1. Overview of Raman Spectroscopy and B&W Tek Portable Instruments and Applications
August 2013

2. Outline Raman Spectroscopy Positioning of Portable Raman vs NanoRam
Instrumentation for B&W Tek Portable Raman Systems
Portable Raman Systems
Sampling and Accessories
Raman Software: BWID, BWSpec and BWIQ
Raman Applications
Hands-on use of iRaman Plus and iRaman EX
Lunch

3. Raman spectroscopy

5. What is Raman Spectroscopy ?
Raman spectroscopy is a form of molecular spectroscopy – the scattering of electromagnetic radiation by atoms or molecules. The Raman signal is an invaluable tool for molecular fingerprinting.
Advantages of Raman Spectroscopy
Little to no sample preparation required
Perform analysis directly through transparent containers
(i.e. plastic bags, glass, etc.)
Enables both qualitative and quantitative analysis
Highly selective
Fast analysis times
Insensitive to aqueous absorption bands
KEY: Sensitivity, S/N, performance/cost, reproducibility, qualitative/quantitative

6. Unchanged Elastic Scattering
Raman Scattering
Raleigh
Unchanged Elastic Scattering
Laser
785 nm Excitation
Raman
Inelastic Scattering
1,000,000 photon vs.1

7. Raman Spectrum
A Raman spectrum is a plot of the intensity of Raman scattered radiation as a function of its frequency difference from the incident radiation (usually in units of wavenumbers, cm-1). This difference is called the Raman shift.
(CC) Aliphatic Ring
(CC) Aromatic Ring
C=C

8. Raman Spectral Information
500
1000
1500
2000
2500
3000
Raman Shift (cm-1)
(CH3)2C=O
CH3CH2OH
(CH3)2S=O
CH3CH2O2CCH3
C6H5CH3
C=O
CH3
Aromatic-H
S=O
CH2
C-O
Aromatic

9. Limitations of Raman Spectroscopy
Weak signal (efficiency ~ 10-8)
Fluorescence interference

10. Raman Diagram
A Raman Instrument consists of a laser, sampling optics (probe), and an optical spectrometer. Because of very weak Raman scattering signals lasers are used as intense excitation sources.

11. Sampling
Fiber optic probes can also be easily adapted to a variety of different sampling chambers
Liquid flow cells
Gas flow cells
Optical microscopes
Direct sampling accessories allow for extreme portability and improve collection efficiency.
Point & Shoot
Vial Holder
Large Bottle Sampler

12. Excitation Wavelengths
532nm
785nm
1064nm
Sesame Seed Oil
Second diagram showing why flour …

13. What can Raman do for you?
Strong Raman Signal
Active Pharmaceutical Ingredients
Alcohols
Antibiotics
Antioxidants
Buffers
Coatings
Diluents
Emulsifiers
Excipients
Flavors
Fragrances
Lubricants
Monomers and polymers
Polyatomic inorganics
Preservatives
Solvents
Vitamins
Weak Raman Signal
Materials that are dark in color
Highly fluorescing molecules
Fillers/binding agents
Glass
Thin-walled plastics
Water
No Raman Signal
Black materials
Metals
Mono-atomic ions

14. Raman can measure through a broad class of packaging
Bottles
Thickness
Amber Glass
< 2 mm
Clear Glass
< 3 mm
High Density Polyethylene (HDPE)
< 1 mm
Teflon FEP
Polystyrene
Vials
Amber and Clear Glass
Bags
Polypropylene (PP)
< 0.1mm
Polyethylene (PE), Low-Density Polyethylene (LDPE)

15. Product positioning

16. Portable Raman Spectrometer
Rugged design
No moving parts for reliability
Small size for on site analysis
Light weight
Fiber probe for easy sampling
Battery option
High performance to cost ratio

17. Raman Spectrometer – Product Line

18. Summary Product Positioning
Compact, handheld, highly portable
Portable – computer required
Rapid material identification in a regulated or non-regulated environment
Laboratory analysis for identification, and general research applications including quantitative analysis
Samples that fluoresce, such as highly colored samples, natural products, and samples with fluorescing impurities
Easy operation and direct results - nonexpert users
Flexible system that can be used by nonexpert users, in QA/QC labs and research applications
Fully integrated all-in-one measurement and analysis tool in touchscreen GUI
Flexibility in setting measurement parameters and in data analysis options

19. Target for Handheld and Portable Raman
NanoRam is purpose-build for ID testing
Portable Raman instruments are versatile for use in research and development applications
The iRaman series is available with different laser excitation to suit particular applications and needs
BWSpec and BWID software used for the data collection
Quantitative analysis can be done with BWIQ
Use of flexible fiber optic probe makes interfacing to different types of samples in lab, process, and field possible

