1

Pfizer Confidential 1

From FID to Structure in 30 minutes?

Anna Codina

ARD, Pfizer Global Research and Development

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INTRODUCTION

Speed

3

Increase Productivity

FAST

FASTER

Maintaining Quality

,FASTEROH, NO!

4

0101

1.41.31.21.11.00.90.80.70.60.50.40.30.20.10

Time (sec)

Computer Assisted Structure

Elucidation (CASE)

Smart

Formula

3D

NMR

WorkbookStructure

ElucidatorN

NH

O

S

O

NO

O

N

N

N

5

Smart Formula 3D

Smart

Formula

3D

High sensitivity, accuracy, isotopic fidelity and resolution

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Smart Formula 3D

Smart

Formula

3D

MS MSMS

Formula Fragments

www.bdal.de/uploads/media/LCGC_March_2008_Appl._Books.pdf

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0101

1.41.31.21.11.00.90.80.70.60.50.40.30.20.10

Time (sec)

Elucidator

Smart

Formula

3D

NMR

WorkbookStructure

ElucidatorN

NH

O

S

O

NO

O

N

N

N

8

Elucidator Workflow

NMR: pp 2D MS: MF

Generate MCD

Generate Structures

From MCD/s

Check MCD

Find the Best

Structure !

Check data/

add information

Data base

9

0101

1.41.31.21.11.00.90.80.70.60.50.40.30.20.10

Time (sec)

Before NMR Workbook

Smart

Formula

3D

Structure

ElucidatorN

NH

O

S

O

NO

O

N

N

N

CASE limiting Step

10

0101

1.41.31.21.11.00.90.80.70.60.50.40.30.20.10

Time (sec)

After NMR Workbook

Smart

Formula

3D

NMR

WorkbookStructure

ElucidatorN

NH

O

S

O

NO

O

N

N

N

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INTRODUCTIONANTIPYRIN

Proof of concept

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NMR Workbook

3 Finish the

peak picking in the

HSQC

1 Create new

project and data

folder

2 Short cut mode: H peak picking

and multiplet analysis

4 Save the project

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Elucidator – Data Grooming

• Molecular formula added

•HSQC parameters: multiplicity edited CH2 positives

•HMBC calibration: mainly 3 bond correlations

•Number of Carbons – Symmetry / number of protons

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MCD check and Elucidation

•Allow bonds between heteroatoms

•No fuzzy generation

•Filter by carbon assignment

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Generate Structure From MCD

8/27636

Database: Generated Moleculesd_{N}(^{13}C) (ListA) (8 pts)d_{N}(^{1}H) (ListA) (8 pts)

8642# in ListA

2

4

6

8

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Time?

• Processing: 1-2 min

• Data grooming: 12 min limiting step!

• Structure Calculation: 8/27636 in 8m 36s

• Total time: 22 min

• Added additional data: 13C, CN_HMBC and COSY ~ 3 h(Calculation time: 2 s)

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INTRODUCTIONFLUCONAZOLE

Increasing Difficulty

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Fluconazole

13C acquired and used to count carbons,

taking into account F-C coupling

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The Clock is Ticking

12/2550 8h and still calculating structures!

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Calculation Stopped!

Data Grooming: 3 h

Structure calculation > 8h

11 h !

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Structure Search

Structure search based on chemical

shifts before starting the elucidation

Is the first step to success!

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Fluconazole

<< 30 min

No need for data grooming

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INTRODUCTIONSILDENAFIL

Increasing Difficulty

Decreasing Amount of Material (100 µg)

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Elucidator Workflow

NMR: pp 2D MS: MF

Generate MCD

Generate Structures

From MCD/s

Check MCD

Find the Best

Structure !

Check data/

add information

Data base

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Training

NMR: pp 2D MS: MF

MCD from fragments

Generate Structures

From MCD/s

Check MCD

Find the Best

Structure !

Check data/

add information

Data base

Data baseData base

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Dunn, P.J et al; Green Chem., 2004, 6, 43-48

C148.06

NH

N

C153.61

C137.73

C124.36

C144.86

N

N

O

CH337.77(ob)

CH227.06(fb)

CH221.58(fb) CH3

13.74(fb)

CH314.18(fb)

CH345.19(ob)

CH245.65

CH245.65

CH253.43

CH253.43

CH264.81(ob)CH

113.14

C123.66

C126.25

CH129.82

CH131.29 C

159.79

N N

S(v = 6)

O

O

O

1. Create Fragments

2. Create MCD from Fragment

3. Calculate Structures

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Incorrect

structure –

substitution

pattern wrong

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Elucidator v.12 Results

CH337.77

CH313.74

CH314.18

CH345.19

27.0621.58

45.6545.65

53.4353.43

64.81

113.14

129.82131.29

153.61

137.73124.36

144.86

148.06

123.66

126.25

159.79 NH

N

N

N

N

N

O

O

OO

S

CH337.77

CH313.74

CH314.18

CH345.19

27.06

21.58

45.65

45.65

53.43

53.43

64.81

113.14

129.82

131.29

153.61

137.73124.36

144.86

148.06

123.66

126.25

159.79

N

H

N

N

N

N

NO

O

O

O

S

Database: Generated Moleculesd_{N}(^{13}C) (ListA) (30 pts) d_{N}(^{1}H) (ListA) (30 pts)

252015105# in ListA

1

2

3

4

5

6

BEST STRUCTURE

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Sildenafil5 min

N

NH

O

S

O

NO

O

N

N

N

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Conclusions

What has changed?

» Synchronisation of spectra

» Good automatic peak picking

» Easier and faster structure elucidator module

» Enhanced NMR and MS instruments

Fast CASE with small amounts of material is now reality

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Acknowledgments

• Don Richards and the SE team – Pfizer

• Steve Coombes – AZ

• Ryan Sasaki – ACD

• Rainer Kuemmerle, Detlef Moskau and Helena Kovacs –Bruker Biospin

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