Chem1122010 GasChromatography

Thepurposeofthisexperimentistodeterminethecompositionofsolutionsthatcontainamixtureofdifferent
alcohols.Theanalysisisperformedusinggaschromatography.

Introduction
Thetermchromatographyappliestotheseparationofchemicalconstituentsinasamplesotheycanbeeither
detectedorutilizedindividually.Gaschromatography(GC)isamethodofseparatingvolatilecompounds
(thosewithahighvaporpressureorarelativelylowboilingpoint)sothattheymaybedetectedindividuallyin
complexmixtures.Compoundsareseparatedbasedondifferencesintheirvaporpressuresandtheirattraction
tosolidmaterialsinsidetheinstrument(agaschromatographorGC).Becausethevaporpressureofagiven
compoundisafunctionoftheintermolecularforcesbetweenmolecules,GCtakesadvantageofdifferencesinat
leastoneofthepropertiesofmatterdiscussedinlecturesandinthetext.

InGC,thesampleisinjectedintotheinstrumentusingasmallsyringe.Thesampleissweptintotheinstrument
usingacarriergas(usuallyHe)wherethesampleisseparatedintoitsindividualchemicalcomponents,called
analytes.Separationisachievedbybothattractiontothestationaryphase(thecoatingontheinsideofthe
column)anddifferencesinvaporpressure.Becausevaporpressurevarieswithtemperature,thetemperatureof
theinstrumentisoftenadjustedduringthechromatographicrun.Adetector,whichisdesignedtosense
analytemoleculesastheyexittheGC,isattheendofthecolumn.

Wewillbeusingathermalconductivitydetector(TCD).Thatmeasureschangesinthepropertiesofthecarrier
gas.Thechangesareduetothepresenceoftheseparatedanalytemoleculesinthecarriergasstream.Because
theanalytemoleculesbinddifferentlytothestationaryphase,theytravelthroughtheGCcolumnatdifferent
rates.Thatis,theyhavedifferentretentiontimesonthecolumn.Asananalyteappearsinthedetector,its
presenceissignaledbyapeak.Thus,agaschromatogramconsistsofaseriesofpeaks,oneforeachofthe
componentsofthesample.Thechromatogramisdisplayedonachartrecorderorcomputerscreen.

Anexampleofachromatogramisshownbelowforamixtureoftwocompounds,AandB.


Thepositionofapeakonthexaxisisameasureofretentiontimeandisafunctionofthestructureofthe
compound.Theyarelabeledonthechromatogramaboveas(tr)Aand(tr)Bforthetwocomponents,AandB.The
areaunderthepeakisafunctionofthatcompoundsconcentrationinthesample.Theareaofthepeakis
measuredbyassumingthepeakhasatriangularshape,withthebasemeasuredbyextrapolatingthesidesof
thepeaktothebaseline(shownaboveasWAandWB).Theareaisthenxheightxwidthatthebase.

Thepercentcompositionofeachcomponentinanunknownsampleisdeterminedusingtheareaunderitspeak
inthechromatogramofthatunknownmixtureandthatcomponentsresponsefactor.Aresponsefactorisa
conversionfactorbetweentheareaunderthecurveforacomponentanditspercentabundanceinthemixture.
Theresponsefactorisdeterminedbycomparingtheareaunderthepeaktothepercentabundanceofa
componentforasolutionofknowncomposition.Becausethesolutionyouusetodeterminethisforeachofthe
fourcompoundsiscomposedof25%ofeachcompound,
25%
response factor =
area under peak

Differentcompoundsaffectthedetectordifferently,soeachcompoundhasitsownuniqueresponsefactor.

Onceyouhavetheresponsefactorsandretentiontimesforthefourcompounds,youcananalyzeyourunknown
samples.Thepercentcompositionofanycomponentinanunknownmixtureisdeterminedbymultiplyingthe
peakareafortheunknownmixturebytheresponsefactorforthatcomponent.

response factor percent

[analyte]sample = area sample
response factor cm3

Besuretoidentifywhichcompoundeachpeakrepresentsbymatchingitsretentiontimetotheknowns.