20. Similar features: Handheld and Portable
Use class 3b laser source and carry warning labels; fully comply with laser safety requirements of 21CFR
ANSI standard safe distance same as NanoRam ~ 30 cm
Have flexibility of different sampling accessories
Adjustable laser power settings
21 CFR11 compliant software available
Data can be exported and further explored and analyzed

21. Some key differences: Software
Portables
Only BWID Pharma is compliant for regulatory and can be used for material release decision
BWSpec software interface suited for laboratory use and analysts
In depth data analysis requires software beyond BWSpec such as BWIQ
Supports customer software development via SDKs
NanoRam
Touchscreen user interface, with methods and libraries is for qualitative analysis
Closed software system
Full audit trail, login with different user levels comply with regulatory needs, and work well in manufacturing and QC environment
Data management is with NID software;

22. Some key differences: Hardware
Portables
External bar code reader option
Fiber-based sampling
External rechargeable battery option
Not IP rated- more suitable for lab environment
Can be packaged in NEMA enclosure for process use
NanoRam
Integrated barcode reader
Non fiber-based sampling
Battery operated
Dust and splash proof (IP64)

23. Operational differences between portable and NanoRam
Little need to set measurement settings
Intensity correction automatically applied
Soft key for laser on/off
Rapid analysis results displayed in GUI
Purpose built for Identification and Verification
i-Raman Series
Set integration time, laser power, averaging
Manually enable intensity correction
Physical laser on/off key
Data or results rapidly displayed – depending on software used
Quantitative analysis options with BWIQ

24. Portable Raman Instrumentation

25. Key Instrumentation for Portable Raman
Stabilized laser: 532nm, 785nm, 1064nm
Multi-element CCD (Charge Coupled Device) array
High Rayleigh rejection fiber optic probe
Quiet detection system
Software: BWID, BWSpec, BWIQ

26. Relative Intensity Correction
In Raman spectroscopy, intensity correction is applied to correct relative intensities against a traceable standard for individual spectrometers
For systems with 785 nm excitation use SRM2241
Enable the correction in the software using a factory-generated correction file (Ratio3 file)

27. Permanent Wavelength Calibration
NIST traceable wavelength calibration ensuring Raman shift accuracy with standards- Tylenol and cyclohexane
Follows ASTM E (2007)

28. i-Raman PLus

29. i-Raman Plus Positioning
Research applications
Quantitative analysis
Final product inspection
Counterfeit detection
PAT process applications – proof of concept and at line applications such as dispensing, mixing, granulation, reaction monitoring, etc.

30. i-Raman® Plus Technical Specifications
BWS465
Detector
Back-Thinned CCD Array
Excitation
532 nm
785 nm mW
< 50mW
< 300mW
Range / Resolution
532 H: cm-1 ~ 614nm
785 S: cm-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

31. i-Raman Series System setup

32. i-Raman® Plus
Higher S/N ratio allows for greater specificity of measurement
Greater range than any handheld allows you to see more C-H Stretching
Comprehensive suite of accessories, software, allow versatility and portability
BWIQ allows development of Quantitative Raman methods BWID allows user to develop libraries for
Final Product Identification
Counterfeit Detection
CH stretching
On-board processing includes:
Averaging
Smoothing
Dark Compensation

33. Signal to noise compared to iRaman Plus

34. i-Raman® EX Technical Specifications
BWS485
Detector
TE Cooled InGaAs
Excitation
1064 nm mW
450 mW max
Laser Power Control
0 to 100%
Range / Resolution
cm-1 ~ 1296nm
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

35. i-Raman® EX
Material fluorescence reduces Raman applicability on a small subset of materials – especially those that are highly colored
Utilize 1064 nm excitation
Reduce fluorescence
Increase productivity vs other techniques
Research grade instrument
provides enhanced material
characterization opportunities

36. i-Raman® EX
Comparative spectra for an Alka Seltzer tablet

37. Raman Accessories

38. Sampling
Fiber optic probes can also be easily adapted to a variety of different sampling chambers
Liquid flow cells
Gas flow cells
Optical microscopes
Variety of sampling accessories allows for portability and improved collection efficiency.
Industrial probe option for process applications

39. Fiber Optic Probe Standoff or direct contact with sample
Excitation fiber is filtered to prevent interference from silica’s Raman signal
Collection fiber is filtered to remove wavelength of the excitation laser
Fibers decrease the throughput of a system

40. B&W Tek Fiber Optic Probes
Lab grade – standard
Industrial grade probe
Standoff or direct contact with sample
Sample form: solid, liquid, gas, powder
Industrial immersive version: for process-type applications

41. confidential
Raman Probe Tip
Black distance regulator is for direct measurement of solid samples
Remove distance regulator if going through vial or quartz cuvette
Some samples may burn, depositing soot on probe tip
Clean probe tip with lint free cloth wetted with isopropyl alcohol

