Benedict's reagent

Question/Answers

MAMMALIAN HEART
question/answers

● A – right atrium
● B - aorta
● C - pulmonary artery
● D – coronary artery
INVESTIGATING VITAMIN C
questions/answers

1 Young fruits are unripe. As the fruits develop, they become ripe.
A student suggested that the vitamin C content of fruits increases as they become ripe.
To test this, the student carried out the following investigation:

unripe apricots, peaches, apples and mangos were obtained

a sample of each fruit was crushed

each crushed sample was placed into a separate test tube

distilled water was then added to each of the crushed samples

each mixture was shaken and filtered to give four extracts, one of each fruit

the vitamin C content of each extract was determined by adding it drop by drop to 1 cm3
0.1% DCPIP solution until this solution changed color

This method was repeated using ripe apricots, peaches, apples and mangos.
State the independent variable and the dependent variable in this investigation.

Independent variable= Ripeness

Dependent variable= Vitamin c content

(ii) State two variables, other than the volume and concentration of DCPIP, that should
be controlled in this investigation?

1. mass of fruit ;
2. volume of (distilled) water ;
3. time for ripening / age of fruit ;
4. extraction method qualified ;
5. temperature ;
6. pH
(iii) Vitamin C is water-soluble. The method used assumes that all the vitamin C from
the fruit is in the extract.

Suggest how this assumption could be tested.

1. {take the residue / eq} (from the filter paper) ;

2. add (distilled) water to it ;
3. test for vitamin C (with DCPIP) to show no decolourisation (to show assumption
is right) / eq

(c) Three more students repeated this investigation.

Describe how you would show the variability of all the data on a new graph.

1. plot the {mean / average} ;

2. plot {standard deviation / standard error / error bars / range bars}

Dietary vitamin C is an antioxidant that may be linked to the risk of cardiovascular

disease (CVD).

(a) Explain what is meant by this statement.

● there is a correlation (between vitamin C in the diet and CVD) (1)

● effect of vitamin C on an aspect of CVD

(b) The vitamin C content of food and drink can be measured using DCPIP.

The concentration of vitamin C in a solution can be determined by finding the volume of

the solution that causes DCPIP to change colour.

The higher the concentration of vitamin C in the solution, the smaller the volume needed
to cause the colour change.

(i) Explain what happens to DCPIP in the presence of vitamin C.

● color change from blue to colourless (1)

● because DCPIP is reduced (1)
Plant foods in the diet contain antioxidants, such as vitamin C. Storage time and
conditions affect the concentration of vitamin C in these foods.

(a) Explain how increasing the quantity of plant foods that contain vitamin C in the diet
may protect against heart disease.

An explanation that includes the following points:

● because (antioxidants / vitamin C) reduce (the quantity of) free radicals (1)
● that cause damage to {cells / tissues / blood vessels / endothelium} (1)
● reducing {plaque / atheroma} formation (1)

(b) The effect of storage time and temperature on the vitamin C content of four foods
was investigated.

(i) Describe a method to allow a quantitative comparison of the vitamin C content of two
of these foods.

● use {equal / controlled} masses of the foods (1)

● use {standard / same} extraction method (1)
● measure out {equal / same} volume of DCPIP solution (1)
● add food extract dropwise until {no colour remains / blue to colourless} (1)
● record {volume / number of drops} used (1)
● explanation of how to calculate vitamin C content / description of calibration of
DCPIP with known vitamin C concentration (1)

(ii) State the independent variable in this investigation.

Types of food / fruit

MEMBRANE PERMEABILITY,INCLUDING EFFECT OF

ALCOHOL PH,SUBSTRATE AND INITIAL RATE
question/answer
Beetroot cells contain a red pigment called betalain. The molecules of this pigment are
too large to pass through intact cell membranes. If the membranes are damaged, the
pigment will leak out of the cells.

An investigation was carried out into the effect of the concentration of two types of
alcohol on the permeability of beetroot cell membranes.

Cylinders were cut from beetroot tissue. These cylinders were rinsed in distilled water.
The cylinders were placed in test tubes containing different concentrations of two
different alcohols: methanol or ethanol.

After five minutes, each cylinder of beetroot tissue was removed. The degree of redness
of the solution in each test tube was then measured using a colorimeter.

