Ethanol Detection Enzyme - Case Study
BACKGROUND
This project was prompted by feedback from the Michigan Department of Agriculture & Rural Development (MDARD). Ethanol has been added as an octane enhancer in some Michigan gasoline for some time, it increases the octane almost 3 Anti Knock Index points. If gasoline contains ethanol, there is a limit of 10% of total volume for conventional fuels. [1] Around 13 billion gallons of fuel ethanol were added to motor gasoline produced in the United States in 2014.[2] Most gasoline in the U.S. contains up to 10% ethanol by volume (E10), also known as the 'blend wall', which all gasoline vehicles can use. Gasoline containing 15% ethanol can only be used by light-duty vehicles with a model year 2001 or newer, and only flex-fuel vehicles can use gasoline with greater than 15% ethanol.
Congress has mandated increasing volumes of ethanol to be added to U.S. fuel to improve U.S. energy independence and also to support a move away from fossil fuels. In addition to these initiatives, more ethanol production from bio-stock in the U.S. means an increase in jobs for U.S. citizens and lower transportation costs since it's produced and used within the country and limits the need for overseas shipping. Ethanol is considered to be greener than gasoline, because corn and other plants absorb carbon dioxide from the atmosphere as they grow. The fuel still releases CO2 when it burns, but the net increase is lower.
There are cons, however, to using ethanol and ethanol blends as a domestic fuel source. Ethanol has a lower energy content than gasoline and therefore delivers less power when burned. This converts to higher fuel consumption and less miles per gallon in a conventional vehicle. Therefore, it is becoming increasingly important to monitor ethanol levels in fuel blends for Quality Assurance and Quality Control measures.
Another market to note when mentioning QA/QC ethanol testing is the alcoholic beverage industry. Alcoholic beverages are made by converting sugars to alcohol through a process called fermentation. Extracts from grapes, barley, and more are fermented, which converts the naturally occurring glucose to ethanol. Monitoring ethanol content is essential during the fermentation process and concentration can affect taste, microbial activity, solubility of other constituents, and is also important for federal and state labeling statutes.
APPLICATION
Most ethanol test kits for on-site analysis of gasoline use densitometry measurement, which are rudimentary disposable test tubes that measure ethanol by the difference in density between ethanol and water. This method requires careful calibration and an eyeball's guess using units printed or marked on the side of the tubes. Considering these circumstances brings a reliability and accuracy concern into question. With an increasing interest and importance in determining ethanol levels in conventional fuels and biofuels, more reliable and accurate test methods need to be employed on-site and in analytical laboratories.
Current quantification methods for ethanol in the alcoholic beverage industry include gavimetric analysis, ebulliometry, HPLC, FTIR, gas chromatography, and IR. Many of these methods described require a large investment in special equipment or rely on methods which are outdated, inaccurate, or not reliable. Many times, distilleries, breweries, and wineries do not have their own analytical laboratories, and therefore must send their samples to an external laboratory and wait for results, which can be very costly. Offering a colorimetric, on-site, easy to use, and cost-effective test kit to these beverage producers will allow them to make more timely decisions and save by doing these testing procedures internally.
PROGRESS
An optimal clone of the enzyme has been chosen and successfully produced in our lab. We've completed numerous assays with ethanol standards, and have shown the enzyme to be effective in accurately quantitatively measuring ethanol concentration. We're testing the enzyme's compatibility with real-world samples, including various alcoholic beverage samples and plan to begin testing fuel samples soon.
What else is on the docket? We're putting our enzymes through the wringer, which includes checking for potential interferences, optimizing test kit ranges, enzyme stability for storage and use, and working temperatures. We're also gathering input from the fuel and alcoholic beverage industries and researching other potential uses for this enzyme. Our photometer team will soon be adding in a function to the software which will enable users to spot-check ethanol with a digital readout on mobile devices, and the ability to track and store data, then export it for further analysis. Have suggestions, questions, or comments regarding this project? Please contact us!
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