CPAC SATELLITE WORKSHOPS 2009
Micro-Reactors andMicro-Analytical
DR BRUNO LENAINKaiser Optical System
RAMAN SPECTROSCOPYA VALUABLE TOOL FOR
PAT APPLICATIONS
History of Raman Spectroscopy
• 1928 C.V. Raman and Krishnan discover “a new type of secondary radiation” - Raman effect.
• Only 1 in 1,000,000 (0.0001%) photons are scattered inelastically.
• In the 1930s, Raman became the principal means of non-destructive chemical analysis.
• 1939-1945 PE develop the first commercial IR.
• After WWII, commercial IR surpassed Raman in this role.
• Advent of lasers in the 1960s revived interest in Raman to some extent. – First Raman renaissance – lasers, photon counting, double monochromators.
His
tor
y of
Ra
ma
n
• 1986: FT-Raman instrument demonstrated.• Late 1980s: notch filters improved the quality of Raman
spectra.• 1990s: dispersive Raman developments including compact
NIR lasers, multi-channel detectors, refinement of Raman microscope, and fiber-optic probes.
• Early 1990’s – Second Raman renaissance – compact integrated dispersive Raman systems and analytical FT-Raman
• Early 1990’s – in-line 24/7 process Raman becomes a reality
History of Raman Spectroscopy
His
tor
y of
Ra
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n
Who is Kaiser?Formed in 1979, as a division of Kaiser Aerospace and Electronics.Headquarters in Ann Arbor, MI, USABecame a Division of Rockwell Collins, Inc in 2000.Our historical Expertise is in the area of holographic optical technology.Since 1990, Kaiser has been a major supplier to the Raman community.The Spectroscopic Products represent the largest single division of Kaiser.We are Global - having offices or representation in North America, Europe, and Asia.
Kaiser Technology Introductions1991 – Holographic Notch Filter1992 – Holographic Laser Bandpass Filter1993 – HoloSpec f/1.8i Imaging Spectrograph1994 – HoloProbe Process Raman Analyzer & HoloPlex Volume Phase Transmission Grating1995 – Universal Fiber Optic Probehead1996 – Confocal Raman Microscope1997 – Internal Calibration Option for Raman Products1997 – Immersion Probe Optics1998 – Raman Calibration Module and Calibration Transfer1999 – RamanRxn Systems Analyzers / HL5000R Modular Systems 2000 – Invictus NIR Laser2000 – HoloReact and HoloMap Software2001 – MR Probe / Pilot Probe Line of Process Probes2002 – HTS System for Wellplate Analysis2003 – MRA & RamanRxn2T analyzers, Pilot-E Probe Line2004 – RamanRxn3L Laboratory Analyzer and Invictus 532 nm Laser2005 – RamanRxn3 ATEX Certified Process Analyzer 2006 - PhAT System for Solid-State Analysis
A HISTORY OF MEETING THE MARKET’S NEEDS & CHALLENGES!
Raman Scatteringfrom Molecular Vibrations
ν 2πc1= (k
μ)½ ν = Vibrational frequencyk = Spring force constantμ = Reduced mass of atoms, m1m2/(m1+m2)
Higher vibrational frequency with stronger chemical bond and lighter atoms
Rayleigh
Anti-Stokes
Stokes
POLARIZABILITY
Raman spectroscopy provides information on the chemical make-up of molecules by observing the vibrational energies of the molecules.
Raman is complementary to mid-IR BUT different intensities and selectivity.
