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Electronicenergylevelchangesandcolor

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ElectronicSpectraofMolecules:TheAbsorptionofUVandVisibleLight
ByWaltVolland
BellevueCommunityCollege
Description

Wavelengthabsorbed

Phenolphthalein

Equipment

Procedure

Chromatography

Data&report

Description
Absorptionoremissionofultravioletorvisiblelightbyamoleculedependsonelectron
transitionsbetweenmolecularorbitalenergylevels,justasabsorptionoremissionof
electromagneticradiationbyanatomisdeterminedbyelectrontransitionsbetweendifferent
energylevelsintheatomandtheEsforthosetransitions.Molecularspectrafollowrules
analogoustotherulesforatomicspectra:energyisabsorbedonlywhentheamountofenergy
providedmatchesthedifferenceinenergy,E,of2energylevels.Whenanelectrongoesfrom
ahighertoalowerenergystate,aphotonofdefinitewavelengthandfrequencyisemitted.
Everyatomormoleculehasacharacteristicelectronicspectrumdependingonitscharacteristic
Es.Becauseofamolecule'sgreatercomplexity,wecanoftenconstructamoleculethatwill
giveaparticularspectrum,ratherthanhavingtojustacceptthespectraavailableaswedowith
atoms.Thispossibilityarisesbecauseoftheinterdependenceofmolecularorbitalenergylevel
valuesforthemolecule,molecularshape,bonding,anddistributionofelectrondensitywithinthe
molecule.Energytransitionsinphenolphthaleinindicator,plantpigments,andisolatedpisystems
willbeexaminedtoillustratethis.
Molecularorbitaltheoryprovidesamodelforthewayelectromagneticradiationinteractswith
molecules.Forexample,anelectroninthepibondingmolecularorbital(MO)ofanalkenecan
beexcitedtoapiantibondingMO.Thisisdescribedasa
transition.

Foranisolatedpibondtheenergyseparation,E,betweenthepibondingandpiantibonding
MO'sislargeultravioletlightwithitslargeenergyandshortwavelengthisneededtoexcitethe
pielectron.Molecularorbitaltheorypredictsthattheenergydifference,E,betweenlevelswill
decreaseifthedoublebondisconjugatedwithanotherdoublebond.(Conjugateddouble
bondshaveonesinglebondseparatingthem.Theyarecoplanarandpielectronscanmove
throughoutthepisystem.)Cogjugationexistswhenseriesofalternatingdoubleandsingle
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bond.Thismeansthatthemoleculehasasinglebondbetweenthetwodoublebonds.Theterm
"conjugated"isusedinchemistrytorefertoaseriesofalternatingsingleanddoublebonds. The
predicteddecreaseinEforconjugatedstructuresisalsoobservedinexperiments.
Hereisageneralrulethatdescribestheeffectofdoublebondsconjugationontheenergy
absorbedbythepisystem.Thegreaterthenumberofconjugatedmultiplebondsina
compound,thelongerthewavelengthofthelightthatthecompoundwillabsorb.This
translatesintoasmallerenergyrequirementforthetransitionfromthehighestoccupiedMOto
thelowestunoccupiedMO.Thisideaisillustratedschematicallyinthefollowingdiagramfor
alkeneswherethetransitionisfor
.Astheconjugationincreases,electronsare
delocalizedovermorespaceandthesystemismorestable.

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Compoundname

Condensedformula

Wavelength
absorbed

Ethene,ethylene

CH2::CH2

171nm

1,3butadiene

CH2::CHCH::CH2

217nm

Trans1,3,5
hexatriene

CH2::CHCH::CHCH::CH2 274nm

carotene

seestructurebelow

425nm

Theplantpigmentcarotenehasthislinestructure.

