Thermo Scientific IONPAC AG2 GUARD COLUMN User manual

PRODUCT MANUAL
IONPAC®AG2 GUARD COLUMN
(4 x 50 mm, P/N SP4834)
IONPAC®AG3 GUARD COLUMN
(4 x 50 mm, P/N 043182)
IONPAC®AG4 GUARD COLUMN
(4 x 50 mm, P/N 035310)
IONPAC®AG5 GUARD COLUMN
(4 x 50 mm, P/N 035396)
IONPAC®CG2 GUARD COLUMN
(4 x 50 mm, P/N 035370)
IONPAC®AS2 ANALYTICAL COLUMN
(4 x 250 mm, P/N SP4831)
IONPAC®AS3 ANALYTICAL COLUMN
(4 x 250 mm, P/N 043181)
IONPAC®AS4 ANALYTICAL COLUMN
(4 x 250 mm, P/N 035311)
IONPAC®AS5 ANALYTICAL COLUMN
(4 x 250 mm, P/N 035395)
IONPAC®CS2 ANALYTICAL COLUMN
(4 x 250 mm, P/N 035371)
QUICKSTART STEPS AND LINKS
Click blue text below to get started.
1. See Section 2, “Operation”. Note operation precautions and chemical purity
requirements.
2. See “Quality Assurance Reports”. Run the Production Test Chromatogram as
a system check.
3. See “Column Care” for column cleanup and long-term storage
recommendations. ©DIONEX Corporation, 1997, 1998, 2002
Document No. 032226
Revision 14
12 November 2002

IonPac AS3, AS4, AS5, CS2 Document No. 032226-14 Page 2 of 14
TABLE OF CONTENTS
SECTION 1 - INTRODUCTION ..................................................................................................... 3
1.1 Overview ................................................................................................................................................................... 3
1.2 Chemicals ................................................................................................................................................................. 3
1.3 Column Storage ........................................................................................................................................................ 3
1.4 Recommended Operating Pressures....................................................................................................................... 3
1.5 Applications .............................................................................................................................................................. 3
SECTION 2 - OPERATION ............................................................................................................. 4
2.1 General Operation Precautions .............................................................................................................................. 4
2.2 Chemicals Required ................................................................................................................................................. 4
2.2.1 Inorganic Chemicals .................................................................................................................................................. 4
2.2.2 Solvents .................................................................................................................................................................... 4
2.2.3 DeionizedWater ........................................................................................................................................................ 4
SECTION 3 - STANDARD CONDITIONS .................................................................................... 5
3.1 Standard Reagents ................................................................................................................................................... 5
3.2 Standard Flow Rates and Back Pressures .............................................................................................................. 6
SECTION 4 - INITIALSTART-UP PROCEDURES ...................................................................... 7
SECTION 5 - TROUBLESHOOTING GUIDE ............................................................................... 8
5.1 High Back Pressure ................................................................................................................................................ 8
5.1.1 Finding the Source of High System Pressure............................................................................................................ 8
5.1.2 Replacing Column Bed SupportAssemblies ............................................................................................................. 9
5.1.3 Bed Support Descriptions ....................................................................................................................................... 10
5.2 High Background or Noise .................................................................................................................................... 12
5.2.1 Preparation of Eluents ............................................................................................................................................. 12
5.2.2 A ContaminatedAnion Trap Column ...................................................................................................................... 12
5.2.3 AContaminated Guard orAnalytical Column ......................................................................................................... 12
5.2.4 Contaminated Hardware .......................................................................................................................................... 12
5.2.5 A Contaminated Self-Regenerating Suppressor (SRS™) or the MicroMembrane™ Suppressor (MMS) ............... 13
5.3 Poor Peak Resolution ............................................................................................................................................ 13
5.3.1 Lossof Column Efficiency ...................................................................................................................................... 13
5.3.2 Poor Resolution Due to Shortened Retention Times .............................................................................................. 13
5.3.3 Loss of Front End Resolution ................................................................................................................................. 14
5.4 Spurious Peaks ...................................................................................................................................................... 14

