Listoftopics

1)whatisspectroscopy
2)whatisthebasicprincipleofNMR
3)whatischemicalshift
4)factorsaffectingchemicalshift
5)whatisintegrationofapeakinNMR
6)spinspincoupling
7)firstvssecondorderspectra
8)PopleNotation
10)NMRrelaxation
11)T1Relaxation
12)T2Relaxation
13)whydoweneed2DNMR
14)typesof2DNMR
15)whatiscosy
16)whatisNOESY

Whatisspectroscopy?
Spectroscopyinvolvesthestudyofinteractionofradiationwithmatter

WhatisthebasicprincipleofNMR
TheprinciplebehindNMRisthat
nucleihavespindenotedbyI,nucleusofspinIwillhave2I+1possibleorientations.
Intheabsenceofanexternalmagneticfield,theseorientationsareofequalenergy.Ifa
magneticfieldisapplied,thentheenergylevelssplit.
Ifanexternalmagneticfieldisapplied,anenergytransferispossiblebetweenthe
lowerenergytoahigherenergylevel.
Theenergytransfertakesplaceatawavelengththatcorrespondstoradiofrequencies
andwhenthespinreturnstoitslowerlevel,energyisemittedatthesamefrequency.
Thesignalthatmatchesthistransferismeasuredinmanywaysandprocessedin
ordertoyieldanNMRspectrumforthenucleusconcerned.

Thereare4levelsofinformationtobegainedfromprotonNMRoranyrelatedtodifferent
aspectsofthespectrum:
1.HowmanytypesofH?
Givenbyhowmanybasicgroupsofsignalsarethere
2.HowmanyHofeachtype?
givenbytheintegration(relativearea)ofeachgroup.
3.Whichisthegrouptowhichhydrogenisattached?
givenbythechemicalshiftofeachgroup
4.informationaboutneighbouringgroup
givenbycouplingpatterns.

Whatischemicalshift?
chemicalshiftistheresonantfrequencyofanucleusrelativetoastandardina
magneticfield.
Chemicalshiftisusuallyexpressedinpartspermillion(ppm)byfrequencygivenby

factorsthatcausedifferentchemicalshifts
1)Localdiamagneticshielding
2)Localparamagneticshielding
3)Anisotropy

localdiamagneticshielding
theexternalfieldcauseselectronstocirculateinaplaneperpendiculartothefield
direction.thecurrentsoinducedgeneratesasmalllocalfieldopposedtoBotherefore
shieldthenucleus

localparamagneticshielding:
whenanexternalmagneticfieldappliedalongthezaxisdistortsthewavefunctionof
thegroundstatebymixingintoitsmallfractionoftheexcitedstatewavefunction.inthis
waythefieldpartiallyhelpovercometheenergygapbetweenthehigherexcitedorbital
andcreatesapathfortheelectrontocirculateinxyplane.thisinducedcurrent
generatesamagneticfieldwhichdeshieldthenucleus

Anisotropy:
Thisjustmeans"anonuniform"magneticfield.
Electronsinpisystems(e.g.benzene,alkynes)interactwiththeappliedfieldandare
causedto"circulate"withinthepisystem.Thismotioninducesamagneticfieldthat
causestheanisotropy.
Asaresult,thenearbyprotonswillexperience3fields:theappliedfield,theshielding
fieldofthevalenceelectronsandthefieldduetothepisystem.
Dependingonthepositionoftheprotoninthisthirdfield,itcanbeeithershieldedor
deshielded,

WhatisintegrationofpeakinNMRmean?
AmeasureoftheareaofthepeakswhichisproportionaltothenumberofHthatthe
peakrepresents.


