning of the analysis. Switch to position 2 at time A (see Figures A.1 and B.1) Switch to position 1 on completion of the analysis (typical analysis time: 20 min). Run the chromatograph in accordance with the manufacturers instructions. When injecting the blend ensure the sample loop is completel
39、y flushed by dispensing the total syringe volume taking care not to introduce any air bubbles. An example chromatogram is given in Figure B.1. NOTE The calibration peak elutes at a run time twice that of the backflush valve switch time. This time can vary depending on the sample type and is derived
40、in 6.2. The response factor, F, is calculated using the equation (1): c c A m F (1) where: m c is the mass of the calibration substance used, in grams; A cis the area of the calibration peak, in arbitrary units.Page 7 EN 13273:2001 BSI 06-2001 7 Sampling and preparation of the test sample Sample the
41、 material to be tested in accordance with ISO 607. 8 Procedure 8.1 Determine the optimum backflush time from the calibration carried out in 6.2. 8.2 Weigh (1 0,1) g of the sample to the nearest 0,1 mg into a 25 ml volumetric flask. 8.3 For sulfonic acids only, add one drop of phenolphthalein indicat
42、or (4.3) and then add 2,00 ml of sodium hydroxide solution (4.2). Swirl to dissolve. When completely dissolved, titrate dropwise with sodium hydroxide solution (4.2) to the end point characterized by faint pink colour (total titrant addition typically about 3,2 ml). For the other surface active agen
43、ts, continue to 8.4. Use a burette (5.3). 8.4 Dissolve and make up to volume with methanol (4.1). Allow to stand for a short time such that any precipitate formed settles to the bottom of the flask. 8.5 Load the syringe (5.4) with 1 ml of supernatant from the sample solution. Mount a new syringe fil
44、ter or membrane (5.6). Use this to deliver a filtered sample portion for use in the analysis. Where possible, this should be done by direct delivery into the injection valve, via the needle (5.5). 8.6 Analyse the sample according to the conditions given in 6.3. When injecting the sample ensure the s
45、ample loop is completely flushed by dispensing the total syringe volume taking care not to introduce any air bubbles. NOTE 1 An example chromatogram is given in Figure B.2 and Figure B.4. If the valve was not used, a chromatogram as shown in Figure B.3 should result. NOTE 2 The operation of the back
46、flush valve should be automated. 9 Calculation and expression of results The content of non-ionic substances w NISof the sample expressed in grams per 100 g is calculated using the equation (2): 4 100 s s NIS m F A w (2) where: A sis the area of the peak of non-ionic substances, as used for A c(see
47、6.3) in arbitrary units; F is the response factor of the calibration solution (6.3); m s is the mass of the sample (8.2) in grams. Report the result to the nearest 0,001 g/100 g.Page 8 EN 13273:2001 BSI 06-2001 10 Precision 10.1 Repeatability limit The absolute difference between two independent sin
48、gle test results, obtained using 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 not exceed the repeatability limit, r, in more than 5 % of cases. From the data given in annex C, the relative repeat
49、ability limits are: 12 % of the mean value at a level of 2 % to 3 % on active matter; 19 % of the mean value at a level below 1 % on active matter. 10.2 Reproducibility limit 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 not exceed the reproducibility limit, R, in more than 5 % of cases. From the data given in annex C the relative reproducibility limi
