1. Flow Analysis Methodologies
Recent Advances Part 7
Dom Hebrault, Ph.D.
Principal Technology and
Application Consultant
May 16th 2012

2. Today’s Agenda
 Continuous Flow Chemistry - Analysis Challenges
 ReactIR™ In Situ IR Spectroscopy
 Case studies:
- A Visual, Efficient, Method to Optimize Reaction Conditions: Case study on a Doebner
Modification
- Safer Use and Monitoring of Hazardous Substances: A General, One-Step Synthesis
of Substituted Indazoles using a Flow Reactor and a FlowIR
- Troubleshooting and Improving Product Quality of a Grignard Batch Process in a 6-
Step Drug Synthesis

3. Continuous Chemistry - Analysis Challenges
Today: Limited availability of convenient,
specific, in-line monitoring techniques
 Chemical information
- Continuous reaction monitoring superior to traditional sampling for offline
analysis (TLC, LCMS, UV, etc.)
→ Stability of reactive intermediates
→ Rapid optimization procedures
 Technical knowledge
- Dispersion and diffusion: Side effects of continuous flow – must be
characterized

4. In-Line IR Monitoring
 Monitor Chemistry In Situ, Under Reaction Conditions
- Non-destructive
- Hazardous, air sensitive or unstable reaction species (ozonolysis, azides etc.)
- Extremes in temperature or pressure
- No interference from bubbles, solid, color,…
Attenuated Total Reflectance (ATR)
Spectroscopy

5. In-Line IR Monitoring
 Real-Time Analysis, “Movie” of the reaction
- Track instantaneous concentration changes (trends, endpoint, conversion)
- Minimize time delay in receiving analytical results

6. In-Line IR Monitoring
 Determine Reaction Kinetics, Mechanism and Pathway
- Monitor key species as a function of reaction parameters
- Track changes in structure and functional groups

7. In-Line FTIR Micro Flow Cell in the Laboratory
ReactIRTM Flow Cell: An Analytical Accessory
for Continuous Flow Chemical Processing
Internal volume: 10 & 50 ml
Up to 50 bar (725 psi)
-40 → 120 ºC
Wetted parts: HC276, Diamond/Silicon & Gold
Multiplexing
Spectral range 600-4000 cm-1
Carter, C. F.; Lange, H.; Ley, S. V.; Baxendale, I. R.; Goode, J. G.; Gaunt, N. L.; Wittkamp, B. Org. Res. Proc. Dev. 2010, 14, 393-404

8. FlowIR: Flow chemistry and beyond…
FlowIRTM: A New Plug-and-Play
Instrument for Flow Chemistry and
Beyond
9-bounce ATR sensor
(SiComp, DiComp) and head
Internal volume: 10 & 50 ml
Up to 50 bar (725 psi)
-40 → 120 ºC
Small size, no purge, no
Spectral range 600-4000 cm-1 alignment, no liquid N2

9. Rapid Analysis of Continuous Reaction Optimization
Optimization of a Doebner Modification of
Knoevenagel Reaction in a Continuous
Mode + CO2
 Introduction
Vapourtec R2+/R4
Can reaction optimization and conditions
screening be conducted inline?
How does dispersion affect fraction
collection? FlowIRTM
On-the-fly reaction optimization with
inline FTIR analytics
Vapourtec – Flow Chemistry Solutions – Mettler Toledo collaboration project, U.K. 2011, White Paper

10. Rapid Analysis of Continuous Reaction Optimization
 Results
100°C, 10’
Reference spectra of 4 main components
150°C, 10’ 120°C, 20’
3 main/unique bands
Cinnamic acid
(772cm-1) 4.5 h
Malonic
Acid
(1729cm-1) 80°C, 10’ 120°C, 10’ 100°C, 20’ 100°C, 30’
Benzaldehyde
(828cm-1)
7 reaction “plugs”, on-the-fly variation of
residence time and temperature (1:1.1
benzaldehyde/malonic acid ratio)
Few hours experiment only
Vapourtec – Flow Chemistry Solutions – Mettler Toledo collaboration project, U.K. 2011, White Paper