SPINSPINCOUPLING

Itistheinteractionbetweenthespinmagneticmomentsofdifferentnuclei.InNMR
spectroscopyitgivesrisetomultipletpatterns

Splittingpatternofpeaks,ingeneral,followthetreediagrampattern
Incaseisthegivennucleusiscoupledtonequivalentnucleicertainpealsoverlap
whichresultsindoubleortripletheareaunderthepeakdependingonthenumberof
peaks
The(n+1)Ruleanempiricalruleusedtopredictsplittingpatternofpeaksin1Hand
13CNMRspectra

Itstatesthatifagivennucleusiscoupledtonnumberofnucleithatareequivalent,the
multiplicityofthepeakisn+1.
Incasethegivennucleusiscoupledtononequivalentnucleiitleadsto2^ndifferent
peaks

Mechanismofspinspincoupling

Onenuclearspin

Intheabsenceofnuclearspin,electronspinsarepairedaccordingtothePauli
principle.Theupdown'anddownup'stateshavethesameenergy

Ifonenuclearspinispresent,thetwospinstatesnolongerhaveexactlythe
sameenergy.
Theinteractionbetweennuclearandelectronspinsisnamedthehyperfine
interactionandactstolowertheenergyofthespinthatispairedwiththe
nuclearspin

Twonuclearspin

Intheaboveimage,twodifferenttransitiontakesplaceforBnucleusinABbond
Fromfavouredgroundstate.Inthisarrangement,Bnucleusisinitslowestenergy
stateandallofthenuclearandelectronicarepaired,resultinginlowestenergyforthe
system.ThesecondoptionisnucleusofatomAandBareparallelresultinginhigher
energysystem
NowlettheBnucleusundergotransitionandinvertitsspin.theexcitedstatethat
resultsfromthehigherenergygroundstatehasalowerenergybecauseallitsnuclear
andelectronicspinsarepaired,whiletheexcitedstatethatresultsfrommorestable
groundstateisofhigherstatebecauseitsspinsarenotpaired
ThusweseetwodifferenttransitionsforBnucleusdependingonthespinofA,asa
result,weseeadoubletinthespectrum

Relationbetweencouplingconstatntandchemicalshift
Firstorderspectraisobservedwhenthereisweakcouplingthatisdifferencein
chemicalshiftisfargreaterthanthecouplingconstant
Secondorderspectraarespectrawhichrequiremathematicalanalysisfor
interpretation.Generallyobservedwhensystemisweaklycoupledthatisthedifference
inchemicalshiftbetweentwogroupissimilarinmagnitudetothecouplingconstant,j

WhatisSpinsystemnotationorPopleNotation

Itisconvenientshorthandnotationtodescribethetypeofspinsystem.
Eachchemicallydifferenttypeofprotonisgivenaletter:A,B,Candsoforth
Ifagrouphastwoormoreprotonofonetype,theyaredenotedbysubscriptsasinA2
orB3
Protonsofsimilarchemicalshiftvaluesareassignedlettersthatareclosertoone
anotherinthealphabet,suchasA,B,C.
Protonsofwidelydifferentchemicalshiftareassignedlettersfarapartinthealphabet,
suchX,Y,XvsA,B,C.
InatwoprotonsystemthatexhibitsfirstorderiscalledanAXsystem
InatwoprotonsystemthatexhibitssecondorderiscalledanABsystem
Inatwoprotonsystemthatexhibitsidenticalchemicalshiftandaremagnetically
equivalentisdenotedbyA2
Inatwoprotonsystemthatexhibitsidenticalchemicalshiftandarenotmagnetically
equivalentisdenotedbyAA
ifthreeprotonsareinvolvedandtheyhaveaverydifferentchemicalshift,aletterfrom
themiddleofthealphabetisused,usuallyMegAMX.
ifthreeprotonsareinvolvedandtheyhaveverysimilarchemicalshiftitisdenotedby
ABC

ExaminingthesystemoftwoprotonHAandHBonadjacentcarbonatoms
weexpecttoseetwodoubletswitheachofequalintensityusingN+1rule.
Butweseetwodoubletonlywhenchemicalshiftislargecomparedtothecoupling
constant
IncaseofHAandHBidenticalchemicalshiftandaremagneticallyequivalent
(A2)weseeonlyonepeak
Forallothercasesasadifferenceinachemicalshiftbycouplingconstantbecomes
smallerandsmallerthesplittingpatterncontinuouslychangesi.etheintensityofthe
innerpeakscontinuouslyincreasesinintensitywhiletheouterpeakofthedoublet
decreasesinintensity.

