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    Biology Exam - Paper 6 Revision Notes: Enzyme Activity Lab

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    James
    The document provides revision notes for the Biology exam on several topics: 1) Enzyme activity lab using potato strips and hydrogen peroxide with controls. 2) Chemical tests for starch, sugars, proteins and fats. 3) Photosynthesis labs on chlorophyll, light, and carbon dioxide using iodine tests. 4) Transpiration rate measured with a potometer. 5) Plant response experiments on gravity and light. 6) Osmosis experiments using potato strips in solutions.

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    Biology Exam - Paper 6 Revision Notes: Enzyme Activity Lab

    Uploaded by

    James
    172 views15 pages
    The document provides revision notes for the Biology exam on several topics: 1) Enzyme activity lab using potato strips and hydrogen peroxide with controls. 2) Chemical tests for starch, sugars, proteins and fats. 3) Photosynthesis labs on chlorophyll, light, and carbon dioxide using iodine tests. 4) Transpiration rate measured with a potometer. 5) Plant response experiments on gravity and light. 6) Osmosis experiments using potato strips in solutions.

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    The document provides revision notes for the Biology exam on several topics: 1) Enzyme activity lab using potato strips and hydrogen peroxide with controls. 2) Chemical tests for starch, sugars, proteins and fats. 3) Photosynthesis labs on chlorophyll, light, and carbon dioxide using iodine tests. 4) Transpiration rate measured with a potometer. 5) Plant response experiments on gravity and light. 6) Osmosis experiments using potato strips in solutions.

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    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    172 views15 pages

    Biology Exam - Paper 6 Revision Notes: Enzyme Activity Lab

    Uploaded by

    James
    The document provides revision notes for the Biology exam on several topics: 1) Enzyme activity lab using potato strips and hydrogen peroxide with controls. 2) Chemical tests for starch, sugars, proteins and fats. 3) Photosynthesis labs on chlorophyll, light, and carbon dioxide using iodine tests. 4) Transpiration rate measured with a potometer. 5) Plant response experiments on gravity and light. 6) Osmosis experiments using potato strips in solutions.

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    Biology Exam - Paper 6 Revision Notes

    Enzyme activity lab​:


    2H​2​O​2​ (l) ​→​ 2H​2​O (l) + O​2​ (g)
    This reaction can be catalysed by an enzyme (catalase) or by a non-biological
    catalyst (Manganese IV oxide)
    1. Put 3cm​2​ of hydrogen peroxide in a test tube.
    2. Add fresh potato strips and shake gently.
    3. Keep your thumb on top of the test tube, or use a stopper, to retain the gas.
    4. Do the “glowing splint” test ​→​ the splint relights
    Positive control: repeat the original experiment using manganese IV oxide ​→​ bubbles
    of oxygen form
    Conclusion: the reaction happens because of a catalyst
    1​ negative control: repeat the original experiment using ​boiled​ potato strips ​→
    st​

    nothing happens
    Conclusion: enzymes denature when they are at high temperatures
    nd​
    2​ negative control: repeat the original experiment using water instead of hydrogen
    peroxide ​→​ nothing happens
    Conclusion: hydrogen peroxide is the substrate
    3​rd​ negative control: repeat in a cold environment, the effervescence should be
    slower
    Conclusion: enzymes don’t work as well in the cold

    Chemical tests​:
    -​starch​: add a few drops of ​iodine solution​, a positive result (i.e. starch is present)
    is a ​deep blue-black​ colour, a negative result is orange.
    -​reducing sugars​ (e.g. glucose): ​Benedict’s reagent​, then the mixture is heated for
    2 to 3 minutes. Positive result is an ​orange/brick-red​ colour, negative result is blue
    (the colour of the Benedict’s reagent).
    -​proteins​: add a few drops of ​Biuret reagent​, a positive result is a ​mauve/purple
    colour.
    -​fats​: the ​emulsion test​: ethanol is added to the mixture, this is poured into a test
    tube with an equal number of distilled water, a positive result a milky-white emulsion
    forms.

    Investigating the factors ​necessary​ for


    photosynthesis​:
    Chlorophyll​ – with a variegated leaf...
    1) Leaf is boiled in water for 2 minutes: to break down cell walls, denature the
    enzymes and allow for easier penetration by ethanol.
    2) Warmed in ethanol until leaf is colorless: to extract the chlorophyll, which
    would mask observation (you need to see a color change), chlorophyll is
    soluble in ethanol but not water.
    3) Dipped in water briefly: to soften leaf
    4) Leaf is placed on a white tile and iodine is added: if starch is present the color
    will be blue-black, if it is absent it​ ​will be orange-brown, this is shown against
    the white tile.

    Light​: de-starch the plant by placing it in a dark cupboard or box for 48 hours, so that
    there is no starch in the leaves. Then you can:
    A) Clip a black paper onto both sides of the leaf to make a strip
    B) Make an air-tight bag around a leaf with soda lime (absorbs CO​2​) in it.
    C) Make an air-tight bag around a leaf with hydrogencarbonate solution (provides
    CO​2​) in it.
    Then remove leaf and:
    1. Leaf is boiled in water for 2 minutes: to break down cell walls, denature the
    enzymes and allow for easier penetration by ethanol.
    2. Warmed in ethanol until leaf is colorless: to extract the chlorophyll, which
    would mask observation (you need to see a color change), chlorophyll is
    soluble in ethanol but not water.
    3. Dipped in water briefly: to soften leaf
    4. Leaf is placed on a white tile and iodine is added: if starch is present the color
    will be blue-black, if it is absent it​ ​will be orange-brown, this is shown against
    the white tile.
    Carbon dioxide:
    Required materials

    ● 2 Potted plants
    ● 2 Bell-jars
    ● A Candle
    ● Dish containing Caustic soda
    ● Petroleum jelly
    ● Glass sheets
    ● Iodine solution to test leaves for starch

    Estimated Experiment Time


    Approximately 10 minutes to set up the apparatus and 8-12 hours to carry out the
    observations
    Step-By-Step Procedure

    ● 1.​ Take two young potted plants.


