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p. 10 Featured Article “As Good as It Gets?” The state of separation sciences
p. 12 In Your Words We hear from you! Our Edge readers share their stories
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p. 14 Featured Phenomenex Solutions Ion chromatography, proficiency testing, and personalized method support
Solving the Puzzle: Undependable Performance for Semi-Volatiles
• QCperformancetestcriteriaexposed• Thepyridineeffect–shapeandresponsePages 3-6
Overcoming Common Challenges of PAH MethodsPages 7-9
ENVIRONMENTAL
EDGE2013: Volume 2
Fueling Your Soil and Water Analysis
PhenomenexgivesyouTheEdgewithinnovativeenvironmentalproducts,tips,andarticles.
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Get The Environmental Edge!DuringarecentwebinarQ&Asession, Icaughtmyselfsmilingwhenourteamwasaskedafewever-sofamiliarques-tionslike“HowcanIimprove[insertactivecompoundname]’speakshape/response?”and“HowcanaGCcolumnstayinertwhileanalyzing“crappy”samples?”NowIassureyou,mysmilewasnotoneofjoyathearingthestrugglesofanenvironmentalanalyst,butratheritwasoneofamazementatthecommonalityoftheobstaclesourcustomersfacedayinanddayout.Yet,environmental laboratories faceUNIQUEchallenges,distinct fromanyother typeof lab.Let’sbehonest,Idon’tseemanysludgesamplesonapharmaceuticallabbench.Thesenastysamplematricescontainsomeofthebroadest ranges of compounds, spanning all types of function-alities,activities,andboilingpoints.
We’vegrowntocallthesestruggles“productivitythieves”be-cause they cost labs time and money (often undocumented coststhatnevershowuponanyspreadsheetreport).Whatifwechangedourperspectiveontheseproductivitythieves?In-steadof seeing themasobstacles,what ifwe view themasopportunities for innovation?
That’s just what this volume of the Environmental Edge dis-cusses.Wefirsttakealookatacommonproductivitythiefthatsoundsalittlesomethinglike,“MyGCcolumnsforBase/Neu-tral/Acids(BNA)mayworkgreatonce,butIcanneverquitetellwhatI’llgetwhenIinstallmyback-up.”Thisproblemariseswhenyourlab’sperformancecriteriaaremorestringentthanamanufacturer’sQCtest.Onpage3,wetakealookintohowatargetedchemistrypairedwithatargetedQCtestcanprovidereproducibleperformancefromonecolumntothenext–ensuringinertnessevenforthose“crappy”samplesyoutest.Nextonpage7,we’lltakeonthetaskofextractingPolycyclicAromaticHydrocarbons(PAHs)fromdrinkingwater,whilesimultaneouslyremovinghumicacids,whichcanbeproblematicinterferents.You’llalsoreadaboutinterestingde-velopmentsinionchromatographyofproficiencytesting.
Finally,weintroduceyoutoacoupleofnewsectionsofTheEdgestarringYOU,ourwonderfulreaders.Wewillswitchrolesasyoubecometheauthorandshareyourstories,experiences,andevenyourpuzzlingtroubleshootingquestions.Asalways,[email protected].
Sincerely,
Kali TudelaEnvironmentalMarketingManager
Whatifwechangedourperspec-tive on these productivity thieves? Instead of seeing them as obsta-cles,whatifweviewthemas opportunities for innovation?
IntroductionEnvironmental chemistshave longstruggledwith theanalysisofsemi-volatilesbyEPAMethod8270D.Thismethodcontainsanex-ceptionallylongcompoundlist,whichmaynumberover200ifadd-ingAppendixIXorclient-specifiedanalytes.Avariedspectrumoffunctional groups is present, from reactive acidic phenols to tailing basicamines,aswellasneutralcompoundslikepolynucleararo-matichydrocarbons(PAHs).Theseanalytesaretypicallyextractedfromextremelydirtymatrices,leadingtocontaminantbuild-upandeventuallydecreasedsystemperformance.Activecompoundslikepentachlorophenol (PCP), 2,4-dinitrophenol, benzidine, and pyri-dineoftenexhibittailingpeaksanddecreasedresponsesasare-sult.Toremovesuchcontaminationandrejuvenateperformance,columnsrequirefrequenttrimmingorconditioning,whichadjustsanalyte retention times and adds to the headache of routine system maintenance.
Throughextensiveresearchandtesting,Phenomenexhasdevel-opedadeactivationprocess,Enviro-Inert™ Technology, designed to combat these issues and provide optimized performance for difficultsemi-volatilesanalysis.Equippedwiththistechnology,theZebronZB-SemiVolatilesGCcolumn features supreme inertnessespecially for acids,bases, andpesticideswhilemaintaining thesameselectivityandretentionexpectedfroma5%phenyl-arylenephase.Toeliminatetheguessworkoftenencounteredusingtypi-calcolumnsforsemi-volatilesmethods,thiscolumnhasbeende-signed to perform under conditions even more demanding than re-quiredbytheEPA.InadditiontotherigorousQCtesteveryZebronGCcolumnmustpassforefficiency,retention,bleed,andactivity,eachZB-SemiVolatiles column isQC testedwithmany stringentcriteria specifically implemented to overcome challenges faced in EPA8270D.
More Sensitive Probes for Column Activity Testing
To ensure that the GC system is performing consistently, EPA8270D sets system suitability requirements that the instrumentmustpassusingaspecific“TuneMix.”Thismixcontainsamassspectrometertuningcompound(DFTPP)andacollectionofactivecompounds:PCP(anacid),benzidine(abase),andDDT(areactivepesticide).Thesystemmustmeetminimumperformancerequire-
mentsforthismix,whichareoutlinedinTable 1, before processing analyticalsamples.ThoughtheTuneMixisrequiredbytheanalyti-calmethod,GCcolumnshavetypicallynotbeenQCtestedusingthismix.Withoutsuchscreening,analystsmusttryacolumnwithnoguaranteeofitsperformancespecificallyfortheirmethods.