What is Raman?%
Tran
smitt
ance
Ram
an In
tens
ity
IR Transmission Spectrum
20
40
60
80
Raman Spectrum
1
2
3
4
1000 2000 3000 4000 Wavenumbers (cm-1)
Comparison to FT-IRInfrared
Absorption Dipoles
O-H, N-H, C=O
Sample preparation Non-aqueous samples
RamanEmissionPolarizability
C=C, Aromatics
Hardly any sample preparationAqueous samples,
Comparison to NIRNIR
Absorption OvertonesNo sample preparation necessaryProcess measurementsUnresolved information
RamanEmissionFundamental informationNo sample preparationProcess measurementsHigh spectral density
Iλ=σLCI
Iλ = Raman intensityσ = Raman cross sectionL = PathlengthC = ConcentrationI = Instrument parameters
Analytical Raman Spectroscopy
Sample
Why Raman?• Composition and Structural Information with Fiber
Optic Sampling • Raman is a specific and selective technique providing
well resolved information leading to…BETTER PROCESS UNDERSTANDING
• Flexible sampling (remote sampling)• In-situ – Eliminate Grab samples, Sampling through
containers• Measurement of various types of samples (liquids,
slurries, pastes, solids, powders, etc.)• Ease of use – no longer requires an expert• Robust, stable, low-maintenance instruments
Why Raman ? – Chemical Specificity
90 mg TabletAvicel Powder (Excipient)Acetaminophen Powder (API)
Majority of signal from Crystalline API
Majority of signal from Excipient
Raman Bands Are Sharp
NIR Bands Are Broad
Comparing NIR and Raman
anhydrous
monohydrate
Raman: Clearly identifiable bands are observed for both the monohydrate and anhydrous forms.
NIR: Spectrum dominated by free water! The free water limits quantification of the forms during the process induced transformation (PIT).
PAT Process Induced Transformations (PIT)
RamanRxnApplications
Phosphorus Trichloride
Microwave Reactions
Polymer Opportunities
Heterogeneous Polymer Production
Polymer Microstructure
Polymerization Reactions
Polymer Films & Fiber Monitoring
Titanium Dioxide Manufacture
Chlorosilane Intermediates
Catalysis
Carbon Applications
Semiconductor Applications
Aqueous Solution Analysis
Crystallization Polymorphic Forms
Catalytic Hydrogenation
Grignard Reaction
RamanRxn1 Microprobe Applications
Polymer Science
HTS – Wellplate Analysis
Failure Analysis & Troubleshooting
Biomedical and Biochemistry
Forensic Science
Geology & Geochemistry
Pharmaceuticals
Carbon Applications
Semiconductor Applications
Inorganic Dissolution
Chemical Imaging
SOME OF THE MAIN INNOVATIONS
• LASER TECHNOLOGY• DETECTOR TECHNOLOGY (multi-channels)• OPTICAL DEVICES: Transmission gratings• OPTICAL FIBERS COUPLING• SAMPLING PROBES• COMPUTERS: data manipulations
NIR ExcitationEliminates Most Fluorescence
• Before 1986, most Raman used visible excitation
• Today, the majority of Raman analyzers for pharma applications use 785-nm excitation with dispersive spectrographs.
500 1000 1500 2000 2500 3000
Vitamin B12
532 nm Excitation
785 nm Excitation
Instrument Sensitivity0.1% Acetone in Water, 4 Replicates Overlaid, 785 nm Laser
700 740 780 820 860 900 Raman Shift, cm-1
Inte
nsity
S/N:Peak to baseline = 171Height reproducibility = 164
Laboratory Immersion Sampling
Fermentation
Slurry
Basic of Immersion Sampling
Working Distance
DepthOf Field
Lens WindowRaman back to collection
optics
Excitation Laser in
Common Immersion Sampling Considerations
SolutionOptically Move Working Distance and Depth of Field
Particulates / BubblesReduces ThroughputDecreases Signal/Noise
Sample surface
Conjugated PinholeDiaphragm
Beam combiner
Detector
Below surface Z plane
Above surface Z plane
Excitation laser beam
Raman Microprobe:Confocal Option
Confocal Raman Option
Use an aperture to define an area in space where signal is collected from.
AdvantagesAllows depth profilingFacilitates analysis of fluorescent
species
Disadvantages / ProblemsRelies on small RI change between
air and sampleDrastically reduces total signal for
transparent samplesRequires high alignment tolerances
(historically in direct coupled systems)
Process Development Production
Product Development
Discovery
Raman AnalyzerReaction Analysis
Raman AnalyzerReaction Monitoring
RamanAnalyzer
Process Control
RamanMicroscope
Chemical Imaging
Discovery to Production
SOME APPLICATIONSOF RAMAN
Pharmaceutical Industry
• API Development: Discovery, characterization, synthesis, crystalisation
• API Primary Production: Syntesis on pilot reactors
• Pharmaceutical Development: Blending, Tableting, Coating
• Pharmacetical Secondary Production:
RamanRxn HTS SystemHigh Throughput Screening
Raman well-plates and micro-reactors analysis allows rapid screening ( polymorphs, drug discovery candidates, catalysts etc)
Multimode excitation and low magnification objectives minimize laser damage to sensitive samples
Data analysis allows for fast sample identification.
