Thermo Scientific Dionex IonPac AS32-Fast-4mm Analytical User manual

For Research Use Only. Not for use in diagnostic procedures.
6
Dionex IonPac AS32-Fast-4µm Columns
065763Revision01 •March 2019
User Manual

Thermo Scientific
ProductManual for Dionex IonPac AS32-Fast-4µmColumns
Page 2 of 42
065763-01 For Research Use Only. Not for use in diagnostic procedures.
Product Manual
for
Dionex IonPac AS32-Fast-4µm Analytical Columns
4 ×150 mm (Item # 303151)
2 ×150 mm (Item # 303153)
Dionex IonPac AG32-Fast-4µm Guard Columns
4 × 30 mm (Item# 303152)
2 × 30 mm (Item# 303154)

Thermo Scientific
ProductManual for Dionex IonPac AS32-Fast-4µmColumns
Page 3 of 42
065763-01 For Research Use Only. Not for use in diagnostic procedures.
© 2019 Thermo FisherScientific Inc.Allrights reserved.
All trademarks are thepropertyofThermo FisherScientific Inc.and its subsidiaries.
Thermo FisherScientific Inc.providesthisdocumentto itscustomerswith a productpurchase touse in the product
operation.This documentis copyright protectedand any reproductionofthe whole orany part ofthis documentis
strictly prohibited,except with the writtenauthorization ofThermo FisherScientific Inc.
The contents ofthis documentare subject to change without notice.Alltechnicalinformation in this documentis
for reference purposes only. Systemconfigurations and specifications in this documentsupersedeall previous
information received bythe purchaser.
Thermo FisherScientific Inc. makes no representations that this documentis complete,accurate orerrorfree and
assumes no responsibility and will not be liable for any errors, omissions, damageor loss that might result from
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Sale shallgovernallconflicting informationbetweenthetwo documents.
Revision History:
Revision 01,March 2019; OriginalPublication.

Thermo Scientific
ProductManual for Dionex IonPac AS32-Fast-4µmColumns
Page 4 of 42
065763-01 For Research Use Only. Not for use in diagnostic procedures.
Safety and Special Notices
Makesure youfollowtheprecautionarystatementspresentedin this guide.The safety and other
specialnoticesappearin boxes.
Safety and specialnotices includethe following:
Indicatesa potentiallyhazardoussituationwhich,ifnotavoided,could resultindeathor
seriousinjury.
Indicatesa potentially hazardoussituationwhich,ifnot avoided,could result indamage
toequipment.
Indicates a potentially hazardous situation which, if not avoided, mayresult in minor or
moderate injury. Alsousedto identify a situation or practice that may seriously damage
the instrumentbut willnot causeinjury.
Indicatesinformationofgeneral interest.
IMPORTANT
Highlights informationnecessaryto prevent damage tosoftware, loss of data, or invalid
test results; or mightcontain information that is critical for optimal performanceof the
system.
Tip
Highlightshelpfulinformationthatcanmakeataskeasier.
SAFETY
!
WARNING
!
CAUTION
!
NOTE
!

Contents
Thermo Scientific
ProductManual for Dionex IonPac AS32-Fast-4µmColumns
Page 5 of 42
065763-01 For Research Use Only. Not for use in diagnostic procedures.
Contents
1. Introduction.........................................................................................7
1.1 Dionex IonPac AS32-Fast-4µm/Dionex IonPac AG32-Fast-4µm
Column Packing Specifications....................................................................................................................7
2. Installation..........................................................................................9
2.1 Thermo Scientific Dionex High Pressure Ion Chromatography Systems .........................................9
2.2 SystemRequirements...................................................................................................................................10
2.3 SystemVoid Volume.....................................................................................................................................10
2.4 Column Start-Up...........................................................................................................................................10
2.5 Dionex IonPac Guard Columns.................................................................................................................10
2.6 Sample Concentrators..................................................................................................................................11
2.7 The Injection Loop........................................................................................................................................11
2.8 Dionex Anion Dynamically RegeneratedSuppressor Requirements ...............................................12
2.9 Dionex Anion Chemically RegeneratedSuppressor Requirements..................................................12
2.10 Using Dionex Displacement Chemical Regeneration (DCR)
with the Chemical Suppression Mode ......................................................................................................12
3. Operation..........................................................................................13
3.1 General Operating Conditions...................................................................................................................13
3.2 General Operational Precautions..............................................................................................................14
3.3Quality Assurance Reports.........................................................................................................................14
3.4 Chemical Requirements and Solvent Compatibility ............................................................................14
3.5 Eluent Preparation........................................................................................................................................15
3.5.1 Eluent Generation..........................................................................................................................................15
3.5.2 Making Eluents that Contain Solvents .........................................................................................................15
3.5.3 Manually Prepared Eluents...........................................................................................................................16
4. Example Applications.........................................................................17
4.1 Isocratic Elution With andWithout a Guard Column........................................................................18
4.2 Analysis of Polythionates.............................................................................................................................19
4.3 Analysis of Aromatic Sulfonates, Pigments, and Dyes .........................................................................20
4.4 Analysis of Perchlorate in High Chloride and Sulfate Matrices........................................................22
4.5 Analysis of Persulfate...................................................................................................................................24