Internal testing revealed that benzidine, used to assess a col-umn’sbasicactivity,wasnotasufficientlysensitiveprobetofullycharacterizesmallamountsofsuchactivity.Analystsusinggen-eral purpose columns may pass benzidine peak skew require-mentsyetstillfailcalibrationcurvesforotheractivecompounds.Tocombatthis,analystsoftenaddextraconcentrationlevelstotheircalibrationcurves.Thisapproachallowslowlevelstobeig-nored because five passing data points may still be attained at highlevels.Whilethisachievesthegoalofpassingacalibrationcurve,itconsequentlyrevealspotentialinconsistenciesinalab’slow-level detection data and also adds time and complexity toanalreadychallengingmethod.Toovercome these issues,ZB-
EPA 8270D Method Criteria
Criteria EPA RequirementPhenomenex Requirement
Pyridine
PeakResponse: NotSpecified ≥0.6*
Pentachlorophenol
PeakResponse: NotSpecified ≥0.3
PeakSkew: ≤2.0 ≤2.0
Benzidine
PeakSkew: ≤2.0 ≤2.0
DDT
Breakdown: <20% <20%
Injection: 50ngorlessoncolumn 0.2ngoncolumn
*Requirementof0.5for60mx0.25mmx0.25µmdimensions
Table 1.SummaryofZB-SemiVolatilesPerformanceRequirements
Increased column activity can lead to poor acid/base sensitivity and analyte misidentification, which are common hindrances to the accuracy of semi-volatile methods. This is most apparent when ana-lyzing compound lists with various types of functional groups and reactivity characteristics, as in EPA Method 8270D. This study explores the inertness of leading GC columns used for semi-volatiles methods and presents the advantages afforded by using columns designed and QC tested specifi-cally for EPA 8270D.
Improved Inertness and QC Testing Standards for EPA Method 8270D with Zebron™ ZB-SemiVolatiles GC Columnsby Kory Kelly and Kristen Parnell
Semi- Volatiles
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IfPhenomenexproductsinthisguidedonotprovideatleastanequivalentseparationascomparedtootherproductsofthe same phase and dimensions, send in comparative data within45daysandkeeptheproductforFREE.
4 5
Table 2.QCtestconditionsforZB-SemiVolatiles
Conditions used for all columns evaluated
Dimensions 30meterx0.25mmx0.25µm
Injection Split100:1@175°C,1µL
Carrier Gas Hydrogen@40cm/sec(constantpressure)
Oven Program 40°Cfor2minto300°C@15°C/minfor3.5min
Detector FID@325°C
Sample ZB-SemiVolatilesQCTestMixAnalytes are 20 ppm in dichloromethane1.Pyridine2.Pentachlorophenol3.DFTPP4.Benzidine5.DDT6.DDD
Liner SingleTaperwithWoolinMiddle
Inlet Seal Easy Seals™InletBaseSeal
Septa PhenoRed™-400InjectorSepta
Results and DiscussionAchromatogramfortheZebronZB-SemiVolatilesQCTestMix isshowninFigure 1onaZB-SemiVolatilescolumn.Thefirstanalytepeakinthechromatogramispyridine.Thisactiveaminetailswhena column has activity andwill result in a lower peak height andlowerRFvalue.Inthischromatogram,theRFforpyridineis1.06(ZB-SemiVolatilesminimumrequirementis0.6).Thesecondpeak,PCP,would disappearwhen activity is present. This column stillshows an RF of 0.37 (ZB-SemiVolatilesminimum requirement is0.3).ThefinalpeakinthechromatogramisDDT.ThisisanactivepesticidethatbreaksdownintoDDDandDDEinthepresenceofcolumnactivity.TheZB-SemiVolatilescolumndisplayslowactivityasnobreakdownproductsarevisibleinthischromatogram.
The second column tested was aRestek® Rxi®-5ms column with thechromatogram shown in Figure 2.Forthiscolumn,pyridineshowsex-cessivetailingandhasanRFof0.34.ThisdoesnotmeettheminimumRFrequirement of 0.6, and may leadto difficulties in passing calibration curves due to lower sensitivity forbases.WhilethePCPresponsefac-torpassesourminimumrequirementof 0.3, this Rxi-5ms column wouldstill not pass our QC requirementsdue to the failing activity towardbases.
ThethirdcolumntestedwastheRestekRxi-5SilMSasshowninFigure 3.Thiscolumnshowedbetterresponseforpyridinecom-paredtotheRestekRxi-5mswitharesponsefactorof0.53.How-ever, thisRF isstillhalf thatof theZB-SemiVolatilescolumnanddoesnotmeetminimum requirement of 0.6.Additionally, the in-creaseinpyridineresponsewasachievedbyexchangingresponseofacidsforpyridine.TheresponseofPCPhasnowdecreasedto0.26,whichleavesphenolssubjecttogreateractivityandthepos-sibilityof failingcalibrationcurvesordecreasingcolumn lifetime.Thoughan improvementovertheRxi-5ms,thiscolumnstilldoesnotpassminimumQCrequirements.
The fourthcolumn testedwasanAgilent®HP-5msUltra InertasshowninFigure 4.ThiscolumnhadthelowestpyridineresponseofallcolumnstestedwithanRFof0.28,whichfailstherequirement
SemiVolatilescolumnsarefirsttestedwithaQCmixthatensurespassingperformanceforefficiency,bleed,retention,andactivity.ThecolumnsarethenrequiredtopassasecondQCtestusingacompositemix includingtheEPA8270DTuneMixandpyridine,amoreactivebasethanbenzidine.Passingcriteriaforthisnew,more sensitive mix (the ZB-SemiVolatiles QC Test Mix) dictateminimumrequirementsthataretypicallyunmonitoredbyGCcol-umnmanufacturers.
