Innovations in FTIR and UV-Vis:

Solutions for your Chromatography

Success

Agilent Technology and Science Symposium, Houston, 06/27/2013

Michelle TombaProduct SpecialistMolecular Spectroscopy

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Outline

• Infrared spectroscopy - FTIR– Cary 630 FTIR – true innovation!

• Why you won’t be missing your old bench • Sampling technologies & Software• Applications

–FTIR microscopy – eye opener• Applications

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Outline

• UV-Vis/NIR Spectroscopy– Cary 60 & 8453 – efficient UV-Vis

• When sample size does not matter anymore • Flexibility

– New platform for solids – nothing better…• Principle

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Agilent Growing Molecular Spectroscopy Portfolio

WHY FTIR?

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70 u

m

70 um

Chemical Image

Image @ 3295 cm-1 - Nylon 3500 3000 2500 2000 1500 1000

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Image @ 1285 cm-1, poly phthalate, ~4 microns

Why most chemists are avoiding FTIRIssues related to instrumentation

• Not stable (vibration, conditioning, humidity…)• Alignment of optics• Requirement for purge gas• Bulky• Long acquisition time (minutes)

Issues related to sample preparation• KBr pellets, NaCl plates -> long, costly, non-

reproducible, humidity• Liquid cells -> tedious, tricky handling• Difficult to run unstable samples

Issues related to software• For spectroscopists– requirement for data

manipulation

Solution implemented:• Dissolve/heat/extract and go to LC/GC/MS

Why most chemists used to avoid FTIRIssues related to instrumentation

• Not stable (vibration, conditioning, humidity…)• Alignment of optics• Requirement for purge gas• Bulky• Long acquisition time (minutes)

Issues related to sample preparation• KBr pellets, NaCl plates -> long, costly, non-

reproducible, humidity• Liquid cells -> tedious, tricky handling• Difficult to run unstable samples

Issues related to software• For spectroscopists– requirement for data

manipulation

Solution implemented:• Dissolve/heat/extract and go to LC/GC/MS

FTIR benefits: example of applicationsIdentification (major, minor, impurities)

• Incoming material testing• Finished products• Packaging, o-rings• Forensics/out-of-lab• Surfaces (identification of coatings)

Quantitation• Solid substrates: polymer ratios, API• Liquids: food, additives, alduteration, solvent additives…• Surfaces (cleanliness, thickness, aging/oxidation…)

Molecular information• Molecular changes• Synthesis• Kinetics

Cary 630 FTIR – Innovation on all aspectsSolving problems through innovation…

630 EngineDialPath TumblIR

Di, Ge, ZnSe ATRDiff/Spec Reflectance Transmission

Agilent Confidential

Cary 630 FTIR – Anywhere and with ease of useSize, reliability, consistency and robustness

• Toxic API (ex: anticancer drugs)

• Air sensitive compounds• Explosive/Hazard risks

Cary 630 with ATR: Identification of unknowns

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Calcium OxalateUric Acid

MethanolSalicylic acidMethyl salicylate

Medical

Synthesis

• As styrene (PS) concentration in SBR increases, polymer material becomes harder, but less elastic

• To develop a quantitative method for the level of PS available, 4 standards were measured with increasing levels of PS– 0% (maroon), 5% (red), 23% (green),

45% (blue)

• Peak height absorbance of 700 cm-1 band was used for developing quantitative method

1031

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69945pct styrene butadiene copolymer num 199, cary 630 atr rep1_2011-06-29t20-25-27(1)

995

964

911

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Cary 630 with ATR: QA/QC of SBR Polymer

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DialPath Changes How Liquids are Analyzed by FTIR• Qualitative (raw material ID) and

quantitative (impurity levels determination) analysis on a SINGLE accessory – no need for disassembling of traditional liquid transmission cell to change pathlengths

• Viscous liquids analyzed without blockage and leakage issues typically associated with traditional variable transmission cells

• Handles viscous liquids and volatile solvents equally well

• As easy to use as ATR, but more sensitive with 30-1000 micron pathlengths available

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250mL Fill Volume

3mL Fill Volume

Cary 630 with DialPath Sampling TechnologyAs easy for liquid measurement as using ATR, and with the added benefit of variable and extended pathlength