HTS wells distribution
HTS – Raman of Solvates (AN 311)
950 900 850 800 750 700 650
Raman Shift (cm–1)
Inte
nsity
Anhydrate
Isopropanol Solvate
Acetone Solvate
Ethanol Solvate
Methanol Solvate
In Situ Polymorphic transformation
Polymorphs may have Different Properties- i.e. Solubility, Dissolution Rate, Stability, or Bioavailability.Raman is able to Discriminate between Polymorphs because Different Crystal Forms Provide Intensity and Frequency Changes in the Raman Spectrum.The Raman Technique can be Applied without Sample Preparation and Allows for Non-Destructive and In-SituMeasurements.
Raman Is Perfect for In-Situ Optimization and Process Monitoring of Pharmaceutical Actives
Sampling: Non-contact or Immersion probesfiber optical cable (2~200 m)
Raman: On-Line Instrumentation
Characterization of Progesterone
Form II
Form I
100 110 120 130 140Temperature (°C)
Endotherm
Form I
Form II(121.2° C)
(129.1° C)
(5° C/ Min.)
XRD patterns DSC curvesCrystal Forms I and II
Raman Spectra of Progesterone
Raman Shift (cm-1)
1600 1650 1700
0 1000 2000 3000
Form II
Form I
Δ 5 cm-1
For this Study the C=O Stretching Vibration was used to Quantitate Form I and Form II Polymorphs. Form I @ 1662 cm-1. Form II @ 1667 cm-1.
Crystal Forms I and II
Polymorphic Transformation at 45° C
Raman Shift (cm –1)16801660
Form I @1662
Form II @1667
1670
15
35
55
75
95
0 20 40 60 80Transformation time (min)
Con
c. o
f For
m
Form I
Form II(45° C)
Crystallizations were monitored over the temperature range from 5 to 45° C .Slurry: 2 grams Progesterone (25ml Organic Sol.) added to 500ml H2O .Temperature control and stirring were provided by a LabMax automated lab reactor.Polymorph concentration was determined from the C=O stretch band center position.Raman measurements were made in-situ with the RamanRxn1.
10
30
50
70
90
1661 1663 1665 1667
C=O Peak Position (cm-1)
Form
I C
onc.
(wt%
)
Calibration:% Progesterone in Slurry
Catalytic Hydrogenation Reaction with
In Situ RamanRxnArne Zillian, Solvias (Novartis)
RamanRxn: Application Note
Catalytic Hydrogenation Reaction
Intermediate (hydroxylamine) is a potential thermal safety hazardPreferred pathway excludes the intermediate species
ReactantIntermediateProduct
Catalytic Hydrogenation Reaction
ReactantIntermediateProduct
Catalytic Hydrogenation Reaction
Summary:Raman provides a clear understanding of nitro-compound hydrogenation to primary amino-compounds.
Raman spectroscopy was used to examine the mechanistic and kinetic properties of the reaction.
In-situ measurements were possible even in the presence of heterogeneous catalyst.
Solvent subtraction was unnecessary using the RamanRxn1 Analyzer.
Catalytic Hydrogenation Reaction
Monitoring Monomer Distillation Production
Elmer Lipp and Ronda Gross, Dow Corning
RamanRxn: Application Note
Chlorosilane Production Chemistry
Si (s) + MeCl (g) Me2SiCl2 , MeSiCl3 , Me3SiCl
Me4Si , SiCl4
MeHSiCl2 , Me2HSiCl , HSiCl3
Cat, Heat
Me2SiCl2
Hydrolyze[ -Si(Me2)O- ]
Polydimethylsiloxane
Desired Plant Benefits?