Standardized Performance Criteria for Troublesome Compounds
The EPA method calculates analyte response factors based on peakareaincomparisontotheinternalstandard,whichdoesnotindicatepeakshapeperformance(abroadpeakandasharppeakcouldbothhavethesamepeakareas).Becausepoorpeakshapenegativelyimpactssignal-to-noiseratiosandquantitation,QCtest-ing forZB-SemiVolatilesmeasuresamoreaccurateperformanceindicator – response factors based peak heights as opposedto peak areas. Specific response factors are not defined for themethod– theEPA8270D requirement states that acompound’sresponsemust fallwithinwhat is typically expected for that an-alyte. However, to provide analysts with a consistent measureof performance, Phenomenex sets minimum response require-ments in theQC test every ZB-SemiVolatiles columnmust pass (Table 1).Inthisstudy,minimumresponserequirementsaresetat0.6forpyridineand0.3forPCP.Noothercolumnmanufacturerhaspreviouslysetrequirementsforthislevelofperformance.
Improving Calibration Success at Low-Level Concentrations
ZB-SemiVolatilescolumnsmustdemonstratesatisfactoryQCper-formanceatlevelsthattypicallygiveanalyststhemosttrouble–thelowendofacalibrationcurve.EPAmethod8270Drecommendsaconcentrationof“50ngorless”(1.0µLsplitlessinjectionofa50ppm solution). Due to the activity in typical columns, labs ofteninjectthemaximum50ngtoimprovetheirchancesofpassingcali-brations – higher concentrationswillsaturateanycolumnactivity,al-lowingmoredatapointstorespondnormally.TheQCmixforZB-Semi-Volatiles is injectedmuch lowerat20 ppm to more accurately assess true column activity. Additionally,labs often attempt to improve sen-sitivity by using splitless or even pulsed splitless (as opposed to split) injections to introduce themost sample onto the column. Toensure their performance at ex-tremely low detection levels, ZB-SemiVolatiles columns are tested usingasplit injectionwitha100:1splitratio(effectively0.2ngoncol-umn–250timeslessthanthemaximumallowed50ng).
ExperimentalConditionsusedforallcolumnsare listedinTable 2 and reflect theQC conditions used for every ZB-SemiVolatiles column. Allcomparisonsshownwereperformedwithinthesametimeframebythesameanalystonthesameinstrument.Toaccuratelyandconsistentlymeasurepeakintensitiesandtoaccountforanymi-nor differences in sample volumes, the internal standard method ofcomparisonwasusedwith the inertcompoundDFTPP.Peakheights of the analytes of interest were compared to the peakheight of the internal standard. This ratio results in a responsefactor(RF)assimilarlycalculatedinEPAMethod8270D.
Figure 3.QCTestMixonRestekRxi-5SilMS
0 2 4
1
2
34
5
6 8 10 12 14 16 18 20 minRetention Time
Inte
nsi
ty (p
A)
11
13
15
17
19
21
23
25
20577
9
Poor PCP response RF = 0.26
Poor response RF = 0.53
App
ID 2
0577
Figure 2.QCTestMixonRestek®Rxi®-5ms
0 2 4
1 2
3 45
6 8 10 12 14 16 18 20 minRetention Time
Inte
nsi
ty (p
A)
11
13
15
17
19
21
23
25
20575
9
Peak tailingRF = 0.34
App
ID 2
0575
Figure 4.QCTestMixonAgilentHP-5msUltraInert
0 2 4
1 2
34
6
5
6 8 10 12 14 16 18 20 minRetention Time
Inte
nsi
ty (p
A)
11
13
15
17
19
21
23
25
20575
9
DDT breakdown
App
ID 2
0574
Figure 5.QCTestMixonAgilentDB-5msUltraInert
0 2
1
2
34
5
4 6 8 10 12 14 16 18 20 minRetention Time
Inte
nsi
ty (p
A)
11
13
15
17
19
21
23
25
20576
9
Poor response RF = 0.66
Poor PCP response RF = 0.20
App
ID 2
0576
Figure 1.QCTestMixonZebronZB-SemiVolatiles
1
2
3 4
5
Inte
nsity
(pA
)
11
13
15
17
19
21
23
25
20573
0 2 4 6 8 10 12 14 16 18 20 minRetention Time
9
No DDT breakdown
Strong PCP response RF = 0.37
Good response and peak shape for pyridine! RF = 1.06
App
ID 2
0573
ZB-SemiVolatilescolumnsare tested using a split injectionwitha100:1split
ratio...250 times less than themaximumallowed...
of0.6.Thoughthiscolumnperformedpoorlyforbases,itdidhavethehighestRFforacidswithaPCPRFof0.40.ThisRFisslightlybetter than that of ZB-SemiVolatiles, but occurs at the expenseofpoorresponseforbases.Inaddition,thiswastheonlycolumntestedthathadsignificantbreakdownofDDT(12.4%).Thoughthisvalueisbelowthe20%requiredbythemethod,thiswastheonlycolumntestedthatshowedsuchactivity.
The final column tested was an Agilent® DB-5ms Ultra Inert asshown inFigure 5. Thiscolumnhadgood response forpyridinewithanRFof0.66andwastheonlyothercolumntomeetthemini-mum0.6RFrequirementforpyridine.Asseenwithothercolumns
tested,thisresponsecomeswithatradeoff.TheDB-5msUltraInerthadthelowestRFforPCPofanyofthecolumnstested(0.20).Thisfailstherequirementof0.3.
Ofallcolumns tested,onlyZB-SemiVolatileswouldhavepassedtheminimumperformancerequirementsforbothpyridineandPCPresponse as summarized in Table 3.Thoughallcolumnsdisplayedsimilarpeakshapesforbenzidine,varyinglevelsofcolumnactiv-itygreatlyaffected the responseofmoreactivebasicpeaks likepyridine.Dramaticdifferences inperformanceareapparentwheninspectingpyridinepeakshapeandsignal intensityas illustratedin Figure 6.
