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Enzyme Biosensor

Enzyme Biosensor. Detection of glycerides in biodiesel. Team Members: Ben Wilson Matt Yourek Megan Templeton. Mentor: Dr. Tom Hess. Client: Dr. Dev Shrestha. Opportunity Statement.

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Enzyme Biosensor

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  1. Enzyme Biosensor Detection of glycerides in biodiesel Team Members: Ben Wilson Matt Yourek Megan Templeton Mentor: Dr. Tom Hess Client: Dr. Dev Shrestha

  2. Opportunity Statement Design, fabricate, and test a handheld biosensor that uses a modified pH meter combined with an immobilized enzyme electrode to detect the amount of free and total glycerol in a sample of biodiesel.

  3. Current Industry Standard Currently biodiesel makers use a gas chromatograph to measure the amount of glycerol within their biodiesel. While the GC provides a very accurate measurement, it is a time consuming process to prepare and test a sample.

  4. Similar Products Blood glucose meters Blood pressure sensors i-Spec Q-100 - uses impedance spectroscopy technology, which is similar to infrared spectroscopy

  5. Client Needs Small Light Fast response time Simple to use Accurate Easy to calibrate Reusable Strong/Durable Inexpensive Sensitive to small changes in pH User friendly

  6. Technical Specifications Dimensions ≤ 8”x 6”x 5” Weight of about 2 lbs Response time of 10 minutes or less Able to test four samples in one hour Error margin of 5% of concentration measurement Can withstand repeated drops from 5ft Does not bend or flex Storage for electrode Result is automatically displayed in large numbers Total of five buttons or less Visual or audio signal when measurement is complete Audio signal on button push Large buttons that take < 3.5lbf to push Matte/textured finish for added grip Battery operated Takes user < 5 minutes to change electrode Automatic zeroing upon button push Visual or audio signal for low battery One electrode can be used up to 50 times Costs less than $300 to fabricate

  7. Instrument Design 1 Pros: Very similar to pH meter Easy to store membrane Cons: Higher chance of damage to electrode Could experience bad connection between electrode and instrument

  8. Instrument Design 1

  9. Instrumental Design 1

  10. Instrument Design 2 Cons: • Would need to be more durable/sealed • More complicated to wash and store membrane Pros: Simple to use Doesn’t need to be held

  11. Possible Membranes Gelatin-Lipase membrane - cited in literature research paper PVA membrane - found during membrane research PVDF membrane - found during membrane research

  12. Membrane Design 1 Gelatin-Lipase Membrane Pros: Easy to make Similar testing published in papers Cons: Published paper does not include all information Membrane falls apart in storage buffer and water Does not give reliable mV readings

  13. Membrane Design 2 PVA Membrane Pros: Mechanically strong and flexible Resistant to oil and solvent Has ionic conductivity that allows easy flow of electrons Used in a biosensor to detect glycerides Cons: Complex process to create membrane Water-soluble, so it might dissolve during reaction

  14. Membrane Design 3 PVDF Membrane Pros: Resistant to acids, bases, and solvents Mechanically strong Low cost Cons: Requires very advanced equipment/techniques A detailed process to create Used for the growth of neurons, not typically used with a biosensor

  15. Experimental Data Tested both pure canola oil and a biodiesel sample using the gelatin-lipase membrane The following graphs are the results of millivolt change during the testing process

  16. Experimental Data Peak to peak average = 22 mV Pure canola oil sample

  17. Experimental Data Peak to peak average = 18 mV Biodiesel 30 minute sample (Part 1)

  18. Experimental Data Peak to peak average = 18 mV Biodiesel 30 minute sample (Part 2)

  19. Experimental Data Using the estimated peak to peak value for each test, as well as the glyceride percent concentration we can obtain the following two point calibration

  20. Microcontroller Option 1 Arduino UNO with ATmega328 Pros: Easy to use/program Comes in design kits Lots of memory LCD screen comes with wiring instructions for Arduino Cons: New programming language to learn Minimum voltage input of 6V Slower clock rate Problems could be made worse by mass of online information

  21. Microcontroller Option 2 PICAXE 14M2 Pros: Familiar programming language Faster clock rate Dr. Wolbrecht teaches a class that uses them Cons: Manufactured and distributed from UK Not much programming help/information Less memory

  22. Design Suggestions Instrument - Design 1, external electrode design Membrane - N/A Microcontroller - Option 1, Arduino UNO with ATmega328 LCD screen - hacktronics LCD screen, comes with Arduino wiring Housing material - ABS 3D printed plastic

  23. Further Experimentation Creation and testing of PVA and PVDF membranes Effect of multiple layers of cellulose acetate solution on reaction time and membrane preservation Relation of mV amplitude to concentration percent

  24. Detailed Schedule Nov 30 – Dec 9: Testing of gelatin-lipase membrane Dec 2: Snapshot Dec 6: Design review Christmas Break Jan 17 – Jan 25: Testing of PVA membrane Jan 26 – Feb 7: Testing of PVDF membrane Feb 8 – Feb 24: Finishing touches on design and ordering of any parts Feb 27 – Mar 9: Fabrication Spring Break Mar 19 – Apr 26: Validation Apr 27: Engineering Expo

  25. Budget Materials for Membranes: $561 Misc. Chemicals: $500 Microcontroller: $90 LCD Screen: $11 Misc. Electronics: $50 Housing: $300 Total: $1512

  26. Questions? Ben Wilson – Mechanical Enginner Matt Yourek – Biological and Agricultural Engineer Megan Templeton – Biological and Agricultural Engineer

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