IonPac AS3, AS4, AS5, CS2 Document No. 032226-14 Page 3 of 14
SECTION 1 - INTRODUCTION
1.1 Overview
Thismanualdescribeshowtopreparethiscolumnforoperation.Theinitialstart-upprocedureusuallyconsistsofinstallingacolumn
in the Chromatography Module, flushing it with deionized water to remove the storage solution, and equilibrating it with eluent.
1.2 Chemicals
Thechemicalsusedtomakeeluents,standards,regenerantsandpost-columnreagentsmustbeofveryhighpurityinordertoobtain
reliable, consistent and accurate analytical results. Before starting an analysis, consult Section 2.2, “Chemicals Required,” for
chemicalpurityrequirements.
1.3 Column Storage
If a column will not be used within one week after testing, prepare it for long-term storage. Flush the column first with deionized
water and then with the storage solution specified in Section 3, “Standard Conditions,” of this manual. Cap both ends securely,
using the plugs supplied with the column.
1.4 Recommended Operating Pressures
Operatingacolumnaboveitsrecommendedpressurelimitcancauseirreversiblelossofcolumnperformance.SeeTable2,“Standard
Flow Rates and Back Pressures,” for standard and maximum back pressures.
1.5 Applications
For help in selecting the correct column for your application, contact the nearest DIONEX Office (see, “DIONEX Worldwide
Offices”).
Alwaysrememberthatassistanceisavailableforanyproblemthatmaybeencounteredduringtheshipmentoroperation
of DIONEX instrumentation and columns through the DIONEX North America Technical Call Center at 1-800-
DIONEX-0(1-800-346-6390)orthroughanyoftheDIONEXOfficeslisted in, “DIONEXWorldwideOffices.”

IonPac AS3, AS4, AS5, CS2 Document No. 032226-14 Page 4 of 14
SECTION 2 - OPERATION
2.1 General Operation Precautions
2.2 Chemicals Required
Obtaining reliable, consistent and accurate results requires eluents that are free of ionic and spectrophotometric impurities.
Chemicals,solventsand deionized water usedtoprepareeluentsmust be of thehighestpurityavailable.Low trace impurities and
lowparticlelevels in eluents alsohelptoprotect your ion exchangecolumnsandsystem components. DIONEX cannotguarantee
propercolumn performancewhenthe qualityofthechemicals,solventsandwaterused toprepareeluents hasbeencompromised.
2.2.1 Inorganic Chemicals
ReagentGrade inorganic chemicals should always be used toprepare ionic eluents. Whenever possible, inorganic chemicals that
meetorsurpassthelatestAmericanChemicalSocietystandardforpurityshouldbeused.Theseinorganicchemicalswilldetailthe
purity by having an actual lot analysis on each label.
2.2.2 Solvents
CAUTION
Eluents used on columns containing low cross-linked resins must contain less than 5% organic solvents.
Ifsmallamountsofsolventsareaddedtoioniceluentstomodifytheionexchangeprocessorimprovesamplesolubility,thesolvents
usedmustbefreeofionicimpurities.However,sincemostmanufacturersofsolventsdonottestforionicimpurities,itisimportant
that the highest grade of solvents available be used. Currently, several manufacturers are making ultrahigh purity solvents that
are compatible for HPLC and spectrophotometric applications. These ultrahigh purity solvents will usually ensure that your
chromatographyisnotaffectedbyionicimpuritiesinthesolvent.CurrentlyatDIONEX,wehaveobtainedconsistentresultsusing
High Purity Solvents manufactured by Burdick and Jackson and Optima®Solvents by Fisher Scientific.
2.2.3 Deionized Water
Thedeionizedwater usedtoprepareeluents shouldbeDegassed Type I Reagent Grade Water with aspecificresistanceof 18.2
megohm-cm.Thedeionizedwatershouldbefreeofionizedimpurities,organics,microorganismsandparticulatematterlargerthan
0.2µm.BottledHPLC-GradeWater(withtheexceptionofBurdick&Jackson)shouldnotbeusedsincemostbottledwatercontains
an unacceptable level of ionic impurities. Finally, thoroughly degas all deionized water prior to preparing any eluents.
CAUTION
< 5% Organic Solvent in the Eluent
Filter and Degas Eluents
Filter Samples