11. Rapid Analysis of Continuous Reaction Optimization
 Results
Development of an in-situ real time assay
method
- ReactIR algorithm: iC Quant and iC IR
- Simple univariate model (trans-
cinnamic acid 772 cm-1 with 2 baseline
points)
(0.1-0.5M)
Trans-cinnamic acid
(772 cm-1)
“Proof of concept” univariate model
Limited number of datapoints
Model used to predict concentration
Vapourtec – Flow Chemistry Solutions – Mettler Toledo collaboration project, U.K. 2011, White Paper

12. Rapid Analysis of Continuous Reaction Optimization
 Results 1:2
Development of an in-situ real time assay
method: 1:1.2
- Application to the previous screening
- 100°C, 20’ to 30’ represent an optimum
at (1:1.1 benzaldehyde / malonic acid
ratio)
[M]
120°C, 20’
3.5 h
100°C, 10’
0.35
150°C, 10’
1: 1.1 1:1.5 1:2
0.30 Steady state
0.25
0.20 Variation of benzaldehyde / malonic acid:
0.15
- From 1:1.1 to 1:2 (100°C, 20’)
0.10 - No significant improvement
- Real time FTIR provides confirmation of
80°C, 10’ 120°C, 10’ 100°C, 20’ 100°C, 30’
steady state and concentrations in the
plug
Vapourtec – Flow Chemistry Solutions – Mettler Toledo collaboration project, U.K. 2011, White Paper

13. Rapid Analysis of Continuous Reaction Optimization
 Conclusions
No issue with CO2 bubble
Faster, more efficient, optimization
Provides a picture of flow dispersion,
helps enhance separation and off-line
analysis
 Compared to former in-house solutions
Can be heated, cooled, pressurized
Wide spectral range and high sensitivity
Turn-key, affordable, space efficient
Vapourtec – Flow Chemistry Solutions – Mettler Toledo collaboration project, U.K. 2011, White Paper

14. Safer Use and Monitoring of Hazardous Substances
Vapourtec R2+/R4
A General, One-Step Synthesis of
Substituted Indazoles using a Flow
Reactor and a FlowIR
 Introduction FlowIRTM
Time-efficient and safe production of
small amounts of pharma-relevant
fragments
Reduce inventory of hydrazine under
“forced” conditions in flow mode Real time monitoring of concentrations
NH2
N
O2N CHO O2N
• indazole
+ H2N
NH2
• azine
F F
Hydrazine Hydrazone
• hydrazone
Faster optimization of conditions
O2N NO2 O2N
• reagent excess
• temperature
N N
+ N
N
• residence time
F F
Azine (minor) Indazole (major)
Rob C. Wheeler, Emma Baxter, Ian B. Campbell, and Simon J. F. Macdonald GlaxoSmithKline, Stevenage, U.K.; Organic Process Research and
Development, 2011, 15 (3), 565–569; Vapourtec – Flow Chemistry Solutions – Mettler Toledo collaboration project, U.K. 2011, White Paper

15. Safer Use and Monitoring of Hazardous Substances
 Results
Indazole
Screening (7 experiments in 2.5 h):
hydrazine excess, temperature, and
residence time 1:1.2, 150°C
15’
Intermediate
1:1, 150°C
15’
1:1.2, 100°C
15’
1:1, 100°C Azine
1:1, 50°C 15’
15’
1:1.2, 50°C
15’
1:1, 25°C No reaction at 25°C
15’
Hydrazone only at 50°C: 1st step is faster
No full conversion of hydrazone even at
150°C
0.2 eq. excess hydrazine: 4% more
indazole
2.5 h
Rob C. Wheeler, Emma Baxter, Ian B. Campbell, and Simon J. F. Macdonald GlaxoSmithKline, Stevenage, U.K.; Organic Process Research and
Development, 2011, 15 (3), 565–569; Vapourtec – Flow Chemistry Solutions – Mettler Toledo collaboration project, U.K. 2011, White Paper

16. Safer Use and Monitoring of Hazardous Substances
 Introduction
Is 150°C still too low?
Temperature more efficient than
increase of residence time
1:1.2, 200°C
15’
1:1.2, 150°C 1:1.2
30’ 200°C
1:1.2, 150°C 5’
5’
Integration of ReactIR software (iC IR)
with Flow CommanderTM software
Facilitates automated experiment
optimization
Allows accurate sampling of plugs for
fraction collection and analysis
Rob C. Wheeler, Emma Baxter, Ian B. Campbell, and Simon J. F. Macdonald GlaxoSmithKline, Stevenage, U.K.; Organic Process Research and
Development, 2011, 15 (3), 565–569; Vapourtec – Flow Chemistry Solutions – Mettler Toledo collaboration project, U.K. 2011, White Paper