Faster Response Time
An in-situ Measurement•Ease-of-Sampling, Remote, Fibers
Reduce Material Handling
Reduce Cost-of-Ownership•Maintenance, Materials
Analytical Requirements
Measure Chlorosilane Mixtures Ranging from >95% to <5%Precision of Current Method (~0.1%)Analysis Cycle Time < 5 minutesContinuous, Unattended Operation on Multiple Sampling PointsEasy to Implement and MaintainPlant Distributed Control System (DCS) Interface via ModBus
Chlorosilane Reference Materials
100 200 300 400 500 600 700 800 900
Raman Shift, cm-1
Inte
nsity
Me2SiCl2
MeSiCl3
Chlorosilane Reference Materials
0 500 1000 1500 2000
Raman Shift, cm-1
Inte
nsity
Me3SiCl
MeHSiCl2
*
*
Sampling Options
Pilot Plant Trial Production Installation
Immersion Probe Non-ContactOptic
Stream Stream
Mk II ProbeHead
System Control Diagram
Monitoror
Control
DistributedControlSystem
Fiber-OpticData Links
Data InputModBus
RamanAnalyzer
ProductionSampling
Points
0
20
40
60
80
100
11:00 12:00 1:00 2:00 3:00 4:00 5:00 6:00
Time of day
Con
cent
rati
on (%
)
Me3SiClGCRaman
MeHSiCl2 GCRaman
Chlorosilane Distillation –Start-up
The Process Raman Analyzer Provided the Desired Results
1. Detection Limits for Chlorosilanes ~ 0.1%
2. Analysis Cycle Time < 5 minutes
3. Automated, easy-to-use with Little Maintenance
4. System Controlled through Plant DCS
5. Simple Sampling Handling
6. Multiple Sampling Points with One Analyzer
Conclusions
PhAT (Pharmaceutical Area Testing) Raman System Representative Sampling of
Inhomogeneous Materials
SpecificationSpot Size: 3 mm (std, 6 mm opt)Depth of Field : +/- 12mmLaser power: 200mWMultiple Fiber CollectionLow Energy Density
6 mm
3 mm
MR Probe
x10
Tablet Analysis with the large spot illumination
• solves some major limitations of Raman for quantitative tablet analysis.– Representative Sampling…the 3 to 6-mm laser spot
size allows a much greater portion of a static sample to be interrogated in a single measurement.
– Reproducible Sampling…the depth of field provided by this probe design eliminates the sensitivity of the Raman response to small changes in sample placement from one measurement to the next.
• The superior reproducibility allowed by the large probed volume promises better results for quantitative analysis by Raman.
Quantitative In-Line Monitoring of Tablet
Coating
Arwa El Hagrasy, Shih-Ying Chang, Divyakant Desai and San Kiang
Bristol-Myers Squibb Company Am. Pharm. Rev., 9, 40-45 (2006)
On-Line System using PhAT - Coating
Abbreviated Raman spectra of Spectra from Tablets at Different Stages of Coating.
COATING Conventional Small Spot Size
y = x - 4·10-6
R2 = 0.9732
0
20
40
60
80
100
0 20 40 60 80 100Predicted
Obs
erve
d
COATING- PhAT - Large Areay = x - 2·10-6
R2= 0.9992
0
20
40
60
80
100
0 20 40 60 80 100Predicted
Obs
erve
d
AirHeadTM Gas-Phase Probe
Direct Insertion / Multipass Probe designSealed Optical Design100°C temp/ 650 psiFiber lengths up to 30 metersConstructed of SS 316
Gas-Phase Installation example
Cross FittingCell
AirHead
Sample Flow
Sapphire Window
Focusing Optics
Reflector
0
10000
20000
30000
40000
50000
60000
70000
0 500 1000 1500 2000 2500 3000 3500 4000
N2
O2
H2O
~300 ppmCO2
Lab Ambient Air Signature, 532 nm (Dark subtracted only, not intensity calibrated)
CO2 in Air
H2
CO2
CH4
CO
H2
*
Raman Gas Phase – Petrochemical Sample
Message – “The New Standard in High Performance, Quantitative, Versatile Sampling,
Transportable Raman Analyzers”
RamanRxn2 & RamanRxn2 Hybrid Analyzers
RamanRxn2
RamanRxn2 Hybrid
From laboratory to process
KEY FEATURESCertified to ATEX Standards
CalCheckTMOutstanding Precision allows use of PLS chemical models and calibration transfer between instruments
AutoCalTMReliable operation in severe operating environment
785nm NIR Laser• Low Fluorescence Background
• Multichannel OperationSequential 1 to 4 Channels
Raman Analyzer: Multi ChannelFiber Optics up to 300 m !
