1、Designation: D 5297 95 (Reapproved 2004)Standard Test Methods forRubber Chemical AcceleratorPurity by High PerformanceLiquid Chromatography1This standard is issued under the fixed designation D 5297; the number immediately following the designation indicates the year oforiginal adoption or, in the c
2、ase of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. Asuperscript epsilon (e) indicates an editorial change since the last revision or reapproval.1. Scope1.1 These test methods cover the determination of the purityof present commercially availabl
3、e rubber chemical acceleratorsin the range from 80 to 100 % by high performance liquidchromatography (HPLC) using ultraviolet detection and exter-nal standard calculations.1.2 Expertise in HPLC is necessary to the successful appli-cation of these test methods.1.3 This standard does not purport to ad
4、dress all of thesafety concerns, if any, associated with its use. It is theresponsibility of the user of this standard to establish appro-priate safety and health practices and determine the applica-bility of regulatory limitations prior to use.2. Referenced Documents2.1 ASTM Standards:2D 3853 Termi
5、nology Relating to Rubber and RubberLaticesAbbreviations for Chemicals Used in Com-poundingD 4483 Practice for Evaluating Precision for Test MethodStandards in the Rubber and Carbon Black ManufacturingIndustriesD 4571 Test Methods for Rubber ChemicalsDetermination of Volatile MaterialD 4936 Test Met
6、hod for Mercaptobenzothiazole Sulfena-mide Assay by Reduction/Titration2.2 ISO Standard:3ISO 6472 Rubber Compounding IngredientsAbbreviations3. Terminology3.1 Definitions:3.1.1 external standard calculationa method of calculat-ing the percent composition by measuring the area of theanalyte peak, mul
7、tiplying by a response factor, and dividing bythe sample concentration. All components are assumed to beresolved from the component of interest.3.1.2 lot samplea production sample representative of astandard production unit, normally referred to as the sample.3.1.3 specimenalso known as the test por
8、tion, it is theactual material used in the analysis. It must be representative ofthe lot sample.3.2 Abbreviations: The following abbreviations are in ac-cordance with Terminology D 3853 and ISO 6472:3.2.1 MBTSBenzothiazyl disulfide.3.2.2 MBS2-(morpholinothio)benzothiazole.3.2.3 CBSN-cyclohexyl-2-ben
9、zothiazolesulfenamide.3.2.4 TBBSN-t-butyl-2-benzothiazolesulfenamide.3.2.5 DIBSN,N8- diisopropyl-2-benzothiazolesulfena-mide.3.2.6 DCBSN,N8 - dicyclohexyl-2-benzothiazolesulfe-namide.3.2.7 DPGdiphenylguanidine.3.2.8 DOTGdi-o-tolylguanidine.4. Summary of Test Methods4.1 A specimen is dissolved in the
10、 appropriate solvent and afixed loop volume is analyzed by isocratic HPLC using athermostated C18 reversed phase column for materials 3.2.1-3.2.6 and a silica normal phase column for materials 3.2.7 and3.2.8, and an ultraviolet (UV) detector. Peak areas are deter-mined using a chromatographic integr
11、ator or laboratory datasystem with the amount of analyte being determined byexternal calibration.5. Significance and Use5.1 These test methods are designed to determine the purityof rubber chemical accelerators.1These test methods are under the jurisdiction of ASTM Committee D11 onRubber and is the
12、direct responsibility of Subcommittee D11.11 on ChemicalAnalysis.Current edition approved Dec. 1, 2004. Published December 2004. Originallyapproved in 1992. Last previous edition approved in 2000 as D 5297 95 (2000).2For referenced ASTM standards, visit the ASTM website, www.astm.org, orcontact ASTM
13、 Customer Service at serviceastm.org. For Annual Book of ASTMStandards volume information, refer to the standards Document Summary page onthe ASTM website.3Available from American National Standards Institute (ANSI), 25 W. 43rd St.,4th Floor, New York, NY 10036.1Copyright ASTM International, 100 Bar