Toreview:Youtellwhichcompoundsareinasamplebywhentheyaredetectedandhowmuchispresentby
theareaunderthepeak.Youmeasureretentiontimesandresponsefactorsusingknownsolutions,andthen
usethatinformationtoanalyzeunknownsolutions.

SamplestoAnalyze:

Fourcompoundsareinvolvedinthisexperiment:pentanol,hexanol,heptanol,andoctanol.

KnownSamplesareprovidedofpurepentanol,hexanolandoctanolandamixtureof25%eachofpentanol,
hexanol,heptanolandoctanol.

Avarietyofunknownsamplesthatcontainmixturesofthesefourcompounds.

Procedure:

Your instructor will assist you with the operation of the GC.

1. Your instructor will provide you with three Unknown Samples, which are mixtures of the four possible alcohols.
You will also receive three Pure Solutions, each containing one of the possible compounds (pentanol, hexanol
and octanol).

2. Inject 10 L of Standard into the B injector on the right of the GC.

3. Inject each of the three Pure Solutions into the GC. Inject 5 L volumes for these known samples that contain
just one compound. Collect the chromatograms on the chart paper. At the start of each run, mark on the chart
paper where the run started. The chart paper moves at a rate of 1 cm/minute. At the end of each run, label the
chromatogram with the sample name, injection volume, and other parameters.

You should now know what the retention time is for each of the your alcohols. By comparing the
three Pure Solution chromatograms with the chromatogram of the Standard, you can identify the
peaks. The peak for heptanol will be the one in your Standard that does not correspond to any of
the three Pure Solution peaks!

4. Analyze the three Unknown Samples by injecting 10 L of each into the GC. Your Unknown Samples may
contain one, two, or three of the compounds.

CalculationsandReport
Thepurposeoftheexperimentistodeterminethecompositionoftheunknownsolutions.Thosecompositions
makeuptheconclusionofyourreport.Thingsyouneedtodo:

1.Determinetheretentiontimesofthefourcompounds.Thoseforpentanol,hexanolandoctanolcanbe
determineddirectlyfromthechromatogramsperformedontheirpuresamples.Usethechromatogramofall
fourcompoundstoidentifythepeakforheptanolanddetermineitsretentiontime.

2.Usingthechromatogramthatcontainedallfourcompounds,calculatearesponsefactorforeachcompound
bycomparingitspercentcompositiontotheareaunderitspeak.Theunitsoftheresponsefactorare
percent/cm%.

3.Makeatableofretentiontimesandresponsefactorsforthefourcompounds.

4.Foreachunknownsample:
a.identifywhichcompoundsarepresent.
b.usetheareaundereachpeaktodeterminethepercentcompositionforeachcompoundpresent.
c.Makeatablelistingthecompoundspresent.Foreachcompoundpresent,listtheareaunderthepeak
andthecalculatedpercentcomposition.

GasChromatographyREPORTFORM NAMES:

KnownCompounds

Compound RetentionTime,min Area,cm3 ResponseFactor,%/cm3

Pentanol
Hexanol
Heptanol
Octanol

Unknown#____
Component Retentiontime CompoundName Area,cm3 PercentComposition
Component1
Component2
Component3

Unknown#____
Component Retentiontime CompoundName Area,cm3 PercentComposition
Component1
Component2
Component3

Unknown#____
Component Retentiontime CompoundName Area,cm3 PercentComposition
Component1
Component2
Component3

PostLabQuestions:
1.Twodifferentcommonmethodsofseparatingcompoundsusingacolumndependonboilingpointandon
molecularsize.Incolumnsdependingonboilingpoint,thecompoundwiththelowestboilingpointspendsthe
mosttimeinthegasphase,whereitcanmove.Thelowestboilingcompoundthereforemovesdownthecolumn
fastest.Inthemethodusingmolecularsize,calledsizeexclusionchromatography,smallermoleculesgetstuck
inporesinthecolumnwhereaslargermoleculescantfitandfloatonby.Therefore,thelargermoleculesmove
downthecolumnfaster.Toanswerthis,youneedthestructuresandboilingpointsofthefourcompounds.

Whichofthesetwoismorelikelytobeusedinthisexperiment?

2.Examineyourdataandexplainanyfacetsifthereareanythatyousee.