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Sterilization Device for Liquid Chromatography Solvents

Develop a device to prevent bacterial contamination in UPLC columns, extending their lifespan without altering chemical compositions. Initial design concepts, UV probe feasibility, filter and pump selection, UV block design, cost analysis, and recommendations for improvement.

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Sterilization Device for Liquid Chromatography Solvents

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  1. Design Team Nick Roulleau, Michael Vose Michael Racette, Michael McKay Sterilization Device for Liquid Chromatography Solvents Advisor Professor Mohammad Taslim

  2. Introduction • Background • Problem Statement • Past Art • Design Requirements • Design Concepts • Prototype Design • Component Analysis • Recommendations

  3. What is Liquid Chromatography? A substance comprised of components A and B is dissolved in a solvent and enters the analytical column, where it is separated

  4. Basic Components of an HPLC System From http://www.waters.com/WatersDivision

  5. Problem From http://www.waters.com/WatersDivision

  6. Design Goal To mitigate the risk of blockage at the inlet frit due to bacterial contamination and extend the useful life of the UPLC column.

  7. Existing Solutions • In-line filters • Guard columns and cartridges • Pre-filtration of samples and mobile phase liquids

  8. Product Requirements • Mandatory: • Must be adaptable for use worldwide • Must extend the useful life of columns • Must meet safety standards (ISO, UL and CE) • Must operate for 1 year w/o user intervention • Desirable: • Should be able to filter two bottles simultaneously • Should meet customer acceptance criteria • Low-maintenance • Easy to use • Cost

  9. Constraints • Cannot change the chemical composition • Of the solvent • Of the sample • Cannot create risk of causing pump cavitation • Cannot hinder bottle accessibility • Cannot negatively impact system resolution

  10. Initial Design Concepts UV Probe Pump/filter--Cap enclosure Pump/filter--External enclosure

  11. Preliminary Design – UV Probe • Inexpensive • Simple Design

  12. Why Not Use Ultraviolet Radiation as a Primary Solution? • Degradation of organic solvent modifiers (Low Risk) • Degradation of aqueous additives (Low Risk) • User safety from UV-C exposure (Medium Risk) • UV can inactivate but not remove bacteria

  13. Filter Sizing • How many bacteria could be generated per year? • Logarithmic growth: • Assuming worst case • 100% replicating • Short generation time • Neglecting lag phases and cell death • Filter capacity = 107 CFU/cm2

  14. Filter Sizing With logarithmic bacterial growth, filter area becomes exceptionally large in a short period

  15. Current DesignExternal Filter Enclosure with UV Dual-head brushless DC pump UV lamp with multiple sterilization lines Pall AcroPak 200 filters

  16. Filter Selection • Membrane with material compatibility • Sufficient capacity to contain 1 year of inactivated bacteria

  17. Pump Selection Micro-diaphragm pump • Dual pump heads • Ability to run dry • DC brushless motor for long life

  18. Pump Pressure Requirements • Pump must deliver sufficient differential pressure (Δp) to move fluid through filter • Darcy’s equation for porous media: L = membrane thickness p1 = pump-side pressure p2 = outlet pressure Q = flow rate k = permeability constant for filter A = effective filter area (EFA) µ = viscosity

  19. UV Block Design-Initial Concept

  20. UV Block Design A dA • 99.99% inactivation requires a UV dose of at least 40 mJ/cm2 for nearly all species of bacteria • Dose is a function of the irradiance (mW/cm2) and time of exposure (in seconds) Dose = Irradiance x time

  21. UV Block Design

  22. UV Block Design 13 loops necessary with an 18W UV bulb and thin wall FEP tubing

  23. Test Planning Pump Sensor Device Column • Verification Test • Does the Device Meet Design Requirement? • Pump Particle-Laden Water from Bottles With and Without Device • Compare Backpressure and/or Flow Rate

  24. Test Results Backpressure was reduced by 28% when our device was used

  25. Cost Analysis • Developed target costs by estimating: • Annual costs without the assistance of our device (excluding operational costs) • Savings in material costs by implementing our device • Potential savings for high-end users = $44,000 • Minimum estimated annual savings = $600 • Target production cost = $500 • Target prototyping cost = <$1500

  26. Recommendations for Further Development • Improve manufacturability of the design • Simplify tubing system • Smaller pump • Custom filter size • Analyze effectiveness of UV with microbiological testing

  27. Summary • Introduction to liquid chromatography • The problem and its source • Requirements of a good solution • Design considerations • Prototype design and analysis • Recommendations

  28. Questions???

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