14、r Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.5.2 Since the results of these test methods are based on anintegrated peak area, it is assumed that all analytes of interestare resolved from interfering peaks.6. Interferences6.1 Components co-eluting with the analyte of i
15、nterest willcause erroneous results; thus it is required that the system becapable of providing a minimum of 10 000 theoretical plates.7. Apparatus7.1 Liquid Chromatograph, consisting of the following:7.1.1 Precision chromatographic pump,7.1.2 Variable wavelength UV detector,7.1.3 A method for therm
16、ostating the column at 35 6 1C,for example, a column oven or water jacket, and7.1.4 A fixed loop injector with a nominal volume of 10mm3(L) or less.7.2 HPLC Column:7.2.1 A C18 (ODS) reversed phase column packed withspherical, totally porous monomolecular 5-m particles ca-pable of providing 40 000 th
17、eoretical plates per metre. (Aminimum of 10 000 plates is required for this analysis.) Thiscolumn should be reserved for this analysis.7.2.2 For materials 3.2.7 and 3.2.8, use a silica normal phasecolumn packed with spherical, totally porous 5-m particlescapable of providing 40 000 theoretical plate
18、s per metre. (Aminimum of 10 000 plates is required for this analysis.) Thiscolumn should be reserved for this analysis.7.3 Integrator/Data System, capable of determining abso-lute amounts of analyte of interest by means of integration ofdetector output versus time.7.4 Analytical Balance, capable of
19、 measuring within 60.01mg.8. Reagents and Materials8.1 Acetic Acid, glacial.8.2 Acetonitrile, HPLC grade.8.3 Chloroform, AR grade.8.4 Ethanol, HPLC grade.8.5 Ethanolamine.8.6 n-Hexane, HPLC grade.8.7 Methanol, HPLC grade.8.8 Water, HPLC grade.9. Calibration and Standardization9.1 Aprimary standard o
20、f known purity is used to determinethe response factor for each analyte.TEST METHOD ASULFENAMIDEACCELERATORPURITY10. Procedure10.1 Chromatographic Conditions:10.1.1 Determine the mobile phase composition and theflow rate by adjusting the chromatographic parameters for theparticular column chosen. Th
21、e mobile phase consists of theappropriate mixture of HPLC grade acetonitrile and HPLCgrade or equivalent water, both containing 0.001 M glacialacetic acid or less depending on the particular column chosen.(HPLC grade methanol may be added to the acetonitrile/watereluent to achieve the necessary sepa
22、ration for DIBS andMBTS.)10.1.2 For the analysis of the sulfenamides, adjust the flowrate and mobile phase composition to provide a capacity factor,k8, in the range from 4 to 6 for the analyte of interest, and aminimum resolution, Rs, of 2 between the MBTS impurity andthe analyte of interest.NOTE 1D
23、ifferent liquid chromatography columns may exhibit differ-ent elution characteristics. Suggested chromatographic starting parametersfor analysis are as follows:PercentH2OAPercentAcetonitrileAPercentMethanolAFlow rate(cm3/min)DCBS 5 95 0 2.5CBS 20 80 0 2.0TBBS 30 70 0 1.7MBS 45 55 0 1.4DIBS 15 0 85 1
24、.0AContaining 0.001 M glacial acetic acid.10.1.3 The capacity factor, k8, is defined as the retentiontime of the analyte, tA, minus the retention time of anunretained solute (solvent peak), to, divided by to:k8 5 tA2 to!/to(1)10.1.4 The resolution, Rs, is a function of the capacityfactor, selectivit
25、y, and the theoretical plates of the column:Rs5t22 t1!1/2 tw11 tw2!(2)where:t1,t2= retention times of the analyte and MBTS, andtw1,tw2= peak widths at 10 % of the peak height.10.2 DetectorMonitor the absorbance of all componentsat 275 nm. The detector sensitivity should be set to 1absorbance unit fu