42. Cleaning & Care of Fibers
confidential
Cleaning & Care of Fibers
End caps should be placed on fibers when not in use
Leave the fiber probe connected to minimize the risk of contamination of the fiber ends
Do not bend or coil fibers too tightly
Clean fiber ends before connecting to ports for best results
Cleaning is especially critical for Raman probes
Any debris will be “burned” onto fiber tip
Fiber must be re-polished to fix problem

43. Portable Video Microscope Sampling System (BAC151A)
Compatible with all BWT Raman probes
Precise target and focusing
Good for precise measurement of small sample volume
Digital camera and LED illuminator
Coarse and fine XYZ adjustment
Standard objectives from 10x to 100x
Bright and dark field illumination for various sample surfaces
Low cost and portability

44. B&W Tek Raman Probe Holder (BAC150)
Compatible with BWT Raman probes
Coarse and fine XYZ adjustment
Z-axis adjustment allows for laser focusing on the desired plane

45. B&W Tek Enhanced Raman Cuvette Holder (BAC100A)
Precise focusing
Three-point locking mechanism
Increase Raman signal up to 3 times in comparison to a standard cuvette holder
Standard 10x10 mm (inside dimension) cuvette
Liquid or powder sample

46. B&W Tek Liquid Sample Flow Cell
Compatible with BWT Raman probes
On-line process monitoring
Kalrez® O-ring, chemical resistant
Offers high throughput and stability
Quartz or sapphire window options

47. IQ/OQ available for i-Raman series
OQ includes the performance qualification on installation
BWID software has function for performance test
Successfully executed at global pharmaceutical companies

48. Raman Software

49. Chemometric (Spectral) Analysis
The use of mathematical and statistical methods for analysis of chemical or spectral data
Correlation analysis for material identification and/or verification (Qualitative)
Multivariate analysis and complex systems with large data sets (Quantitative / Qualitative)
Uncover underlying trends in the data and detect outliers,
Develop models based on data similarity for classification
Develop quantitative models to predict response in unknown data sets
Allows for real time monitoring and control

50. B&W Tek Software
BWID™ Pharma and BWID ™ Standard: Qualitative analysis by library searching
BWID ™ Pharma developed for use in regulated environment
BWSpec™: general purpose spectrometer software for data acquisition and basic data manipulation and viewing
BWIQ™: chemometric software for quantitative analysis classification and prediction

51. Spectral Analysis Examples
PLS Regression Modeling for the Development of concentration curve for quantitative prediction.
Correlation Based Library Searching for Unknown Material Identification using HQI
Measure 20 representative spectra
Build method based on PCA and establish threshold p-value (typically 0.05)
Test Method 
PC1
PC2
Method Development for verification of “known” materials using multivariate classification modeling to determine p-value.

52. BWID™ Pharma Software
21 CFR Part 11 compliant software with electronic records and signatures
Operator/Developer/Adminstrator users
User-defined and commercial third-party libraries can be used
One click Identification or Verification of sample spectra
Reporting functions for analysis and instrument diagnostics

53. BWID™ - PHARMA User Levels
Operator:
Run sample identification
Perform diagnostic tests
Print results for recently performed ID analysis and performance test
Electronically sign an ID analysis record
Change his/her own password
Developer:
Create/modify a data library
Setup an operation preset method
Configure performance test parameters
Configure ID analysis report
Select, view, and print a report for any ID analysis and performance test
Electronically sign and approve an ID analysis record
Configure graphical user interface (GUI) settings
Configure licensed instruments
Administrator:
Manage user accounts:
View user accounts
Add/lock/unlock/reset/disable a user account
Modify user account properties
Backup/restore user accounts file
Configure access policy

54. Spectrum Search: Identification
Results immediately when run sample or on stored data
MATCH/NO MATCH results
Ability to add new spectrum to library after search

55. Spectrum Search: Verification
Results immediately when run sample or on stored data
To verify must choose product name
PASS/FAIL results
Ability to add new spectrum to library after search

56. BWID™ Audit trail

57. Instrument Diagnostics
Performance test interface and tracking of results
Performance tests include:
Test of USB instrument connection and dark scan
Noise test
Peak position using standard

58. e-signature
Signature for review, approval, rejection available based on user level
e-signature added to printed report

59. BWSpec™
General purpose spectrometer software

60. BWSpec™ Software Interface

61. BWIQTM: Chemometric Software
Classification and Regression
Range of methods and computational algorithms that provide accuracy and speed with reduced memory requirements
State-of-the-art pretreatment methods for spectral smoothing and baseline correction
Enhanced multivariable methods
Vector processing for non-linear datasets