A CB D
Conclusions• Raman Instrument Reality has now caught up with
Theoretical Benefits.• A range of applications & utilities have been
demonstrated already.• Raman can facilitate process understanding and be a
great partner in PAT implementation, it allows to:• OPTIMIZE, CHARACTERIZE, MONITOR AND
CONTROL from
RESEARCH TO MANUFACTURE
Acknowledgements Who’s Getting Value ? – Pharma/PAT
“A Valuable Technique for Polymorph Screening”, C. Anderton, Eur. Pharm. Rev., 68-74 (2004) – GSK“Statistical Analysis of Difference in the Raman Spectra of Polymorphs”, S.M. Mehrens, U.J. Kale, and
X.Qu, J. Pharm.Sci, 94, 1353-1367 (2005) – Pfizer & Oakland University“Multivariate vs. Univariate Quantitation of Polymorphs in Drug Product by Raman Spectroscopy”, F.
LaPlant, FACSS, 181, Oct (2004) – Pfizer“Crystallization Monitoring by Raman Spectroscopy: Simultaneous Measurement of Desupersaturation
Profile and Polymorphic Form in Flufenamic Acid Systems”, Y.Hu, J.K. Liang, A.S. Myerson, and L.S.Taylor, ACS, March (2005) – Purdue University and IIT
“Adventures in Crystallization and Polymorphism”, M. Mitchell, MT Users Forum, March (2005) - Pfizer“Real-time Monitoring of Tritium Gas Reactions using Raman Spectroscopy” J. R. Heys, M. E. Powell,
and D. E. Pivonka, J. Labelled Compounds and Radiopharma, 47, 983-995 (2004) – AstraZeneca“Real-Time In Situ Raman Analysis of Microwave-Assisted Organic Reactions.” D.E. Pivonka and J.R.
Empfield, Appl. Spectrosc., 58, 41-36 (2004) – AstraZeneca“Use of a TG/DTA/Raman System to Monitor Dehydration and Phase Conversions”, A.S. Bigalow Kern,
W.J. Collins, R.T. Cambron, and N.L. Redman-Furey, J. Test & Eval., in press (2005) – Procter & Gamble Pharmaceuticals
“Process Development with In Situ Raman spectroscopy”, G. Zhou and Z. Ge, IFPAC, I-029, Jan 13 (2005) – Merck
“Real World Applications of PAT During Pharmaceutical Product Development”, S. Arrivo, EAS, Nov (2004) – Pfizer
Acknowledgements Who’s Getting Value ? – Pharma/PAT
“Raman Spectroscopy: A PAT Tool for Quantitative Assessment of Tablet Potency” J. Johansson, J. Eriksson, S. Folestad, and B. Lagerholm, FACSS, 508, Oct (2004) –AstraZeneca
“Effectively using PAT in a Process Development Environment to Expedite Processing in a Pilot Plant Facility”, C. Ray, R. Wethman, and J. Wasylyk, PAT, 17-20 (2005) – BMS
“Raman Spectroscopy for Quantitative Monitoring of Solid Dosage Manufacturing Process” S-Y Chang, A.El Hagrasy, W. Early, H. Guo, D. Li, S. Kothari, S. Paruchuri, and V. Nesarikar, IFPAC, I-142, Jan 13 (2005) – BMS
“Quantitative Model Development for Inline Raman Monitoring of Polymorph Transformation and Model Transferability”, D. Jayawickrama, IFPAC, I-163, Jan 13 (2005) - BMS
“The Use of Process Analytical Technologies in Multipurpose Plants”, M. Warman, Pittcon, March (2005) – Pfizer
“Development and Implementation of a Quantitative On-line Raman Method for Pharmaceutical In-Process Monitoring”, R. Wethman, MT Users Forum, March (2005) –BMS
“In-Line Monitoring of Hydrate Formation during Wet Granulation using Raman Spectroscopy”, H.Wikstrom, P.J.Marsac and L.S.Taylor, J. Pharm Sci, 94, 209-219 (2005) – Purdue University
“Comparison of Techniques for In-line Monitoring using Raman Spectroscopy.”, H. Wikstrom, I.R.Lewis and L.S. Taylor, Appl. Spectrosc., 59 July (2005) – Purdue University