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The world leader in serving science

Dr. J. Mark Dixon

NMR Product Marketing Manager

Thermo Fisher Scientific

Advantages of the picoSpin 80 NMR Spectrometer in Pharmaceutical Research & Development

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Agenda

• Introducing the Thermo Scientific™ picoSpin™

80 NMR Spectrometer

• NMR analysis in a research laboratory

• Practical challenges facing the chemist in order to obtain

chemical information using NMR

• Solutions to those challenges using the picoSpin 80 NMR

Spectrometer

• Structure Verification/Confirmation

• Quantification/Mixture Analysis

• Reaction Monitoring

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Our Family Has Grown

• picoSpin 80 NMR • 82 MHz Larmor Frequency

• Replaceable Capillary Cartridge

• Small Sample Volume

• Compact & Portable

• Remote Operation

• Web Browser Interface

• picoSpin 45 NMR • 45 MHz Larmor Frequency

• Replaceable Capillary Cartridge

• Small Sample Volume

• Compact & Portable

• Remote Operation

• Web Browser Interface

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Minimal Maintenance

Ease of Use

Larger Field Strength

Greater Shift Dispersion

Higher Sensitivity

Easier Access to

Richer Information

The picoSpin 80 Value Proposition

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Larmor Frequency: 82 MHz

1H or 19F NMR of Liquids

Compact & Portable: 19 kg (43 lbs)

Total Weight

Chemical Shift Resolution: 18 ppb

Replaceable Capillary Cartridge

40 µL Sample Volume

Controlled by web browser

No Cryogens, Compressed Gas, etc.

Standard Lab Environment

All Solid-State Technology, No Moving Parts

picoSpin 80 Spectrometer Features

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Why Now?

• Nd2Fe14B magnets

• Microcoil NMR experiments by A.G. Webb et al., U. of Ill.

• Advances in Rf (Tx and Rx) Electronics… (Mobile Phones)

Magnetic Materials Micro Coil Technology Radiofrequency Electronics

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1H NMR Analysis In A Research Lab

• Go-to technique for structure confirmation • Often used daily for rapid decision making

• Used extensively for structure elucidation • Powerful tool when used in tandem with other techniques, e.g. LC/MS

• Ideal method to study mixtures & kinetics of reactions • Reaction monitoring in real time can unveil mechanistic behavior

• Excels in determination of stereo- & regio-chemistry • Look “inside” the molecule at topological information

• Good predictive tools available • Educationally useful, or as a helping hand

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Typical 1H Spectrum @ 82MHz

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Practical Challenges Using High-Field NMR

• High-field instrument - good ; Remote location - bad • Could be in another building, or even on another site

• Central facilities are expensive, and popular • That combination usually leads to an over-subscribed service

• Calendar-based operation is inefficient when busy

• Long wait times impact productivity • Robotic systems help but queues grow quickly at peak times

• Even one hour can mean a half-day lost due to lab safety restrictions

• Real-time experiments need exclusive access • Magnet time is precious, yet shared instrument sits idle waiting for the

next sample

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Solutions Using The picoSpin 80

• 82 MHz offers rich chemical information content • Disproportionately so compared to drop in field strength from 300+MHz

• And also when compared to lower magnetic field strengths

• Proximity of instrument = precious time re-gained • No trekking between buildings in the rain, use time productively

• One instrument per 10 chemists is better than one per 100+ • Shorter wait times, even when longer acquisition time taken into account

• Lightweight : can be hand-carried around the lab or between labs

• Reaction monitoring made simple • Syringe aliquot from reactor, inject into pS80, press ‘Run’ – No delay

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Ibuprofen 1H Spectrum @ 82MHz and @ 42MHz

82MHz

42MHz

Note the

complete

resolution of

H-14

1,2,3,5 7

14

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13,15

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TMS

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Structure Verification/Confirmation

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Did I Make What I Think I Made?

• Most chemical reactions affect few functional groups • Can manifest as obvious changes to 1H spectrum

• Looking for identifying marker(s) as proof of synthesis

• Simple example : hydrolysis of isobutene • Markovnikov’s Rule applies

OR H2O

[H2SO4]

0ºC

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1H Spectrum of Starting Material : Isobutene

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1H Spectrum of Favored Product : t-Butanol

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1H Spectrum of Disfavored Product : 2-Methylpropanol

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Easily Distinguish Products From Starting Material

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Quantification/Mixture Analysis

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• NMR signal area is proportional to concentration

• Relative Quantification • Relative proportion of compounds in mixture

• Normalize signal areas – determine mass ratios (%w/w)

• Easy and free with NMR

• Absolute Quantification • Accurate determination of the purity levels of active pharmaceutical

ingredients (APIs), and drug analysis

• Pharmaceutical compound libraries

• Forensic analysis

• Food sciences

Quantitative NMR (aka qNMR)

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Quantification: Simple Example

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Quantification: Simple Example

3 H’s

1 area “unit”

2 H’s

3 area “units”

200mM

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Quantification: Easily Calculate Relative & Absolute

Lidocaine :

0.33 units per ‘H’

CH2Cl2 :

1.53 units per ‘H’

Relatively :

4.6 to 1 mixture

or

82.2% CH2Cl2

17.8% Lidocaine

Lidocaine : 200mM

Absolutely :

4.6 to 1 mixture

920mM CH2Cl2

200mM

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Reaction Monitoring

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Example Experimental Procedure

Acetic acid 3-Methylbutanol HCl

250 µL 200 µL 1 drop

5.3 mmol 2.3 mmol Conc.

• Weigh and mix starting materials

• Add acid, start timer

• Inject 40 µL aliquot into picoSpin 80 capillary cartridge

• Start NMR data acquisition

• Set delay and scans for multi-hour acquisition

• Save individual spectra

• Process data off-line

-H2O

H+

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Predicted Spectrum Of All Components Combined

Starting Materials Product

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Track Highlighted Signals To Monitor The Chemistry

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Stacked Spectral Display Of All Data

time axis

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Stacked Spectral Display Of All Data

time axis

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A Snapshot Every So Often Tells The Story

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In Summary

Today, we have

• Explored the advantages of the picoSpin 80

NMR spectrometer in a Research Laboratory

• Explored the richness of 80 MHz NMR using

• Structure Verification

• Quantitative NMR

• Reaction Monitoring