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Thereare11doublebondsinthealternatingdoublebondsinglebondsystemincarotene.
Thedoublebondsareessentiallycoplanarandtheelectronsaredelocalizedovera22
carbonchain.
Thewavelengthabsorbedbycaroteneisintheblueregionofthevisiblespectrum.Whenwe
lookatcarotene,no"bluewavelengths"reachoureyesbecausethecarotenemoleculeis
absorbingthese.Therestofthewavelengthsinthevisibleregion(minustheblue)reachour
eyesandcarotenelooksorangetous.Remember,thecoloroflightabsorbedbythemolecule
isnotthecolorwesee.Itistheonewedon'tsee!
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ObservedColorof
Compound

ColorofLight
Absorbed

Approximate
WavelengthofLight
Absorbed

Green

Red

700nm

Bluegreen

Orangered

600nm

Violet

Yellow

550nm

Redviolet

Yellowgreen

530nm

Red

Bluegreen

500nm

Orange

Blue

450nm

Yellow

Violet

400nm

Absorptionoremissionofultravioletorvisiblelightbyamoleculedependsonelectron
transitionsbetweenmolecularorbitalenergylevels,justasabsorptionoremissionof
electromagneticradiationbyanatomisdeterminedbyelectrontransitionsbetweendifferent
energylevelsintheatomandtheEsforthosetransitions.Molecularspectrafollowrules
analogoustotherulesforatomicspectra:energyisabsorbedonlywhentheamountofenergy
providedmatchesthedifferenceinenergy,E,of2energylevels.Whenanelectrongoesfrom
ahighertoalowerenergystate,aphotonofdefinitewavelengthandfrequencyisemitted.
Everyatomormoleculehasacharacteristicelectronicspectrumdependingonitscharacteristic
Es.Becauseofamolecule'sgreatercomplexity,wecanoftenconstructamoleculethatwill
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giveaparticularspectrum,ratherthanhavingtojustacceptthespectraavailableaswedowith
atoms.Thispossibilityarisesbecauseoftheinterdependenceofmolecularorbitalenergylevel
valuesforthemolecule,molecularshape,bonding,anddistributionofelectrondensitywithinthe
molecule.Energytransitionsinphenolphthaleinindicator,plantpigments,andisolatedpisystems
willbeexaminedtoillustratethis.
Phenolphthaleinisanotherexampleoftheeffectofmolecularshapeondelocalizationofpi
electrons.Phenolphthaleinmoleculeiscolorlessinacidsolutionanditspisystemisnonplanar.
Theelectronsinthedoublebondsare"localized.Theexcitationenergyislargeandinthe
ultravioletrange.Themoleculelosesprotonsinbasicsolutionsandformsaminus2anion.This
negativeionisflatandthepisystemisconjugated.Intheplanarionthepielectronsinthe
elevendoublebondscandelocalizeoverthe3rings,theCOO1carboxylgroupandthe
oxygens.Theanionimpartsacolortosolutionsbecausetheenergyseparationbetweenthepi
andpiantibondinglevelsissmallerandmatcheslightinthevisiblerange.

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Inthefirstpartofthisexperimentyouwillobservethisdifferenceincolorforphenolphthalein
anditsanion.
Inthesecondpartofthisexperimentyouwillextractchlorophyllfromgreenleavesandthenuse
chromatographytoseparatechlorophyllafromchlorophyllb.Youmayalsoseethe
separationofcaroteneandotherplantpigments.Youwillbeaskedtoexplainthedifferent
colorsofthe2chlorophyllsbyexaminingtheirstructuresandidentifyingthetypeofchangein
molecularorbitalenergylevelscausedbythedifferenceinstructure.
Paperchromatographyisaseparationtechniquethatanyonewhoeverspilledcoffeeorteaona
pieceofpaperhasseen.Thesolventwetsthepaper,creepsalongcarryingsolutesalongwithit.
Thedifferentdissolvedmaterialsmovewiththesolvent,butatdifferentratesbecausethepaper
attractsthesolutesdifferently.Youwillusethisbehaviorinasystematicwaytoseparate
pigmentsinchlorophyll.Youwillmakeanextractofleafpigmentsbysoakingplantleavesina
mixtureofcoldacetoneandethanol.Theextractappearstobegreen,butotherpigmentcolors
maybemaskedbythestronggreentint.Youwillusepaperchromatographytoseparateany
pigmentsintheextract.Theseparationoccursbecausethepigmentshavedifferentsolubilitiesin
themobilesolventanddifferentabsorptionaffinities(attractions)tothechromatographypaper.
Theseparationprocesswillgiveachromatogram.