A colorimeter is an instrument that measures the amount of light absorbed or

transmitted by a coloured solution. As the intensity of the colour increases, the
absorbance of light by the solution increases and the transmission of light through the
solution decreases.

(a) (i) Explain why the pieces of beetroot tissue were rinsed in distilled water after being
cut.

1. to remove any {betalain / pigment / stain / dye / juice / colour / redness} ;

2. which would affect the results / so the {results / investigation / experiment} are
valid / make sure any reading is due to the effect of alcohol / to avoid anomalies ;

(ii) Name one variable, other than time, that should be controlled in this investigation.

Describe how this variable could be controlled.

Variable AND How it could be controlled.

1. temperature ;
2. use a water bath which is {thermostatically controlled / (set) at {constant /
fixed} temperature / (set) at quoted temperature (10-50 C)} / incubator ;
3. volume of the alcohol / volume stated (in the test tube) ;
4. suitable volumetric device used ;
5. any aspect of beetroot sample ;
6. suitable method described ;
7. pH ;
8. by using a buffer ;
Before measuring the absorbance of each coloured solution, the colorimeter
was set to zero using alcohol only.

Suggest why this was done.

1. idea of taking account of a {possible variable / alcohol absorbance} ;

2. to ensure that the absorbance (measured) is due to {betalain / eq} (only) /
make results valid / avoid zero error / avoid offset error / for a (valid)
comparison ;

(iii) Describe how this investigation could be modified to measure the

variability in the effects of these alcohols on beetroot membrane
permeability.

1. (experiment / investigation) should be repeated ; 2. under same conditions / at

each concentration ;

(iv) State how the variability in the results should be analysed.

{calculate / plot / use / check overlap} in {SD / error bars /

range bars / standard error} ;

Beetroot cells contain a red pigment called betalain. This pigment does not
leak out of the cells, unless their membranes are damaged.

A student investigated the effect of detergent on beetroot cell membranes.

Discs of tissue were cut from a beetroot. Any betalain on the outside of the
discs was removed.

One disc was placed into each of five test tubes, containing 0.2% detergent
solution.

The five tubes were left for 30 minutes at 20 °C.

The discs were then removed carefully.

Betalain had leaked from the discs and formed a red solution in each test
tube.

Light was shone through this solution. The percentage of the light
transmitted through the solution was recorded.
The same procedure was repeated using detergent concentrations of 0.0%,
0.5%, 1.0% and 2.0%.

(a) (i) State the independent variable in this investigation.

concentration of detergent

(ii) State two abiotic (environmental) variables that should be controlled in

this investigation.

• temperature (1)
• pH(1)
• volume of detergent solution (1)

(iii) Choose one of the variables you named in (a)(ii) and state how it could
be controlled. (1)

Variable
How it could be controlled

Temperature: thermostatically controlled water bath / incubator/ac

room/environmental chamber (1)

volume of detergent solution:appropriate measuring device (1)

pH:use of buffer (1)

(iv) Explain why the discs were removed carefully.

An explanation that includes the following points:

● in order to avoid (physical/more) damage to the

discs/membranes/cells (1)
● which would cause more pigment release (1)
● causing the results to be invalid (1)

(v) Explain why five discs were used for each concentration of detergent.

● to calculate standard deviation / measure reliability / variability (1)

● to identify anomalies (1)
Detergent permanently damages beetroot cell membranes.

Explain what will happen to the rate of loss of betalain if these discs are then
transferred to test tubes of distilled water.

● the discs (that were in the detergent) will continue to lose pigment as
membrane (permanently) damaged (1)
● the discs that were in 0.0% detergent solution will have no loss of pigment
as the membranes were not damaged (1)
● discs will lose pigment more slowly/stop because there is less/no pigment left
in the beetroot (1)
● discs which were in higher concentrations of detergent will have slowest rate
of loss, because they have the least pigment left (1)
(a) The diagram shows a colorimeter, which is an instrument that
can measure the amount of light transmitted through a coloured
solution.

coloured filter light source

light detector

transmitted light

The transmission of light through the solution decreases as the

intensity of the colour increases.

(i) Explain how using a colorimeter allows valid measurements of

light transmitted through the betalain solutions.

