Molecular Spectroscopy

Funtamentals
Gustavo Riether

The world leader in serving science

Agenda

• Overview
• What is molecular spectroscopy
• Introduction to eletromagnetic radiation
• Introduction to UV-Vis
• Lambert-beer/colour analysis
• How equipment works
• Sampling
• Introduction to FT-IR
• Vibrations
• Lambert-beer law again
• How equipment works
• Difference between IR, NIR, FIR
• Quant in IR
• Introduction to Raman
• Difference between IR
• Advantages/disadvantages

2
Agenda 2

• iS5/iS5N
• Accessories
• Aplications, demands, capabilities
• iS10
• Accessories
• Aplications, demands, capabilites
• iZ10, Continuum, TGA
• iS50
• Research made easy
• Expandable
• NIR, MIR, FIR
• Modules
• Aplications

3
Agenda 3

• Raman Fundamentals
• DXR2 Smart Raman
• Why microscopy?
• Microscopy
• Types (point and shoot, linear mapping, mapping, imaging)
• Mapping vs. Imaging
• Raman vs IR
• Sampling (raman even surface, IR)
• DXR2 Microscope
• DXR2xi
• Applications
• Hot trends
• Future applications
• FT-IR microscopy
• Continuum, iN10, iN10 MX
• Ultra fast screening
• Applications
• Sample preparation

4
Agenda 5

• NIR
• Equipment (antaris, ingot, blend target, etc)
• NIR modules
• Applications
• UV-Vis & Micro UV
• Equipment (put info about ND too)
• Applications
• IGS
• Fundamentals
• Applications
• picoSpin
• Fundamentals
• Advantages
• Applications

5
Introduction – Molecular Spectroscopy

Electromagnetic Radiation

• UV-Vis to FIR

6
Molecular Spectroscopy

• Spectroscopy:
• Interaction of the light with the matter

• Spectrometer (spectrophotometer):
• Instrument which measures this interaction

• UV-Vis?

• FT-IR:
• NIR?
• MIR?
• FIR?

• Raman?

7
Molecular Spectroscopy: Laying the Foundation

• Overview of Molecular Spectroscopy

• What is it? Why is it useful?

• Set the stage

• The tools
• Getting good data
• Determining what it means

• Get Answers not just Data

• Identify
• Quantify
• Characterize

8
The Power of Molecular Spectroscopy

• Work fast…
• Little or no sample prep, rapid screening
• …on a range of samples…
• Organics, inorganics, films, coatings, paper, fibers,
more
• …and answer fundamental questions
• What is this stuff? How much is in there? In what
form?

• Analyze samples from ‘cradle to grave’

• R & D, process control, QA/QC, failure analysis, waste

9
General ‘Rules’ for Molecular Spectroscopy

Response “Peaks”

Energy
• Energy varies along the x-axis
• Wavelength in nanometers or microns, Wavenumber (cm-1)
• Response is along the y-axis
• Absorbance, transmittance, reflectance
• Different molecules absorb light differently (“Peaks”)
• This is what enables spectroscopy to tell us something!

10
 Basics: UV-Visible Spectroscopy and Color
• “Simplest” technique
• “Dispersive” Spectrometers
• Color analysis

Absorbance
• Qualitative information
• Absorption frequencies
• Quantitative information
• Intensity
Wavelength (nm)

Electrons start in the ground state

• Light can cause the electron to jump to an excited
state
• This is absorption
• And the intensity of the absorption depends on the
number of electrons, so we can extract
quantitative information (Beer’s Law)

11
Intensity (y-axis): Beer’s Law

• Key idea: Peak Height relates to Concentration

• Beer’s Law (Lambert-Beer, Beer-Lambert, Beer-Lambert-Bouguer…)

• Absorbance = Absorptivity * Path Length * Concentration
• A = εlc (or A=abc sometimes)
• If path length and absorptivity are constant, then the absorbance
changes only with concentration

Wavenumber (cm-1)

12
Introduction to Infrared spectroscopy

• Infrared is a form of electromagnetic energy

• Electric and magnetic fields vibrate at a right angle to each other
• In terms of wavelength range, Infrared is the region between
Visible and Microwaves

• Molecules irradiated by infrared light, vibrate in various modes

absorbing energy and heat
• When we are close to a fire or are irradiated by infrared energy our skin
molecules vibrate, which is why we feel the heat

UV

13
Molecular Vibrations

Bending Twisting
+ -
C C C C C

Stretching Deformation
1 00

95

90

85

80

75

70

65

60
%T

55

50

45

40

35

30

25

20

15

4 000 3 500 3 000 2 500 2 000 1 500 1 000

c m -1

14
 Basics: FT-IR Spectroscopy and Composition

• Chemical bonds vibrate like springs

• Absorption of IR light occurs at the specific frequencies of those
vibrations 1 k
Frequency of vibration =
2p 
• So the absorption peaks in the IR spectrum can help identify the
molecule
Unknown Material

Wavenumber (cm-1)

15
Fingerprint Region

• The addition of a methyl group, from ethyl to propyl acetate, does not
significantly change the IR spectrum in the stretching region, but in the
fingerprint one… look how they are different!