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EquipmentandSupplies
Alfoil

13gfreshspinachleaves

2beakers,100mL

ice

graduatedcylinder,10mL

phenolphthaleinindicatorsolution

medicinecup

acetone

3Beraltypepipets

vinegar

erlenmeyerflask,50mL

ethanol

Beraltypepipetwithnarrowstem

5MNaOH

glassjarabout7inchestall(about1
quart)

petroleumether
chromatographypaper

2pencils

plasticwrap

string

2rubberbands

scissors

tape
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Procedure
Pleasereadtheentireprocedurebeforestartingtheexperiment.
Wearyourgogglesatalltimeswhiledoingtheexperiment!
Gatheryourequipmentandchemicals.CoveryourworkareawithAlfoilorwaxedpaperto
catchspillsandspatters.
PreparationforExtractingChlorophyll
Dothisfirst,thendothephenolphthaleinpartoftheexperiment.Finally,finishupwiththe
chlorophyllchromatography.
Collect1to3gramsoffreshspinachleaves.Ifyoucan'tgetspinach,otherfreshgreenleaves
(turnipgreens,broccolileaves)willwork,butyourchromatogrammaybeslightlydifferentfrom
theonedescribedlaterinthisexperiment.Anydarkgreenleavesshouldallowyoutoextract
the2chlorophyllpigments.
Makeanicebathbyplacingalittlecoldwaterandsomeiceinabeaker.
Removethecontainersofacetoneandethanolfromyourkit.Donotopenyet.
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Setthe2containersintheicebathsotheacetoneandtheethanolgetcold.Donotletthewater
getintotheacetoneorethanolcontainers.
PhenolphthaleinMoleculeandAnionColors
Pourabout2mLofwaterintooneofthemedicinecupsfromyourkit.Thereferencemarkson
thecupareadequateyoudonotneedtomeasurethewaterwithyourgraduatedcylinder.
Addoneor2dropsofthephenolphthaleinindicatorsolutiontothewaterinthemedicinecup.
YoucandothiswithoneofyourBeraltypepipetsifthereisno"builtin"dropperinyour
phenolphthaleincontainer.
UseadifferentBeraltypepipettoadd2dropsofvinegartothemedicinecup.Stirwithyour
stirringrod.Noteandrecordthecolorofthesolutioninthemedicinecup.
USECAREWHENPOURINGTHENaOH.ITISCAUSTIC.
Ifthe5.0MNaOHsolutiongetsintoyoureyes,rinsetheeyeballfor15minutesinwarm
runningwater(forceyoureyelidsapartwithyourfingerstokeepyoureyeopenandtheeyeball
exposedtotherunningwater).Contactyourphysicianforadviceifyoureyecontinuestohurt
or"burn".
UseathirdBeraltypepipettoadd2or3dropsof5MNaOHtothecontentsofthemedicine
cup.Stirwithyourstirringrod(noneedtowashtherodbetweensteps).Noteandrecordthe
colorofthemixtureinyourcup
Discardthesolutioninthemedicinecupbypouringitintothedrainfollowedbycoldwater.
PreparationofPigmentExtract