​ because the same {light source / filter / size cuvette} used for all
measurements (1)
​ because zeroing / calibrating (1)
​ because objective measurement (1)
​ because no light lost / gained (1)

(ii) Temperature and pH affect the rate of leakage of betalain from discs of
beetroot.

Describe how each of these variables could be kept constant in this

investigation.

Temperature=use of thermostatically controlled water bath / incubator (1)

pH =use of buffer (1)

(ii) Compare and contrast the effect of these two salts on the loss of betalain
from the beetroot cells.

Similarities

​ both salts affect leakage (1)

​ in both, greatest effect is over a small initial
increase in salt concentration (1)
​
​
​ Differences
​ calcium chloride causes {leakage / intensity of colour} to
decrease, ammonium sulfate causes {leakage / intensity of
colour} to increase, (with increase in concentration) (1)
​ a small initial change in ammonium sulfate concentration causes
a larger change in leakage compared with a small initial change
in calcium chloride concentration (1)
​
​ Explain how you could modify the method to decide if the difference in
colour intensity, at a concentration of 0.002 mmol dm−3, is statistically
significant.
​ repeat readings at 0.002 mmol dm-3 (1)
​ under same conditions (each time) (1)
​ calculate (mean and) standard deviation (1)
LIGHT MICROSCOPE AND MITOSIS
question/answers

(a) (i) Give one reason why each of the following steps was taken.

Step 2 to {stain / dye / colour} the chromosomes (1)

Step 4 to spread / separate the cells out (1)

Step 5 to locate and then magnify (the cells) (1)

(ii) Explain one safety precaution that must be taken when carrying out this procedure?

e.g. rinsing tips in water / wearing gloves / goggles

because hydrochloric acid, acetic orcein is corrosive / irritant

allergy to orcein
(b) Describe how the procedure shown in the flow chart could be used to investigate the
effect of pH on the mitotic index?

● use range of pH solutions (1)

● (in which plants) grown (1)
● (several) root tips (from each pH) (1)
● take cells from same part of root tips (1)
● count number of cells undergoing mitosis and total number of cells (1)
● in several fields of view (1)

(iii) Explain why the mitotic index is affected by pH, as shown by the graph

● enzymes involved (1)

● (enzymes are) protein (1)
● 6.2 is the optimum pH (for these enzymes) (1)
● at pH {below optimum / above optimum} the active site is not the right shape /
enzyme denatures / bonds holding shape change (1)
● enzyme substrate complexes do not form (1)

Animal and plant cells can be observed using a light microscope.

The structure of human cheek cells was investigated.

(a) Describe a safe method to prepare and examine the structure of human cheek cells.

● use of cotton bud (1)

● followed by use of stain/dye (1)
● place cells (on slide) under coverslip (1)
● use of high power of microscope (1)

e.g. bud into disinfectant/sterile/fresh bud/toothpick/wear gloves/ goggles/safe use of

microscope/slides/careful use of bud/stain to prevent injury
(a) Describe a safe method to observe the stages of mitosis in these roots.

● cut root tip

● (root tips) placed in (warm) acid (1)
● (root tips) then placed in named stain (1)
● (root tip placed on a microscope slide and) {macerated / teased / described} /
squashed (1)
● use of high power (on a microscope) (1)
● correct ref to safety issue (1)

e.g. (acetic / ethanoic) orcein / toluidine blue /methylene blue

e.g. rinsing tips in water / wearing gloves / goggles

Describe how this investigation could be modified to determine if the differences

between the mitotic indices are significant.

● each treatment should be repeated (1)

● {all conditions / named condition} should be kept constant (1)
● (mean and) SD calculated / error bars (1)
● look for overlap in SDs / perform t-test (1)

2 Flowering plants produce seeds that can be stored in a seed bank. This is done to
conserve the biodiversity of plants.

(a) (i) Describe how seeds are treated and then stored in a seed bank.

● the water content is reduced (1)

● the seeds are x-rayed (1)
● the seeds are kept at {low temperature / dry} (1)
The longer seeds are stored in seed banks, the less likely they are to germinate. This is
due to ageing.

Ageing of seeds can affect the growth of roots when the seeds germinate. Determining
the mitotic index of roots is one way to study growth of roots. (i) Describe how the
mitotic index of roots can be determined.