16
Other Basic Rules

• Infrared spectroscopy absorptions follow the Beer’s-Lambert law

• Absorbance = a x b x c

• Where…
• a = infrared absorption characteristics of the material
• b = pathlength
• c = concentration
Infrared accessories or sampling techniques which cannot
guarantee constant pathlength, can be used for identification
analysis but not for quantitative or semi-quantitative
determinations. The same sample analyzed with different
pathlength, would result in different absorptions
That is why, to obtain some
quantitative information
from diluted samples at
various concentrations,
pathlength must remain
constant
Double pathlength = twice the
molecules interacting Double concentration, at b=k,
= double absorption equals double absorption

17
Other Basic Rules

• Transmittance scale visually enhances the lower intensity peaks and is

ideal for spectral observations (eye only)
• Absorbance scale is the one to use for peak integration and quantitative
analysis since it is linear with the concentration of the sample

18
FT-IR: NIR vs MIR vs FIR

• NIR - Overtones:
• Macro properties – humidity, proteins, ashes, fat, etc
• Heterogeneous products – feed, flours, grains, etc
• Fibers and probes
• Requires statistics and a large number of samples
• Low specificity

• MIR - Vibrations:
• Micro properties – content (high ppm to %), mixtures, structures and chemical
variations, etc
• High specificity

• FIR – Rotations:
• Oxides, polimorphs, addictives

19
Choose Components to Set Spectral Range

Low
energy
Overtones Fundamentals motions

Raw material id Structure or Inorganics

identity and
polymer
modes

20
IR vs Raman

IR: Excitation: IR source

→ Absorption and vibration of chemical bonds

Pyroelectric detector

Sample

Heat source
Prism or grating
Moving slit

21
IR vs Raman

Raman: Laser → Rayleigh Filter → Raman effect

(sample emission) – Complementar technique

22
Basics: Raman Spectroscopy and Morphology

Sample n=1
First
LASER n=0
Excited
State

Virtual
State

Energy
n=1 Ground
n=0 State

Stokes Rayleigh Anti-Stokes

• A vibrational spectroscopy technique Scattering Scattering Scattering

• Sensitive to vibrational modes of covalent bonds in molecules

• Most sensitive to highly symmetrical vibrations (i.e. C-C, Si-Si)
• Sensitive to anything that changes bond energy (i.e. bond angle)

23
IR vs Raman

IR vs Raman: complement each other

24
Vibrational Spectroscopy Comparison
Infrared
• Absorption
• Senses dipole vibrations
O-H, N-H, C=O
• Sample preparation necessary,
short optical pathlength required
• Limited fiber-optic potential
• Non-aqueous samples
• End groups dominant in spectra
25
Raman
• Emission of scattered laser light
• Senses polarizable vibrations
C=C, Aromatics
• Little or no sample preparation,
measure through transparent
packaging
• Considerable fiber-optic potential
• Aqueous samples
• Molecular backbone more
prominent
• Readily subtractable spectra
What are the main applications of the IR and Raman?

• Analytical Market:
• What is this? How much do I have in this sample? Is it ok? Etc

• Each pure substance have an unique spectrum

• Main applications:
• Identification of compounds and substances;
• Quantification – Concentration, variations, chemical changes;
• Spectral comparison;
• ID Confirmation – Know product;
• Reverse Engineering – Competition analysis;
• Chemical structures elucidation;
• Among others;

26
Advantages of FT-IR and Raman

• Advantages of the FT-IR and Raman:

• Quick technique
• Universal
• Non destructive
• Almost no sample preparation
• Low cost of ownership and operation
• All kinds of samples:
• Solids, liquids and gases
• All shapes
• Reliable and Reproducible results
• Safe