17. Real Time Product Quality Control for Flow Processes
Troubleshooting and Improving Product O OH
Quality of a Grignard Batch Process in a AcOH
Ar
6-Step Drug Synthesis Aldol

O OMgBr O
MeMgBr
Introduction AcOH
Ar OEt Ar Ar
Ketone
Impurity headache PhMe/THF
2-Me-THF
OMgBr OH
MeMgBr
• <10% aldol during development study Ar
AcOH
Ar
• 40% during 1000 L campaign
Alcohol
Challenges and Objectives
• Flow process
• Real time process quality control(*)
• Proof of concept on 40 Kg scale
• Aldol ≤ 1%
• Conversion ≥ 97%
(*) Off-line analysis (HPLC) takes 20-40’
“Leaving the Tap Open…”, Fabrice Odille, AstraZeneca, Continuous Flow Technology in Industry, RSC, York, UK, March 19-21 2012

18. Real Time Product Quality Control for Flow Processes
Toluene
at 730cm-1
 Preliminary results in flow
Reference
spectra
• Eq. MeMgBr: 2 → 1.5 Ester carbonyl
2-Methyl-THF
at 1752cm-1 at 1383cm-1
• Eq. NEt3: 6 → 3.5 Ketone carbonyl
at 1721cm-1
• T°: -10 → 0°C
• Fast reaction < 20 s Product Grignard
Starting
reagent
material
Aldol: 40% → ≈ 1%
Reaction
spectra
Enolate
at 1252cm-1
No ester
starting material
No product
ketone!!
Alfa Laval ART® Plate Reactors
“Leaving the Tap Open…”, Fabrice Odille, AstraZeneca, Continuous Flow Technology in Industry, RSC, York, UK, March 19-21 2012

19. Real Time Product Quality Control for Flow Processes
[Ester]1752cm-1
10%
 Excellent system stability upon flow
rate changes 3%
1%
 Conversion measurement ≥ 97% with
Solvent, 2-Me-THF at 1383cm-1
2-Me-THF 2-Me-THF
2-Me-THF
14 mL/min 1 mL/min Stop flow
7 mL/min qualitative/quantitative peak height(*)
Starting material, ester at 1752cm-1  Conversion measurement ≥ 99%
requires quantitative model
(*) results within +10% versus IPC-HPLC
“Leaving the Tap Open…”, Fabrice Odille, AstraZeneca, Continuous Flow Technology in Industry, RSC, York, UK, March 19-21 2012

20. Real Time Product Quality Control for Flow Processes
Toluene, Grignard at 730 cm-1
Starting material, ester at 1752cm-1
Solvent, 2-Me-THF at 1383cm-1
Start Grignard Switch Increase Grignard Increase ester flow Increase ester, increase Grignard
reagent and off ester to 2 eq., switch rate, decrease increase to 1.1 eq.
ester pumps pump ester pump on Grignard to 0.8 eq. Grignard to 1 eq.
“Leaving the Tap Open…”, Fabrice Odille, AstraZeneca, Continuous Flow Technology in Industry, RSC, York, UK, March 19-21 2012

21. Real Time Product Quality Control for Flow Processes
 Scale-up validation - Lab
• 500 g ketone product
• 4-5 s residence time
• 25 mL/min
• 4-6 h
 Scale-up validation - KiloLab
• 30 kg ketone product
• Same residence time
• 72 mL/min
• 92 h
• Project timeline ≤ one week
“Leaving the Tap Open…”, Fabrice Odille, AstraZeneca, Continuous Flow Technology in Industry, RSC, York, UK, March 19-21 2012

22. Acknowledgements
 Vapourtec Ltd. (U.K.)
- Chris Butters, Duncan Guthrie
 Flow Chemistry Solutions (U.K.)
- Andrew Mansfield
 AstraZeneca, Sodertalje (Sweden)
- Fabrice Odille, Mats Ridemark, Daniel Fahlen
 Mettler Toledo Autochem
- Will Kowalchyk (USA), Jon Goode (U.K.)