IonPac AS2, AS3, AS4, AS5, CS2 Manual Document No. 032226-14 Page 5 of 14
SECTION 3 - STANDARD CONDITIONS
3.1 Standard Reagents
Table 1
Storage Solutions, Eluents and Regenerants
Column Storage Standard Standard
Solution Eluent Regenerant
ANIONANALYTICALANDGUARDCOLUMNS
IonPac®AS2/AG2 100 mM NaOH 3 mM Na2CO3/2 mM NaOH 10X Eluent Concentration
IonPacAS3/AG3 100 mM NaOH 2.2 mM Na2CO3/2.8 mM NaHCO310X Eluent Concentration
IonPacAS4/AG4 100 mM NaOH 2.25 mM Na2CO3/2.8 mM NaHCO310X Eluent Concentration
IonPacAS5/AG5 100 mM NaOH 2.8 mM Na2CO3/2.2 mM NaHCO310X Eluent Concentration
CATIONANALYTICALANDGUARDCOLUMNS
IonPac CS2/CG2 100 mM NaOH 10 mM Oxalic/7.5 mM Citric Acids 10X Eluent Concentration
(pH 4.35)

IonPac AS2, AS3, AS4, AS5, CS2 Manual Document No. 032226-14 Page 6 of 14
3.2 Standard Flow Rates and Back Pressures
Table 2
Standard Flow Rates and Back Pressures
Column Standard Expected Maximum
FlowRate Back Pressure Back Pressure
AnionAnalyticalandGuardColumns
IonPac AS2/AG2 3.0mL/min 600 psi/200 psi 1,500 psi
IonPac AS3/AG3 3.0mL/min 600 psi/200 psi 1,500 psi
IonPac AS4/AG4 2.0mL/min 1,000 psi/300 psi 2,000 psi
IonPac AS5/AG5 2.0mL/min 1,000 psi/300 psi 2,000 psi
Cation Analytical and Guard Columns
IonPacCS2/CG2 2.3mL/min 650 psi/150 psi 1,500 psi

IonPac AS3, AS4, AS5, CS2 Document No. 032226-14 Page 7 of 14
SECTION4 -INITIAL START-UPPROCEDURES
Fortheinitialstart-upofbothanionandcationanalyticalcolumns,preparetheeluentlistedonthetestchromatogram,setthepump
to the proper flow rate and perform the following initial start-up procedures. In the case of a guard or suppressor column, use the
test conditions on the production test chromatogram for the analytical column required for the application.
A. Connect the column directly to the pump with a short piece of tubing.
B. Direct the effluent of the column to a separate waste container.
C. Pump10mL(30mLforpacked-bedsuppressors)ofdeionizedwaterthroughthecolumnatthestandardflowratelisted
in Table 2, “Standard Flow Rates and Back Pressures.”
D. Assemble the system (trap, concentrator, guard, analytical and suppressor columns depending on the application).
E. Pump10mLofthestandardproductiontesteluentthatislistedontheAnalyticalColumnTestChromatogram,through
the System to equilibrate the columns.
F. PerformtheProductionTestChromatogramtotestthecolumnandsystemperformance.Notethatguardandanalytical
columns are fully equilibrated when successive runs have the same solute retention times.
NOTE
IonPac FastAnionAnalytical Column
You must use a 20 µLinjection loop with the IonPac FastAnion
to prevent excessive system band broadening and column overloading.
Maintain the eluent at between pH 2 and 11. Do not use hydroxide eluents.