Contents
Thermo Scientific
ProductManual for Dionex IonPac AS32-Fast-4µmColumns
Page 6 of 42
065763-01 For Research Use Only. Not for use in diagnostic procedures.
5. Troubleshooting.................................................................................25
5.1 High Back Pressure......................................................................................................................................27
5.1.1 Findingthe Source of High SystemPressure...............................................................................................27
5.1.2 Replacing Column Bed SupportAssemblies (4 mmand 2 mmcolumns only)........................................27
5.1.3 Filter Eluent....................................................................................................................................................28
5.1.4 Filter Samples................................................................................................................................................28
5.2 High Background..........................................................................................................................................28
5.2.1 Preparation of Eluents ...................................................................................................................................28
5.2.2 A Contaminated Trap Column......................................................................................................................29
5.2.3 Contaminated DionexCR-TC Column ........................................................................................................29
5.2.4 A Contaminated Guard orAnalytical Column............................................................................................ 29
5.2.5 Contaminated Hardware................................................................................................................................29
5.2.6 A Contaminated Suppressor..........................................................................................................................29
5.3 Inconsistent Retention Times .....................................................................................................................29
5.3.1 Drifting to Shorter Retention Time...............................................................................................................30
5.3.2 Drifting to Longer Retention Time...............................................................................................................30
5.3.3 Oscillating Retention Times..........................................................................................................................30
5.4 Poor Peak Resolution ...................................................................................................................................31
5.4.1 Loss of Column Efficiency...........................................................................................................................31
5.4.2 Analyte Specific Efficiency Loss .................................................................................................................32
5.4.3 Shortened Retention Times...........................................................................................................................33
5.4.4 Loss of Front-End Resolution.......................................................................................................................34
5.5 Spurious Peaks...............................................................................................................................................34
6. Appendix A –Column Care................................................................35
6.1 Recommended Operation Pressures.........................................................................................................35
6.2 Column Start-Up...........................................................................................................................................35
6.3 Column Storage .............................................................................................................................................35
6.4 Chemical Purity Requirements..................................................................................................................35
6.4.1 Inorganic Chemicals......................................................................................................................................36
6.4.2 Deionized Water............................................................................................................................................36
6.4.3 Solvents..........................................................................................................................................................36
6.5 Column Cleanup............................................................................................................................................37
6.5.1 Choosing the Appropriate Cleanup Solution...............................................................................................38
6.5.2 Column Cleanup Procedure..........................................................................................................................39
7. Appendix B –Quality Assurance Reports............................................40
8. Appendix C –Additional Information.................................................42
8.1 General Information on PEEK™ Tubing ...............................................................................................42

1–Introduction
Thermo Scientific
ProductManual for Dionex IonPac AS32-Fast-4µmColumns
Page 7 of 42
065763-01 For Research Use Only. Not for use in diagnostic procedures.
1. Introduction
The Thermo Scientific™Dionex™IonPac™AS32-Fast-4µmcolumn is designed foranalysisof
low polarity anions,polysulfonatedaromatics,aromaticdyes,andpigments.Itsselectivityallows
for sub-ppb quantification of perchlorate in high chloride and sulfatematrices using suppressed
conductivity and mass spectrometry detection. The DionexIonPac AS32-Fast-4µm column is
available in standardbore(4mm i.d.) and microbore (2mm i.d) formats,each 150mm long.The
Dionex IonPac AS32-Fast-4µm column is compatible with pH 0-14 eluents and eluents
containingorganic solventsfrom0–100% in concentration.The DionexIonPac AS32-Fast-4µm
column can be used with any suppressible ionic eluent that does not exceed the capacity of the
Thermo Scientific DionexAnionDynamically RegeneratedSuppressor600(DionexADRS 600).
The Dionex IonPac AS32-Fast-4µm column has nominal efficiency for perchlorate of at least
7500 plates/column using standardoperatingconditions.
1.1 Dionex IonPac AS32-Fast-4µm/DionexIonPac AG32-Fast-4µmColumn Packing
Specifications
ResinCharacteristics:
Nominal Particle Size: 4 µm (Analyticaland Guard column)
ParticleCross-linking: 55%
Ion exchange capacity: 126 µeq per4 × 150 mm column
31.5 µeq per2 × 150 mm column
25 µeq per4 × 30 mm column
8 µeq per2 × 30 mm column
Functional Characteristics:
FunctionalGroup: Alkanolquaternary ammoniumion
Hydrophobicity: Ultralow
*AnalyticalandGuard Column resin composition:supermacroporous ethylvinylbenzene polymer
cross-linkedwith divinylbenzene