    ● 2.​ Apply petroleum jelly on two glass sheets.
    ● 3.​ Place the potted plants on these glass sheets.
    ● 4.​ On one glass sheet, along with the potted plant place a burning candle.
    ● 5.​ In the other, place a dish containing caustic soda.
    ● 6.​ Cover them with the bell jars.
    ● 7.​ Leave undisturbed for a few hours and test the leaves from each pot for the
    presence of starch.

    Note

    ● Potted plants must ideally have been kept in the dark to make the leaves
    starch-free before including them in this experiment.
    ● The petroleum jelly makes the bell jars airtight.

    Investigating what happens when ​varying​ the factors


    affecting photosynthesis​:
    NOTE: this diagram is from an IGCSE paper 6 (and the “gas” is oxygen and the
    pondweed is in water)
    Light intensity​: (NOTE: I copied this from “model answer”) First a lamp is
    placed as close as possible to the apparatus, then the experiment is repeated several
    times, each times with the lamp further away from the apparatus. Heat from the bulb
    is prevented from affecting the result by placing a clear glass sheet between the lamp
    and the apparatus, and the pond weed used is left for several minutes in each new
    light intensity to allow it to adjust to new conditions before rate is measured.
    Carbon dioxide​:​ vary the amount of hydrogen carbonate in the solution, this
    supplies the plant with carbon dioxide for photosynthesis (light intensity and
    temperature are controlled variables)

    Temperature:​ ​set up the apparatus in several different-temperature


    environments
    The effect of temperature on plant growth
    Investigating transpiration rate​: the potometer (page 91)
    1. Leafy shoot must be cut, the apparatus filled and the shoot fixed to the potometer,
    all under water to prevent air locks in system. Capillary tube must be horizontal
    otherwise the bubble will move because of its lower density.
    2. Allow plant to equilibrate (5min) before introducing air bubble. Measure rate of
    bubble movement at least 3 times, and use reservoir to return bubble to zero each
    time. Record air temperature and find mean of readings.
    3. Find out leaf area to calculate rate of water uptake per unit of leaves.
    Variables: temperature, air humidity, light intensity and wind (using a fan vs. no fan, I
    guess).

    Investigation: how plants respond to gravity


    (geotropism)
    1) Freshly germinate broad bean seedlings inside a glass jar, the seed is held by a
    roll of moist clotting paper.
    2) Seedlings are allowed to grow for a further five days, with the jars placed a) the
    right way up b) upside down and c) on its side.
    In each case the roots will turn to go downwards, and the shoot turns to grow
    upwards, (diagram page 162).

    Investigating the light sensitive region


    (phototropism)
    1) There are three groups of coleoptiles (oat shoots). A) Has its tips removed, B) tips
    are covered and C) are untreated.
    2) The coleoptiles are measured, and lengths recorded.
    3)They are put in light proof boxes with one gap which only allow light to enter
    laterally (from the side).
    4) They are measured 2-3 days later, and new lengths are recorded.
    Untreated coleoptiles will grow the most.
    Other experiments:
    Mark rings around the shoot, all the same length away from each other. After growth
    towards the sunlight, the spaces will increase in different areas showing where
    growth has taken place.
    (All of these experiments on phototropism are on pages 160-161 with diagrams).

    Osmosis, hypertonic, hypotonic and isotonic


    solutions & determining the concentration inside a
    cell
    Remember:
    In an ​isotonic solution​: concentration of solute outside cell = concentration inside
    cell ​→​ no change in size
    In a ​hypertonic solution​: concentration of solute outside cell > concentration inside
    cell ​→​ cell shrinks
    In a ​hypotonic solution​: concentration of solute outside cell < concentration inside
    cell ​→​ cell swells

    Potato strips are placed in solutions of different concentrations (you have to know the
    concentrations). They are measured in mass before and after being in the solutions.
    The solution in which the potato strip does not change in mass is the isotonic
    solution, but it is unlikely that the solution will be exactly the right concentration, so
    you have to plot a graph and find what the concentration is. Concentration on the
    x-axis and mass change in the y-axis (note: the line will go above and below the
    x-axis since some of the potato strips will shrink). Where the line intersects the x-axis,
    there is no mass change so that is the concentration inside the potato.

    Demonstrating osmosis​:
    This is like the potato strip experiment but it simply demonstrates that osmosis
    occurs, it does not determine the concentration of the cells
    The flaming nut experiment​:
    Investigating Insulation:
    Concentration of Vitamin C (ascorbic acid) in fruit
    juices
    Finding the xylem in a cross section of a stem and
    root
    Demonstrating that respiration uses oxygen and
    produces CO​2

    or
    Demonstrating the effect of pH on enzyme activity
    (I don’t think this is the kind of lab that you have to remember)

    OR

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