62. BWIQ: DEG in glycerin B&W Tek i-Raman Plus, 785nm Laser
16 samples with diethylene glycol (DEG) in glycerin
DEG concentration from 0 – 48.3%
Integration time 20 second, 300 mW laser power

63. Raman Applications

64. Portable Raman Applications
Product contamination (i.e. alcohol, DEG in glycerin)
Counterfeit drug identification
Forensic analysis
Food and agriculture industry applications
Process Analytical Technology (PAT)
QC: 100% incoming raw material identification
Environmental analysis
Geology and Gemology
Minerals and rocks analysis
Gemstone identification and examination
Art and Archaeology
Biomedical Analysis

65. Applications in the Pharmaceutical Industry
Identification
Verification of the identity of incoming raw materials.
Identification and analysis of counterfeit drug products.
Research
Polymorph screening; material identification; structural elucidation;
Process Analysis
Real time quantitative analysis for process analytical technology (PAT) such as blending, titration, reaction monitoring and polymorphic transition monitoring.

66. Verification of Incoming Raw Materials
Pharmaceutical manufacturing facilities are moving toward 100% inspection of incoming raw materials, to support cGMP and PIC/S guidelines.
Raman is ideal because % of common pharmaceutical ingredients are Raman active. Additionally Raman can measure through common packaging materials such as glass and plastic.
Handheld Raman spectrometers are commonly used to provide on the spot "Pass/Fail" decision to verify the identity of incoming raw materials.

67. Polymorphic screening by Raman
Mannitol has at least three polymorphic forms: alpha, beta, and delta. The beta form is considered to be the most stable
The vapor-induced transition from the less stable δ form to β form can be monitored by Raman spectroscopy and with dynamic vapor sorption
DVS Case Study 615
In-situ Monitoring of a Moisture-Induced Polymorphic Transition using Raman Spectroscopy and Gravimetric Vapor Sorption
Majid Naderi, Jiyi Khoo, Manaswini Acharya and Dan Burnett
Surface Measurement Systems Ltd.
Raman spectra taken at 2-hour intervals for δ D-mannitol transition to β form
DVS Case Study 615:
Majid Naderi, Jiyi Khoo, Manaswini Acharya and Dan Burnett, Surface Measurement Systems Ltd.

68. 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
Raman Spectroscopy is currently being use for not only identification of counterfeit drug products but also to analyze the quality and purity
For example the FDA is currently using portable Raman spectrometers for the identification of glycerin contaminated with DEG.
From Forbes 7/25/2012
Roger Bate, a scholar at the American Enterprise Institute, has spent six years researching fake and otherwise inferior medicines, and his field work has been pulled together in a worthy new book, Phake: The Deadly World of Falsified and Substandard Medicines.  He estimates, probably conservatively, that more than 100,000 people are killed worldwide by dangerous drugs every year.  And that statistic does not take into consideration the incalculable morbidity and misery caused by such products.
Bate describes the widespread distribution of fakes, noting that they are more likely in the poorest locations, whereas substandard drugs — those legally but poorly made — are almost never found in countries with average annual per capita income above $20,000.  Bate and his colleagues sampled thousands of drugs from 22 cities and found that roughly 12 percent failed quality tests and were substandard, degraded or counterfeit.
Bate’s work is more than a detailed analysis; it is also a revelatory first-hand account of the counterfeit drug trade.  His adventures in Nigeria and India are fascinating and his exposure of a Middle Eastern counterfeit ring is alarming, not least because the most recent examples of fake drugs in America probably originated in the Middle East.  An interesting tidbit in Bate’s book is that criminals in the Middle East sold fake drugs in Iraq that were purchased with our tax dollars.
In his book, Bate proposes solutions to this pernicious, worldwide problem.  Among them is the introduction of an international treaty to penalize counterfeits.  He acknowledges that this will take a while to implement, but he calls it a necessary condition to combat the fake drug trade.  Bate also suggests track-and-trace technologies and the deployment of handheld spectrometers to spot fakes. Although these are expensive, he explains how they are used even in impoverished Nigeria and argues that if the Nigerians can do this, then so can other richer nations.  (There are other, easy-to-use high-tech devices for this kind of “molecular tracking” as well, which via a unique “barcode” determined by ratios of radioisotopes found naturally in the drug, can ascertain whether any given batch of drug is from one or another legitimate manufacturing site.)
Bate argues persuasively that ”[u]ntil international bodies clean up their act and stop blurring the boundaries between safe medicines and sub-standard copies and until there are harsh local and international penalties for manufacturing and carrying counterfeits, the pirates will continue to get away with murder.”  Literally.
Best 785

69. Applications in Forensic Analysis
Nondestructive Narcotic Drug identification
Explosives Identification:
Exact Chemical Compositions of Material
(i.e. PETN, RDX)
Binding Agents Within Explosive Materials
Identification and Analysis of Toxic Solvents and Chemical and Bio-warfare Agents
Trace Forensic Evidence Analysis:
Including fibers, fabrics, pigments, inks, paint chips, etc.