1–Introduction
Thermo Scientific
ProductManual for Dionex IonPac AS32-Fast-4µmColumns
Page 8 of 42
065763-01 For Research Use Only. Not for use in diagnostic procedures.
Table 1 Dionex IonPac AS32-Fast-4µm/Dionex IonPac AG32-Fast-4µm Operating
Parameters
Column(s)
Typical Back
Pressure
psi (MPaa), 30°Cb
Standard
Flow Rate
mL/min
Maximum
Flow Rate mL/minc, d
Dionex IonPac AS32-Fast-
4µm,
4mm Analytical column
~ 2500 (17.24)
1.0
1.0
Dionex IonPac AG32-Fast-
4µm,
4mm Guard column
~ 500 (3.44)
1.0
1.0
Dionex IonPac AS32-Fast-
4µmand AG32-Fast-4µm,
4mm columns
~ 3000 (20.68)
1.0
1.0
Dionex IonPac AS32-Fast-
4µm,
2mm Analytical column
~ 2200 (15.17)
0.25
0.25
Dionex IonPac AG32-Fast-
4µm,
2mm Guard column
~ 500 (3.44)
0.25
0.25
Dionex IonPac AS32-Fast-
4µmand AG32-Fast-4µm, 2
mm columns
~ 2700 (18.62)
0.25
0.25
aNote:1 MPa= 145.04 psi
bTotalbackpressureat standard flowrates
cIn all cases,flowrate should not result in pressuresover5,000 psi
dWhen developinga method formultiplecolumn formats orlong term, widespreaduse,it is
important to operatewithin the maximum flow rate listed above to ensurea reproducible
method.
Forassistance,visit UnityLab Servicesonlineatwww.unitylabservices.com.
FromtheU.S., call theCustomerCareCenterforDionexProductsat 1-800-346-6390.
OutsidetheU.S.,callthe nearest ThermoFisherScientificoffice.
Exceeding the maximumflow rateslisted intheabovetable candisrupt theuniformity of
the packingofthecolumnbedand irreversibly damagetheperformance ofthecolumn.
NOTE
!
WARNING
!

2–Installation
Thermo Scientific
ProductManual for Dionex IonPac AS32-Fast-4µmColumns
Page 9 of 42
065763-01 For Research Use Only. Not for use in diagnostic procedures.
2. Installation
Read theinstrumentmanuals.Thismanualassumes that youare using Thermo Scientific
Dionex instrumentationandare familiarwiththeinstallationand operationoftheThermo
Scientific Dionex IonChromatograph(IC).If youdo not understand theoperationofthe
system, take the time to familiarizeyourself with the various system components before
beginningananalysis.
The proper configuration of an Ion Chromatography System(ICS) is dependent on thecolumn
format. Although they can typically use the same system, the useof a 2 mm or 4 mm column
requires different set up to ensure optimumperformance. The selected format and analysis type
will affect the type of pump recommended. A gradientpump is designed to blend and pump
isocratic, linear, or gradient mixtures of up to four mobile phase components at precisely
controlled flowrates.An isocratic pump is forapplicationsnotrequiring gradient ormulti-eluent
proportioning capabilities. Forhighpressureapplications (over3000psi),theuseof high-pressure
consumables,components,and fittings arerequired.
2.1 Thermo Scientific DionexHigh Pressure Ion Chromatography Systems
A minimum of a Dionex High Pressure Ion Chromatography System(HPIC) is recommended
when running DionexIonPac 4µm columns due to the higherbackpressuresgeneratedat typical
operational flow rates. Systems should havethe capability to operateup to at least 5000 psi.
StandardICsystems,withan upperlimit of3000 psi,areinsufficient forpropercolumn operation.
All systems should allow 100% metal-free operation to prevent column damage. This includes
pump heads andallflow paths.
Careshould alwaysbe takennot to exceedthemaximumoperatingpressureofthesystem
components. ICS systems with lower backpressure capabilities are not recommended as
reducedflowratesmayresult inlossofperformance.
Contactyourlocalrepresentative forinformationonhow to customize yoursystemto
yourapplicationneeds.
NOTE
!
WARNING
!
NOTE
!