● fixing root tip described (1)

● stain used (1)
● description of maceration / squashing (1)
● view through high power microscope (1)
● count of number of dividing cells and divide by total number of cells (1)

PLANT MINERAL DEFICIENCIES / IONS

question/answer

● nitrate / sulfate / ammonium (1)

 ● magnesium / nitrate (1)
● calcium (1)

ii) Devise an experiment, which will produce valid data, to investigate the deficiency of a
named inorganic ion on the growth of a plant.

● control of {species / type / age} of plant (1)

● control of temperature / soil pH /light intensity / time of growth (1)
● complete solution described and solution without named ion (1)
● measurement of dependent variable described (1)
● replication to measure variability / find SD (1)

Pieces of plant tissue, placed in a solution of salt (sodium chloride) in water, may gain or
lose mass or remain unchanged.

(a) Explain the changes that occur when the mass increases.

​ there is an intake of water (1)

​ (down a) water potential gradient outside to inside / (from) high WP to low WP / (from) low salt
concentration (outside) and a high salt concentration (inside) (1)

by osmosis /described (1)

 ​ cut pieces of same {size / surface area / potato} (1)
​ find mass of each piece before and after soaking (1)
​ soak pieces in solutions for {same length of time / stated time} (1)
​ blot dry before weighing (1)
​ repeat for each solution (1)
​ description of how percentage change calculated (1)

The enzyme invertase breaks down sucrose into its two component monosaccharides.
(a) (i) Describe the structure of sucrose.

glucose and fructose joined by glycosidic bond (1)

(ii) The effect of pH on the initial rate of the reaction catalysed by invertase was
investigated at 25 °C.

State the independent variable in this investigation

PH

(iii) Suggest why the temperature was kept at 25 °C.

​ temperature affects {action / reaction rate / number of (S and E) collisions} of enzyme (1)
 ​ (25°C) {may be / is} optimum (for invertase) / gives rate which is not too fast or slow / (1)

(iv) Name one non‐biological (abiotic) variable, other than temperature, that should be
kept constant in this investigation.

invertase / sucrose concentration (1)

(v) Describe how you would keep the variable you have named in (a)(iv) at a constant
value.

​ dissolve a stated mass of {invertase / sucrose} in a stated volume of water / use a stock solution
of {sucrose / invertase} (1)
​ use the same volume of this solution for each pH (1)

(vi) Explain why the initial rate of this reaction was determined.

the rate slows down as {sucrose is used up / sucrose becomes limiting} (1)

so (rates / results} cannot be compared (1)

(vii) Suggest how the initial rate of this reaction could be determined.

​ measure the mass of {sucrose broken down / products formed} at intervals over time (1)
​ plot a graph of mass (y) against time (x) (1)
​ find the gradient of the graph at its start (1)

Similarities:

​ both show an optimum (1)

​ both have same / similar activity at 70 C (1)
 ​ Differences
​ optimum is at a lower temperature for immobilised than non-immobilised (1)
​ temperature affects immobilised enzymes more (1)

● description of the use of all three solutions to grow onions (1)

● description of a relevant biotic factor kept constant (1)
● description of a relevant abiotic factor kept constant (1)
● appropriate measurement of {plant parts / plant} made at start and finish (1)
● after {same/ specified / stated} time (1)

e.g. age, variety (ignore species) NOT size as it is DV

e.g., light/ temperature / pH of solution

e.g. height, length, mass

Accept if it is clear all started at same height/mass etc. / if measure difference or increase in height / mass
etc.

if time quoted must be a week minimum

Describe the information that is needed to decide if any of the differences seen are
statistically significant.

● grow several plants in each solution (1)

● calculation of (means and) SDs (1)
 ● check for overlap (of means plus and
minus SD) for each difference (1)
● carry out a statistical test (1)

1. nitrates are needed for {proteins / amino acids / DNA / RNA / nucleic acids /
ATP / eq} ;
2. magnesium is needed for chlorophyll / eq ;
3. calcium is needed for the {middle lamella / calcium pectate / pectin / eq} ;

TENSILE STRENGTH
questions/answers

In an investigation, the tensile strength of fibres from five plant species was determined.
The breaking force was measured by finding the mass in grams that would break the

fibre.

(a) Describe a valid method that could be used to compare the breaking force of these
fibres.