70. Illicit Drug Analysis
Portable Raman spectrometers are commonly used for the Identification of a frequently encountered illicit street drug.
In the example to the right we are showing the spectrum of a confiscated white powder which was analyzed using correlation analysis against a spectral library of common illicit drugs and positively identified at cocaine with a HQI of

71. Identification of Explosives
Portable Raman spectrometers are also well suited for the identification of explosives and hazardous materials.
Typically surface enhanced Raman spectroscopy (SERS) is utilized for this application because it allows for the detection of trace levels of explosives.
For example in a recent publication it was shown that PETN could be detected at concentrations as low as 5 pg.
SERS Spectra of PETN
0.2 g
200 pg
5 pg
Talk about SERS in more detail.
SEM image of SERS substrate used in the measurement.
S. Botti et al., J. Raman Spectrosc. 2013, 44, 463–468

72. Product Contamination – Methanol-Laced Spirits
Over the past several years an alarming trend has become evident that there are serious issues with contaminated alcohol within the EU, and in particular Eastern Europe.
Studies have shown that the maximum tolerable concentration of methanol in alcoholic beverages with about 40% alcohol is about 2% (v/v) by volume.
In September of 2012 when the Czech Republic banned the sale of hard liquor after 20 people died from the consumption of methanol-laced spirits.
After an exhaustive study of different screening tools the Czech Republic turned to the use of portable Raman spectroscopy as the screening tool of choice for the identification and quantification of methanol in contaminated spirits.
Methanol, a potent toxicant in humans, occurs naturally at a low level in most alcoholic beverages without causing harm. However, illicit drinks made from “industrial methylated spirits” [5% (v/v) methanol:95% (v/v) ethanol] can cause severe and even fatal illness. Since documentation of a no-adverse-effect level for methanol is nonexistent in the literature a key question, from the public health perspective, is what is the maximum concentration of methanol in an alcoholic drink that an adult human could consume without risking toxicity due to its methanol content? Published information about methanol-intoxicated patients is reviewed and combined with findings in studies in volunteers given small doses of methanol, as well as occupational exposure limits (OELs), to indicate a tolerable (“safe”) daily dose of methanol in an adult as 2 g and a toxic dose as 8 g. The simultaneous ingestion of ethanol has no appreciable effect on the proposed “safe” and “toxic” doses when considering exposure over several hours. Thus, assuming that an adult consumes 425-ml standard measures of a drink containing 40% alcohol by volume over a period of 2 h, the maximum tolerable concentration (MTC) of methanol in such a drink would be 2% (v/v) by volume. However, this value only allows a safety factor of 4 to cover variation in the volume consumed and for the effects of malnutrition (i.e., folate deficiency), ill health and other personal factors (i.e., ethnicity). In contrast, the current EU general limit for naturally occurring methanol of 10 g methanol/l ethanol [which equates to 0.4% (v/v) methanol at 40% alcohol] provides a greater margin of safety.
Source: doi: / A.J. Paine and A. D Dayan , Hum Exp Toxicol November (11)

73. Example using Methanol-Laced Coconut Rum
CH3 bending vibration at 1013 cm-1 increases with methanol concentration. A PLS regression method can be developed to readily measure concentration of MeOH in alcoholic beverage by portable Raman spectroscopy

74. Applications in Art & Archeology
Portable Raman spectroscopy is widely used for the analysis of paintings, ceramics, statues (surface coatings), and other artifacts.
The flexibility of fiber optics in conjunction with the non-destructive & non-contact nature of Raman allows measurements to be taken in-situ.
This application has become so popular that in 2001 a biennial international conference was formed solely dedicated to the subject of the use of Raman spectroscopy in art and archeology.
Painting on cathedral ceiling, pigments on a budha, cave paintings, old artwork, coatings on sculptures

75. Portable Raman Microscopy of Ancient Pigments
Analysis of pigments on the ceiling of a cathedral in Spain using a portable Raman spectrometer connected to a tripod-mounted video microscope for precision alignment.
Courtesy of M.J. Ayora Cañada y A. Dominguez Universidad de Jaén
Lead oxide (Pb3O4)
Cinnabar (HgS)

76. Analysis of metal corrosion
Raman spectroscopy was used at the Guggenheim Museum in Bilbao Spain to analyze environmental effects on the corrosion of seven steel sculptures from Richard Serra.
The figures below show the presence of lepidocrocite, magnetite, and iron sulfate from Richard Serra’s Inverse Blind Point sculpture, as well as a SEM image of the surface confirming the results of the Raman analysis
K. Castro et al., COST Action D42 Extended Abstract. 2011, 113–116