2–Installation
Thermo Scientific
ProductManual for Dionex IonPac AS32-Fast-4µmColumns
Page 10 of 42
065763-01 For Research Use Only. Not for use in diagnostic procedures.
2.2 System Requirements
Dionex IonPac columns are designed to be run on DionexIon Chromatographs equipped with
suppressed conductivity detection. We recommend the use of ferrules and fittings rated with a
pressure of >5000 psi. The use of Thermo Scientific Dionex IC PEEK ViperTM fittings is
recommended to achieve consistent low dead volume connections and ensure optimum
chromatographic performance. DionexIC PEEK Viper fittings are available in convenient kits
for Thermo Scientific DionexIntegrionTM systems with conductivity detectors (Item# 088798)
and DionexICS-6000 systems with conductivity detectors(Item# 088803).
2.3 System Void Volume
When using 2 mm columns, it is particularly importantto minimize systemvoid volume. The
systemvoid volume should bescaled down to at least 1/4 of the systemvolume in a standard 4
mm system. For best performance, all of the tubing installed between the injection valve and
detectorshould be 0.005" i.d.PEEK tubing.In orderto reduce systembackpressureat higherflow
rates,0.010" i.d. PEEK tubing may be used forconnections beforetheinjection valve,but peak
efficiency will be compromised if used for postinjector connections, which may also result in
decreased peak resolution. Minimize the lengths of all connecting tubing and remove all
unnecessaryswitchingvalvesandcouplers.
2.4 Column Start-Up
The column is shippedusing 4mM KOH as thestorage solution.Flushthecolumn to waste with
the QAReluentforat least 30minutes beforeattaching thecolumn outlet tothe suppressor.Install
thecolumn in thecolumn module andtestthe column performance underthe conditionsdescribed
in the QAR. Continue making injections of the test standard until consecutive injections of the
standardgive reproducible retention times.Equilibrationis complete when consecutiveinjections
of thestandardgive reproducibleretentiontimes.
IMPORTANT
Whenmakinganytubingconnections,firstturnoffthepump.Thiswillavoidanyslippage
of theferruleunderhighpressureconditions.
2.5 Dionex IonPac Guard Columns
A DionexIonPac Guard Column is normally usedwith theDionexIonPac AnalyticalColumn.A
guard is placed before the analyticalcolumn to prevent sample contaminantsfromdamaging the
analyticalcolumn.It is easiertocleanorreplace the guard column thanit is theanalyticalcolumn.
Placing a guard column in front oftheanalyticalcolumn will causeretention times toincreaseby
approximately 20% under isocratic test conditions. Replacing the guard column at the first sign
of peakefficiency lossordecreased retentiontime will prolong the life ofthe analyticalcolumn.

2–Installation
Thermo Scientific
ProductManual for Dionex IonPac AS32-Fast-4µmColumns
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065763-01 For Research Use Only. Not for use in diagnostic procedures.
2.6 Sample Concentrators
The function of a concentrator column is to strip ions of interest froma measured volume of a
relatively clean aqueous sample matrix. This process “concentrates” the desired analyte species
onto the concentrator column, lowering detection limits by 2-5 orders of magnitude. The
concentratorcolumn is used in lieu ofthe sample loop at the startofthe analysis.
Dionex Concentrator columns or a Dionex IonPac Guard Column can be used for trace anion
concentration work with DionexIonPac columns. A pump is used to load the sample onto the
concentrator column in the OPPOSITE direction of the eluent flow. Once concentration is
complete,the eluent flowis thendirectedthroughtheconcentratortotheanalyticalcolumn.When
using concentration techniques,careshould be takennotto overloadtheconcentratorcolumn by
concentrating an excessive amount of sample. If an excessive amount of sample is used,
inaccurateresults may be obtained. It is possible during the concentration step for the stronger
binding polyvalent ions to elutetheweakly retainedions fromthe concentratorcolumn.Formore
detailed information on sample concentration techniquesforhigh sensitivityworkand a detailed
discussion of concentration techniques, refer to the appropriateconcentrator manualfor your
application.
Dionex IonPac Concentrator Columnsare designed for use with specificeluentsystems.
Use onlyconcentratorcolumnsdesignedfortheeluentsystemyouareusing.
2.7 The Injection Loop
2.7.1.1 The 2 mm System Injection Loop, 2 –15 µL
For most applications on a 2 mm analytical system, a 2 –15 µL injection loop is sufficient.
Generally, you should not inject more than 12.5 nanomoles of any one analyte onto a 2 mm
analytical column. Injecting larger numbers of moles of a sample can result in overloading the
column which can affect the detection linearity. For low concentrations of analytes, larger
injection loops can be used to increase sensitivity. The DionexIonPac AS32-Fast-4µm 2 mm
columns require a microbore systemconfiguration. Install an injection loop one-fourth or less
(<15 µL) of the loop volume usedwith a 4mm analyticalsystem.
2.7.1.2 The 4 mm System Injection Loop, 10 –50 µL
For most applications on a 4 mm analyticalsystem, a 10 –50 µL injection loop is sufficient.
Generally, you should not inject more than 50 nanomoles of any one analyte onto the 4 mm
analytical column. Injecting larger numbers of moles of a sample can result in overloadingthe
column which can affect the detection linearity. For low concentrations of analytes, larger
injection loopscan beused toincreasesensitivity.
CAUTION
!

