IR microscopy at Merck KGaA - bruker.com QM/LIMS Support & Projects ASI ... ICP-MS XPS HPLC-MS XR F ... Imaging area 600 µm x 600 µm Franziska Burkert, Merck KGaA, ...

IR microscopy at Merck KGaA

Franziska Burkert Franziska Burkert

Ettlingen, 13th November 2013

Agenda

Merck KGaA1

2 Central analytical services (CAS)

Special sample analytics in CAS3

Franziska Burkert, Merck KGaA, 13th November 20132

Special sample analytics in CAS3

4 FT-IR microscopy in troubleshooting analytics

FT-IR imaging for Research & Development 5

6 Summary

Milestones in the history of Merck

1668

Friedrich Jacob Merck (1621-1678) buys the Angel Pharmacy (Engel-Apotheke)

1827

Emanuel Merck (1794-1855) startsproduction on an industrial scale

Merck is the oldest pharmaceutical and chemical com pany in the world


Merck is a leading pharmaceutical, chemical and life sciencecompany that conducts its operations in four divisions:

– Merck Serono: Prescription medicines of both chemical andbiological origin in specialist indications

– Consumer Health: Over-the-counter pharmaceuticals forpreventive health care and self-treatment of minor ailments

Merck at a glance

preventive health care and self-treatment of minor ailments

– Performance Materials: High-tech performance chemicalsfor applications in fields such as consumer electronics, lighting,coatings, printing, plastics, and cosmetics

– Merck Millipore: Products and services for the life science industry, used in research and analytical laboratories as well as in pharmaceutical manufacturing

� Approximately 39,000 employees in 66 countries

� Merck manages its operating activities under the umbrella of Merck KGaA, which was listed on the Frankfurt Stock Exchange in1995 and admitted to the DAX® in 2007

� Around 30% of the total capital is publicly traded, while the Merck family, as general partner, indirectly holds around 70%Franziska Burkert, Merck KGaA, 13th November 20134

Pützer Tower and Pyramidat Darmstadt headquarters

www.merckgroup.com

Central analytical services (CAS)

SO-A(Site Operations-

Analytics)

SO-ASAnalytical Services

SO-ABBioanalytical

ChemistryAssistant

Assistant

AssistantSO-AC

Analytical Chemistry

Physical Methods &Chromatography & Mass Bioprocess Analytics Molecule Characterization

55 Franziska Burkert, Merck KGaA, 13th November 2013

ABB-1Biochemistry,Immunology

ABB-2Microbiology

ABB-3Life Science Analytics

ABB-4Molecular- andCellular Biology

ABB-5Biomolecule Analytics

ABC-6Quant. Bioanalytics

ABC-7Molecule Analytics

ABC-8Phys. Chem. Prop.,

XRD

ABC-9Bio NMR &

NMR Spectroscopy

ASRReporting & Laboratory

Services

ASSSampling & CAS

QM-Support

ASLSAP QM/LIMS

Support & Projects

ASIIT-Support &

Development, PAT

ACC-1Impurity Profiling

ACC-4Mass Spectrometry &Gas Chromatography

ACC-5Chromatography Methods

ACC-6OLED-Analytics

AC9Titration,

Special Analytics

ACP-2Element Analytics

ACP-7Therm. Analysis,

Molec. Spectroscopy

ACP-8Particle Charact., SEM,

Troubleshooting Analytics

Physical Methods &Spectroscopy (SO-ACP)

Chromatography & MassSpectrometry (SO-ACC)

Bioprocess Analytics(SO-ABB)

Molecule Characterization(SO-ABC)

ACC-3Liquid Chromatography

� Troubleshooting analytics� Identification of impurities: fibers, particles, discolorations ...

� Root cause analysis

� Investigations planned, coordinated and documented by SPA team

Special sample analytics in CASTasks


� Microspectroscopy for Research & Development � Chemical imaging

� Surface-sensitive analyses

� Time-dependent and temperature-dependent analyses

� Clean room (nominally class 10.000)

� Sample preparation in laminar flow box (class 100) inside the clean room

� Four microscope systems located in the clean room

Special sample analytics in CASThe clean room


Special sample analytics in CASThe microscopes


IR microscopy in troubleshooting analytics


Troubleshooting analytics

� Occurrence of unknown substances � during production

� quality control

� customer complaint

� ...� ...

� Impurities in manufacturing processes and products� Unexpected & unwanted occurrence

� Unknown identity & risk

� Analytical method unclear, often various analytical methods necessary for identification

� Short response time important


Troubleshooting analyticsTypical work flow

Customer

Sample preparation

Documentation1H-NMREI-MS

Special Sample Lab


IR/Raman microscopy

Sample preparation(e.g. filtration)

Optical microscopy

Electron microscopy/EDS?

Organic

Inorganic

UV/VIS

ToF-SIMS

ICP-MS

XPS

HPLC-MS

XRF…

GC-MS

EI-MS

lab core competence coordination of other methods

Fast, high spatial resolution, non destructive

Troubleshooting analyticsParticle isolation

� Isolation of impurity

� Preparation on sample holder

� Characterization by optical � Characterization by optical microscopy � Documentation of structure, size, color,

number of particles ...