26、ll scale (AUFS).10.3 Integrator/Data SystemThe integrator settingsshould be adjusted to give a full-scale response to 1 absorbanceunit (AU).10.4 Standard PreparationWeigh at least 50 mg to thenearest 0.01 mg of the standard in a 50-cm3volumetric flaskand dilute to volume with acetonitrile. Adjust th
27、e standardconcentration if necessary by serial dilution with acetonitrile togive a maximum absorbance (peak height) between 0.4 and 0.8AU (the linear range of the chromatographic system). Thestandard must be analyzed within4hofbeing diluted.NOTE 2Preparation of StandardsThe analytical standards are
28、pre-pared by multiple recrystallizations of the sulfenamides. Dissolve 100 g ofthe sulfenamide in 200 cm3of analytical reagent (AR) grade toluene withslight warming. Add2gofactivated carbon and stir for 30 min. Filter thehot solution by gravity and cool in an ice/acetone bath. Filter the crystalswit
29、h suction. Repeat this crystallization. Dissolve the analyte crystals fromthe second toluene crystallization in hot methanol, cool in an ice/acetonebath, and filter with suction. Repeat the alcohol recrystallization and dryat low pressure at 50C overnight. The procedure can be repeated until thedesi
30、red purity is obtained. The purity of the standard is estimated bygradient HPLC analysis of the impurities and differential thermal analysis(DTA). The purity of the standard should be reestimated by HPLC of theimpurities every 90 days. The standard should be stored at 5C or lower.Volatile matter and
31、 free amine content can be measured using TestMethods D 4571 and D 4936, respectively.10.5 Test PreparationTo ensure specimen homogeneity, 5g of the lot sample should be ground with a mortar and pestle.D 5297 95 (2004)210.6 Analysis:10.6.1 Weigh at least 50 mg to the nearest 0.01 mg of thespecimen i
32、nto a 50-cm3volumetric flask. Dissolve in acetoni-trile (a sonic bath is recommended) and dilute to volume withacetonitrile. Adjust the concentration, if necessary, by serialdilution with acetonitrile to give a maximum absorbance within10 % of the standard absorbance. Filter through a chemicallyresi
33、stant filter with a nominal pore size less than or equal to 0.5m. Analyze within4hofdilution. Chromatograph thestandard and measure the area.TEST METHOD BBENZOTHIAZOLEACCELERATORPURITY11. Procedure11.1 Chromatographic Conditions:11.1.1 Determine the mobile phase composition and theflow rate by adjus
34、ting the chromatographic parameters for theparticular column chosen. The mobile phase consists of theappropriate mixture of HPLC grade acetonitrile and HPLCgrade or equivalent water, both containing 0.001 M glacialacetic acid or less depending on the particular column chosen.11.1.2 For the analysis
35、of the benzothiazoles, adjust the flowrate and mobile phase composition to provide a capacity factor,k8, in the range from 4 to 6 for the analyte of interest, and aminimum resolution, Rs, of 2 between the MBTS impurity andthe analyte of interest.NOTE 3Different liquid chromatography columns may exhi
36、bit differ-ent elution characteristics. Suggested chromatographic starting parametersfor analysis are as follows:PercentH2OAPercentAcetonitrileAPercentMethanolAFlow rate(cm3/min)MBT 65 35 0 2.0MBTS 20 80 0 2.0AContaining 0.001 M glacial acetic acid.11.1.3 The capacity factor, k8, is defined as the r
37、etentiontime of the analyte, tA, minus the retention time of anunretained solute (solvent peak), to, divided by to:k8 5 tA2 to! / to(3)11.1.4 The resolution, Rs, is a function of the capacityfactor, selectivity, and the theoretical plates of the column:Rs5t22 t1!1/2 tw11 tw2!(4)where:t1,t2= retentio