2–Installation
Thermo Scientific
ProductManual for Dionex IonPac AS32-Fast-4µmColumns
Page 12 of 42
065763-01 For Research Use Only. Not for use in diagnostic procedures.
2.8 Dionex Anion Dynamically Regenerated Suppressor Requirements
A Dionex Anion Dynamically Regenerated Suppressor should be used for applications that
requiresuppressedconductivitydetection.
Solventcontainingeluentsshould beusedintheAutoSuppressionExternal WaterMode.
For4 mm DionexIonPac AS32-Fast-4µmcolumns,use aDionexADRS600Anion Dynamically
Regenerated Suppressor(4mm, Item# 088666).
For2 mm DionexIonPac AS32-Fast-4µmcolumns,use aDionexADRS600Anion Dynamically
Regenerated Suppressor(2mm, Item# 088667).
For detailed information on the operation of the Dionex Anion Dynamically Regenerated
Suppressor, see Document No. 031956, the “Product Manual for the Dionex DRS 600.”
2.9 Dionex Anion Chemically Regenerated Suppressor Requirements
A Thermo Scientific Dionex Anion Chemically Regenerated Suppressor (DionexACRS 500)
may be used insteadofa DionexADRS600 forapplications thatrequiresuppressedconductivity
detection. Usea 4 mmDionexACRS 500 (4 mm, Item# 085090) with the 4 mm DionexIonPac
AS32-Fast-4µmcolumn. For 2 mm operation,usethe 2mm DionexACRS 500 (2 mm, Item#
085091). Thesesuppressors are compatible with all solvents andconcentrations with which the
systems andcolumnsarecompatible.
For detailed information on the operation of the Dionex Anion Chemically Regenerated
Suppressor, see Item # 031727, the “Product Manual for the Dionex Anion Chemically
Regenerated Suppressor 500, the Dionex ACRS 500”.
2.10 Using Dionex Displacement Chemical Regeneration (DCR) with the Chemical
Suppression Mode
The Dionex Displacement Chemical Regeneration (Dionex DCR) Mode is recommended for
chemical suppression using sulfuric acid and the Dionex Anion Chemically Regenerated
Suppressor (DionexACRS 500). See the DionexDCR kit manual, Document P/N 031664, for
details.
Use propersafety precautionsinhandlingacidsand bases.
CAUTION
!
SAFETY
!

3–Operation
Thermo Scientific
ProductManual for Dionex IonPac AS32-Fast-4µmColumns
Page 13 of 42
065763-01 For Research Use Only. Not for use in diagnostic procedures.
3. Operation
3.1 General Operating Conditions
The following conditions should be usedasa starting point towardsmethoddevelopment.
Table 2 General Operating Conditionsfor the Dionex IonPac AS32-Fast-4µm Column
Sample Volume:
2 mm: 2.5 µL Loop
4 mm: 10 µL Loop
Column:
2 mm: DionexIonPac AS32-Fast-4µm2 mm AnalyticalColumn +DionexIonPac
AG32-Fast-4µm 2 mm Guard Column
4 mm: DionexIonPac AS32-Fast-4µm4 mm AnalyticalColumn +DionexIonPac
AG32-Fast-4µm 4 mm Guard Column
Eluent:
4.0 mM KOH (QAR eluent)
Eluent Flow
Rate:
2 mm: 0.25 mL/min
4 mm: 1.0 mL/min
DRS Suppressor:
2 mm and 4mm:
DionexAnion Dynamically Regenerated Suppressor,DionexADRS600, AutoSuppression
RecycleMode
or CRS
Suppressor:
2 mm and 4mm:
DionexAnion Chemically Regenerated Suppressor,DionexACRS500
CRS Regenerant:
50 mN H2SO4
Expected
Background
Conductivity:
<1 µS
Long-term
StorageSolution
(> 1 week):
Eluent
Short-term
StorageSolution
(< 1 week):
Eluent
Forassistance,visit UnityLab Servicesonlineatwww.unitylabservices.com.
FromtheU.S., call theCustomerCareCenterforDionexProductsat 1-800-346-6390.
OutsidetheU.S.,callthe nearest ThermoFisherScientificoffice.
NOTE
!