IonPac AS3, AS4, AS5, CS2 Document No. 032226-14 Page 8 of 14
SECTION5-TROUBLESHOOTING GUIDE
ThepurposeoftheTroubleshooting Guide is to help you solve operatingproblemsthatmayarisewhileusingDIONEXcolumns.
Formoreinformation on problemsthatoriginatewiththe Ion Chromatograph(IC)orthesuppressor, refer totheTroubleshooting
Guide in the appropriate operator’s manual. If you cannot solve the problem on your own, call the nearest DIONEX Office (see,
“DIONEXWorldwideOffices”).
5.1 High Back Pressure
5.1.1 Finding the Source of High System Pressure
Totalsystempressure when using specific DIONEX columns is flow rate dependent.Referto Table 2, “Standard Flow Rates and
Back Pressures,” for specific flow rates and back pressures for the column system that your are operating. If the system pressure
ismorethan20%higherthanthetotalbackpressureestimatedfromthebackpressureslistedinTable2,“StandardFlowRatesand
BackPressures,”itisadvisabletofindoutwhatiscausingthehighsystempressure.ThesystemshouldbeusedwithaHigh-Pressure
In-LineFilter(P/N035331)fortheeluentswhichispositionedbetweenthepumppressuretransducerandtheinjectionvalve.Make
sure you have one in place and that it is not contaminated.
A. Make sure thatthe pump is set to the correct eluentflow rate. Higherthanrecommendedeluentflowrateswillcause
higher pressure. Measure the pump flow rate if necessary with a stop watch and graduated cylinder.
B. Find out what part of the system is causing the high pressure. Itcouldbeapieceoftubingthathaspluggedorwhose
wallsarecollapsed,aninjectionvalvewithapluggedport,acolumnwithparticulatespluggingthebedsupport,aplugged
High-Pressure In-Line Filter, the suppressor or the detector cell.
To find out which part of the chromatographic system is causing the problem, disconnect the pump eluent line from the injection
valveandturnthepumpon. Watch the pressure; it should not exceed 50psi.Continueaddingthesystem’scomponents(injection
valve, column(s), suppressor and detector) one by one, while watching the system pressure. The pressure should increase up to
amaximumwhenthecolumn(s)areconnected.ThismaximumsystempressurecanbeestimatedfromthedatainTable2,“Standard
Flow Rates and Back Pressures.” No other components should add more than 100 psi of pressure. If high system back pressure
isfoundtobeduetoaspecificsystemcomponent,refertotheappropriatemanualforcleanuporreplacementofproblemcomponent.

IonPac AS3, AS4, AS5, CS2 Document No. 032226-14 Page 9 of 14
5.1.2 Replacing Column Bed Support Assemblies
If the column inlet bed support is determined to be the cause of the high back pressure, it should be replaced. To change the inlet
bed support assembly, refer to the following instructions, using one of the two spare inlet bed support assemblies included in the
ShipKit.
A. Disconnect the column fromthe system.
B. Carefully unscrew the inlet (top) column fitting,using twoopenendwrenches.
C. Remove the bed support. Turn the end fitting over and tap it against a benchtop or other hard, flat surface to remove
the bed support and seal assembly. If the bed support must be pried out of the end fitting, use a sharp pointed object
suchasapairoftweezers,butbecarefulthatyouDO NOTSCRATCHTHEWALLSOF THE ENDFITTING.Discard
the old bed support assembly.
D. Place a new bed support assembly into the end fitting. Makesure thattheendof the columntubeisclean and freeof
anyparticulatemattersothatitwillproperlysealagainstthebedsupportassembly.Usetheendofthecolumntocarefully
start the bed support assembly into the end fitting. Part numbers for bed supports and other DIONEX components can
befoundin,“DIONEXProductSelectionGuide.”
CAUTION
Ifthecolumntubeendisnotcleanwheninsertedintotheendfitting,particulatemattermayobstructapropersealbetween
theendofthecolumntubeandthebedsupportassembly.Ifthisisthecase,additionaltighteningmaynotsealthecolumn
but instead damage the column tube or the end fitting.
E. Screw the endfitting back onto the column. Tightenitfingertight,thenanadditional1/4turn(25inxlb).Ifthecolumn
continuestoleak, contactyournearestDIONEX Office(see,“DIONEXWorldwide Offices”).
F. Reconnect thecolumn tothesystem andresume operation.
NOTE
Replace the outlet bed support ONLY if high pressure persists after replacement of the inlet fitting.

IonPac AS3, AS4, AS5, CS2 Document No. 032226-14 Page 10 of 14
5.1.3 Bed Support Descriptions
The following bed support assemblies are supplied with the different DIONEX Column Lines:
ColumnLine Assembly Component
Description P/N Descriptions
4-mmHPIC(Tefzel®) 035843 Bed Support
030838 O-Ring
FLOW
O-RING
BED SUPPORT