AWACSS (EVK1-CT-2000-00045)ENVSENS, Lecce, Italy, 2002 AWACSS Dr. Dale M. Willard, Universität...

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SAWACSS (EVK1-CT-2000-00045) ENVSENS, Lecce, Italy, 2002

AWACSSAWACSS

Dr. Dale M. Willard, Universität Tübingen (Gauglitz Group)Contract #: EVK1-CT-2000-00045

Duration: 3 Year, RTD Project (Starting March, 2001)Visit: http://barolo.ipc.uni-tuebingen.de/awacss

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Automated Water Analyser Computer Supported System

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MotivationMotivation

More Effective Water Management More Effective Water Management Requires:Requires:

• Cost-effective, continuous monitoring.Cost-effective, continuous monitoring.

• Network monitoring for trend analysis Network monitoring for trend analysis and early warning.and early warning.

• Automated and rugged equipment.Automated and rugged equipment.

Pollution of ground, surface and Pollution of ground, surface and river water byriver water by

• AgricultureAgriculture

• Industry & MiningIndustry & Mining

• TrafficTraffic

• Boating, Sports, TourismBoating, Sports, TourismToday's State of Monitoring:Today's State of Monitoring:

• Standard analytical methods utilised, Standard analytical methods utilised, such as HPLC and MS.such as HPLC and MS.

• Time consuming, trained technicians, Time consuming, trained technicians, automation very difficult.automation very difficult.

• Ever increasing list of contaminants.Ever increasing list of contaminants.

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Goal: Develop a cost-effective, on-line, water monitoring Goal: Develop a cost-effective, on-line, water monitoring biosensor that will measure a variety of small organic pollutants biosensor that will measure a variety of small organic pollutants

in short-time with remote control and surveillance.in short-time with remote control and surveillance.

Expanded multi-analyte analysis capability allowing for up to 32 simultaneous analyte measurements. Immunochemical reagents are being developed for a host of small organic water pollutants.

Novel design approaches to the optical detection and fluidics including miniturized integrated optics and microfluidics. Reduced reagent volumes. Limits of detection matching EU prescribed requirements (~1 ppb).

Intelligent remote surveillance and control that will allow for unattended continuous monitoring. Instruments will be rugged and easy to handle by untrained personnel.

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Monitoring of EstuariesOrigin of Pollution

Monitoring of CoastsDirection of Pollution

Monitoring of Side StreamsOrigin of Pollution

Monitoring ofIndustrial PlantsContamination

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Methodology: Immunoassay coupled with Total Internal Methodology: Immunoassay coupled with Total Internal Reflection FluorescenceReflection Fluorescence

Laser

IO-ChipWaveguide

Detection

PC

SamplesWaste

SignalCollection

Fluidics

Analyte derivative

Analyte

Antibody

Fluorophor

Analyte derivative

Analyte

Antibody

Fluorophor

Analyte

Antibody

Fluorophore

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Flow

Transducer

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Flow

Remaining free antibodies arecaptured and detected.

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Flow

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FlowSamples are pre-incubatedwith labelled antibodies.

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AWACSS TechnologyAWACSS Technology

• Miniaturisation

(μL volumes)• Quick Analysis

(ca. 15 min.)• Multi-analyte Detection

2D Array Detection

32 Analysis Spots

Microfluidics

Integrated Optics

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Active Areas of WorkActive Areas of Work

InstrumentDesign

Immuno-chemistry

MarketSurvey &Tutorial

WaterMonitoring

Software& Network

IntegratedOptical

Chip

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Instrument DesignInstrument Design

1st Year Goal: Design and order the equipment necessary to assemble the AWACSS instruments.

Progress: Microfluidics designed with preliminary testing: small volumes & Poiseuille flow.

Designed or selected pumps, valves, lasers, detectors, power supplies, filters, circuit boards, computers.

Autosampler chosen.

Preliminary testing of electronics.

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Integrated-Optical (IO)-Chip DevelopmentIntegrated-Optical (IO)-Chip Development

Goal: Develop IO-Chips capable of simultaneously illuminating 32 analysis windows in a miniaturised format.

Advantages:

• Single mode waveguides allow for efficient and consistant analysis window illumination.

• Compact dimensions allow for microfluidic design and reduce sample & reagent volumes.

• Spatially defined but resolved analysis windows allow efficient detection and suppress signal crosstalk.

