You are on page 1of 34

A SEMINAR ON: ADVANCES IN NMRCORRELATION SPECTROSCOPY

4/21/2012

Prepared By Mr Nandan V. Shinde .Damle M.pharm Q.A.(1st sem)

Guided By Mrs.M.C

1

4/21/2012

NMR SPECTROSCOPY
Based upon measurement of absorption of electromagnetic radiation in the radiofrequency region of roughly 4 to 600 MHz  Nuclei of certain atoms are involved in the absorption process  In order to cause nuclei to develop the energy states required for absorption to occur, it is necessary to place analyte in an intense external magnetic field.

2

THE BASIC IDEA OF nD NMR

4/21/2012

For a single uncoupled proton resonance

P: Preparation E: Evolution M: Mixing D: Detection

Amplitude modulation of a singlet resonance as a function of t1 (signal decays due to spin relaxation)

FT t2

FT t1

3

will produce 2D peaks at their corresponding chemical shift offsets in both dimensions (2D spectra are in fact 3D images). Contour Plot 4 . of offsets nA and nX.The concept can be expanded to 3 and n dimensions 4/21/2012 two-dimensional (2D) experiment threedimensional (3D) experiment In the previous slide two-dimensional experiment. a sample containing two uncoupled spins A and X.

A necessary condition for the magnetization magnetization transfer to takes place during the mixing pulse (M).4/21/2012 If spins A and X are J coupled. Diagonal Peak Cross-Correlation Peak 5 . As a result. during the evolution time t1 the magnetization will evolve into anti-phase components. giving origin to a cross-correlation peak between them. each magnetization (A and X) will not only modulate with its own resonance frequency but they will also modulate with the resonance frequency of the coupled spin.

and spin topology filtration reduce the complexity. Relayed correlation. Both avenues can lead to three-dimensional spectroscopy.4/21/2012 Extensions of the standard COSY experiment. 6 . and multiple quantum spectroscopy (MQS) increase the information content. while exclusive correlation (E. multiple quantum filtering (MQF).COSY). total correlation spectroscopy (TOCSY).

HMQC. Multiple quantum. TOCSY. Hetero TOCSY . Correlations Through Chemical Bonds: -Homonuclear ( COSY. although 2D NMR is not limited to these nuclei only. NOESY-HSQC. NOESY -HSQC) Correlations Through Space: -Homonuclear .ADVANCE METHODS FOR STRUCTURE ELUCIDATION OF SMALL MOLECULES 2D NMR Spectroscopy We will focus mainly in 1H and 13C nuclei at natural abundance. NOESY) -Heteronuclear (HSQC.

DQF-COSY Correlating coupled homonuclear spins. Typically used for correlating protons coupled over two or three bonds but may be used for any high-abundance nuclide. The basic COSY experiment.Through Chemical Bond 2D Homonuclear Chemical Shift Correlations : COSY-90 Correlating coupled homonuclear spins. Additional information on magnitudes of coupling disavtgtants may be extracted from 2D peak fine structure. 4/21/2012 8 . Typically used for correlating protons coupled over two or three bonds. Higher-resolution display than basic COSY. Singlets suppressed.

9 . Typically used for correlating protons coupled over two or three bonds but may be used for any high-abundance nuclide. Vicinal and geminal coupling relationships can be differentiated in some cases. Delayed COSY Correlating coupled homonuclear spins through small couplings. Often used to identify proton correlations over many bonds (>3). Reduced 2D peak structure over basic COSY. hence also known as longrange COSY.4/21/2012 Contd…. COSY-β Correlating coupled homonuclear spins.

10 . Powerful technique for analyzing complex proton spectra. INADEQUATE Correlating coupled homonuclear spins of low natural abundance.4/21/2012 Contd… TOCSY Correlating coupled homonuclear spins and those that reside within the same spin system but which may not share mutual couplings. Employs the propagation of magnetization along a continuous chain of spins. Typically used for correlating adjacent carbon centers at natural abundance but has extremely low sensitivity.

primarily 13C– 13C..4/21/2012 Contd. but employing 1H excitation and detection for sensitivity improvement. 11 . ADEQUATE Correlating coupled homonuclear spins of low natural abundance.

4/21/2012 THE COSY SPECTRUM OF m-dinitrobenzene 12 .

4/21/2012 THE COSY SPECTRUM OF 1-Chloro 2-Propanol 13 .

Nitro propane 14 .4/21/2012 COSY Spectrum of 2.

2) To determine each & every proton in beta sheet of the pancreatic trypsin. 15 .4/21/2012 Applications: 1) To determine structure in organic and biological samples.

HETCOR shows coupling between protons and the carbons to which they are attached.4/21/2012 HETCOR Hetronuclear Correlation Spectroscopy The 2D NMR Spectra that displays 13C-1H shift correlations are called HETCOR. 16 .

2 3.4-3.9 3.4/21/2012 THE HETCOR SPECTRA OF 1-Chloro 2-Propanol Chemical shifts δH δC 1.5 20 67 51 17 .

4/21/2012 HETCOR Spectrum of 2 Nitropropane No δ1H δ13 C 1 2 1.66 79 18 .56 21 4.

1 2 δ13 C 23 25 δH 0.92 1.4/21/2012 HETCOR SPECTRUM FOR ISO-PENTYL ACETATE NO.69 3 4 6 37 63 20.1 2.52 4.09 1.04 19 .

4/21/2012 INADEQUATE Incredible Natural Abundance Double Quantum Transfer Experiment Correlating coupled homonuclear spins of low natural abundance. Typically used for correlating adjacent carbon centers at natural abundance but has extremely low sensitivity. 20 .

4/21/2012 21 .

