
www.defiant-tech.com
3
Calibration
Calibration
It isnecessarytocalibratetheFROGforadesiredapplication.Acalibrationmustbe
openedintheEllvinSoftwarefortheFROGtocorrectlyanalyze samples.
Calibrationfiles canbecreated(withanalysisdatafromchemicalstandards)or
loaded(ifsomeoneelse calibratedtheFROG)withthesoftware.Defiant
recommendsre-calibratingtheFROGatleasteverythree weeksforthemost
accurateanalyses.
Loading anExisting CalibrationFile
IftheFROGwas calibratedbysomeoneelse,thecalibration filewill mustbeloaded
alongwiththesoftware.ThecalibrationfilecanbefoundontheUSBDrivelocated
inthezipperpouchintheFROG’scase.Toloadacalibration filecompletethefol-
lowingsteps:
1. LocatethefileontheUSBDrive.Itisafileendingin.ana.Copyand pastethe
filetoalocationonthecomputerthatiseasytofind.
2. InthecalibrationtaboftheEllvinGCSoftwareclickon theopenicon tothe
rightofthe“analytefile”box,locatethefilethatwas copiedandpastedand
openit (Figure12).
3. Verifythatthetargetanalytes arelistedundertheanalytes list(Figure13).
4. IftheFROGwas calibratedatadifferentelevation thantheoneitwill beused,
adjustretentiontimes.
Creating a CalibrationFile
Tocreateacalibration filedo thefollowing:
1. Createanewcalibrationfilebyclickingonthe“new”icon totherightofthe
calibrationfilebox(Figure14).
2. Labeland saveyournewcalibration file.
3. Add youranalytesbyclickingon the“new”icon underneaththeanalytelist
(Figure13).Typeintheanalytenameand alias (whichwill appearon thedis-
playwhenrunninginstandalonemode)and selectOK.Repeatthisstepfor
eachtargetanalyteuntil all desiredanalytes havebeenadded.
Generating and AddingData to a Calibration
Tocreateacalibration,aseries ofdifferentknownconcentrationsamples mustbe
run ontheFROG.Defiantrecommendsrunningfivecalibrationstandardsfrom
lowesttohighestconcentration.Refertolatersectionsregardingrunningsamples
and analyzingthedata.Thestepstogenerateand adddatatoacalibrationareas
follows:
1. Run5knownsamples ontheFROGstartingwiththelowestconcentrationsand
endingwiththehighestconcentration,notingwhichconcentrationcorresponds
toeachrun.
2. Analyze oneofthesamplerunsintheanalyzetabofthesoftware(see page9
forhowtoanalyze data).
3. Intheresultstableundertheanalyze tab,highlighttheanalyteyouwouldlike
toaddcalibration dataforand enterintheknownconcentration
4. Withthedesiredanalyteselectedintheresultslist, rightclick on thehighlight-
edrowandselectthecorrectanalytefromthelist.Selectyes whenthesoftware
asksifyou wanttocopythepeakstothecurrentcalibration,thissendsthedata
tothecalibration file.Repeatsteps3and 4forall analytes ateachconcentra-
tionrun.
CalibrationCurveand Equations
Once datahas beenaddedtoacalibration,Ellvinwill createacalibrationcurvefor
thetargetanalyte(s).Figureshowsanexamplecurve.Thecalibration curvedisplays
thedegree ofcorrelation (R²value)betweenanalytestandardsofvaryingconcentra-
tionsandtheinstrument’sresponse tothem.Thecorrelation iscalculatedas botha
linearequationandasecond-orderquadraticequation.Theusermayviewtheequa-
tionsandchart foreitherPeakArea correlation orforPeakHeightcorrelation.Ellvin
uses these parameterstocalculateconcentrationsforfuturesamples.Notethatfor
samplescreening,aonepointcalibrationmaybeused,thispointshouldbethesame
concentrationoftheaction levelforthescreening.
Figure12:Openinga CalibrationFile.
Figure14:Creatinga NewCalibrationFile
Figure15:Addinga NewAnalytetoa CalibrationFile
Figure13:AnalytesList and
addingnewanalytes.
Figure16:SendingDatatoa CalibrationFile
Figure17:CorrelationEquationsandCalibrationCurve
Home
Whenadjusting retentiontimes, the
newvaluegoeshere.