Targeting Material characterization by

Raman

Dr Enrique Lozano

Sales Director MMS E&A

Raman is a technique largely used for mat.

Char. mostly due to the fingerprinrt

characteristics materials show under this

technique, plus other considerations:

• Capability of quantification

• Confocality potential

• Fiber compatible

• Many sources of light

• Complement other techniques

IR and Raman - Complementary Excitation spectrum

Absorption spectrum

Raman Pros and Cons

Applicable to aqueous solutions

Less demanding to optics

Easy sampling for solids

Weak signal (efficiency ≈ 10-6)

Fluorescence interference

Mid-IR NIR reflectance Raman

Mechanism: absorption by

fundamental molecular

vibrational modes

Molecular Specificity: high

Signal Strength: strong

Energy: 600-4000 cm-1

Sampling: direct material

contact required, glass and

water appear opaque; can

rarely see through container

materials

Hardware: probes extremely

expensive, and short in length,

size and weight

Mechanism: absorption by

harmonics of fundamental

vibrational modes of X-H bonds

(e.g., N-H, O-H)

Molecular Specificity: low

Signal Strength: weak

Energy: 4000-12,500 cm-1

Sampling: non-contact, but

close proximity to material

required (<5 mm)

Hardware: longer probes

possible (meters); quartz optics;

dispersive, interferometry

common

Mechanism: inelastic

scattering by vibrational,

rotational, low frequency

molecular modes.

Molecular Specificity: high

Signal Strength: weak

Energy: 65-4000 cm-1 shift

Sampling: container

interference is mitigated by

accessories

Hardware: CCD detection;

quartz optics; long probes

common (kilometers in some

cases)

Raman Diagram

Because of very weak Raman scattering signals lasers are used as intense excitation sources.

Bench Top

Large size

$50K to >$150 K

Resolution: 0.1 to 5 cm-1

Expensive parts

Portable

Compact, small footprint

$10 K - $45 K

Resolution: 3, 5, 10 cm-1

Low maintenance

Bench Top Raman vs. Portable Raman

Portable Raman Spectrometer

Rugged design

Small size

Light weight

Fiber probe

Battery option

High performance to cost ratio

Pharmaceutical Industry

Pharmaceutical manufacturing facilities

are moving toward 100% inspection of

incoming raw materials.

Confirm the content of each container is

verifiable at the molecular level.

Raman spectra can be acquired through

transparent packaging materials, bags and

bottles.

"Pass/Fail" decision

Support CGMP and 21 CFR Part 11

Inspection of Raw Materials in

Pharmaceutical Manufacturing

Rapid Screening of Protein

Therapeutics Products

US FDA used B&W Tek MiniRam™ series in their pilot program of rapid Screening of Protein Therapeutics Products in the fields.

Insulin methods: to distinguish insulins with different origins and formulations, as well as insulins that have been modified by external stresses such as heat and agitation

Heparin sodium methods: to detect the presence of over-sulfated chondroitin sulfate in API

Advantageous for process control

Effectively maintaining safe supply

of protein-based pharma products

Manufacturing

Raman spectral change during

HSWG (High Shear Wet Granulation)

polymorph transformation

New Raman bands for form B

appear and for form A disappear

Material Characterization

DVS water sorption and desorption cycle (a.) and Raman spectra (b.) for β D-mannitol.

Chemical Industry

0 2000 4000 6000 8000 10000

5000

10000

15000

20000

25000 Top to Bottom

0-1 min

1-2 min

2-3 min

3-4 min

4-5 min

NOA63 Under 375nm Laser Curing

~ 2960cm-1

Re

lative

In

ten

sity

Raman Shift (cm-1)

5 mW 375 nm 30 s

Epoxy Cure Monitoring

0 1 2 3 4 5

1000

1200

1400

1600

1800

2000

NOA63 Under 375nm Laser Curing

Re

lative

Pea

k In

ten

sity

Time of Cure (min)

Evolution of 2960cm-1 peak

intensity with curing time

Polystyrene identification

Process Application

Si (s) + MeCr (g)

Me2SiCl3,

Me2SiCl2,

Me2SiCl,

MeHSiCl2,

Me2HSiCl,

HSiCl3,

Me4Si,

SiCl4

TiO2·H2SO4·H2O → TiO2·H2SO4 →TiO2 (nanophase) → TiO2 (anatase)

→ TiO2 (rutile)