4/21/2012 22 .

4/21/2012 23 .

replaces the single mixing pulse of the basic COSY experiment. The spin-lock mixing time. a priori . so it does not suffer from cancellation of anti-phase peaks under conditions of low digital resolution or large linewidths. In these instances.TOtal COrrelation SpectroscopY – TOCSY (Former HOHAHA) 4/21/2012 .A second significant feature of TOCSY that contrasts with COSY is that it utilizes the net transfer of in-phase magnetization.Number of transfer steps associated with each crosspeak not known. 24 . m.Correlates all protons within the same spin system . The TOCSY sequence. this feature makes TOCSY the more sensitive of these two methods.

Sample: Chentobiose Solvent: CDCl3 Spectrometer: AVANCE 400 Probehead: Inverse Broadband with z-Gradients TOCSY 4/21/2012 B 1 A 1 A Anomeric H B Anomeric H B 1H NMR A 1H NMR 25 .

Employs detection of high-sensitivity nuclide. e. 19F. 31P (an „inverse technique‟). so it is better suited for crowded spectra but can be more sensitive to experimental imperfections. e. 31P (an „inverse technique‟). 1H. Provides improved resolution over HMQC.g. 19F. Employs detection of high-sensitivity nuclide.4/21/2012 Through Chemical Bond 2D Heteronuclear Chemical Shift Correlations : HMQC (Heteronuclear Multiple Quantum Correlation) Correlating coupled heteronuclear spins across a single bond and hence identifying directly connected nuclei. Experimentally robust sequence.g. most often 1H–13C. 1H. HSQC (Heteronuclear Single Quantum Correlation) Correlating coupled heteronuclear spins across a single bond and hence identifying directly connected nuclei. 26 . well suited to routine structural characterization.

Most valuable in correlating 1H–13C over two or three bonds. 1H.4/21/2012 HMBC (Heteronuclear Multiple Bonds Correlation) Correlating coupled spins across multiple bonds. 31P (an „inverse technique‟). HETCOR (Heteronuclear Correlation) Correlating coupled heteronuclear spins across a single bond. H–X–Y Triple-resonance methods for correlating protons and a heteratom (Y) whilst using a second heteroatom (X) to either relay the correlations or edit the correlation spectrum. typically 13C. e.g.nuclide. so may find use when this is critical. so has significantly lower sensitivity than inverse techniques. Essentially HMQC tuned for the detection of small couplings. Employs detection of the lower. Powerful tool for linking together structural fragments. otherwise superseded by above methods. Benefits from high resolution in 13C dimension. 27 . Employs detection of high-sensitivity nuclide. 19F.

Observes steady-state or equilibrium NOEs generated from the saturation of a target. for which NOEs are positive. 28 .4/21/2012 Through Space 2D Homonuclear Chemical Shift Correlations : NOE difference Establishing NOEs and hence spatial proximity between protons. but may fail for mid-sized molecules (zero NOE). NOESY(2D or 1D) Establishing NOEs and hence spatial proximity between protons. Suitable for „small‟ (Mr << 1000) and large molecules (Mr > 2000) for which NOEs are positive and negative respectively. Suitable only for „small‟ molecules (Mr << 1000). Observes transient NOEs generated from the inversion of a target. Estimates of internuclear separations can be obtained in favourable cases.

Suffers from low sensitivity but 1H detected variants can help. 1H–13C. NOEs are positive for all molecular sizes. but is prone to interference from other mechanisms so requires cautious interpretation. for example. Estimates of internuclear separations can be obtained in favourable cases. Suitable for any molecule but often essential for mid-sized molecules. 29 . Can provide useful stereochemical information when homonuclear NOEs are insufficient or inappropriate.4/21/2012 ROESY(2D or 1D) Establishing NOEs and hence spatial proximity between protons. HOESY Establishing heteronuclear NOEs and hence spatial proximity between different nuclides. Observes transient NOEs in the rotating-frame.

meaning separate resonances are observed for each exchanging species.4/21/2012 EXSY (2D) Qualitative mapping of exchange pathways in dynamic systems when exchange rates are slow on the NMR chemical shift timescale. Quantitative data on exchange kinetics can be obtained in favorable cases. 30 .

Silverstein.S.Gary Lampmann. Published by : „Books Cole‟ Pg no:.4/21/2012 REFERENCES: I.http://www. Stanley Crouch.wikipedia. George Kriz.intermnet. http://www. Webster . 2007. P.6th Edition . 406-409 III.http://en. Douglas Skoog. “Instrumental Analysis”.h tm VII.queensu. Robert M.es/inteRMNet/doc/chem843-5.250-275 IV.Pavia . 540-550 31 .org/wiki/Correlation_spectroscopy V.342-350. Published By :Cengage lerning Pg No:. “Introduction To Spectroscopy” 3rd edition.chem.551-594 II.ca/facilities/NMR/nmr/webcourse/cosy. Francis X.ua.Donald J. Published By :Willey Publications Pg No:. India Edidition.“Spectrometric Identification of Organic Compounds” . James Holler .Kalsi .PDF VI. “Spectroscopy of Organic Compounds” 6th edition Published By: New Age Publication Pg No:.

A. Q.C.4/21/2012 My Sincere thanks to…. Mohali) & ALL MY BATCHMATES/COLLIUGES 32 . M.NIPER. Mr.O..J .Kshirsagar Sir (H. Damle Madam (Vice-Principal) Mr. Department) Mr. S. Somnath G. Mrs. Abhijeet Ladake Student M.D. 3rd Sem.A. Kandekar Junior research fellow (Chemistry Department .pharm Q.

4/21/2012 THANK YOU!! 33 .

4/21/2012 Any Questions? 34 .