27
 Microspectroscopy

• Combine FT-IR or Raman • Point and shoot

with microscope + +

+
+

• Probe content, homogeneity, • Line maps

contaminents • Cross-Sections

• Examine layers, adhesives

• Area maps
• Chemical Images

28
Future Reach

• Inorganics
• Far-infrared

• Polymorphism
• FT-Raman or Raman

• Deformulation
• TGA-IR
• GC-IR
• Multi-component search

29
How FT-IR Works

Michelson Interferometer
Fixed mirror

Beamsplitter
IR
Source

l 0 -l

Moving mirror

Detector

30
 Time Domain to Frequency Domain

Moving Mirror

• One mirror moves…

• …one doesn’t
• This causes the two beams to
‘interfere’ when they recombine
• Watch how the detector signal
(bottom) varies with mirror movement Fixed Mirror
(top)…

• The result is a Time versus Detector

Signal (Intensity) plot

Detector Signal

31
The FTIR Interferogram

4 frequencies 10 frequencies Broadband source

32
Raw Data to Final Spectrum

Background
Volts

Single Beam
2200 2150 2100 2050 2000
Data points
1950 1900
Spectrum
4000 3000 2000 1000
Wavenumbers (cm-1)

Interferogram Single Beam

%T
Ratio
Single Beam
Volts

3000 2000 1000

Wavenumbers (cm-1)

Sample
4000 3000 2000 1000
Wavenumbers (cm-1)
2400 2300 2200 2100 2000 1900 1800
Data points

33
Inside the FT-IR Spectrometer

Interferometer

Laser
(reference)

Sample
Compartment

Detector

Infrared Source

Thermo Scientific™ Nicolet™ iS™10 FT-IR Spectrometer

34
Sampling Measurements

• Transmission
• Absolute reference measurement
• Analyzes entire sample
• Qualitative or quantitative
Io=0o • Sample preparation can be difficult

• Reflection
• Specialized optical modules
• Liquids, solids, gels or coatings
• Analyzes all or part of the sample
Io=Ro • Qualitative
• Minimum sample preparation

35
FT-IR Reflection Measurements

• Attenuated total reflectance (ATR)

• Solids and liquids
• Surface analysis

D S
• Diffuse reflectance (DRIFTS)
D D
D • Solids/powders
• Bulk analysis

• Specular reflectance
• Reflective solids or placed on mirror
• Surface or bulk analysis
Io=Ro

36
FT-IR Reflection Measurements: Principles

• Attenuated total reflectance (ATR and HATR)

• Beam focused into crystal
• Evanescent wave 1-2 microns deep
• Sample and crystal contact
• Surface analysis
• Useful for 99% of the samples

D
D S • Diffuse reflectance (DRIFTS)
D
D • Beam focused into sample cup
• Scattered light recollected
• Sample mixed with KBr
• Bulk analysis

• Specular reflectance
• Sample must have reflective layer or be on mirror
Io=Ro
• Beam reflected off sample at varying angles
• Bulk or surface analysis

37
FT-IR Reflection Measurements: Variables

• Attenuated total reflectance (ATR and HATR)

• Angle of light
• Refractive index of crystal and sample
• Wavelength of light

D
D S
D
• Diffuse reflectance (DRIFTS)
D • Mixing (KBr)
• Particle size and light scatter

• Specular reflectance
• Angle of light
• Effective penetration depth
Io=Ro
• Polarization of light

38
Transmission: The Archetype Method

• Simple Concept: Light passes through

• Transmission = best quality

• Prep takes time, skill Polymer - saturated

absorbance

Additive -
absorbance on scale

50 micron thick
pressed film

39
Attenuated Total Reflectance – A T R
The Choice for: • Packaging
• Corrosive liquids
• Polymers
• Inorganic materials
• Rubbers
• Rigid samples
• Carbon black (Ge plate)
• Metal coatings (Reflectance
• Pharmaceuticals plate)
• Resins • Concrete and building
• Paints materials
• Inks Pressure
device tip
• Paper coatings
0.75 Polycarbonate Pellet, by ATR
0.70

0.65 Sample Material

0.60

0.55
Crystal mount
0.50
Polycarbonate disk
0.45
ATR crystal
Log(1/R)

0.40
by ATR
0.35

0.30

0.25

0.20

0.15

0.10

0.05
-0.00
3500 3000 2500 2000 1500 1000
Wavenumbers (cm-1)
Infrared beam

40
Properties of Some Common ATR Crystals

ATR Spectral Refractive Depth of Penetration (µ)

Material Range (cm-1) Index (at 45º & 1000 cm-1) Uses

Good for most samples,

especially strong
Germanium 5,500 - 675 4 0.66
absorbing samples, such
as dark polymers

8,900 - 1,500 &

Silicon 3.4 0.85 Resistant to basic solutions
360-120

Very resistant to acidic

AMTIR 11,000 - 725 2.5 1.77
solutions

ZnSe 15,000 - 650 2.4 2.01 General use

Good for most samples.

