CPAC Webinar Feb. 2009Process Spectroscopy and

Optical SensingBrian Marquardt Ph.D.Director – Applied Optical Sensing LabApplied Physics LabUniversity of WashingtonSeattle, Washington 98105

Process Raman Applications Pharmaceuticals Food quality and safety Polymers/coatings Fermentation/biotech Cellular/tissue Oil/fuels/petrochemicals Oceanography/environment Challenges

Reproducible sampling Fluorescence

Quantitative Raman = Effective Sampling

• no moving parts• sapphire spherical lens• constant focal length and sample volume• probe is ALWAYS aligned when in contact with sample• effective sampling of liquids, slurries, powders, pastes and solids• high sampling precision allows it to be used effectively to monitor dynamic mixing systems (powder/slurry/particle)•improved measurement precision leads to robust multivariate calibration of process Raman data

CPAC developed, patented and licensed Raman ballprobe

Real-time FermentationMonitoring

Image from Purves et al., Life: The Science of Biology, 4th Edition

Yeast Fermentation Process

Analysis of a Batch Fermentation Process

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Raman Shift (cm-1)

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nsity

Fermentation Raw Raman Spectra

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Fermentation Raw Raman Spectra

• 10 second acquisition, 20 accumulations, sample every 10 minutes • Analysis was run continuously for 8 days

Raw Raman Data for Fermentation Batch Reaction (8 day run)

Fluorescence

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-500

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Raman Shift (cm-1)

Flu

ore

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rre

cte

d I

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ity Fermentation #3 Raman Fluorescence Corrected Spectra

Raman Data After Fluorescence Correction Algorithm Applied

Brian Marquardt - confidential

Raman Shift (cm-1)

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nsi

ty

Maltose

Ethanol

Maltose

3D Plot of fluorescence corrected Raman fermentation data

Raman Analysis and Optical Trapping of Single Cells

Raman Microscope and Instrument Raman Microscope and Microchip

Micro-reactors and Raman lead to improved understanding and control

O

NO

NOO

ON

O

O

NO

N

O

O

O

NO

NOO

+ ++ +H2SO4

HNO3

2-nitrotoluene 4-nitrotoluene 2,4-dinitrotoluene 2,6-dinitrotoluene3-nitrotoluene

Raman Shift (cm-1) 500 1000 1500 2000

2,4-dinitrotoluene

2,6-dinitrotoluene

2-nitrotoluene

3-nitrotoluene

4-nitrotoluene

nitric acid

sulfuric acid

toluene

• without residence time module• flow rate: 0.89 ml/min (residence time ~ 5 min)

PCA Analysis on data after mixing:1st PCA scores Increase in reaction yield after each temperature step

Real-time Understanding and Control

CPAC/FDA/Corning MicroReactor

Goal: to improve reaction development and optimization through the use of continuous glass microreactors, NeSSI and analytics

Funded by the FDA to demonstrate the benefits of improved reactor design, effective sampling and online analytics to increase process understanding (QbD)

QbD Project began November 2008

What is NeSSI?• Industry-driven effort to

define and promote a new standardized alternative to sample conditioning systems for analyzers and sensorsStandard fluidic interface

for modular surface-mount components

Standard wiring and communications interfaces

Standard platform formicro analytics

What does NeSSI Provide Simple “Lego-like” assembly

Easy to re-configure No special tools or skills required

Standardized flow components “Mix-and-match” compatibility between vendors Growing list of components

Standardized electrical and comm. (Gen II) “Plug-and-play” integration of multiple devices Simplified interface for programmatic I/O and control

Advanced analytics (Gen III) Micro-analyzers Integrated analysis or “smart” systems

NeSSI Gas Generation System

Fully automated gas generation system for sensor calibration:

1.4 Stage dilution, able to produce and maintain gas concentrations of 100% to 0.1%(1000 ppm) from standard bottle gas2.Fully calibrated, automated system with set and forget capability

Automated Circor NeSSI Gas/Vapor System

N2

O2

Mixed Gases

Mass FlowControllers

Small Optical SensorsOxygenMoistureAmmoniaHydrogenCommon Solvents

AlcoholsEstersAmines

Chlorinated OrganicsOrganic Hydrocarbons

(BTEX)Carbon Dioxide

(in development)Hydrogen Sulfide

(in development)

• vapochromic chemistry• optical response to analytes

• simple design• reversible response• low power• inexpensive• fast response times• high quantum efficiency• long term sensor stability• sensitive to a variety of analytes• wireless communication• battery powered

Vapochromic Humidity Sensor

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per10.Mas

per20.Mas

per30.Mas

per50.Mas

per70.Mas

per80.Mas

Elements:Slope:Offset:Correlation:RMSEC:SEC:Bias:

60.9998240.0075850.9999120.3423230.374996

-1.272e-06

Measured Y

Predicted Y- Measurment time – 100 ms- 3 reps per concentration

Sensor response to O2 Gas

Pre

dict

ed O

2 %

Calculated O2 %

R2 = 0.990, 3 PCRMSEC = 1.0744

20 replicates at each concentrationConcentration range: 0 -100% Oxygen

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ty (

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Low Concentration Dissolved O2 Calibration

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Y Predicted 1

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easu

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Samples/Scores Plot of CTFOS5320to1300

R2 = 0.9972 Latent VariablesRMSEC = 0.67861

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31

R2=0.997, 2 PCs, RMSEC= 0.67861

5 replicates at each concentrationConcentration range: 1 μmol/L - 39 μmol/L

Pre

dic

ted

[O

2] (

μm

ol/

L)

Measured [O2] (μmol/L)1 μmol/L = 32.5 ppb

39

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39 μmol/L

1 μmol /L

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LIBS: Remote Elemental Analysis

• Remote elemental analysis with no sample preparation

• Fiber-optic delivery or long range delivery of laser by telescope for remote analysis

• Laser-induced plasma ablates and super heats samples to provide elemental spectral data

Potential Applications Analysis of metal complexes in food, cellulosic biomass,

pharmaceuticals and fermentation apps. Determination of ionic and inorganic species in a variety of

chemical/production processes Glasses, ceramics, zeolites, alloys, corrosion analysis

Quantitative analysis of catalyst composition for screening and development

Couple with vibrational techniques to develop a hyphenated technique (Raman/LIBS) to define both organic and inorganic analytes in a process system

Acknowledgements

CPAC Washington Tech. Center National Science Foundation National Institute of Health, Charlie Branham and Wes Thompson Many current and past CPAC sponsors