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Oil/lubricants

Environmental

Dialpath – Raw Material IdentificationSpectra collected with 30um DialPath setting

glycerol

propylene glycol

dipropylene glycol

triacetin

Spectra, are then searched against reference spectra in a library to produce matches scaled from 0 to 100 (with 100 being a perfect match)

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Dialpath – Determination of impurities

Spectra collected with 75um DialPath setting

ethylene glycol in glycerol diethylene glycol in glycerol

R2=0.9895 R2=0.9745

Limit of detection – 0.04% Limit of detection – 0.02%

This fits well with FDA limits of <0.1% impurities in glycerol

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Intuitive: Routine FTIR must have flexible, simple to use (intuitive) software

Software guides the user through

the selected method. Also recognizes the

correct sampling interface to guide proper sampling techniques and

cleaning.

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FTIR Microscopy: spatial and chemical information

2.2 um

IR Image @ 1000 cm-1 (10 microns)

FTIR Microscopy

Transmission, reflection, ATR

Near IR, Mid IR, Far IR

Spatial and Chemical heterogeneity

FTIR Imaging of Sampled Fingerprint

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3351.670 12.5502927.470 0.116

1643.730 0.159Row = 22 Col = 89

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Image created based on 1640 cm-1, amide I (protein) absorbance

SD00223 MILK-PROTEIN HYDROLYSATE PEPTONE

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COCAINE HCl in KBr

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Detection of illicit drugs on Fingerprint

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COCAINE

ATR FTIR imaging in forensic science, Agilent

CONTINUOUS INNOVATIONS INUV-VIS SPECTROSCOPY

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UV-Vis benefits: example of applications

QA/QC – raw materials and chemical analysis

Quantitative formulation studies

Stability studies

Drug binding assays

Kinetics measurements and reaction monitoring

8453 Key Features…• Photodiode array detector for complete spectrum capture• Fast full spectrum kinetics• Multi-component analysis• Small footprint – open sample compartment

Cary 60 Key Features…• Fiber optics for improved workflow• Analysis of small volumes• Xenon flash lamp for low ongoing costs of ownership• Photometric range (4.0 Abs with cuvette)

for turbid solutions

UV-Vis Spectroscopy in the Laboratory

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Cary 60 with Fiber optic Flexibility for increased throughput

Fiber Optics

QA/QC on raw materials and finished product in manufacturing

Chemical identification or study of chemical processes

Wavelength Scans – What is it?

Kinetics – what processes are occurring?

Wavelength Scans – What is it?

Concentration – How much?

Cary 60 - Fiber Optic Probes

Replaceable Tips Various selection of pathlengths Cleanable and reusable.MicroprobesDiameter: < 4 mm10mm (2-5mm option) pathlength

Cary 60 UV-Vis with Fiber Optics Microprobe for small samples at low temp

Fiber Optics for improved workflow- Take the instrument to the sample and improve workflow- Measure samples in situ: hot, cold, radioactive, noxious- Full UV and Vis spectrum – no expensive quartz cuvettes

needed- No compromise in data quality:

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f(x) = 0.0197656250762911 xR² = 0.9997749096739

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Cary 60 UV-Vis – Nano/Microvolumes Accurate spectro measurementsSmall volume measurements -

Traycell- Save precious sample – measure 0.5 to 10 µL- Accurate and reproducible results - Excellent linear dynamic range- Flexibility – no additional dedicated system

required

mg/mL

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April 22, 2023Confidentiality Label

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• Remote (<1.5m from instrument) diffuse reflection measurements! • Fiber optic accessory with movable detector• 1.5 mm light patch – inbuilt CCD allows visualization of position• Needs fiber optic coupler (G6865A – for Cary 60)• Understand the sample surface porosity ie small beam

Requested by the Art Restoration Community

Cary 60 – Surface measurementsNon-destructive testing

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Innovation for Advanced Surface MeasurementsNothing similar: %T, %R, angle, polar.Cary UMS

Analyze virtually any sample.

Measure transmittance at any angle, and measure absolute reflectance without moving the sample - at any polarization.

Perform reflection, and transmission measurement at variable angles on the Cary UMS – unattended.

Questions?

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