FOOD2000: Food Science Antioxidants in Fruits and Vegetables - Engineering Assignment Help

Assignment Task :

Task:

Recorded Video Script:

• Hello and welcome to another practical in the FOOD2000 practical series. Today we are measuring antioxidant activity using the Ferric Reducing Antioxidant Power.

• Assay or commonly known as FRAP Assay.

• For this exercise you will have 4 x 2mL Eppendorf tubes containing approximately 50mg of either freeze-dried broccoli sprout powder or acai powder.

• Begin by pipetting 1500uL of distilled water into each Eppendorf tube containing your broccoli and acai powder.

• Next, mix the sample on a vortex for a few seconds each.

• On the left is an unmixed sample and on the right is a sample mixed using the vortex. It is important to disperse the sample in the water to aid in extraction of the antioxidants. Course Co-ordinator: Semester 1, 2021 Dr Deirdre Mikkelsen (d.mikkelsen@uq.edu.au) 2

• After vortexing, place the four Eppendorf tubes containing your samples in a water bath set to 37 degrees C for 15 minutes. Place a rack on top of your samples to stop them from floating.

• Every 3 minutes take the Eppendorf tubes out of the water bath and vortex them. Make sure to wipe the tubes with paper towel before vortexing.

• After 15 minutes has elapsed, transfer the tubes to a centrifuge. Remember to balance the tubes in the centrifuge and secure the lid. The samples will be centrifuged at 14,000g for 5 minutes.

• After centrifugation carefully remove your Eppendorf tubes. You can see in this image the separation between the particles at the bottom of the Eppendorf tube and the liquid suspended above. The liquid is called the supernatant and the particles are referred to as the pellet.

• Separate the supernatant from each Eppendorf tube as shown, be careful when pouring to not disturb the pellet. If you are not comfortable pouring out the supernatant, you can you use a pipette. Make sure to label the supernatant as the water-soluble fraction and DO NOT discard the pellet.

• Next, using a 5000uL pipette, transfer acetone to each pellet.

• After you have added acetone to each pellet, repeat the previous steps for vortexing, incubating and centrifuging to acquire an acetone supernatant. Separate the acetone supernatant into its own tubes and label them acetonesoluble fraction. You should now have 8 Eppendorf tubes in total.

• In this image you can see our 8 Eppendorf tubes containing the supernatant from both our water and acetone extraction.

• Next, we can prepare the standard curve. Starting with a stock solution of 1mM ferrous sulphate prepare the following solutions by referring to the table in your lab manual (table is in step 14). Transfer the correct amount on ferrous sulphate and water into each cuvette as indicated by the table.

• Using parafilm cover the top of each cuvette and invert them to mix.

• Now that all our standards and samples are prepared, we can prepare to add the FRAP reagent. First, using a 1000uL pipette, transfer 100uL of each standard in sample into fresh cuvettes as shown in the table (step 19).

• Next add 3000uL of FRAP reagent and 300uL of distilled water to each cuvette. You will notice an instant colour change after adding FRAP reagent as shown in the video.

• After you have added your FRAP reagent and distilled water to each cuvette, cover the cuvettes with parafilm and invert to mix. Write the name of each standard and sample on top of the parafilm to keep track of each cuvette

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