Semi-Volatiles(cont’d) Semi-Volatiles(cont’d)
Peak tailingRF = 0.28
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IntroductionPolycyclicaromatichydrocarbons(PAHs)areby-productsofburning fuel and have been classified as carcinogenic, mu-tagenic,andteratogenic.Thesepollutantshavebeenfoundtobepresentinfoodsaswellasdrinkingwater,whichhavecausedtheEPAtoregulatetheallowedlevelsofPAHsunderEPAmethod 550.1. Under the EPAmethod, PAHs are ex-tractedfromwatersamplesusingasimpleC18solidphaseextraction (SPE) sorbent. However, this sorbent simultane-ouslyextractshumicacidswhichcancauseinterferencesinsubsequentanalysis.Humicacidsareproducedasorganicmatter decays and are thereforemajor constituents of soilandwater,meaning they arepresent in almost all samplestestedforPAHs.
InanefforttoimprovetheanalysisofPAHs,PhenomenexhasdevelopedStrataPAH,aSPEsorbentthattargetsPAHsandsimultaneouslyremoveshumicacidsfromthesample.PAHscan thenbeelutedwhile humicacids remainbound in thesorbent.NotonlydoesStrataPAHproducecleanerextractsbutitalsoproduceshigherrecoveriesofPAHsascomparedtotheEPA550.1method.
Experimental Conditions100µLofPAHstandardsinacetonitrile,obtainedfromUltraScientific,Inc.,werespikedinto100mLofwater/acetonitrile(75:25).ThepreparedPAHsampleswerethenpassedthroughtheStrataPAHsorbentasdirectedinthemethoddescribedin Table 1.Aftertheextraction,sampleswereblowndowntodrynessundernitrogengasat35°C.Thesampleswerethenreconstitutedin100µLofdichloromethaneandinjectedontoa Zebron™ZB-5msGCcolumn.Aseparatestudywasper-formedtoensurethattheStrataPAHsorbentwascapableofremovinghumicacidsfromwatersamples.Usinghumicacidstandards, obtained from International Humics SubstancesSociety(IHSS)a1Lsolutioncontaining1mg/Lofstandardinwaterwasmade.Usingthesameextractionprocedurespeci-fied in Table 1,100mLofthesolutionwaspassedthrougha1.5g/6mLStrataPAHtubewhileanother100mLof thesolutionwaspassedthrougha1g/6mLStrataC18-Etubeasacontrol.StrataC18-Ewasselectedasacontrolsorbentbecause C18 SPE sorbent is specified in EPA 550.1. The
1
2
3
4
5
Table 1. StrataPAHSPEProtocol
Strata PAH 1.5 g/ 6 mL SPE Tubes (Part Number 8B-S130-7CH)
Condition: - 10 mL Dichloromethane- 20 mL Methanol- 20 mL D.I. Water
Load: - 100 µL PAH standards (100 µg/mL in Acetonitrile) spiked into 100 mL Water/Acetonitrile (75:25)
Wash: - 5 mL Methanol/D.I. Water (50:50)
Dry: - 15 seconds under 10” Hg vacuum
Elute: - 6 mL Dichloromethane
Improved Recoveries of Polycyclic Aromatic Hydrocarbons (PAHs) as Defined in EPA 550.1 and Simultaneous Removal of Humic Acids from Water Using Strata® PAHby Zeshan Aqeel, Seyed Sadjadi, and Erica Pike
PAHs
StrataPAHtargets PAHs and simultaneously
removes humic acids from the sample
effluents from each sorbentwere blown down to dryness undernitrogengas at 35°C andwere reconstituted in 250µLof 5mMAmmonium formatebuffer and250µLofmethanol. 5µLof eachreconstitutedsamplewasthenanalyzedusinganAgilent®1200SLHPLCcoupledtoanABSCIEX™4000QTRAP®(withTurboV™ ion source)usingaKinetex®C82.6µmcore-shellcolumnforanalysis.
Polycyclic aromatic hydrocarbon compounds (PAHs) are effectively extracted from water samples while humic acids, which often interfere with chromatographic separation, are removed from the sample using a SPE sorbent, Strata® PAH. It was also found that Strata PAH provides consistent, high recoveries of all 16 analytes listed under EPA Method 550.1.
Ordering InfomationZebron™ ZB-SemiVolatiles GC Columns ID (mm) df (µm) Temperature Limits (°C) Part No.
20-Meter
0.18 0.18 -60 to 325/350 7FD-G027-08
0.18 0.36 -60 to 325/350 7FD-G027-53
30-Meter
0.25 0.25 -60 to 325/350 7HG-G027-11
0.25 0.50 -60 to 325/350 7HG-G027-17
60-Meter
0.25 0.25 -60 to 325/350 7KG-G027-11
Easy Seals™ Inlet Base Seals Part No. Description Unit
Standard, single groove for splitless applications, 0.8 mm dia. inlet hole
AGO-8619 Easy Seals Inlet Base Seal, Gold Plated for Agilent GCs 2/pk
AGO-8620 Easy Seals Inlet Base Seal, Gold Plated for Agilent GCs 10/pk
PhenoRed™-400 Injector SeptaPart No. Description Diameter Unit
PhenoRed-400 GuideRight™ Injector Hole Septa
AGO-7916 PhenoRed-400, rated to 400 ˚C 3/8 in. (9.5 mm) 50/pk
AGO-7917 PhenoRed-400, rated to 400 ˚C 7/16 in. (9.5 mm) 50/pk
ConclusionLeadingGCcolumnsusedforEPAMethod8270Dweretestedtodeterminehowtheywouldperformto thestringent requirementsoutlinedintheZebronZB-SemiVolatilesQCtestprocess.DesignedtoprovideGCanalystswithaguaranteeofperformancespecifi-callyfortroublesomesemi-volatilesmethods,thisprocesscharac-terizes both acidic and basic activity and sets previously unmoni-toredrequirementsforGCcolumnperformancethatareevenmorestringent than required by EPA 8270D.Minimum peak responserequirementsforpyridine(0.6)andPCP(0.3)serveasametricforcharacterizing both basic and acidic column activity, which arebothpresentinmethodslikeEPA8270D.Columnswithhigherini-tialpeakresponsescanbeexpectedtomaintainperformanceovertime,providinglongerlifetimesforthemethod.Additionally,higherresponsesallowrunsatlowerdetectionlevels,improvingthesen-sitivityoftheanalysis.