Diamond 25,000 - 100 2.4 2.01 Extremely caustic or hard
samples

41
Failure Analysis with ATR

1157
0.6 Waxy surface of residue diamond Orbit ATR

1729
• Residue collected on filter

1099
1022
1450
Abs
0.4

1632
1535
1379

578
membrane

687
0.2
• Analyzed by direct contact with
Clean margin of membrane
Diamond ATR 0.4
• Membrane + residue

Abs
0.2
• Subtract membrane
• Search 0.4 Subtraction Result
1.5 Subtraction Result:Waxy surface of sample1 by diamond Orbit ATR
1.4 Ethyl Acrylate Homopolymer - Coatings Technology Library
0.3

Abs
1.3
0.2
1.2
1.1 0.1
1.0
4000 3000 2000 1000
0.9
Wavenumbers (cm-1)
Absorbance

0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1 Ethyl Acrylate Homopolymer
4000 3000 2000 1000 Coatings Technology Library
Wavenumbers (cm-1)

42
Inorganic Residue on Turbine

0.35

0.30

1082
0.25
Small clump ~ 1 mm

2921

1417
1738
0.20
3206

2852
0.15
Absorbance

612
0.10

0.05

0.00

-0.05
Ammonium sulfate
-0.10
reference spectrum

3000 2000 1000

Wavenumbers (cm-1)

43
Black Rubber Samples on Ge

• Carbon black has

high index
• Ideal for Ge
• Multiple
components
• Base polymer
• Silane slip-aid

44
Smart™ iTR/iTX Mid-IR Germanium

• Effective Analysis
• Tires
• Cables, belts
• O-Rings, gaskets
• Automotive components

• Useful Techniques
• SmartTM iTR/iTX with germanium
• TGA-IR (co-polymers and fillers quant analysis)

• Will a Diamond ATR be useful?

• Yes - for all compounding materials (before black blending)
• No - not for their finished products

45
 Multi-reflection ATR

• More sensitivity for weakly absorbing bands

• Constant path length
• Low maintenance system – cleaning easy
• Can handle highly viscous samples
• Flow and temperature controlled versions available

0.4 12 reflections
10 reflections
0.3
Absorbance

1 reflections
Containment Trough
Liquid Sample
0.2

0.1
Crystal
1400 1380 1360 1340
Wavenumbers (cm-1)

46
Measuring FAME in biodiesel blends

 ARK allows crystal plate changes to fine

tune path length
• 60° for high concentrations
• 45° for low concentrations
 Simple calibration and quantitation

Ester peaks

Calibration

47
Cadmium Additive using Far-IR

• Masterbatch analyzed using built-in ATR on the iS50

• the ATR accessory can collect spectra in the Far-IR region, down to 80 cm-1

48
Extended Range ATR: Far-Infrared

 Far IR using the diamond ATR

• Shows that it contains cadmium pigment

49
Mid-Far Infrared: Single Beamsplitter or Combination

50
Advanced ATR Correction: Comparison

Transmission

AdvATR Corr

• Advanced ATR Correction Applied

Transmission • No major differences in transmission, corrected

AdvATR Corr

51
Positional Isomers of a Drug Compound

Acetamidophenol isomers
0.50 **2- ac etamido phn eol
2-acetamidophenol 2
0.40

0.30
Abs

0.20

0.10

0.40 **3- ac etamido phe nol

2-acetamidophenol 3
0.30
Abs

0.20

0.10

0.00
0.40 **4- ac etamido phe nol
2-acetamidophenol
0.30 4
Abs

0.20

0.10

0.00
400 0 350 0 300 0 250 0 200 0 150 0 100 0 500
W av enu mber s ( c m- 1)
Typical ZnSe ATR limit Typical KBr limit

52
Transmission and ATR

• The #1 and #1 Techniques!

• Generally
• Transmission better for Quantitative
analysis
• ATR requires less sample preparation
• Both are scalable to Microscopy
Thermo Scientific Nicolet iS50 FT-IR
Spectrometer

No Compromise

53
The Rest is Up to Physics!

• Molecules absorb specific frequencies

• Chemically similar materials absorb in the
same range
• Esters: around 1750 cm-1
• Carbonyls: around 1600 cm-1
• Hydrocarbons: around 3000 cm-1
• Information extracted
• Identification
• Quantity
• Kinetics

54
Fundamentals to Future

• Molecular Spectroscopy answers basic questions

• Color, Composition (Qualitative and Quantitative)

• Usable in any environment

• Laboratory, On-line, Mobile Labs

• Extendable
• Hyphenation, multi-range

55
 Break

Next topic: Our IR Solutions – iS5; iS10; iS50

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