NMR Spectroscopy: Principles and Applications. Nagarajan Murali 2D NMR Heteronuclear 2D Lecture 7
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1 NMR pecroscop: Principles and Applicaions Nagarajan Murali D NMR Heeronuclear D Lecure 7
2 Heero Nuclear D-NMR Two dimensional NMR can be used o correlae NMR signals arising from differen nuclei such as 3 C, 5 N, 3 P ec wih he aached proons. We have alread seen D-ediing eperimens like APT, NEPT, and DEPT ha ransfer coherences from H o 3 C. ince he correlaed nuclei are of differen pe usuall we will represen one species as spin and he oher as spin.
3 HETCOR - eperimen A COY eperimen when correlaing wo differen pe of nuclei is called Heeronuclear Correlaion (HETCOR eperimen. The pulse sequence is shown below. Usuall, spin is H and he shifs of which in w is correlaed wih spin, he aached heero nuclei, ha is deeced in he period whose chemical shif appear in w dimension.
4 HETCOR - eperimen pin coherences are ecied b he firs spin 90 o pulse ha labels is offse in (period A. The subsequen spin echo sequence (period B convers he -spin in-phase coherence ino -spin ani-phase coherence wih respec o spin. The subsequen wo 90 o pulses effec he coherence ransfer o ani-phase -spin coherence (period C which is convered ino -spin in-phase coherence b he spin echo sequence (period D and deeced during.
5 HETCOR - eperimen is insrucive o go hrough he spin evoluion o conras HETCOR from COY. ( J cos( sin(, ( cos( cos( sin( sin( cos( J cos( J cos( J cos( J cos( sin( cos( sin( sin( J sin( J sin( J sin( J A he end of period C
6 HETCOR - eperimen ince we deec onl spin, he -spin coherence is he onl relevan erm during period D and period. sin( sin( sin( J sin( J sin( J (,J ( n he -spin coherence evolve wih is own offse. The coupling o spin is removed b he decoupling field. Also noe ha in also here is no evoluion of he coupling erm onl he offse of spin- is labeled. A he end of period D or a he sar of.
7 HETCOR - eperimen The schemaic D specrum can be represened as below. sin( sin( J sin( J A peak a in w and in w appear. is he cross peak equivalen of he COY specrum. There is no diagonal peak as we are correlaing wo differen nuclei.
8 HQC - eperimen n HETCOR eperimen, he magneiaion sars from spin and ransferred o spin and he spins are deeced. Thus, here is a signal enhancemen of g /g as we have seen in he D lecure of NEPT and DEPT schemes. sin( sin( J sin( J nsead of deecing he low g nucleus, we can sar as in HETCOR and frequenc label -spin coherences in and deec -spin b an era NEPT back ransfer, we ge a new eperimen called Heeronuclear ingle quanum Correlaed pecroscop (HQC. We no onl ge he g /g advanage, bu also addiional higher sensiivi in he raio (g /g s 3/ b deecing he spins. N g g 3/
9 HQC - eperimen The HQC pulse sequence is given below: (a The NEPT sequence (par A +B creaes -spin aniphase coherence wih respec o spin and he -spin coherence is frequenc labeled during (period C. The final wo 90 o pulses convers he -spin aniphase coherence o -spin aniphase coherence and he deecion of spins sar immediael. n (b afer period D anoher spin echo is added o refocus he spin coherence so ha decoupling can be used. Period D+E is known as reverse NEPT.
10 We sar a he end of period A HQC - Eperimen sin( sin( cos( ( sin( cos( } sin( cos( { sin( cos( sin( cos( sin( cos(, J J J J J J J J J
11 HQC - Eperimen We sar a he end of period D sin( J Period sin( J sin( J E cos( J sin( J cos( cos( cos(
12 D-HQC-ummar ( J 4J ( J cos( J sin( J,( w s cos( ws sin( ws cos( ws in( ws
13 Eamples - HQC H- 3 C (naural abundance muliplici edied HQC of disaccharide is shown below. D=/J invers XH (black responses relaive o XH and XH 3 (red
14 D-Heeronuclear Muliple Quanum Coherence pecroscop (HMQC J ( J Decoupling RF Muliple Quanum Coherence cos( J sin( J,( w s ( J cos( ws sin( ws cos( ws in( ws
15 Eamples - HMQC H- 3 C (naural abundance HMQC of Menhol is shown below.
16 Long Range Correalion- Heero Nuclear Mulibond Correlaion (HMBC n boh HQC and HMQC he delas are se based on one-bond J coupling consan values. f we wan o observe long range couplings across muliple bonds hen heir values are much smaller and var over a wide range. n HMQC, here are wo delas and he peak inensi hus depend as sin ( J B ploing his funcion for various coupling consans we can opimie he dela o observe long range coupling and hence he remoe cross peaks in a D specrum. uch an eperimen is known as HMBC.
17 Long Range Correalion- Heero Nuclear Mulibond Correlaion (HMBC n HMBC eperimen, we look for onl he long range coupling peaks and suppress he direc peaks ha we see in a HMQC or HQC specrum b using he sequence below. The dela is se o /( J and is he usual long dela corresponding o long range coupling. The one bond coupling generae is own aniphase coherence ha is convered ino MQC b he firs 90o pulse on he -spin. The eperimen is done wice once wih his pulse along (-ais and again along (--ais and added o suppress he direc peak. 3. 5ms 0*3.5ms 6. 5ms J *60
18 Eamples - HMBC H- 3 C (naural abundance HMBC is shown below.
19 HMQC and HMBC A pical HMQC (Lef and HMBC (Righ of H- 3 C correlaion specra will look as below.
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