Of the columns tested, only ZB-SemiVolatiles hadhigh andbal-anced responses for both acids and bases. Other columns hadgood responses for bases but sacrificed the response for acids, or vise versa. TheHP-5msUltra Inertwas also theonly columnthat exhibitedanyactivity forDDT.The improved resultsonZB-SemiVolatilesdemonstratethetruevalueofchoosingaGCcolumndesigned,deactivated,andQCtestedtodeliverhighlyinertperfor-mancespecificallyforsemi-volatilesmethods.
Figure 6.Overlaidpyridinepeakforallcolumnstested
Intensity(p
A)
9.00
11.00
13.00
15.00
17.00
19.00
2.5 2.7 2.9 3.1 3.3 3.5 3.9
ZB-SemiVolatiles
DB-5ms Ultra Inert
Rxi-5Sil MS
Rxi-5ms
HP-5ms Ultra Inert
3.7 min
Table 3.SummaryofResponseFactors(RF)forpyridineandpentachlorophenol(PCP)oneachofthecolumnstested.
Column Pyridine RF PCP RF Result
Zebron™ ZB-SemiVolatiles 1.06 0.37 Pass
Restek®Rxi-5ms 0.34 0.34 Fail
RestekRxi®-5SilMS 0.53 0.26 Fail
Agilent®HP-5msUltraInert 0.28 0.40 Fail
AgilentDB-5msUltraInert 0.66 0.20 Fail
RFiscalculatedbydividingpeakheightofanalytebypeakheightofDFTPPasinternalstandard.
Semi-Volatiles(cont’d)
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8 9
Results and DiscussionThe GC/MS analysis (resultant chromatogram displayed in Fig-ure 1 shows that recoveriesof the16analyzedPAHswerecon-sistentlyhigher than those reportedusing theEPA550.1 recom-mended method (Figure 2 and Table 2),except fornaphthalenewhichprovedtobedifficultduetoitsvolatility.Inanefforttoreducethe volatility issue, a deuterated form of naphthalene aswell as2-Methylnaphthalenewereanalyzedaswell. Inadditiontohigherrecoveries, theLC/MSdata shows thatStrataPAHwasalso ef-fectiveforremovinghumicacidsfromtheextractedsamples,thusprovidingacleanersampleandlowerbackground(asillustratedbytheLC/MSbaselineinFigure 3)ascomparedtothecurrentextrac-tionmethodrecommendedinEPA550.1
ConclusionStrataPAHeffectively extractedPAHs fromwater sampleswhileproviding benefits such as higher recoveries and improved baseline resolution as compared to the recommended method is outlined in EPAmethod550.1.
References1.JimmieW.Hodgeson,etal.EPAMethod550.1,July1990
2.6 µm Analytical Columns (mm)Phases 50 x 4.6 75 x 4.6 100 x 4.6 150 x 4.6C8 00B-4497-E0 00C-4497-E0 00D-4497-E0 00F-4497-EO
OtherKinetexparticlesizes,phases,anddimensionsareavailable,please visit www.phenomenex.com/kinetex.
Kinetex® Core-Shell HPLC/UHPLC Columns1.7 µm Minibore Columns (mm)Phases 50 x 2.1 100 x 2.1C8 00B-4499-AN 00D-4499-AN
2.6 µm Minibore and MidBore™ Columns (mm)Phases 50 x 2.1 100 x 2.1 50 x 3.0 100 x 3.0C8 00B-4497-AN 00D-4497-AN 00B-4497-Y0 00D-4497-Y0
Ordering Information
Strata® PAH Solid Phase Extraction TubesSorbent Mass Part Number Unit750 mg 8B-S130-WCH 6 mL (30/box)1.5 g 8B-S130-7CH 6 mL (30/box)
Column: Kinetex 2.6 µm C8Dimensions: 50 x 2.1 mm
Part No.: 00B-4497-AN Mobile Phase: A: 5 mM Ammonium acetate
B: MethanolGradient: Time (min)
0 2 6 6.01
B (%)15959515
Flow Rate: 0.4 mL/minTemperature: Ambient
Detection: MS @ 580.4 amu / 536.5 amu (ambient)Backpressure: 210 bar
Sample: Humic Acids from Suwannee River
3.00 3.05 3.10 3.15 3.20 3.25 3.30 3.35 3.40 3.45 3.50 3.55 3.60 3.65 3.70 3.75 3.80 3.85 3.90 3.950.0
5.0e7
1.0e8
1.5e8
2.0e8
2.5e8
3.0e8
3.5e8
4.0e8
4.5e8
5.0e8
5.5e8
5.8e8Intensity, cps
min
Figure 3. EffectiveRemovalofHumicAcids
StrataPAHproduces
cleanerextracts and
also produces higher recoveriesofPAHs
2
1,2
3
4
5
67
8
9
10
11
1213
19651
1415
16
17 18
19 20
4 6 8 10 12 14 16 18 20 22 min
8000000
7000000
6000000
5000000
4000000
3000000
2000000
1000000
Abundance
Figure 1. GCAnalysisofPolycyclicAromaticHydrocarbons(PAHs)
Humic Acids Extract from Strata-PAH
Humic Acids Extract from Strata-C18-E
AppID
19651
AppID
19662
PAHs(cont’d) PAHs(cont’d)
110
90
70
50
30
10
Naphth
alene
Acena
phthale
ne
2-Meth
ylnap
hthale
ne
Acena
phthen
e
Fluoren
e
Anthrac
ene
Phena
nthren
e
Fluoran
thene
Pyrene
Benz[a
]anthr
acen
e
Chrys
ene
Benzo
[b]�uoro
anthe
ne
Benzo
[k]�u
oroan
thene
Benzo
[a]pyre
ne
Inden
o[1,2,3
-cd]pyre
ne
Dibenzo
[a,h]a
nthrac
ene
Benzo
[g,h,i]p
erylen
e
Strata PAH EPA 550.1
Figure 2. PAHPercentRecoveriesfromTapWater1
Zebron™ ZB-5ms GC ColumnsID(mm) df(µm) Temp. Limits °C Part No.10-Meter0.10 0.10 -60 to 325/350 7CB-G010-020.18 0.18 -60 to 325/350 7CD-G010-0815-Meter0.25 0.25 -60 to 325/350 7EG-G010-1120-Meter0.18 0.18 -60 to 325/350 7FD-G010-080.18 0.32 -60 to 325/350 7FD-G010-510.18 0.36 -60 to 325/350 7FD-G010-5325-Meter0.20 0.33 -60 to 325/350 7GE-G010-1430-Meter0.25 0.25 -60 to 325/350 7HG-G010-110.25 0.50 -60 to 325/350 7HG-G010-170.25 1.00 -60 to 325/350 7HG-G010-220.32 0.25 -60 to 325/350 7HM-G010-110.32 0.50 -60 to 325/350 7HM-G010-170.32 1.00 -60 to 325/350 7HM-G010-2260-Meter0.25 0.25 -60 to 325/350 7KG-G010-110.32 0.25 -60 to 325/350 7KM-G010-11