● how fibre fixed (1)

 ● how force applied (1)
● one variable which should remain constant stated (1)
● replicate (the experiment/measurements) (1)
● (and) calculate the mean/SD (1)

‘attached to a retort stand / hanging from a clamp’

e.g. gradual addition of masses / increase tension of a newton metre gradually

e.g. Relative humidity / temperature / length of fibre

(b) To calculate the tensile strength of the fibre, the cross-sectional area has to be
determined.

(i) Devise a method to determine the cross-sectional area of a fibre, using the following
equipment:

● a sharp blade
● a microscope
● a microscope slide and coverslip
● an eyepiece graticule
● a stage micrometer.
● a (transverse) section/layer/slice of the fibre is cut (1)
● ensure section is flat (1)
● graticule calibrated (with stage micrometer) (1)
● diameter measured/found
● and converted to measurement with calibration data (1)
● area calculated using πr2 (1)

(ii) Name the two types of tissue found in fibres from plant stems.

● xylem (1)
● sclerenchyma (1)
 ● fibre variables taken into account (1)
● appropriate environmental variable controlled (1)
● appropriate description of how masses are added (1)
● adding masses until fibre
breaks / measure the mass that
{breaks the fibre / that the fibre can hold before breaking} (1)

ANTIMICROBIAL PROPERTIES
Question/answers
A- • area of zone {with no bacteria / of inhibition}

Ii- • to allow (chemicals in the extract) to diffuse into agar / to stop growth of bacteria (1)

(iii) Describe how the area of a zone of inhibition could be determined.

● measure {diameter / radius} of zone (1)

● apply area = πr2 (1)

(iv) Describe the control disc used in this investigation.

● filter paper discs qualified (1)

● (soaked in) {solvent / water} (1)

(c) Thismethodcanbeusedtosuggestthatanextracthasanantimicrobialeffect. Other factors

can affect the size of the zone of inhibition.

State two factors, other than the antimicrobial effect, that could affect the area of the
zone of inhibition in this investigation.

● {solubility / concentration / volume} of extract / solvent used (1)

● size of molecules in the extract (1)
● rate of diffusion of extract into agar (1)
1 Some plant extracts have been shown to have antimicrobial properties.
(a) Describe a method that can be used to study the antimicrobial properties of a

plant extract.

● description of {bacteria grown (on nutrient agar) / bacterial lawn} / bacterial broth
(1)
● filter paper discs soaked in plant extract (placed on agar) / plant extract placed in
well (cut in agar) / plant extract added to broth (1)
● control variable described (1)
● one safety aspect described (1)
● incubate for suitable time (1)
● clear zone around {disc / well} {measured / scored / turbidity measured (1)

Dependent variable= {diameter / area} of zone of inhibition/ number of colonies (on

agar) / degree of turbidity / transmission/ absorbance / number of cells (1)

Units= mm / mm2/ number (cells) per cm3 / % (transmission) / au (absorbance) (1)

(ii) Describe a safe method, that uses your dependent variable, to obtain the

results shown in the graph.

● description of how bacteria are cultured (1)

● description of how extracts are added to culture (1)
● control of feature of independent variable described (1)
● incubate at suitable temperature for suitable time (1)
● appropriate measurement of DV (1)
● ref to relevant safety measure (1)

Plant extracts can be used to make new drugs for the treatment of infections. In trials to
test these drugs, placebos and double-blind trials are used.

Explain the purpose of each of these.

placebos= it is used as a control / more validity (1)

● to {eliminate} / account for} a psychological effect (of

taking the drug) (1)

double-blind trials= so that {doctor / scientist} does not know if the treatment contains

the (new) drug or not (1)

● to remove bias from results (1)

POLLEN TUBE GERMINATION
QUESTION/ANSWERS

1 The stigma of a flower secretes a solution of sucrose. Pollen grains germinate in this
solution to produce pollen tubes.

The effect of sucrose concentration on the germination of pollen grains was

investigated.

Pollen grains from a single flower were scraped into the cavity of a slide containing
sucrose solution.

After one hour, the slide was viewed through the high power of a microscope.

The number of germinating pollen grains was counted together with the total number of
grains in the field of view.

This procedure was repeated for each sucrose solution, using pollen grains from the
same flower.