3–Operation
Thermo Scientific
ProductManual for Dionex IonPac AS32-Fast-4µmColumns
Page 14 of 42
065763-01 For Research Use Only. Not for use in diagnostic procedures.
3.2 General Operational Precautions
The following precautions shouldalwaysbe adheredto whenusingDionexIonPaccolumns:
•Samples and manually preparedeluentsshould always be filteredanddegassedto protect
thesystemand column fromparticulatesandensurea stable background.
•Eluents and samplesusedshould be used within the allowable limits forthecolumn.
•Do not exceed the operationalpressureofthe system.
•Takecarenot to exceed themaximum operationalflowrateand pressureofthe column.
If the pressure approached the maximum allowed pressure,reduce theoperationalflow
rate.
Table 3 Operational Limits for the Dionex IonPac AS32-Fast-4µm Columns
Eluent pH
Between 0and 14
Sample pH
Between 0and 14
Maximum Flow Rate for 2 mm Columns
0.25 mL/min
Maximum Flow Ratefor 4 mm Columns
1.0 mL/min
Maximum Operating Pressure
5,000 psi(34.47MPa)
3.3 Quality Assurance Reports
Each column is qualified to ensure it meets specifications. Example copies of these Quality
AssuranceReports(QARs)canbe found in AppendixB –Quality AssuranceReports.TheQAR
suppliedwith the column should be usedas aguideto ensuresystemperformance.
3.4 Chemical Requirements and Solvent Compatibility
Chemical purity can influence separation performance. Only chemicals of the highest purity
shouldbeused.Referto Section 6.4,“ChemicalPurity Requirements”formoredetails.
Some solvents may be usedforcleaningoreluent modification.
Adding solvent to the aqueous eluent canreducethe peak responseby up to half due to
increased eluent viscosity, decreased ionization of organic acids, and lower peak
efficiencies.Therefore,onlyusesolventintheeluentwhenneededforimprovedresolution
of analytesofinterest.
NOTE
!

3–Operation
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ProductManual for Dionex IonPac AS32-Fast-4µmColumns
Page 15 of 42
065763-01 For Research Use Only. Not for use in diagnostic procedures.
Table 4 Typical HPLC Solvents for Cleaning and Use with Dionex IonPac Columns
Solvent
Maximum Operating Concentration
Acetonitrile
100%
*Higherconcentrations may onlybe usedforlimited durationapplicationssuch as column clean-
up at pressures <4000 psi.
Use ofa Dionex AERS500eoperatedintheAutoSuppressionExternalWaterMode is
recommendedwhenusing eluentscontainingorganicsolvents.RefertotheSuppressor
ProductManualforoperationlimitationsintheelectrolyticmode(poweron).
3.5 Eluent Preparation
3.5.1 Eluent Generation
It is recommended thatDionexIonPac columns are used with DionexHPICSystems equipped
with a Thermo Scientific Dionex Eluent Generator Cartridge. The use of eluent generation
providesa more stablebaseline,improvedperformance,andincreased reproducibility compared
to theuseofmanually preparedeluents.
The DionexEluent Generator is used to automatically produce eluents either isocratically or as
gradients fromdeionized water. Please refer to the Dionex EG manual for information on the
operationofthe Dionexeluent generators.
OnlyDionex EGC500cartridgesrated for5,000psi orhighershouldbeusedwhenrunning
Dionex IonPac 4µm columns due to the higher backpressures generated at typical
operationalflowrates.
3.5.2 Making Eluents that Contain Solvents
Mixing solvents with water should be done on a volume to volume basis. For example, if a
procedure requires an eluent of 40% acetonitrile, prepare the eluent by adding 400 mL of
acetonitrile to an eluent reservoir.Then add 600 mL of deionized wateror eluent concentrateto
theacetonitrile in thereservoir.Using this procedure to mix solventswith waterwill ensure that
a consistent,truevolume/volume eluent is obtained.Premixing waterwith solvent will minimize
thepossibility ofoutgassing.
Whenpurging ordegassing eluentscontaining solvents,do not purgeor degastheeluent
excessivelysinceit ispossiblethatavolatilesolvent canevaporateorbe “boiled” off from
the solutionunderreduced pressure.
CAUTION
!
CAUTION
!
NOTE
!