Recommended GC AccessoriesPart No. Description Dimensions UnitAG0-4680 Split/Splitless FocusLiner™
Single Taper with wool4 x 78.5 x 6.3 mm 5/pk
AG0-8620 Easy Seals™ Inlet Base Seals, Gold Plated
Single Groove 0.8 mm 10/pk
AG0-4696 PhenoRed™-400 Injector Septa 7/16 in (11 mm) Diameter 50/pk
Table 2. PAHPercentRecoveriesfromTapWater1
Strata PAH % EPA 550.1 %
Naphthalene 58.05 72.8
2-Methylnaphthalene 86.98 --
Acenaphthalene 95.86 64.1
Acenaphthene 97.47 67.1
Fluorene 91.50 72.5
Anthracene 102.1 63.3
Phenanthrene 103.1 59.5
Fluoranthene 101.1 80.7
Pyrene 99.45 80.7
Benz[a]anthracene 89.97 78.1
Chrysene 94.70 73.1
Benzo[b]fluoranthene 82.25 65.9
Benzo[k]fluoranthene 86.08 74.9
Benzo[a]pyrene 82.42 70.0
Indeno[1,2,3-cd]pyrene 70.33 74.0
Dibenz[a,h]anthracene 74.63 64.7
Benzo[g,h,i]perylene 76.70 67.3
Column: Zebron™ ZB-5msDimensions: 30 meter x 0.25 mm x 0.25 µm
Part No.: 7HG-G010-11 Injection: Split 15:1 @ 310 °C, 1 μL
Carrier Gas: Helium @ 1.4 mL/min (constant flow)Oven Program: 140 °C to 240 °C @ 15 °C/min to 275 °C @ 4 °C/min to 320 °C @ 10 °C/min
for 5 minDetector: MS @ 270 °CSample: 1. D8-Naphthalene
2. Naphthalene 3. 2-Methylnaphthalene 4. Acenaphthalene 5. Acenaphthene 6. Fluorene 7. Anthracene 8. Phenanthrene 9. Fluoranthene 10. D10-Pyrene 11. Pyrene 12. Benz[a]anthracene 13. Chrysene 14. Benzo[b]fluoranthene 15. Benzo[k]fluoranthene 16. D12-Benzo[a]pyrene 17. Benzo[a]pyrene 18. Indeno[1,2,3-cd]pyrene 19. Dibenz[a,h]anthracene 20. Benzo[g,h,i]perylene
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The Phenomenex WaySince our founding in 1982, Phenomenex has been a leader inseparationsciencewiththevisionofresearching,developing,andcommercializing exciting new—and potentially revolutionary—technologiesandmakingthemreadilyavailabletothebroadchro-matographycommunity.
Sohowdoyoufind“TheNextBigThing?”InadditiontotheworkofourownR&Dteam,werecognizethatagreatwaytogetaglimpseinto the possible future of chromatography is to have a pulse on the cutting edge research emerging from our colleagues at top uni-versities,researchinstitutes,andindustrialprograms.AparticularlyusefulvenuetoobtainsuchaviewisattheannualGCxGC-ISCCmeeting.IrecentlyattendedthiseventinPalmSprings,California–the10thGCxGCSymposiumand37thInternationalSymposiumonCapillaryChromatography. Iwould like toshare justa few in-sights from that meeting to illustrate the amazing future potential of chromatography,whilealsoprovidingaperspectiveonhowPhe-nomenexviewstheseearlyresearchdevelopmentsasbutonestepintheprocessofdevelopingusefulcommercialproducts.
As Good As It Gets?Editorial: Environmental applications
Multi-dimensional chromatography is beingmore commonly ap-pliedtohighlycomplexenvironmentalsamples.Notableatthecon-ferencewerepapersandpostersonmulti-dimensionalchromatog-raphyfortheanalysisoftoxicnaphthenicacidsinoxidizedoilspills,the comprehensive analysis of halogenated organics (including all theStockholmConventionpersistentorganicpollutants),andtheanalysis of complex PAHmixtures (including parent, alkyl, nitro,oxy,thio,chloroandbromo-PAH’s),tonamejustthreeexamples.
Nano-worldOne of the most interesting fields of current study involves chro-matography at the nano scale, in terms of either particle size and columndimension. Interestingneweffectsarebeingseenat thisscale (suchasthepressure-flow-particlesizerelationship),whicharecounterintuitive.Tocitebutoneexample:acapillaryLCcolumnpackedwith0.125micronsilicaparticleswithaplateheightof15nanometers. Ina3cmcolumn,that’sa lotof theoreticalplates—you do the math!
The Journey ContinuesSo,what’sthetakeawayhere?Well,asexcitingasthesedevelop-mentsare fromascientificstandpoint, theyarea longway frompractical commercial products that can be routinely and consis-tentlyimplemented.Theyarefullofpromise,buttheyarenotop-timum solutions. This is to be expected. Leading edge researchisseldomoptimum,or itwouldn’tbe leadingedge. It revealstheunexpected;itpointstheway;itshowswhat’spossible.But,itwilltakealotmoreworktoputthesetoolsinthehandsoftheeverydaychromatographer and permit the generation of consistent, reliable analyticalresults.