38、n times of the analyte and MBTS, andtw1,tw2= peak widths at 10 % of the peak height.11.2 DetectorMonitor the absorbance of all componentsat 275 nm. The detector sensitivity should be set to 1absorbance unit full scale (AUFS).11.3 Integrator/Data SystemThe integrator settingsshould be adjusted to giv
39、e a full-scale response to 1 absorbanceunit (AU).11.4 Standard PreparationWeigh at least 50 mg to thenearest 0.01 mg of the standard in a 50-cm3volumetric flaskand dilute to volume with acetonitrile for MBT and chloroformfor MBTS. Adjust the standard concentration if necessary byserial dilution with
40、 acetonitrile to give a maximum absorbance(peak height) between 0.4 and 0.8 AU (the linear range of thechromatographic system). The standard must be analyzedwithin4hofbeing diluted.NOTE 4Preparation of StandardsThe analytical standards may beprepared by multiple recrystallizations of the benzothiazo
41、les. The purity ofthe standard is estimated by gradient HPLC analysis of the impurities anddifferential thermal analysis (DTA). The purity of the standard should bereestimated by HPLC of the impurities every 90 days. The standard shouldbe stored at 5C or lower.11.5 Test PreparationTo ensure specimen
42、 homogeneity, 5g of the lot sample should be ground with a mortar and pestle.11.6 Analysis:11.6.1 Weigh at least 50 mg to the nearest 0.01 mg of thespecimen into a 50-cm3volumetric flask. Dissolve MBT inacetonitrile and MBTS in chloroform (a sonic bath is recom-mended) and dilute to volume with acet
43、onitrile for MBT andchloroform for MBTS. Adjust the concentration, if necessary,by serial dilution with acetonitrile to give a maximum absor-bance within 10 % of the standard absorbance. Filter with achemically resistant filter with a nominal pore size less than orequal to 0.5 m. Analyze within4hofb
44、eing diluted.Chromatograph the standard and measure the area.TEST METHOD CGUANIDINE ACCELERATORPURITY12. Procedure12.1 Chromatographic Conditions:12.1.1 Determine the mobile phase composition and theflow rate by adjusting the chromatographic parameters for theparticular column chosen. The mobile pha
45、se consists of theappropriate mixture of HPLC grade n-hexane, ethanol, andmethanol containing 0.01 M ethanolamine or less depending onthe particular column chosen.12.1.2 For the analysis of the guanidines, adjust the flowrate and mobile phase composition to provide a capacity factor,k8, in the range
46、 from 6 to 8 for the analyte of interest.NOTE 5Different liquid chromatography columns may exhibit differ-ent elution characteristics. Suggested chromatographic starting parametersfor analysis are as follows:Percentm-HexanePercentEthanolAPercentMethanolAFlow rate(cm3/min)DPG 91 4 5 2.0DOTC 91 4 5 2.
47、0AContaining 0.01 M ethanolamine.12.1.3 The capacity factor, k8, is defined as the retentiontime of the analyte, tA, minus the retention time of anunretained solute (solvent peak), to, divided by to:k8 5 tA2 to! / to(5)12.2 DetectorMonitor the absorbance of all componentsat 240 nm. The detector sens
48、itivity should be set to 1absorbance unit full scale (AUFS).12.3 Integrator/Data SystemThe integrator settingsshould be adjusted to give a full-scale response to 1 absorbanceunit (AU).12.4 Standard PreparationWeigh at least 50 mg to thenearest 0.01 mg of the standard in a 50-cm3volumetric flaskand d
49、ilute to volume with 45/55 (v/v) ethanol/methanol.Adjustthe standard concentration if necessary by serial dilution withD 5297 95 (2004)3m-hexane to give a maximum absorbance (peak height) be-tween 0.4 and 0.8 AU (the linear range of the chromatographicsystem). The standard must be analyzed within4hofbeingdiluted.NOTE 6Preparation of StandardsThe analytical standards are pre-pared by multiple recrystallizations of the guanidines. The purity of thestandard is estimated by gradient HPLC analysis of the impurities anddifferential