3–Operation
Thermo Scientific
ProductManual for Dionex IonPac AS32-Fast-4µmColumns
Page 16 of 42
065763-01 For Research Use Only. Not for use in diagnostic procedures.
Alwaysdegasand storealleluentsinplasticeluentbottlespressurizedwithhelium.Only
heliumcanbeusedto purgeanddegasioniceluentscontainingsolvents,sincenitrogenis
solubleinsolventcontainingeluents.
Acetonitrile (ACN)hydrolyzesto ammonia andacetatewhenexposedto basic solutions.
To prevent eluent contamination from acetonitrile hydrolysis, always add acetonitrileto
basic aqueous eluents by proportioning the acetonitrile into the basic eluent with the
gradientpump.Keep theacetonitrileinaseparateeluentbottlecontainingonlyacetonitrile
andwater.
Neveraddtheacetonitrile directlyto thebasiccarbonateorhydroxideeluentsolutions.
3.5.3 Manually Prepared Eluents
Eluents should be stored in plastic bottles under a heliumatmosphere to ensure contamination
free operation and proper pump performance(nitrogen can be used if eluents do not contain
solvents).Contaminationfromcarbondioxide whenbasic eluentsareexposedtotheaircan cause
performance variability such asretentiontime shifts.
NOTE
!
NOTE
!
SAFETY
!

4–Example Applications
Thermo Scientific
ProductManual for Dionex IonPac AS32-Fast-4µmColumns
Page 17 of 42
065763-01 For Research Use Only. Not for use in diagnostic procedures.
4. Example Applications
Forassistance,visit UnityLab Servicesonlineat www.unitylabservices.com.
FromtheU.S., call theCustomerCareCenterforDionexProductsat 1-800-346-6390.
OutsidetheU.S.,callthe nearest ThermoFisherScientificoffice.
The chromatograms in this section were obtained using columns that reproduced the Quality
Assurance Report on an optimized Ion Chromatograph. Different systems will differ slightly in
performance due to slight differences in column sets, systemvoid volumes, liquid sweep-out
times of different components,and laboratorytemperatures.
Ensure thatyour systemis properly configured and that all of the eluents have been made from
high purity reagentsand deionized water.Allwaterused in the preparation of eluentsshouldbe
degassed, deionized water with a specific resistance of 18.2 megohm-cm. For chemical purity
requirements,see Section 6.4,“ChemicalPurityRequirements”.Afterrunning synthetic standards
to calibrate your system, you may find that real sample matrices foul your columns. For this
reason, it is always advisable to use a guard column to protect the analytical column. If column
performance deteriorates and it is determined that theguard or the analytical column has been
fouled, refer to the column cleanup protocols in “AppendixA –Column Care”. If your sample
matrices are relatively low in ionic concentration,you may be able to increase the sensitivityof
yoursystemby using sample concentrationtechniques.
NOTE
!

4–Example Applications
Thermo Scientific
ProductManual for Dionex IonPac AS32-Fast-4µmColumns
Page 18 of 42
065763-01 For Research Use Only. Not for use in diagnostic procedures.
4.1 Isocratic Elution With and Without a Guard Column
Isocratic elution of anions on the DionexIonPac AS32-Fast-4µm Analytical Column has been
optimized utilizing a hydroxide eluent. By using this eluent, mono- and divalent anions can be
isocratically separated and quantitated in a single injection.TheDionexIonPac AS32-Fast-4µm
Analytical Column should always be used with the Dionex IonPac AG32-Fast-4µm Guard
Column.Retention times will increase by approximately 20% when a guard column is placedin-
line priorto the analyticalcolumn.
Figure 1 Dionex IonPac AS32-Fast-4µm Column (4 x 150 mm) With and Without Guard
Column
Figure 2 Dionex IonPac AS32-Fast-4µm Column (2 x 150 mm) With and Without Guard
Column
Column: See chromatogram
Eluent: 4 mM KOH
Flow Rate: 1.0 mL/min
Inj. Volume: 10 µL
Temperature: 30˚C
Detection: Suppressed Conductivity,
Dionex AERS 500, 4mm
AutoSuppression, recycle mode
Suppressor Current: 10 mA
Peaks: mg/L
1. Fluoride 1.25
2. Bromide 5.0
3. Iodide 10.0
4. Perchlorate 7.5
5. Sulfate 5.0
0 2 4 6 8 10 11
0
11
1 2 3 4
5
Minutes
µS
1 2 3 4
5
Dionex IonPac AS32-Fast-4µm 4 x 150 mm
Dionex IonPac AG32-Fast-4µm 4 x 30 mm +
Dionex IonPac AS32-Fast-4µm 4 x 150 mm
Column: See chromatogram
Eluent: 4 mM KOH
Flow Rate: 0.25 mL/min
Inj. Volume: 2.5 µL
Temperature: 30˚C
Detection: Suppressed Conductivity,
Dionex AERS 500, 2mm
AutoSuppression, recycle mode
Suppressor Current: 3 mA
Peaks: mg/L
1. Fluoride 1.25
2. Bromide 5.0
3. Iodide 10.0
4. Perchlorate 7.5
5. Sulfate 5.0
Dionex IonPac AS32-Fast-4µm 2 x 150 mm
Dionex IonPac AG32-Fast-4µm 2 x 30 mm +
Dionex IonPac AS32-Fast-4µm 2 x 150 mm
0 2 4 6 8 10 12
0
10 1 2 3
4
5
Minutes
µS 1 2 34
5