The history of chromatography has been an amazing story of continual scientific advances. This
progress is commonly attributed to advances in analytical instrumentation. However, the parallel
advances in separation chemistry—the media and the columns—have played a critical role in
such progress. Beginning with packed GC columns and gravity flow LC, the field has progressed
to today’s fast and efficient capillary GC columns and UHPLC core-shell technology. Much like
Moore’s Law in solid state electronics, chromatography has witnessed a continuing, exponential
decrease in plate height with a corresponding growth in column performance.
by David C. Kennedy, PhD
Seen at the Show
Into Another DimensionMulti-dimensionchromatographywaspioneeredinthe1990’sandismostfamiliarintheformofGCxGC.Atthemeeting,aGCxGC-TOFMS separationwas described that attained a peak capacityof 6,000peaks in a 7-minute run. This is approaching the theo-reticallimitof4,300peaksina1-minuterun.Thepowertounravelcomplexmixtureswasalsodemonstratedinanotherpaperwhereacomplexmixtureofolefinmonomersandpolymerswasanalyzedbypyrolysis-GCxGC,resultinginachromatogramwith14,983dis-tinctpeaks.WithGCxGCbecomingmoremainline,someresearch-ersaremovingontoGCx2GC,LCxLC,andhybridtechniquessuchasLCxGC–andevenLCxGCxGC.Theclearimpression:chroma-tographystillhasalongwaytogobeforeitspotentialisexhausted.
Share Your Thoughts!Our customers inspire us and fuel our innovation, so please share your questions and celebrate your recent revelationsaboutenvironmentalanalyses.
Youjustmaylandyourselfaspotasthefeaturedcontributorinthenextedition of The Edge!
I heard several of the conference presenters say that their research wasnotoptimum.Theyhadusedanoff-the-shelfchromatographiccolumn(orevenahome-madecolumn)todemonstrateanexcitingnewphenomenon.Theyacknowledgedthatnewandbetterchro-matographicsupportsandconfigurationsweregoingtobeneededinordertoextractthefullbenefitoftheeffecttheyhaddiscovered.Thatkindofobservationgetsme(andthePhenomenexR&Dteam)very excited because that’s what Phenomenex does best. Wethriveontakingthe“couldbe”andtransformingitintonew,reliablechromatographicsolutions.Inaword:wePhenomenize!Therefore,welookforwardtocollaboratingwiththesecuttingedgeresearch-ersasdiscoveries like thesepave theway fornewPhenomenexproducts and chromatographic applications that you can readily implementinyourlaboratory.
David Kennedy has a PhD in analytical chemistry and is a 40-year veteran of the environmental industry with sequential ca-reers as a research chemist, environmental laboratory manager, instrument company executive, and management consultant.
We thrive on taking the “could be” and transforming it into new, reliable chromatographic solutions.
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Ourcustomersinspireus,youdriveustocontinuallydiscovernewso-
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relate to us all?
MyearlyelutersonaGCmethodare
displaying poor peak shape. What
suggestions do you have for getting
sharperpeaksinthebeginningofmy
chromatogram?
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In Your Words Your Questions, Answered.
Answer:
Early elutingcompoundssometimeshavepoorpeakshapesdependingonmethodconditions.Someana-lytesmaynotfullyfocusontothecolumnwellbecausethe initial temperaturemaybe toohigh.Polaritymis-matchcanalsooccurifasolventdoesn’tdissolvewellinthephase.Thesolvent‘pools’onthecolumnphaseand is pushed down the column by the carrier gas.Some compounds travel with the solvent pools andmayleadtotailingorsplitpeaks.
Here are some suggestions to minimize early eluting peak distortion:
Use a split injection. This limits the amount of solvent that gets onto the column and reduces howmuch analyte dissolves in pooled solvent. Thesplitinjectionalsoreducespeakwidthsbecausethehigherflowthroughtheinletmovescompoundsfromtheinlettothecolumnfaster.Thiswillimprovethepeakshapeconsiderably.Thedownside is that the splitwill sacrificesensitivity.Ifyouhavesensitivitytospareoramoreconcentratedsample,thismaybeanoption.
Decrease the injection volume. This also reduces the amount of solvent thatisintroducedontothecolumn.Italsoreducestheamountofanalytesandthereforewillalsosacrificesensitivity.Lowertheinitialoventempera-ture. Thiswill forceanalytes to condensemore into thephaseand limitsanalytemigrationduring injection.This isknownasanalyte focusing.Thedownsideof this isslightly longercycletimesbecausecoolingtoa lowertemperaturemaytakelonger.
Use a pressure pulsed injection. Thiswillincreasetheflowontothecol-umn,butonlytemporarily.Thishelpsfocustheanalytesandresultsinanar-rowerpeakshape.Usuallyapulsedinjectionwillbeabout10-15psiabovethenormalpressureforthedurationoftheinjection.Afterward,thepressureandflowreturntonormal.EarlyelutingcompoundscaneluteevenearlierandasolventdelaymayneedtobereducedifusingaMSdetector.
Use a GC guard column.Whenusedwithvolatiles,thisissometimescalledasolventgap.Itcanhelptoseparatetheanalytesfromthesolventandre-ducesthesolventpooleffect.Insomeinstances,itcanalsoextendcolumnlifetimebecausetheguardcanbetrimmedorreplacedwhencontaminatedbydirty sampleswhichwouldotherwisesacrifice theperformanceof thecolumn.
Increase the GC column film thickness.Athickerfilmwillbetterdissolvebothvolatileanalytesandresultinlongerretentionforearlyelutingpeaks.Athickerfilmwillalsodissolvegreateramountsofsolventandhelpminimizedistortionduetosolventmismatch.
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Ready to Answer?Visit www.phenomenex.com/InYourWords
Youranswerjustmightappearinthenextvolume of the Environmental Edge!