77. Applications in Geology and Mineralogy
Portable Raman spectrometers are ideal for the identification of gemstones and minerals, including polymorphs and isomorphs.
Non-contact, non-destructive sampling allows for analysis of precious or scarce samples, unlike other techniques such as LIBS.
Anti-counterfeiting of precious, such as identification of diamond from zircon
Images Courtesy of Prof. Rull University of Valladolid

78. Raman evaluated for Geological survey on Mars
Before testing for life on other planets, feasibility studies are done on barren areas of the Earth. One such place is Rio Tinto in Spain, where conditions are analogous to Mars, where portable Raman spectrometers were evaluated for the joint NASA/EAS 2018 Mars rover mission.
Images Courtesy of Prof. Rull University of Valladolid

79. Analysis of Garnet Gemstones
Garnets are a class of silicate minerals which include a number of varieties with the general form X3Y2(SiO4)3.   
Raman spectroscopy’s high selectivity allows for the differentiation of the different garnet varieties. Andradite and grossular fall into the ugrandite group of garnets (calcium in X site), while spessartine falls into the pyralspite group (aluminum in Y site).    

80. 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.

81. Process Analytical Technology
Near Infrared Spectroscopy for PAT NIR2011
Process Analytical Technology
The use of rapid off-line, at-line, on-line or in-line analyzers to obtain analytical data giving the following benefits:
Faster (real-time or near real-time)
Data collected more often
Higher precision data
Provides the pulse of a process
One example is Raman spectral change during HSWG (High Shear Wet Granulation) polymorphic transformation of a drug substance.
Raman bands for Polymorph form B appear and for form A disappear during the process
NIR Katherine Bakeev

82. Epoxy Cure Monitoring
5 mW 375 nm 30 s

83. Process Application: TiO2 phase transitions
TiO2·H2SO4·H2O  TiO2·H2SO4 TiO2 (nanophase)  TiO2 (anatase)  TiO2 (rutile)

84. Polymer Reaction Monitoring
C=C
Breathing mode (for normalisation)
styrene -> polystyrene
Follow the conversion of the vinyl band
Henryk Herman, Ph.D. – Gnosys Global

85. Food & Agriculture Industry
Measuring chain length and extent of saturation of fatty acids in edible oils
Meat product quality analysis
Product contamination
SERS analysis of food contaminants including bacteria, antibiotics, dyes, etc.
Analysis of components in grain kernels
Raw material identification/verification for the food and beverage industries

86. Contamination of extra virgin olive oils
To improve profit margins there are many instances of the adulteration of extra virgin olive oil (EVOO) with light olive oil or other cheaper ingredients
A feasibility study was performed with 8 samples spiked with % and measured on the i-Raman EX
BWIQ software used to develop a quantitative model for light oil in EVOO
2 factor PLS regression over full range of raw Raman spectral data
2 factor PLS model
Error ~ 1.6%

87. Detection of Melamine in Gluten Using SERS
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).
Structure and SEM image of a KlariteTM SERS active gold substrate

88. Raman Analysis of Carbon Nanotubes
RBM – Radial breathing modes, which probe the lattice structure of the CNT allowing for the calculation of tube diameter.
D Band – Disorder band, measures the degree of amorphism of the CNT.
G Band – Tangential Mode, measures the degree of crystallinity (diamond like structure) of the CNT.
RBM G D
MWCNT
SWCNT

89. Applications in the Biomedical Diagnostics
Raman spectroscopy is becoming more pervasive in biomedical diagnostics because of the demand for near real time and minimally invasive analysis. Applications include: biopsies, cytology, drug efficacy studies, histopathology, surgical targets and treatment monitoring.
Some of the most active research areas are the analysis of abnormalities in tissue samples such as brain, arteries, breast, bone, cervix, embryonic media; and the identification of biomarkers for early stage detection of various diseases.
Raman has also been used to investigate blood disorders such as anemia, leukemia and thalassemias (inherited blood disorder), as well as understanding cell growth in bacteria, phytoplankton, viruses and other micro-organisms.
There are a number of important functional groups related to biomedical testing, which have characteristic Raman frequencies. Tissue samples include components such as lipids, fatty acids and protein all of which have vibrations in the Raman spectrum. The most significant spectral regions include:
X-H Bonds (e.g. C-H stretches): cm-1 region
Multiple Bonds (e.g. N≡C): cm-1 region
Double Bonds (e.g. C=C, N=C): cm-1 region
Complex Patterns (e.g. C-O; C-N and bands in the fingerprint region): cm-1 region

