IPPT NMR Talk slides - Introduction of NMR Spectroscopy and Its Applications

7/27/2019 IPPT NMR Talk slides - Introduction of NMR Spectroscopy and Its Applications

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Innovation with Integrity

Introduction of NMR Spectroscopy

and Its ApplicationsAdvanced Medical and Dental Institute (IPPT)

Bruker Malaysia, 10-9-2013


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What is NMR represents?

Nuclear Magnetic ResonanceSpectroscopy


What does NMR detects?

Nucleus !!!

NMR detects


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Nuclear Magnetic Resonance (NMR)

Spectroscopy:Any nuclues ??

NMR Spectroscopy


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NMR Spectroscopy

Nuclei with non zero spin

NMR active nuclei !!!


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OperatorConsoleSuperconductingMagnetwithProbe

HostComputer

NMR Spectrometer


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N

S

N

S

N

S

N

S

N

S

N

S

N

S

N

S

N

S

N

S

N

S

N

S

N

S N

S

N S

NMR Spectroscopy (for I= nuclei)


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NMR Spectroscopy (1H Nuclei)

Higher energy state

Lower energy state

aligned against/

opposed to the field

aligned with thefield

External magnetic field, Bo

In 14.10 TSuperconducting magnet

The proton nucleiprecess at Larmorfrequencies (veryclose to 600 MHz)

Resonance frequency, = B0


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Resonance frequency, =

B0


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At 14.10 Tesla

p

pp

pp

p

pp

p

p

1H nuclei


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Resonance frequency, =

B0


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At 14.10 Tesla

p

pp

pp

p

pp

p

p

600.13 MHz

600.13 MHz

600.13 MHz

600.13 MHz

600.13 MHz

600.13 MHz

600.13 MHz


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p

pp

p

p

p

pp

pp

1H nuclei

pp

p

pp

p

pp

p

ppp

1H nucleiAny different?

Any different?

1H nuclei


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At 14.10 Tesla


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External magnetic field, Bo

Higher energy state

Lower energy state

aligned against/opposed to the field

aligned with thefield

NMR Spectroscopy (1H Nuclei)

Radio frequency pulse



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What type of energy source does the

spectrometer used?


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In 14.10 T Superconducting magnet(600.130 MHz)

NMR Spectroscopy

Radio frequency pulse Radio frequency pulse


Environment 1 Environment 2 Environment 3

600.1320 MHz 600.1440 MHz 600.1562 MHz

Larmor frequency 1

Larmor frequency 2

Larmor frequency 3


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To excite a given nucleus

Carrier frequency (SFO1)

A pulse excites all the magneticnuclei in the molecule simultaneously.

time

RF

tp 90


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Mixture of FIDs

Free Induction Decay (FID)


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Hz 02000300035004000 2500

At 14.10 Tesla

Mixture of FIDs

Fourier transformation

Fourier transformationFourier transformation

Fourier transformation


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Time domain FID


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Frequency domain Spectrum


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1D NMR Spectrum

Chemical shift,

Chemical shift, is measured along thebottom of the spectrum on delta scale (inunits of parts per million, ppm).

It is the position in an NMR spectrum.


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Chemical shifts differences


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1D and 2D Electrophoresis


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How to distinguish between the

1D and 2D NMR Spectra?


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(3.58,79.9)

(4.78,107.3)(5.01,107.3)

(2.08,56.7)

Chemical shifts = Coordinates

2D NMR Spectrum


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ppm 4.344.354.374.38 4.36

H 4.35H 4.37

H 4.35H 4.36

(4.37,54.1)(4.36,54.4)

(4.35,50.8)

(4.35,60.2)

2D NMR Spectrum H and C

3D T i l i t


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F3 (ppm)

F1 (ppm)

F2 (ppm)

H (0 - 14)

N (0 - 40)

C (0 - 220)

N(c)

N(b)

N(a)

H

C

3D Triple resonance experiment on15N,13C labeled of proteins


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3D NMR Spectrum (H, N,C)


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and Its ApplicationsAdvanced Medical and Dental Institute (IPPT)

Bruker Malaysia, 10-9-2013


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Any kind of sample?

Pure substance or mixture ?

NMR Samples


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NMR Samples

Single substance ???


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NMR spectrum of Single Substance

Simple Molecule


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Complex molecule 1


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Complex molecule 2


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Complex molecule 3


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NMR Samples

Mixture ???


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NMR spectrum of a Mixture

Plant Extract 1

Plant Extract 2


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Trends in Biotechnology

Plant Extract 2


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NMR spectrum of a Mixture

Human Urine


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Why NMR technique is important?

NMR Applications Overview

What are the latest NMRapplications?