Progress: Protype IO-Chips have been designed and fabricated. Preliminary tests are now being carried out.

W a v e g u id e In le t

A n a ly sis W in d o w s (3 0 0 m x 1 .5 m m )

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Software and NetworkSoftware and Network

Local station

Local Stations

• Multi-analyte detection

• Data acquisition

• Data evaluation

• Local station control

Control Station

• Global survey and early warning

• Global trend analyses

• Local station control

• Detection limit control

prescribed by EU Directives Local station

Progress:• Measurement station software modules are selected and requirements are defined.• The first generation software simulator is complete.• The network design is defined.

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ImmunochemistryImmunochemistry

Transducer

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Flow

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Flow

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Flow1) Synthesis and Isolation of Antibodies and Analyte Derivatives.• Polyclonal from Livestock• Antibodies Isolated with ELISA

2) Assay Development.• Binding efficiency• Cross-reactivity• Fluorescently labelled

3) Transducer Spotting & Final Testing.• Spotting protocol established• Final Testing will conclude project

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Targeted Analytes under DevelopmentTargeted Analytes under Development

Herbicides2,4-DAtrazineSimazineIsoproturonDiuronFenuronMethoxuronAlachlorPropanilTrifluralinGlyphosate

FungicidesPentachlorophenolPentachlorobenzene

InsecticidesChlorpyrifosEndosulfan

Endocrine Disrupting CompoundsSteriod Estrogens: Progesterone, EstradiolBisphenol AIrgarolNonylphenol

AntibioticsSulphonamideTetracyclines

Suspected CarcinoginsPyreneTrichloroethyleneBTEXDEHP

Industrial Wastes/Chemical MarkersMTBESulphonate DetergentsCaffeine

Assays in Phase 2

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Water MonitoringWater Monitoring

IIQAB, Spain(Llobregat River)

TZW, Germany(Rhine River)

EI, Slovak Republic(Nitra River)

Goals:• Cataloguing and monitoring local water quality using standard methods (HPLC). Data helps community and project establish trends and identify problems.• Establishing monitoring sites for final instruments.• Defining requirements of end-users, validating design issues, and testing final instruments.

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Water Monitoring ProgressWater Monitoring Progress

• Cross-referencing exercise was performed and standardised protocols for sampling, storing, shipment, and extraction developed.

• Target sites for field deployment: Saint Joan Despi (Llobregat River, Spain), Worms (Rhine River, Germany), Novaky Chemical Plant (Nitra River, Slovak Republic).

• Data Collection: 12 measurement sites, 36 compounds9 compounds recorded significant levels

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21.12.1998 09.07.1999 25.01.2000 12.08.2000 28.02.2001 16.09.2001 04.04.2002

c in

µg

/L Karlsruhe

Mainz

Düsseldorf

Atrazine Conc. in Rhine over 3 Years

• Matrix Effect Studies:

Turbity, Temperature,pH, Ionic Strength

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jan 2001 dec 2001

Temperature of Rhine through Calender Year

5 °C5 °C

25 °C25 °C

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Other ResultsOther Results

Market Analysis

• Survey conducted of European and American water monitoring programs.

• Survey found total micropollutant water sample examination market to be >30 million €. Information broken down according to regions and compounds.

• Water Framework Directive 2000/60/EC is found as bench mark legal act. (13 AWACSS compounds are listed as "dangerous substances".)

Multimedia Tutorial

• Efficient Training

• Cost-savings

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PartnersPartnersEberhard-Karls-University Tübingen: Project Management and Surface Immunoassay Development

Central Research Laboratory: Instrument Hardward Design

Siemens: Electronics, Internet Communication, and Market Analysis.

Optoelectronics Research Centre: Integrated Optical Chips

Environmental Institute, Kos: Instrument Design and Field Tests-Slovakia

King‘s College London: Immunochemistry

Institute of Chem. & Environ. Chem. IIQAB: Instrument Design and Field Tests-Spain

DVGW-Technologiezentrum Wasser: Instrument Design and Field Tests-Germany

Funding: This Project is funded under the 5th Framework Programme: Energy, Environment, andSustainable Development. Visit: http://barolo.ipc.uni-tuebingen.de/awacss.

AWACSS (EVK1-CT-2000-00045)ENVSENS, Lecce, Italy, 2002 AWACSS Dr. Dale M. Willard, Universität...

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