Process Application

B&W Tek i-Raman Plus, 785nm Laser

16 samples with 0 – 48.3% diethylene glycol (DEG) in glycerin

Develop PLS model with standard error of < 0.5%

DEG concentration from Integration

Raman spectra of DEG/Glycerin samples:

measured on iRaman Plus

www.bwtek.com

Quantitative analysis of olive oils at

1064nm. iRaman EX Extra virgin olive oil (EVOO) samples

were spiked with small amounts of

light olive oil

8 samples with 0.21- 19.15% were

measured on the i-Raman EX

BWIQ software used to develop a

quantitative model for light oil in

EVOO

Feasibility of this measurement

shown with limited sample set

www.bwtek.com 21

PLS regression model for light oil in EVOO

2 factor PLS

regression over

full range of raw

Raman spectral

data

Counterfeit Drug Identification According to the World Health Organization’s estimates, ~10-15% of

the world’s drug supply (and about 1% in the US) is counterfeit - at

a value of about $200B in 2010

Deployment of handheld and portable Raman spectrometers can help

rapidly spot fakes

Raman is beneficial for

Drug Identification Control: Purity and Quality

Counterfeit Drug Analysis

23 Best 785

http://www.forbes.com/sites/henrymiller/2012

/07/25/fake-and-flawed-medicines-threaten-

us-all/

www.bwtek.com

Security

Legal highs analysis - counterfeits

1,500

2,500

3,500

4,500

5,500

6,500

0 500 1000 1500 2000 2500

Ram

an

in

ten

sit

y (

arb

itra

ry u

nit

s)

Wavenumbers (cm-1)

Citalopram reference Citalopram test1

Citalopram - Antidepressant

Counterfeit analysis

2,000

2,500

3,000

3,500

4,000

4,500

5,000

5,500

6,000

6,500

7,000

0 500 1000 1500 2000 2500

Ram

an

in

ten

sit

y (

arb

itra

ry u

nit

s)

Wavenumber (cm-1)

Sertraline reference Sertraline test 1

Sertraline.- Antidepressant

Counterfeit analysis

0

2,000

4,000

6,000

8,000

10,000

12,000

14,000

16,000

0 500 1000 1500 2000 2500 3000

Ram

an

in

ten

sit

y (

arb

itra

ry u

nit

s)

Wavenumbers (cm-1)

Aspirin reference Aspirin test 1

Aspirin: analgesic (reduce pain), antipyretic (reduce fever), anti-inflamatory

and anti-platelet

Street drugs, Cocaine

Cocaine vs Tetracaine

(legal local anesthetic)

Amphetamine

(pure)

Amphetamine (cut),

strong fluorescence

Street drugs, amphetamine

ingredient in “fake” "ecstasy" tablets

α-Pyrrolidinopropiophenone (α-PPP)

Stimulant drug

Methanol in Alcoholic beverages Methanol is a potent toxicant in humans that can cause severe and even fatal illness.

Studies have shown that the maximum tolerable concentration (MTC) of methanol in alcoholic beverages

with about 40% alcohol is about 2% (v/v) by volume. (A.J. Paine and A. D Dayan , Hum Exp Toxicol

November 2001 20 (11) 563-568)

Having a means to rapidly measure the level of methanol in an alcoholic drink that an adult human could

consume without risking toxicity due to its methanol content s an important public health safety

consideration

The Raman spectrum of Methanol is strong and can be used to minimize risk of harm

www.bwtek.com 31

Methanol in flavored rum

CH3 bending vibration at

1032 cm-1 increases with

methanol concentration.

Can readily measure

concentration of MeOH in

alcoholic beverage by

Raman

Dilution of beverages with

water can also be

measured by Raman

www.bwtek.com 32

Art, Geology and Gemology

Minerals/Rocks/Water Analysis B&W Tek i-Raman was used in European Space Agency's ExoMars

mission to the Red Planet:

Before testing for life on other planets, it's useful to practice on barren

areas of the Earth. One such place is Rio Tinto in Spain, where conditions

are analogous to Mars.

In situ techniques: samples are examined at the site, instead of being

removed and taken to a lab for analysis.

Results are gathered much faster.

Avoid contamination that can occur

during a return journey.

Deteriorated Civil Building Stones

B&W Tek i-Raman was used to measure aragonite in a violet-coloured piece of shell in sample (a); gypsum in sample (b)

Gemstone Identification

Diamond or Zircon?

Raman spectra of diamond and zircon are distinctly different

Diamond shows only one very strong and sharp Raman band at around 1328 cm-1, which corresponds to the C-C stretching mode.

Zircon shows multiple Raman bands at around 349, 431, 967 and 1002 cm-1, which correspond to the Si-O bending mode and stretching mode.

Examination of Inclusions in Minerals

Raman spectrum of a ruby sample when the excitation laser light is focused in the vicinity of one of its inclusions (a)

Raman bands at 710 and 1085 cm-1 match well with the Raman bands of calcite (b)

The inclusion substance of the ruby is calcite

This ruby sample is natural without undergoing heat treatment.

Paint and Pigment Analysis

B&W Tek i-Raman was used for analysis

of blue pigment, lazurite, on a Buddha statue.

Surface Enhanced Raman

Spectroscopy (SERS)

Metal Plasmons Enhancement: Molecules adsorbed on metal

400 800 1200 1600 2000

0

10000

20000

30000

40000

50000

60000

Re

lative

In

ten

sity

Raman Shift (cm-1)

Surface Enhanced Raman Spectrum

of Benzenethiol Measured with MiniRam

Detection of Melamine in Gluten Using SERS

Structure and SEM image of a

KlariteTM SERS active gold substrate

Average SERS spectra (n = 4) acquired from

extracts of wheat gluten containing different

concentrations of melamine: 2.0% (a), 1.0% (b),

0.5% (c), 0.1% (d), 0% (e).