90. Assessment of Axillary Lymph Nodes in Breast Cancer with Raman Spectroscopy
Studies by researchers in the UK have shown that Raman spectroscopy can detect differences in tissue composition, and in particular a wide range of cancer pathologies.
Using PCA-LDA data processing to evaluate the Raman spectra of lymph nodes they have confirmed that the technique can match the sensitivities and specificities of current histopathological techniques.
(principal component analysis- linear discriminant analysis)
Images Courtesy of Jonathan Horsnell,
Gloucester Royal Hospital Department of Biophotonics

91. Applications in Polymer Science
Portable Raman spectroscopy helps to meet needs of the polymer, additive, compounding and masterbatch industries, by allowing for an audit trail to certify the link between the quality of raw materials and finished product.
Raman has many applications in both QA and QC including incoming material inspections, measurement of polymer grade, blend ratios, additives, and ageing.
It can aid in the optimisation of formulations for desired properties and performance by predicting:
Processing properties – MFI, liquid viscosity
Thermal properties – glass transition, melting point
Physical properties – density, mechanical modulus and strength, impact strength
Fire properties – UL scores, LOI, flame retardants

92. Polymer Crystallinity
These bands are related to crystalline and amorphous content, (as well as the interphase)
Dr. Henryk Herman - GnoSys Global

93. Polymer Identification

94. Environmental Science
Water pollution detection using SERS technology
Identification of contaminants in water
Petrochemical analysis
Identification and analysis of sediments in water

95. Environmental Science
Calcium Carbonate condensation in water system

96. In Situ Contaminant Study (quoted)
Field-portable Raman spectrometer reported under DOE
Waste site measurement conducted
Spectral database of over 200 contaminants

97. Thank you for your attention
B&W Tek, Inc.

98. BACK UP SLIDES

99. Energy Diagram for Raman Scattering
Stokes Raman
Rayleigh
Anti-Stokes Raman
Fluorescence
Electronic States
Raman Scattering:
Independent of laserλ
Vibrations → shifts
Intensity ∝ laser power
Raman efficiency10-8
Virtual State
Vibrational States
Ground State

100. Quantum Energy Scheme for Photon Scattering
The Raman effect comprises a very small fraction (about 1 in 107) of the incident photons.
7/28/2011

101. Information From Raman Spectroscopy

102. Components of a Raman Spectrometer
Laser
Narrow Linewidth
Small Form Factor
Low Power Consumption
Extremely Stable Power Output
Spectrometer
High Resolution
Low Noise
Small Form Factor
Low Power Consumption
Glacier T
BWN-OEM

103. Importance of Rayleigh Rejection
Since the Rayleigh scattering can be up to 10,000,000 times stronger than the Raman scattering, it is imperative that it be filtered out.
The quality of the Long-pass filter will determine the wavenumber cut-on of the spectrometer (e.g. 175cm-1or 65cm-1)
The anti-Stokes scattering does not provide any additional information (except in thermal studies) so it is also filtered out.

104. CCD Detector Arrays Linear CCD Arrays Back Thinned CCD
Single Row of Pixels
Mid Light Level Applications
Low Cost
Low Dynamic Range
High Read Noise
Mid Readout Speed
Back Thinned CCD
Multiple Rows of Pixels
Low Light Level Applications
Higher Cost
High Dynamic Range
Low Read Noise
Slow Readout Speed
Revolution in the evolution
phosphor coatings on front-illuminated CCDs

105. Stabilized Laser
Unstable Laser Diode
VBG Stabilized Laser Diode
VBG stabilization results in the correct center wavelength and avoids the phenomenon of “mode hopping”, this insures that the resultant shift is the same each measurement. It is also essential that the laser has a narrow bandwidth laser since resolution of the Raman peaks is directly correlated to the linewidth of the laser .
VBG = Volume Bragg Gratings; this slide is related to Clean Laze patent
Raman Spectrum
Raman Spectrum

106. Optimized Fiber Optic Probe Design
Optimized 6-8 orders Rayleigh rejection
65 cm-1 cut on version
Excitation fiber filtered to prevent silica’s Raman signal
On probe trigger option

107. Raman Pros and Cons Applicable to aqueous solutions
Less demanding to optics
Easy sampling for solids
Weak signal (efficiency  10-6)
Fluorescence interference

108. Mid-IR NIR reflectance Raman
Mechanism: absorption by fundamental molecular vibrational modes
Molecular Specificity: high
Signal Strength: strong
Energy: cm-1
Sampling: direct material contact required, glass and water appear opaque; can rarely see through container materials
Hardware: probes expensive and fragile; typically short in length, and inflexible
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: ,500 cm-1
Sampling: non-contact, but close proximity to material required (<5 mm)
Hardware: longer fiber optics possible (meters); quartz optics; dispersive, interferometry common
Mechanism: inelastic scattering by vibrational, rotational, low frequency molecular modes.
Energy: cm-1 shift
Sampling: container interference is mitigated by accessories
Hardware: CCD detection; quartz optics; long fiber optics common (kilometers in some cases)