whatisNMRrelaxation?
NMRrelaxationistheprocessesbywhichanexcitedmagneticstatereturnstoits
equilibriumdistribution.
Ithastwocomponentlongitudinalandtransverserelaxation

T1Relaxation(longitudinalrelaxation)

T1isthetimeconstantforregrowthoflongitudinalmagnetisation(Mz)
Requiresenergytransferfromspinstoenvironment("lattice")
Energytransferisstimulatedbyfieldfluctuatingcausedbymolecularmotionofa
nearbyprotonorelectronneartheLarmorfrequency

Graphindicatingprocessbywhichthenetmagnetisationreturnstoitsinitialmaximumvalue
(Mo)

T2RelaxationorTransverserelaxation
T2isthetimeconstantfordecay/dephasingoftransversemagnetisation
Mayoccurwithorwithoutenergytransfer
AnythingcausingT1relaxationalsocausesT2relaxation,butT2relaxationcanoccur
withoutT1relaxation.
MajorcausesofT2onlyrelaxationincludedephasingbystaticlocalfield

Graphindicatingthedephasingoftransversemagnetisation

Whydowerequire2DNMR
1)Itgivesusawaytoknowwhichproton/carbonarecorrelatedwitheachother
2)Ithelpsresolveoverlapofcomplexstructurebycorrelatingchemicalshift

2DNMRBasics
Thepreparationtime
Timerequiredtoallowtheexcitednucleitogetbacktheirequilibriumstate
Theevolutiontime
Duringtheevolutiontime,thespinsystemisevolvingundertheeffectofdifferent
factors,eachcoherenceevolvesatitsowncharacteristicfrequencyasafunctionofthe
chemicalshiftandofthescalarcouplingofthecorrespondingnucleus.
TheMixingtime
Itismadeofapulsesequencewhichachievestransfersofmagnetisationdifferent
frequenciesarecorrelated.
Thedetectiontime
Theacquisitionofthemodulatedsignaltakesplaceduringthedetectionperiod.The
sequencewejustdescribeddoesnotconstitutebyitselfa2DNMRexperiment.

typesof2DNMR
1)Homonuclear
Throughbond:COSY,TOCSY,2DINADEQUATE,2DADEQUATE
Throughspace:NOESY,ROESY
2)Heteronuclearcorrelation
OnebondcorrelationHSQC,HMQC
LongrangecorrelationHMBC

(ihavecoveredonlyCOSYANDNOESY.IalsohaveabasicideaofTOCSYANDROESY.
RestIamyettocover)

WhatisCOSY
COSY(CorrelatedSpectroscopY)isausefulmethodfordeterminingwhichsignals
arisefromneighbouringprotons.
Correlationsappearwhenthereisspinspincouplingbetweenprotonsandwhere
thereisnocoupling,nocorrelationisexpectedtoappear.
Thismethodisveryusefulwhenthemultipletsoverlap

WhatisNuclearOverhausereffect?
Whenaspecificnucleusismagneticallyexcitedanditsneighbourisatequilibrium,
relaxationoccursbetweenthetwonucleiBecauseofwhichtheintensityofthenucleus
thatwasatequilibriumchanges.

WhatisNOESY
InNOESY,thenuclearOverhausercrossrelaxationbetweennuclearspinsthat
happensduringthemixingperiodisusedtoestablishthecorrelationsnucleithatare
spatiallycloseratherthanthosethatarethroughbondcoupledtoeachother.
NOESYisusefulfordeterminingwhichsignalsarisefromprotonsthatarecloseto
eachotherinspaceeveniftheyarenotbonded.
OneapplicationofNOESYisinthestudyoflargebiomolecules