AWACSS Automated Water Analyser Computer Supported System Dr

AWACSS Automated Water Analyser Computer Supported System Dr. Dale M. Willard, Universität Tübingen (Gauglitz Group) Contract #: EVK 1 -CT-2000 -00045 Duration: 3 Year, RTD Project (Starting March, 2001) Visit: http: //barolo. ipc. uni-tuebingen. de/awacss AWACSS (EVK 1 -CT-2000 -00045) ENVSENS, Lecce, Italy, 2002

Motivation Today's State of Monitoring: • Standard analytical methods utilised, such as HPLC and MS. • Time consuming, trained technicians, automation very difficult. • Ever increasing list of contaminants. AWACSS (EVK 1 -CT-2000 -00045) Pollution of ground, surface and river water by • Agriculture • Industry & Mining • Traffic • Boating, Sports, Tourism More Effective Water Management Requires: • Cost-effective, continuous monitoring. • Network monitoring for trend analysis and early warning. • Automated and rugged equipment. ENVSENS, Lecce, Italy, 2002

Goal: Develop a cost-effective, on-line, water monitoring biosensor that will measure a variety of small organic pollutants 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. AWACSS (EVK 1 -CT-2000 -00045) Monitoring of Side Streams Origin of Pollution Monitoring of Estuaries Origin of Pollution Monitoring of Industrial Plants Contamination Monitoring of Coasts Direction of Pollution ENVSENS, Lecce, Italy, 2002

Methodology: Immunoassay coupled with Total Internal Reflection Fluorescence Detection Signal Collection Laser PC IO-Chip Waveguide Fluidics Antibody Fluorophore AWACSS (EVK 1 -CT-2000 -00045) Samples are pre-incubated with labelled antibodies. Flow Remaining free antibodies are captured and detected. * Analyte derivative * Samples Waste Transducer ENVSENS, Lecce, Italy, 2002

AWACSS Technology • Miniaturisation (μL volumes) • Quick Analysis (ca. 15 min. ) • Multi-analyte Detection 2 D Array Detection 32 Analysis Spots Microfluidics Integrated Optics AWACSS (EVK 1 -CT-2000 -00045) ENVSENS, Lecce, Italy, 2002

Active Areas of Work Instrument Design Integrated Optical Chip Immunochemistry Water Monitoring Software & Network AWACSS (EVK 1 -CT-2000 -00045) Market Survey & Tutorial ENVSENS, Lecce, Italy, 2002

Instrument Design 1 st Year Goal: Design and order the equipment necessary to assemble the AWACSS instruments. Progress: Microfluidics designed with preliminary testing: small volumes & Poiseuille flow. Autosampler chosen. Preliminary testing of electronics. AWACSS (EVK 1 -CT-2000 -00045) Designed or selected pumps, valves, lasers, detectors, power supplies, filters, circuit boards, computers. ENVSENS, Lecce, Italy, 2002

Integrated-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. AWACSS (EVK 1 -CT-2000 -00045) ENVSENS, Lecce, Italy, 2002

Software and Network Local Stations • Multi-analyte detection • Data acquisition • Data evaluation • Local station control 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. Control Station • Global survey and early warning • Global trend analyses • Local station control • Detection limit control prescribed by EU Directives AWACSS (EVK 1 -CT-2000 -00045) Local station ENVSENS, Lecce, Italy, 2002

Immunochemistry 2) Assay Development. • Binding efficiency • Cross-reactivity • Fluorescently labelled * Flow * 1) Synthesis and Isolation of Antibodies and Analyte Derivatives. • Polyclonal from Livestock • Antibodies Isolated with ELISA Transducer 3) Transducer Spotting & Final Testing. • Spotting protocol established • Final Testing will conclude project AWACSS (EVK 1 -CT-2000 -00045) ENVSENS, Lecce, Italy, 2002

Targeted Analytes under Development Herbicides 2, 4 -D Atrazine Simazine Isoproturon Diuron Fenuron Methoxuron Alachlor Propanil Trifluralin Glyphosate Endocrine Disrupting Compounds Fungicides Pentachlorophenol Pentachlorobenzene Insecticides Chlorpyrifos Endosulfan Antibiotics Sulphonamide Tetracyclines Assays in Phase 2 AWACSS (EVK 1 -CT-2000 -00045) Steriod Estrogens: Progesterone, Estradiol Bisphenol A Irgarol Nonylphenol Suspected Carcinogins Pyrene Trichloroethylene BTEX DEHP Industrial Wastes/Chemical Markers MTBE Sulphonate Detergents Caffeine ENVSENS, Lecce, Italy, 2002

Water Monitoring TZW, Germany (Rhine River) IIQAB, Spain (Llobregat 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. AWACSS (EVK 1 -CT-2000 -00045) ENVSENS, Lecce, Italy, 2002

Water Monitoring Progress • Cross-referencing exercise was performed and standardised protocols for sampling, storing, shipment, and extraction developed. Atrazine Conc. in Rhine over 3 Years • Data Collection: 12 measurement sites, 36 compounds 9 compounds recorded significant levels • Target sites for field deployment: Saint Joan Despi (Llobregat River, Spain), Worms (Rhine River, Germany), Novaky Chemical Plant (Nitra River, Slovak Republic). Temperature of Rhine through Calender Year 25 °C • Matrix Effect Studies: Turbity, Temperature, p. H, Ionic Strength AWACSS (EVK 1 -CT-2000 -00045) 5 °C ENVSENS, Lecce, Italy, 2002

Other 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 AWACSS (EVK 1 -CT-2000 -00045) ENVSENS, Lecce, Italy, 2002

Partners Eberhard-Karls-University Tübingen: Project Management and Surface Immunoassay Development Central Research Laboratory: Instrument Hardward Design Institute of Chem. & Environ. Chem. IIQAB: Instrument Design and Field Tests-Spain DVGW-Technologiezentrum Wasser: Instrument Design and Field Tests -Germany Environmental Institute, Kos: Instrument Design and Field Tests. Slovakia King‘s College London: Immunochemistry Optoelectronics Research Centre: Integrated Optical Chips Siemens: Electronics, Internet Communication, and Market Analysis. Funding: This Project is funded under the 5 th Framework Programme: Energy, Environment, and Sustainable Development. Visit: http: //barolo. ipc. uni-tuebingen. de/awacss. AWACSS (EVK 1 -CT-2000 -00045) ENVSENS, Lecce, Italy, 2002