4–Example Applications
Thermo Scientific
ProductManual for Dionex IonPac AS32-Fast-4µmColumns
Page 19 of 42
065763-01 For Research Use Only. Not for use in diagnostic procedures.
4.2 Analysis of Polythionates
The following chromatograms demonstratethe separationofpolythionates on theDionexAS32-Fast-4µmcolumn.
Figure 3 Analysis of Polythionate Samples Using the Dionex IonPac AS32-Fast-4µm
Column (4 x 150 mm)
Figure 4 Analysis of Polythionate Samples Using the Dionex IonPac AS32-Fast-4µm
Column (2 x 150 mm)
Column: Dionex IonPac AG32-Fast-4µm (4 x 30 mm) /
Dionex IonPac AS32-Fast-4µm (4 x 150 mm)
Eluent: 4 mM KOH from 0 to4 minutes,
4-25 mM KOH from 4 to 10 minutes,
25 mM KOH from 10 to 15 minutes
Flow Rate: 1.0 mL/min
Inj. Volume: 10 µL
Temperature: 30˚C
Detection: Suppressed Conductivity,
Dionex AERS 500, 4mm
AutoSuppression, recycle mode
Suppressor Current: 62 mA
Peaks: mg/L
1. Dithionate 25.0
2. Trithionate 25.0
3. Tetrathionate 25.0
Dithionate
Trithionate
Tetrathionate
0 5 10 15
0
14
3
Minutes
µS
2
1
Column: Dionex IonPac AG32-Fast-4µm (2 x 30 mm) /
Dionex IonPac AS32-Fast-4µm (2 x 150 mm)
Eluent: 4 mM KOH from 0 to 4 minutes,
4-25 mM KOH from 4 to 10 minutes,
25 mM KOH from 10 to 15 minutes
Flow Rate: 0.25 mL/min
Inj. Volume: 2.5 µL
Temperature: 30˚C
Detection: Suppressed Conductivity,
Dionex AERS 500, 2mm
AutoSuppression, recycle mode
Suppressor Current: 16 mA
Peaks: mg/L
1. Dithionate 25.0
2. Trithionate 25.0
3. Tetrathionate 25.0
0 5 10 15
0
15
Minutes
µS
3
2
1
Dithionate
Trithionate
Tetrathionate

4–Example Applications
Thermo Scientific
ProductManual for Dionex IonPac AS32-Fast-4µmColumns
Page 20 of 42
065763-01 For Research Use Only. Not for use in diagnostic procedures.
4.3 Analysis of Aromatic Sulfonates, Pigments, and Dyes
The following chromatograms showthe separation ofvarious aromatic sulfonates,pigments,anddyeson the Dionex
IonPac AS32-Fast-4µmcolumn.
Figure 5 Analysis of Aromatic Sulfonates, Pigments, and Dyes Using the Dionex
IonPac AS32-Fast-4µm Column (4 x 150 mm)
Column: Dionex IonPac AG32-Fast-4µm (4 x 30 mm) /
Dionex IonPac AS32-Fast-4µm (4 x 150 mm)
Eluent: 4 mM KOH from 0 to 4 minutes,
4-40 mM KOH from 4 to 10 minutes,
40-80 mM KOH from 10 to 14 minutes,
80 mM KOH from 14 to 20 minutes
Flow Rate: 1.0 mL/min
Inj. Volume: 10 µL
Temperature: 30˚C
Detection: Suppressed Conductivity,
Dionex AERS 500, 4mm
AutoSuppression, recycle mode
Suppressor Current: 198 mA
Peaks mg/L
1. p-Toluenesulfonic acid 25
2. 2-Naphthalenesulfonic acid 25
3. 4-Sulfobenzoic acid 25
4. 1-Naphthol-4-sulfonic acid 25
5. Naphthalene-disulfonic acid 25
6. Naphthalene-trisulfonic acid 25
7. FDC Yellow #5 25
8. FDC Yellow #6 25
9. FDC Red #40 25
0 5 10 15 20
0
4
1
2
3
4
5
6 7
8
9
Minutes
µS