Workinawellventilatedareaawayfromanyopenflamesorhotsurfaces(stove,space
heater,waterheater,etc.).Acetoneandethanolconverttovaporveryeasily,especiallythe
acetone.Boththeliquidandthevaporareflammable!
Labelyourerlenmeyerflask"pigmentextractmixture".
Cuttheleavesintotinypiecesusingyourscissors.Thesmallerthepieces,thebetterthecontact
betweenpiecesofleafmaterialandtheextractionsolvent.
Placethesmallleafpiecesintheerlenmeyerflaskandadd5mLofcoldethanol.
Thenadd5mLofcoldacetone.Thereisnoneedtowashyourgraduatedcylinderbetweenthe
ethanolandacetone.
Quicklycovertheerlenmeyerflaskwithapieceofplastic.Holdtheplasticwrapsnuglyagainst
theneckoftheflaskwitharubberband.Thishelpskeepthesolventfromevaporatingwhilethe
pigmentsarebeingextractedfromtheleaves.
Dumpthewateroutofthebeaker.Keeptheice.Settheerlenmeyerflaskintheiceinthe
beaker.
Storebothinacool,wellventilatedareaforabout24hours.Donotputthebeakerand
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erlenmeyerinyourrefrigerator.Storeawayfromfood,hotsurfaces,andopenflames.
Thesolventwillextract(dissolve)pigmentsfromtheleafmaterial.Thenyoucandothe
chromatographytoseparatethedifferentpigments.
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Chromatography
Assembleachromatographychamber.Youneedaglassjarthatisabout7inchestall,apiece
ofplasticwraptocoverthetopofthejar,andalargerubberbandtogoaroundtheneckofthe
jartoholdtheplasticwraptightlyinplace.Arecycledjellyjar,instantcoffeejar,orjustabout
anyquartjarwilldo.
Obtainapieceofchromatographypaperthatmeasuresabout3cmby15cm.Handleonlythe
edges.Trynottotouchthesurface.Oilsfromyourfingerscanaltertheabsorptivepropertiesof
thepaperandtheseparationmaynothappennormally.
Usearulerandapenciltodrawalineonecentimeterfromthebottom3cmedgeofthepaper.
DONOTUSEINK!Inkwilldissolveinthesolventandruintheseparationprocess.Thisisthe
linewhereyouwillplaceaspotoftheextract.
Lightlymarktheedgesatonecentimeterdistancesalongtheedgesofthepaper,lengthwise.
Thishelpsyoukeeptrackoftheleadingedgeofyoursolvent(thesolventfront)asit"climbs"
thestripofpaper.
Usethetipofyourscissorstopokeasmallholeabout1/2cmfromthetop3cmedgeofthe
paper.Insertapieceofstringorthreadthroughthehole.Tapetheendsofthestringtothe
middleofapencilorpen.Thepapershouldhangfreelyfromthepencilwhenyouholdupthe
pencil.
Hangthepaperinthecenterofyourglassjarbyplacingthepencilacrossthemouthofthejar.
Rotatethepenciltowindthestringaroundituntilthebottomofthepaperjustclearsthebottom
oftheglassjar.Thepapershouldnottouchanypartofthejar,noteventhebottom.