� Decision which methods will be usedfor further characterization

12 Franziska Burkert, Merck KGaA, 13th November 2013

Troubleshooting analyticsFT-IR microscopy

� FT-IR spectroscopy is viable for identification of most organic and many inorganic materials

� Direct structural information from characteristic group frequencies


characteristic group frequencies

� Fast computer-aided identification: search in spectral databases

� High sensitivity compared to other methods:

Particle size 100 x 10 x 10 µm3

Particle mass ~ 10 ng

After dissolution in 100 µL solvent � concentration ~ 100 ppb

Troubleshooting analyticsLUMOS – IR-Microscope

� Measurement with micro-ATR

� Ge-crystal, tip 100 µm diameter

� Fast measurement (< 1 min)

� High quality FT-IR spectra of� High quality FT-IR spectra ofindividual particles


Troubleshooting analyticsIdentification of unexpected impurities with LUMOS microscope


Combination with element composition/distribution from SEM/EDS mapping enables a reliable identification of single particles.

The subsequent allocation of impurity source within hours safes cost, enables expert handling of complaints and avoids compensation claims.

IR imaging for R&D


IR microscopy for R&DHyperion 3000 & accessories

Franziska Burkert, Merck KGaA, 13th November 201317Linkam Stage

IRRAS

Macro-ATR

� Tissue sections of histological samples embedded in paraffin

� FT-IR microscopy in transmission mode� Fast, non-destructive

� Qualitative and quantitative results

FT-IR imaging for R&DParaffin-embedded tissue sections

� Qualitative and quantitative results

� Focal Plane Array (FPA) detector: in each rectangle 64 x 64 pixel � 64 x 64 spectra� 4096 spectra per single measurement (170 µm x 170 µm)

� 5 x 6 FPA mapping � 122800 spectra (850 µm x 1020 µm)

� Visualization of proteins, fats, bones, pharmaceutical drugs, …


FT-IR imaging for R&DChemical imaging using ATR

� Problem: Sample preparation for transmission IR is sometimes not possible

� Solution: FT-IR imaging using ATR techniques

19

Micro-ATRImaging area 32 µm x 32 µm

Macro-ATRImaging area 600 µm x 600 µm


1732

1653

1546

1162

1 . 5

2 . 0

FT-IR imaging for R&DChemical imaging of tissue sections

� FT-IR imaging of cryo tissue sections

� Increased spatial resolution compared totransmission IR measurements

0 . 0

0 . 5

1 . 0

1 . 5

1 0 0 01 2 0 01 4 0 01 6 0 01 8 0 02 0 0 0

W a v e n u m b e r c m - 1

Abs

orba

nce

Uni

ts

20

0

6.51e+03

3.2e+03

2.8e+03

2.4e+03

2e+03

1.6e+03

1.2e+03

800

400

3.6e+03

4e+03

4.4e+03

4.8e+03

5.2e+03

5.6e+03

6e+03

36400 36500

1115.34-1059.79 X-Achse [µm]

21900

22000

22100Y-A

ch

se [

µm

]

0

1.23e+04

6.2e+03

5.4e+03

4.6e+03

3.8e+03

3e+03

2.2e+03

1.4e+03

7e+03

7.8e+03

8.6e+03

9.4e+03

1.02e+04

1.1e+04

36400 36500

1070.90-963.50 X-Achse [µm]

21900

22000

22100Y-A

ch

se [

µm

]

0

2.33e+04

1.15e+04

9.5e+03

7.5e+03

5.5e+03

3.5e+03

1.35e+04

1.55e+04

1.75e+04

1.95e+04

2.15e+04

36400 36500

1715.28-1485.67 X-Achse [µm]

21900

22000

22100Y-A

ch

se [

µm

]

Micro beads Bone Proteins320 µm x 320 µm 102400 spectra


wavenumber

FT-IR imaging for R&DOther examples for ATR-IR imaging

� Chemical imaging of tablets

0

703

360

260

160

460

560

21800

22000

22200

22400

Y-A

ch

se [

µm

]

� Chemical imaging of multilayered materials


36200 36400 36600 36800

Baseform X-Achse [µm]

21600

Summary

� IR analysis is a powerful technique to identifydifferent organic and inorganic substances

� The LUMOS microscope is the perfect tool foridentification of single small particles

� The Hyperion 3000 is a flexible instrument for� The Hyperion 3000 is a flexible instrument fordifferent R&D setups� IR imaging of tissue sections, tablets, packaging materials,

melt extrudates …

� Surface-sensitive analyses (IRRAS)

� Time-dependent and temperature-dependent analyses


IR microscopy at Merck KGaA - bruker.com QM/LIMS Support & Projects ASI ... ICP-MS XPS HPLC-MS XR F ... Imaging area 600 µm x 600 µm Franziska Burkert, Merck KGaA, ...

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