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    Laboratory Effects of Blanching

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    diwata444
    This document provides instructions for conducting an experiment to test the effectiveness of blanching potatoes at different temperatures and times. It describes the objectives, apparatus, reagents, and procedures to prepare potato enzyme extracts after blanching and then assay the extracts for polyphenoloxidase and peroxidase enzyme activity over time using a spectrophotometer. The procedures involve blanching potatoes, preparing crude enzyme extracts, conducting enzyme assays by measuring absorbance changes over 2 minutes, and recording results.

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    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd

    Laboratory Effects of Blanching

    Uploaded by

    diwata444
    208 views3 pages
    This document provides instructions for conducting an experiment to test the effectiveness of blanching potatoes at different temperatures and times. It describes the objectives, apparatus, reagents, and procedures to prepare potato enzyme extracts after blanching and then assay the extracts for polyphenoloxidase and peroxidase enzyme activity over time using a spectrophotometer. The procedures involve blanching potatoes, preparing crude enzyme extracts, conducting enzyme assays by measuring absorbance changes over 2 minutes, and recording results.

    Original Description:

    A laboratory activity in fruit and vegetable processing

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    This document provides instructions for conducting an experiment to test the effectiveness of blanching potatoes at different temperatures and times. It describes the objectives, apparatus, reagents, and procedures to prepare potato enzyme extracts after blanching and then assay the extracts for polyphenoloxidase and peroxidase enzyme activity over time using a spectrophotometer. The procedures involve blanching potatoes, preparing crude enzyme extracts, conducting enzyme assays by measuring absorbance changes over 2 minutes, and recording results.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    208 views3 pages

    Laboratory Effects of Blanching

    Uploaded by

    diwata444
    This document provides instructions for conducting an experiment to test the effectiveness of blanching potatoes at different temperatures and times. It describes the objectives, apparatus, reagents, and procedures to prepare potato enzyme extracts after blanching and then assay the extracts for polyphenoloxidase and peroxidase enzyme activity over time using a spectrophotometer. The procedures involve blanching potatoes, preparing crude enzyme extracts, conducting enzyme assays by measuring absorbance changes over 2 minutes, and recording results.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 3

    Blanching effectiveness Objectives To practice testing for blanching adequacy Apparatus and Instruments 1.

    Visible spectrophotometer and cuvettes 2. Pipettes, 1, and 2ml 3. Test tubes 4. Parafilm 5. Small blender 6. Whatman no.1 filter paper 7. Knife 8. Ice bucket 9. Erlenmeyer flask, 125ml 10. Beaker, 600ml 11. Graduated cylinders, 50ml 12. Forceps 13. Hot plate 14. water baths, 30, 60, and 80C Reagents and Materials 1. Small potatoes (held in refrigerator overnight) 2. Ice (in the ice bucket) 3. Buffer A: sodium phosphate buffer, 0.1M, pH 6.8, containing 0.1M NaF 4. Buffer B: sodium phosphate buffer, 0.1M, pH 6.8 5. Dopa: 4mg/ml in buffer B, pH 6.8 6. Guaiacol solution (1% v/v in 95% ethanol) 7. Hydrogen peroxide (0.5% v/v) Blanching of the raw potatoes 1. Blanch a potato at the assigned temperature (in the proper water bath or in the beaker with boiling water) for the following times 0, 2.5, 5, 10, 20, 30 minutes 2. Immediately put the potato on ice to cool it down Preparation of Crude Enzyme Extract 1. Peel the blanched potato and cut into small pieces 2. Rapidly weigh about 10g of potato and mix with 50ml ice cold buffer A 3. Grind the mixture in a blender for about 1min 4. Filter the mixture with Whatman no.1 paper into an iced 125ml Erlenmeyer flask and hold on ice until needed

    Enzyme Assay For Polyphenoloxidase: 1. Transfer 2.0ml of buffer B and 0.5 ml of dopa solution to cuvettes 2. Set the wavelength on the spectrophotometer to 475nm and zero the instrument against distilled water. Re-zero after each assay. 3. When everything is set, initiate the reaction by adding 0.5ml of the enzyme extract to the prepared cuvet. Invert to mix and begin recording absorbance readings immediately (to do this, one person should make and record the readings while the other watches the clock and indicates when readings should be taken). 4. Take readings at 15s intervals for 2min for each tube. Record all readings in the table. Blanching temperature = C Absorbance recorded every 15 seconds. Time of Blanching time (min) absorbance 0 2.5 5 10 reading 0 15 30 45 1 00 1 15 1 30 1 45 2 00

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    For Peroxidase: 1. Transfer 0.6ml of buffer B and 0.6ml of the enzyme extract to a test tube 2. Incubate for 5 minutes at 30C 3. Transfer the content of the test tube to a cuvet 4. Set the wavelength on the spectrophotometer to 410 nm and zero the instrument against distilled water. Re-zero after each assay. 5. When everything is set, initiate the reaction by adding 1.2ml of the hydrogen peroxide solution and 0.6ml of the guaiacol solution to the prepared cuvet. Invert to mix and begin recording absorbance readings immediately 6. Take readings at 15s intervals for 2min for each tube. Record all readings in the table.

    Blanching temperature = C Absorbance recorded every 15 seconds. Time of Blanching time (min.) absorbance 0 2.5 5 10 reading 0 15 30 45 1 00 1 15 1 30 1 45 2 00

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