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.

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 statues.

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 densitometry, 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 Many of these methods described require a large investment in special equipment or rely on methods which are outdated, inaccurate, or not reliable.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.

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.

The enzymatic oxygen removal system is a patented technology developed by Nicolas Plumere, Wilbur Campbell, and Ellen Campbell. The systems and methods for oxygen removal from aqueous solutions are presented in which a bi-enzymatic reaction sequence recycles and depletes oxygen to extinction, preferably using an oxidase and a catalase as biocatalysts and a carbohydrate as co-substrate. Contemplated systems and methods are particularly advantageous in conjunction with electrochemical reaction systems in which oxygen would adversely interfere with the reaction system.

The present application relates generally to oxygen removal systems for use with biosensors that operate with recombinant reductases, and particularly with nitrate reductases, chlorate reductases, perchlorate reductases, hydroxylamine reductases, bisulfite reductases, and isoquinoline reductases.

Read the full patent details here.