Tapethewoundupstringinplaceonthepencilsoitcan'tunwindwhenyouletgoofit.
Removepaperfromjarbyliftingthepencil.Applythe"pigmentextractmixture"asdirected
below.

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Thisanimationillustrateswhathappensinthe
separationofatwocomponentmixture.The
solventfrontismarkedbytheupperblackline.
Thereisagreencomponentthatmovesfaster
thantheyellowcomponentinthemixture.The
Rfvalueforthegreeniscloserto"1"thantheRf
valuefortheyellowcomponent.

Taketheplasticwrapoffthebeakerof"pigmentextractmixture".UseaBeraltypepipetwitha
verynarrowstemtodrawupaverysmallquantityofpigmentextract.
Placeonedropofthepigmentextractonorjustslightlyabovethepencillineonthe
chromatographypaper.Thepigmentextractspotshouldbeabout3to5mmindiameterwhich
isthesizeofthisspot.

Letthespotdryforatleast30seconds.
Repeatthisproceduretoapply4or5moredropsdirectlyontopofthefirstdrop.ALLOW
THEPIGMENTTODRYBETWEENAPPLICATIONS!Ifyouapplyalargeamountatone
timeorifthespotisstillwetfromthepreviousdrop,thespotwillspreadoveranunmanageable
areaandgivepoorseparationresults.Youneedtohaveseveraldropsofpigmentonthepaper
tomakethecolorsofminorpartsofthemixturevisibleinyourfinishedchromatogram.
Placetheplasticwrapcoverbackontheerlenmeyerflaskofpigmentextract.Youmayneedto
repeatthechromatographyseparationandneedthisextractfortheduplicatetrial.
Prepareamixtureof1mLacetoneand9mLpetroleumether.Thisisthesolventforthe
chromatographyseparation.
Placeenoughacetonepetroleumethersolventinyour"chamber"togiveadepthof1/2cmin
thebottomofyourglassjar.Theliquidlevelmustbelowerthanthepencilstartlineacrossthe
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paper.
Placethe"loaded"chromatographypaperintothechamber.Lowerthepenciluntilthebottom
ofthepaperjusttouchesthesolventinthebottomofthejar.Youwantthebottomofthepaper
togetwetwithsolvent.Youdonotwantthespotofpigmentextracttotouchthesolventatthis
time.
Coverthetopofthechamberwithplasticwrapusearubberbandtoholdthewrapinplace.
Watchthesolventclimbthepaper.
Gaugethespeedofmovementofthesolventfront.Theprogressofthesolventwillbequickat
first.Thepigmentextractspotwillstarttosmearoutasthesolventcarriesthepigmentsaway
fromthestartlineandthechromatogram"develops".
Allowthechromatogramtodevelopfor10minutes,oruntilthesolventfrontcomeswithin1cm
ofthetopedgeofthepaper.Removethepaperfromthechamberbyliftingthepencil.
Ifyouletthechromatogramdeveloptoolong,thesolventfrontwillactuallycometoastop.
Solventmoleculeswillbeevaporatingfromthewetpaperasfastastheyare"climbing"ontothe
paperatthebottom,sothefrontwillstoprising.However,solventmoleculeswillstillbemoving
upthepaper,washingthepigmentalong.Thedifferentcomponentsofthepigmentmixturewill
starttoovertakeoneanotherand"pileup",undoingtheseparationyouachieved.
Allowthepapertodry.Usepencilstosketchtheoutlinesofthecolorsonthepaper.Beginwith
thecolorclosesttothestartlineandlabelthedifferentcolorregions.Thefirstpigmentspot
shouldbeyellowgreenthisischlorophyllb.Thebluegreenpigmentischlorophylla.You
shouldalsosee2yelloworangexanthophyllsinthemiddle.Thedarkerorangespotnearthe
topiscarotene.Xanthophyllsandcarotenesaremembersofthecarotenoidclassofpigments.
Thechromatogramwillfadeonexposuretolightbecauseoflightinducedoxidationreactions.If
youwanttopreserveit,storeitinadarkplace.

DataandConclusions
Phenolphthalein
Whatisthecolorofthephenolphthaleinmoleculeinacidicsolution?
Whatisthecoloroftheanioninabasicsolution?

Whydoyouseeacolorfortheanioninbasicsolution?

WhichpisystemwillhaveasmallerEforthe
ortheanion?
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Whatistheapproximatewavelengthabsorbedbytheanionofphenolphthalein?

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Chromatographyresults
Theseillustrationsforchlorophyllaandchlorophyllbdonotshowthehydrogenatoms
becausetheywouldobscurethecarbonskeleton.Thecarbon,nitrogen,oxygen,and
magnesiumatomsareshown.Singleanddoublebondsareindicated.Thereare12double
bondsinaseriesofalternatingdoubleandsinglebonds.Thisisalowenergy,conjugatedpi
systeminchlorophylla.
ChlorophyllbhasanaldehydegroupinplaceofaCH3groupinchlorophylla.Therestof
thestructureisthesameforthe2molecules.

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Howmanydoublebondsareinthealternatingpisystemforchlorophyllb?

Explainthedifferenceincolorforchlorophyllaandchlorophyllbintermsofthenumberof
doublebondsinthe2molecules,thewavelengthabsorbedandthesizeoftheEthat
correspondstothiswavelength.

Whyisitnecessarytouseaclosedchamberwhenyoudevelopthechromatogramoftheplant
pigmentextract?

Thespotsthatappearinthechromatogramoftheplantpigmentextractdonotalways
completelyseparate.Thetrailingedgeofonecolormayoverlapwiththeleadingedgeofthe
onebehind.Ifthechromatographypaperwererotatedby90o andanotherdevelopment
periodcarriedoutwiththesolventnowmovingatrightanglestoitsoriginaldirection,would
betterseparationbepossible?Explainyouranswer.

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Allrightsreserved1999WaltVolland

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