Don’t Forget!KeepaneyeoutforyouranswerstothisquestionintheIn Your Words
section of Volume 3 of The Edge.
How does your lab inject extra smiles into the job?Ideas: Special music, rituals, superstitions forsuccess,instrumentnicknames
The Edge’sanalyticalexpert,KoryKelly—theTechnicalManagerforEnvironmentalandGCapplicationsatPhenomenex®–providesvalu-abletipsandtricksforenvironmentaltestingsuccesswhilealsoan-sweringyourtechnicalandtroubleshootingquestions.
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FREE Method Optimization Guidance DidyouknowyourlabhasaPhenomenexTechnicalSupportTeamdedicatedtopersonallysupportyourlab’sneeds?Theycollaboratewithchemistseverydaytoimproveproductivityandresultswhileworkingwithintheboundsofestablishedmethods.
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Proficiency TestingGain Insight by Finding Answers to Performance Questions
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Identify Leadership Opportunities“Doesacertainanalystconsistentlysucceedwithaparticular analysis?”
Increase Network Collaboration“Whichlabsareoutstandingperformers?Iwouldliketospreadbestpracticesandtechnicalexpertiseacrossthenetwork.”
AnswerquestionslikethesewithinsecondswithPTManage—theeasyto use, highly customizable tool from Phenova™ that helps you do more thansimplyreportPTdata.Quicklygeneratereportsthatdelivervaluableinsightaboutyourlaboratory’sstrengthsandweaknesses,allowingyoutotakeaproactiveapproachtoimprovingoveralllabperformance.
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Terms and Conditions SubjecttoPhenomenexStandardTermsandConditions,whichmaybeviewedat www.phenomenex.com/TermsAndConditions
Trademarks StrataandKinetexareregisteredtrademarksofPhenomenex.Zebron,EasySeals,PhenoRed,GuideRight,Inferno,MidBoreandPhenexaretrademarksofPhenomenex.PhenovaandPTManagearetrademarksofPhenova,Inc.,anaffiliateofPhenomenex.FocusLinerisatrademarkofSGE.AgilentisaregisteredtrademarkofAgilentTechnologies.RestekandRxiareregisteredtrademarksofRestekCorporation.QTRAPisaregisteredtrademark,ABSCIEXandTurboVaretrade-marksofABSCIEXPte.Ltd.DionexandIonPacareregisteredtrademarksofDionexCorporation.ShodexisaregisteredtrademarkofShowaDenko,K.K.
Disclaimer PhenomenexisnotaffiliatedwithAgilentTechnologiesorRestekCorporation.Comparativeseparationsmaynotberepresentativeofallapplications.
©2013Phenomenex,Inc.Allrightsreserved.
ShodexoffersaninnovativeICcolumnforthesuppressormethodthat improves both the separation speed and resolution of anions in mostmatrices.Withhightheoreticalplates(>5000/mforSulfate),the column easily and efficiently separates organic and inorganic anionssuchasEPAMethod300analytes,acetate,formate,meth-acrylateandoxalate.High loadingandexceptional resistance toloadingcombinewithfeaturessuchasimprovedseparationofthefluoridepeakfromthewaterdip.Thepatent-pendingPVAgelwasalso specifically engineered to optimize the elution behavior of the carbonate ionsoasnot to interferewith thequantitationofEPAMethod300anions.
• Improved,high-speedseparationofEPAMethod300analytes
• Highefficiency,generalpurposeICcolumn
• Fluoridewell-resolvedfromwaterdip
• GreatalternativetoDionex® IonPac®AS4,AS4AandAS14columns
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Shodex Ion Chromatography Columns
Improving the Analysis of Organic Acids and Inorganic Anions Featured Services
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Organic Acids and Inorganic Anions Column: SI-50 4E
Dimensions: 4.0 x 250 mmEluent: 3.2 mM Na2CO3 / NaHCO3
Flow Rate: 0.7 mL/minTemperature: Ambient
Sample: 20 µL 1. Fluoride 2 mg/L 2. Acetate 10 mg/L 3. Formate 2 mg/L 4. Methacrylate 10 mg/L 5. Chloride 3 mg/L 6. Nitrite 5 mg/L
7. Bromide 10 mg/L 8. Nitrate 10 mg/L 9. Phosphate 15 mg/L 10. Sulfate 15 mg/L 11. Oxalate 15 mg/L
Anions Column: SI-90 4E
Dimensions: 4.0 x 250 mmEluent: 1.8 mM Na2CO3
1.7 mM NaHCO3Flow Rate: 2.0 mL/min
Temperature: AmbientSample: 20 µL
1. Fluoride 2 mg/L 2. Chloride 3 mg/L 3. Nitrite 5 mg/L 4. Bromide 10 mg/L
5. Nitrate 10 mg/L 6. Phosphate 15 mg/L 7. Sulfate 15 mg/L
App
ID 1
4176
App
ID 1
4175
Ordering Information IC ColumnsColumn Type/ Part No.
ID x Length (mm) Plate Number Packing Material Functional Group Applications
IC SI-90 4E 4.0 x 250 >5,000 (SO4) PVA Quaternary ammonium Inorganic anions and organic acidsIC SI-90 G 4.6 x 10 (Guard) — — (General purpose)IC SI-50 4E* 4.0 x 250 >14,000 PVA Quaternary ammonium Inorganic anions and organic acidsIC I-524A 4.6 x 100 >2,000 PHM gel Quaternary ammonium Inorganic anionsIC Y-521 4.6 x 150 >3,000 — — Cations (general purpose)IC Y-G 4.6 x 10 (Guard) — — —IC YK-421 4.6 x 125 >2,500 Hydrophilic Polymer Carboxyl Coated Silica Simultaneous separation of monovalent and divalent cationsIC YK-G 4.6 x 10 (Guard) — — —IC T-521 4.6 x 150 >3,000 S-DVB gel Sulfo (H+) Transition metal ions (packed in PEEK column)IC T-G 4.6 x 10 (Guard) — — —*UseICSI-90Gguard.
Within 8 min
Environmental SolutionsPhenomenex®
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