Forensic Analysis

Non-destructive sample

Probe through various

packaging materials

Rapid results

Portability

C4 plastic explosive

TNT

Sodium nitrite fertilizer bomb

BWTEK Raman Family

i-Raman® Plus Technical

Specifications i-Raman® Plus

BWS465

Detector Back-Thinned CCD Array

Excitation 532 nm 785 nm 830 nm

mW < 50mW < 300mW < 300mW

Range /

Resolution

532 S:

175 - 4000cm-1

~ 4.0cm-1 @

614nm

785 S:

175 - 3200cm-1

~ 4.5cm-1 @

912nm 200 - 2300cm-1

~ 4.0cm-1 @ 912nm

Range /

Resolution

532 H:

175 - 3300cm-1

~ 3.0@ 614nm

785 H:

175 - 2700cm-1

~ 3.5cm-1 @

912nm

Detector Detail -2°C TE Cooled Back-Thinned CCD Array

Dynamic Range 50,000:1

Power 5VDC at 5.5 amp

Optional Battery w/ DC only

i-Raman® Plus Higher S/N ratio allows for greater

specificity of materials

Greater range allows you to see

more C-H Stretching

BWIQ allows users to develop

Quantitative Raman methods for:

Mixture analysis

BWID allows user to develop

libraries for

Final Product Identification

Counterfeit Detection

Comprehensive suite of

accessories, software, allow

versatility and portability

CH stretching

Higher S/N and better spectral resolution

(~5 cm-1)

i-Raman® EX Technical Specifications i-Raman® EX

BWS485

Detector TE Cooled InGaAs

Excitation 1064 nm

mW <450 mW

Laser Power

Control 0 to 100%

Range /

Resolution

175 - 2500cm-1

~ 4.0cm-1 @ 614nm

Dynamic Range 25,000:1

Digitization

Resolution 16 bit or 65,535:1

Integration Time 200 μs to >20 minutes

DC Power 12V DC at 6.6 Amps

Battery Optional

i-Raman® EX

Material Fluorescence reduces

applicability on a small subset

of materials

Utilize 1064 nm excitation

Reduce fluorescence

Increase productivity

Research Grade Enhance

material characterization

opportunities

Extra Virgin Olive Oil with 785nm (in Red) and 1064nm (in Blue)

Applications on 1064nm i-Raman EX

Dye: Rhodamine 6G_785vs1064

Applications on 1064nm i-Raman EX

Fiber: Polyester_785vs1064

Applications on 1064nm i-Raman EX

Medicine: Alka Seltzer_785vs1064

Comparison of Sesame oil spectra

Applications on 1064nm i-Raman EX

NanoRam Technical Specifications NanoRam

BWS455

Detector TE Cooled linear CCD Array

Excitation

785nm ± 0.5nm, Stability <0.5cm-1, Linewidth <2.0cm-

1

mW tunable from 0 mW to 300 mW

Laser Power

Control 0 to 100% in 10% increments

Range /

Resolution

176cm-1 to 2900cm-1

~ 9.0 cm-1 @ 785nm

Display High Visibility OLED touch screen, 3.7" size

Barcode

Reader 1D and 2D barcodes

Data Formats .txt, .csv, .spc

DC Power 12V DC at 6.6 Amps

Battery Rechargeable Li-ion Battery, > 5 hours operation

PC Software NID

Computer

Interface Ethernet, Wi-Fi

MiniRam

MiniRam®

BTR111

Detector TE cooled linear CCD Array

Excitation 532 nm 785 nm

mW < 50mW < 300mW

Range /

Resolution

175 - 4000cm-1

~ 15cm-1 @ 614nm

175 - 3200cm-1

~ 10cm-1 @ 912nm

Detector Detail -14°C TE Cooled linear CCD Array

Dynamic Range 300:1

Power 12VDC at 10.8 amp

Optional Battery w/ DC only

Chemometric (Spectral) Analysis

• Correlation analysis for material identification and/or verification

(Qualitative)

• Uncover underlying trends or patterns, classification

Multivariate analysis of complex systems with large data sets (Quantitative)

Develop models to predict response in unknown data sets

The use of mathematical and statistical methods for analysis of chemical or spectral data

BWIDTM BWIQTM

www.bwtek.com

How to choose excitation wavelength

• 532nm

• 785nm

• 1064nm

How to choose suitable sampling device

• Lab grade Raman probe

• Industrial Raman probe

• Flow cell

Portable Raman Applications Pharmaceutical Industry

cGMP (Current Good Manufacturing Process):

PAT (Process Analytical Technology): on-line, at-line process monitoring and control

QC: 100% incoming raw material identification

Counterfeit drug identification

Painting and Pigment

Forensic Analysis

Biomedical Analysis

Surface Enhanced Raman Spectroscopy (SERS)

Thank you!

Questions?

Dr Enrique Lozano

enriquel@bwtek.com

+447730871095