109. BWID™

110. Administrator Create and edit account of all levels
Back up of account information
Set account policy (i.e. password, user name requirements

111. Data acquisition Settings
Set data acquisition parameters including:
Laser power
Integration time
Dark scan settings
Relative intensity correction

112. BWID™ Operation Setup Choose library to search
Search Algorithm settings for:
Method
Preprocessing
Smoothing
First derivative
Second derivative
Minimum HQI
Number of Hits

113. Identification in BWID™
Can be used with ST Japan spectral library or user-defined libraries
License key needed for software activation
Additional license for ST Japan

114. User-Defined Product Input
Barcode Scan
Manual Entry of fields defined in the Operation Presets

115. Analysis Records
Great benefit for system validations and data traceability
Reporting: save, view, and print the analysis report and line for signature

116. Data Archive

117. BWID™ Software Data Data formats
All output files are binary files - noneditable
Data can be saved as *.spc files (Thermo GRAMS)
User-defined and commercial libraries (ST Japan) can be used
Data and libraries can be on server
Requires mapping to server

118. 3rd party Raman libraries for BWID
Library
No. of Spectra
Raman Database - Complete Collection
8694
Polymers & Polymer Additives
929
Food Additives And Food Packaging
1075
Solvents
460
Biochemicals
1900
Aldehydes And Ketones
1079
Alcohols And Phenols
891
Esters, Lactones, and Anhydrides
2935
Hydrocarbons
565
Flavors, Fragrances, Cosmetic Ingredients
1030
Pesticides
468
Semiconductor
371
Forensic
748
Dyes, Pigments, Stains, Indicators
300
Sulfur and Phosphorous Compounds
977
Hazardous Chemicals
1361
Hazardous and Toxic Chemicals
3035
Pharmaceuticals
1172
High Production Volume (HPV) Chemicals
697
Minerals And Inorganic Materials (wavenumber range cm-1)
1419

119. BWIQ™
Chemometrics software for both quantitative and qualitative analysis
File formats supported
BWTek txt, GRAMS spc, csv, MAT
Sample Selection
Random, Kennard-Stone, SP X-Y
Spectral Preprocessing
airPLS Baseline Correction
Smoothing (Savitzky-Golay, FFT, Whittaker)
Derivatives (1st and 2nd; S-G, differential)
Mean centering
Regression algorithms
MLR, PCR, PLS1, PLS2, Support Vector Machine Regression
Classification algorithms
PLS-DA, PCA-MD, SIMCA, SVM-C
adaptive iteratively reweighted Penalized Least Squares (airPLS) [1] is a novel algorithm for baseline correction in Ranam spectroscopy, which does not require any user intervention and prior information, such as peak detection etc.. The method works by iteratively changing weights of sum squares errors (SSE) between the fitted baseline and original signals, and the weights of the SSE are obtained adaptively using the difference between the previously fitted baseline and the original signals. The baseline estimator is fast and flexible.
airPLS basically consists of two aspects:
Penalized least squares algorithm for signal smoothing
Adaptive iteratively reweighted procedure which changes penalized least squares algorithm into a baseline estimator
[1] Zhang, Z. M., Chen, S., and Liang, Y. Z., Baseline correction using adaptive iteratively reweighted penalized least squares. The Analyst 135 (5), 1138.

120. NanoRam Technical Specifications
BWS456
Detector
TE Cooled linear CCD Array
Laser Excitation
785nm
Laser power
tunable from 0 mW to 300 mW
Laser Power Control
0 to 100% in 10% increments
Range / Resolution
176cm-1 to 2900cm-1 ~ nm
Display
High Visibility OLED touch screen, 3.7" size
Barcode Reader
1D and 2D barcodes
Data Formats
.txt, .csv, .spc, .pdf
DC Power
12V DC at 6.6 Amps
Battery
Rechargeable Li-ion Battery, > 5 hours operation
PC Software
NanoRam ID
Computer Interface
USB to Ethernet, Wifi
IP Protection
Class
IP64 - Protection from infiltration of dust and splashing water

121. Inspection of Raw Materials
Pharmaceutical manufacturing facilities are moving toward 100% inspection of incoming raw materials.
NanoRam allows rapid confirmation of the content of each container at the molecular level.
Raman spectra can be acquired in warehouse through transparent packaging materials, bags and bottles.
"Pass/Fail" decision
Supports cGMP and 21 CFR Part 11

122. NanoRam Incoming non-destructive testing of Raw Materials
Reducing Waste in the Dispensing Suite
Plastic Packaging Identification
At line identification of intermediates
Final product identification
Substandard, Adulterants, and Counterfeit Deterrent