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    Home > Biochemistry News > Plant Extracts News > Tribenzene tetraazole chloride (TTC) method determines root system vitality

    Tribenzene tetraazole chloride (TTC) method determines root system vitality

    • Last Update: 2021-01-07
    • Source: Internet
    • Author: User
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    principle

    chlorinated tribenzene tetraazole (TTC) is a redox substance with a standard redox potentior of 80mV, dissolved in water as a colorless solution, but reduced to produce red and insoluble water tribenzene (TTF), as follows:

    -generated TF is relatively stable, will not be automatically oxidized by oxygen in the air, so TTC is widely used as a hydrogen subject for enzyme testing, plant root caused by TTC reduction, can be enhanced by the addition of serum acid, yanhusoic acid, apple acid, and acetate, and is inhibited by propylene dioxide, iodine acetic acid. Therefore, TTC reduction can indicate
    dehydrogenase
    activity, and as an indicator of root dynamics.



    D-photonometer, analysis of
    balances
    (0.1mg sensitivity);
    stration temperature
    box 1 set; research 1 set; 100ml triangular bottle;
    funnel
    ; pipe pipe 10ml 1st, 2ml 1st, 0.5ml 1st; 20ml scale
    test tube
    ; 10ml capacity bottle; small
    culture
    dishes; test tube holder, spoon; quartz sand suitable.


    reagents
    ethyl

    acetylene;

    Sodium sulphate (Na2S2O4, a strong reducing agent, commonly known as insurance powder);

    1% TTC solution: accurately called TTC 1.0g, dissolved in a small amount of distilled water, fixed to 100ml;

    0.4% TTC solution: accurately called TTC 0.4g, dissolved in a small amount of distilled water, fixed to 100ml;

    Phosphoric acid buffer (1/15mol/L, pH7.0);

    1mol/L sulphuric acid: 55 ml of thick sulphuric acid with a volume of 1.84,
    berries with 500 ml of distilled water added to the mixing side
    , diluted to 1000ml after cooling to 1000ml;

    0.4mol / L sodium sodium sephate: said to take sodium sodium sephate (containing 6 crystalline water) 10.81g, dissolved in distilled water, fixed capacity to 100 ml.

    method:

    1. Qualitative determination

    (1) configuration reaction solution Mix 1% TTC solution, 0.4moL/L sodium sulphate and phosphate buffer in 1:5:4 ratio.

    (2) carefully wash the root, remove the ground part from the stem, put the root into a triangular bottle, pour into the reaction fluid, to immerse the root as a degree, put 37 degrees C around the dark place 1h, to observe the coloring situation, the new root tip of a few millimeters and fine side root are obviously turned red, indicating the presence of dehydrogenase there.

    。 2. Quantify the production of the

    (1) TTC standard curve

    absorb 0.25 ml 0.4% TTC solution into a 10 ml capacity bottle, add a little Na2S2O4 powder, shake immediately after the production of red TTTF. Then use ethyl acetate to the scale, shake well. Then take this liquid 0.25ml, 0.5ml, 1.00ml, 1.50ml, 2.00ml in a 10ml capacity bottle, with ethyl acetate fixed to the scale, you get Standard color series containing TTF25?g, 50?g, 100?g, 150?g, 200?g, with blank as a parameter, the light density is determined at 485nm wavelength and the standard curve is drawn.

    (2) said to take the root sample 0.5g, put in a small petri dish (blank test first add sulphuric acid and then add the root sample, other operations are the same), add 0.4% TTC solution and phosphoric acid buffer of the same amount of mixture 10 ml, the root fully immersed in the solution, in 37 degrees C in the dark insulation 1h, and then add 1mol/L sulfuric acid 2 ml, to stop the reaction.

    (3) to remove the root, absorb water and grind with ethyl acetate 3 to 4 ml and a small amount of quartz sand to present TTF. The red proposed liquid into the test tube, with a small amount of ethyl acetate to wash the residue two to three times, all into the test tube, and finally add ethyl acetate to make the total amount of 10 ml, with phosphorescometer at 485nm color, with blank as a paradiver to read out the light density, check the standard curve, find out the reduction of tetrate.

    (4) calculated

    the resulting data into the lower formula, to find out the strength of nitroazole.

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