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NMR Application Overview


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NMR Applications Overview


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pp

Chromatographicseparations andrecrystallization

C ti l S t i M th d


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IR spectroscopy

High resolution MS

NMR spectroscopy

Conventional Spectroscopic Methods

O O

OH

H

H

HO

OHOH

OH

H

O

O1

2

34

56

7

8

9

10

1112

13

14

15

16

18

19

20

21

NMR d M l l I f ti


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NMR and Molecular Information


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O O

OH

H

H

HO

OH

OH

OH

H

O

O1

2

34

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7

8

9

10

1112

13

14

15

16

18

19

20

21

NMR A li ti O i


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NMR and Metabonomics


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NATURE



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Progress into the clinic- Future directions


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metabonomics is used to probe the real-world nature ofbiochemical functionality and is sensitive to both gene andenvironmental influences; it is, therefore, likely to be morepractical than gene-based measurements of responses totherapy.

we affirm that metabonomics has the power to influenceclinical decision-making in the hospital environment forboth medical and surgical treatments.

NATURE 491 (2012) 384392




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NMR-based metabonomics of biofluids offers great promise for the

minimally-invasive screening of cancer related perturbations, at several

stages of disease management, ranging from early detection anddifferentiation from benign conditions to identification of metastatic

disease and surveillance of recurrence.

metabolic profiling of biofluids through NMR spectroscopy will, intime, provide a unique and important tool in the clinical management of

cancer, and have major impacts in areas such as screening, diagnosis

and monitoring of disease progression.

metabonomics studies have great potential in the emerging area ofpersonalized cancer therapy, namely through the identification of

metabolic markers of treatment efficacy or early toxicity, thus assisting

in the development of novel, targeted therapeutics

Progress in Nuclear Magnetic Resonance Spectroscopy 62 (2012) 5174

Progress into the clinic Future directions



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NATURE 491 (2012) 384392




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NATURE 491 (2012) 384392


How Do You Do Metabolomics?


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RT Shim Coils

RT Shim Canon

TXI

Spinner

Sample

RF Coils

Probe


Biofluids NMR


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Human Urine


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Journal of Proteome Research

Biofluids NMR


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Biofluids NMR

Statistical Analysis


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y

PLS OSC PLS

NMR and Kidney toxicity detection


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y y

NATURE REVIEWS | drug discovery

NMR and Metabolic Signatures of Cancer


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Progress in Nuclear Magnetic Resonance Spectroscopy

Ju

ly

2011



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pp

NMR and Food Analysis


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y




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NMR and Drug Discovery


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NATURE REVIEWS | drug discovery

NMR and Drug Discovery-Screening (STD NMR)


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Screening (STD NMR)



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In cell NMR and Protein-proteinInteraction


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Interaction

NATURE Methods



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Nobel Prize in Chemistry 2002


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In 2002, The Royal Swedish Academy of Sciences has decided to

award the Nobel Prize in Chemistry to

Prof Kurt Wthrich"for his development ofnuclear magnetic resonance spectroscopy for

determining the three-dimensional structure ofbiological

macromolecules in solution".

NMR and Protein


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The first protein structure determined by NMR.


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Innovation with IntegrityJ. Mol. Biol.

Protein structure determination categorizedby experimental method


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81638

9955

626 60188

Protein Structure Deposited in Protein Data

Bank as ofJuly 2013

X-ray

NMR

Electron Microscopy

Solid state NMR

Other

10.8 %

88.3 %

y p

The growth in the number of NMR-deducedprotein structure


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0

500

1000

1500

2000

2500

3000

3500

4000

Before 2000 2000-2005 2005-2010 2010-Today

Protein Structure Deposited at Protein Data BankBased on Solution NMR as ofJuly 2013

p

Common labeling strategy for eachclass size of protein


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Innovation with Integrity Journal of Magnetic Resonance 173 (2005) 193207

82 kDa

p

50 residues ~5.5 kDa

80 residues ~9 kDa

180 residues ~20 kDa

NMR techniques and isotope labeling


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Innovation with Integrity J.J. Chou and R. Sounier, Solution Nuclear Magnetic Resonance Spectroscopy, 495-516

15N-edited NOESY, 15N-edited COSYTriple-resonance multidimensional NMR

Small protein ubiquitin (MW= 8.6 kD)


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1H spectrum of ubiquitin


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NMR and Protein-RNA complexes


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Innovation with Integrity Progress in Nuclear Magnetic Resonance Spectroscopy

NMR and Protein-RNA complexes


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Innovation with IntegrityProgress in Nuclear Magnetic Resonance Spectroscopy

Sensitivity of NMR


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Compared with other analytical techniques, NMR issignificantly insensitive in terms of the minimumsample amounts for an analysis.

The limit of detection (LOD) ofNMR 109 - 1011 molLaser-induced fluorescence 1013 mol,

FT IR and Raman spectroscopy 1012

- 1015

molMass spectrometry 1019 mol


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IPPT NMR Talk slides - Introduction of NMR Spectroscopy and Its Applications