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.

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y of

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• 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

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y of

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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