Innovations in FTIR and UV-Vis: S olutions for your Chromatography Success

1. Innovations in FTIR and UV-Vis:Solutions for your Chromatography Success Agilent Technology and Science Symposium, Houston, 06/27/2013 Michelle Tomba Product Specialist Molecular Spectroscopy

2. 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

3. 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

4. Agilent Growing MolecularSpectroscopy Portfolio

5. 70 um 70 um 0.2 Absorbance 0.1 0.0 3500 3000 2500 2000 1500 1000 Wavenumber Chemical Image Image @ 1285 cm-1, poly phthalate, ~4 microns WHY FTIR? Image @ 3295 cm-1 - Nylon

6. Why most chemists are avoiding FTIR Issues 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

7. Why most chemists used to avoid FTIR Issues 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

8. FTIR benefits: example of applications • Identification (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

9. Cary 630 FTIR – Innovation on all aspectsSolving problems through innovation… DialPath TumblIR 630 Engine Transmission Diff/Spec Reflectance Di, Ge, ZnSe ATR Agilent Confidential

10. Cary 630 FTIR – Anywhere and with ease of useSize, reliability, consistency and robustness • Toxic API (ex: anticancer drugs) • Air sensitive compounds • Explosive/Hazard risks

11. Cary 630 with ATR: Identification of unknowns Calcium Oxalate Uric Acid Medical Synthesis Methanol Salicylic acid Methyl salicylate

12. 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 Cary 630 with ATR: QA/QC of SBR Polymer

13. 250mL Fill Volume 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 3mL Fill Volume

14. Cary 630 with DialPath Sampling Technology As easy for liquid measurement as using ATR, and with the added benefit of variable and extended pathlength Oil/lubricants Environmental

15. Dialpath – Raw Material Identification 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) Spectra collected with 30umDialPath setting

16. Dialpath – Determination of impurities ethylene glycol in glycerol diethylene glycol in glycerol R2=0.9745 R2=0.9895 Limit of detection – 0.02% Limit of detection – 0.04% This fits well with FDA limits of <0.1% impurities in glycerol Spectra collected with 75umDialPath setting

17. 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.

18. FTIR Microscopy: spatial and chemical information IR Image @ 1000 cm-1 (10 microns) 2.2 um

19. 7.6 mm 5.0 mm FTIR Imaging of Sampled Fingerprint Image created based on 1640 cm-1, amide I (protein) absorbance

20. Detection of illicit drugs on Fingerprint COCAINE ATR FTIR imaging in forensic science, Agilent

21. Continuous innovations inUV-Vis SPectroscopy

22. 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

23. UV-Vis Spectroscopy in the Laboratory • 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

24. Cary 60 with Fiber opticFlexibility for increased throughput Chemical identification or study of chemical processes QA/QC on raw materials and finished product in manufacturing Wavelength Scans – What is it? Wavelength Scans – What is it? Fiber Optics Kinetics – what processes are occurring? Concentration – How much?

25. Cary 60 - Fiber Optic Probes Replaceable Tips Various selection of pathlengths Cleanable and reusable. Microprobes Diameter: < 4 mm 10mm (2-5mm option) pathlength

26. 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 – noexpensive quartz cuvettes needed • No compromise in data quality: Abs DNA concentration (ng/µl)

27. Cary 60 UV-Vis – Nano/Microvolumes • Accurate spectro measurements • Small 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 Abs 1 Abs mg/mL

28. Cary 60 – Surface measurements • Non-destructive testing Requested by the Art Restoration Community Confidentiality Label • 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

29. Innovation for Advanced Surface Measurements • Nothing 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. Confidentiality Label

30. Questions? 30