The percentage germination of pollen grains was calculated. (a) (i) State the
independent variable in this investigation.

concentration of sucrose (solution)

(ii) State one abiotic (environmental) variable that should be controlled in this

Investigation.

temperature / pH

(iii) Explain why the results might have been affected if the variable you named in (a)(ii)

had not been controlled.

​ an effect of the factor on the germination (1)

​ because enzymes are involved in (pollen germination/metabolism)
(1)
​ description of {metabolism/enzymes} being affected by the factor
These results suggest that the optimum concentration of sucrose for pollen grain
germination is 40 g dm–3.

Explain how the method could be modified to find the actual optimum concentration of
sucrose.

repeat investigation using more sucrose concentrations (1)

between 20 and 60 g dm-3 (1)

You are provided with a solution containing 500 g dm–3 of sucrose .

Devise a procedure to investigate the effect of different concentrations of sucrose on the

rate of growth of pollen tubes, using this sucrose solution.

answer that includes five of the following points:

use a range of at least 5 concentrations of (sucrose) solutions

(1) detail of dilution method using the sucrose solution (of 500 g dm- 3) (1)

at least one control variable named (1)

use 100 ppm boric acid solution (1)

use a microscope and graticule to measure pollen tube length (1)

stated times (for measurements of pollen tube

3 Pollen grains germinate in a solution that contains sucrose and boric acid.
The effect of boric acid concentration on the percentage germination of pollen grains
was studied.
(a) State the dependent variable in this study.

number of grains germinated (1)

(b) In this study, the solutions used were maintained at pH 6. Explain why this pH was
maintained.

● because it might affect {(pollen grain) germination / enzymes (involved in germination)}

(1)
● it is (thought to be) the optimum for {(pollen grain) germination / enzymes (involved in
pollen germination)} (1)

Describe how the results shown in the graph could have been obtained. You are
provided with a 500 mg dm–3 boric acid solution.

● make (100, 200, 300 mg dm-3) boric acid solutions from 500 mg dm-3 solution by dilution
(1)
● control variable identified (1)
● view through high power of microscope (1)
● suitable method for measuring pollen tube length described (1)
● record results every hour for 4 hours (1)

(e) Compare and contrast the effect of boric acid concentration on pollen grain
germination with its effect on pollen tube growth.

Use the information given in this question.

● {both / germination and growth} show optimum at 100 mg dm-3 (1)

● for growth boric acid concentrations above 100 mg dm-3 are inhibitory whereas for
germination all concentrations above 100 mg dm-3 are stimulatory (1)
 ● it {carries / transports} the male {gametes / nuclei / generative nucleus} (1)
● to the ovule / micropyle / ovary / ovum / female gamete (1)
● to {fuse with / fertilise} {ovum / egg (cell) / female gamete / female nucleus / polar nuclei
(1)
● digest tissue of style (1)

(ii) Explain why pollen tubes grow in the same direction from the stigma, after
pollination.

● the tubes grow towards the {micropyle / ovule / ovary / ovum / female gamete} (1)
● {due to chemicals (released by the embryo sac) / it is chemotropic} (1)

(b) The effect of sucrose solutions of different concentrations on the growth of pollen
grains was investigated. The percentage of pollen grains that germinated and the length
of the pollen tubes were measured, one hour after they were placed in the sucrose
solution.

(i) State two dependent variables in this investigation.

● percentage germination / number germinating (1)

● pollen tube length (1)

(ii) Explain why changes in temperature will affect this investigation.

● it involves {enzymes / (chemical) reactions} (1)

● example of how {enzymes / (chemical) reactions} are affected by temperature (1)
(c) A solution containing calcium ions has to be added to the sucrose solutions. These
solutions were made using:

● a 2.0 mol dm−3 sucrose solution

● a solution containing calcium ions and water

(i) Describe how you would make 10 cm3 of a 0.4 mol dm−3 sucrose solution using these
two solutions.

An answer that includes the following points:

● description of method (1)

● calculation of dilution factor / use of C1V1=C2V2 to calculate sucrose solution volume as 2
(cm3) (1)
● stating the volume of sucrose solution and calcium ion solution to be used to make
10cm3 (1)

ii) The calcium ion solution is alkaline.

State how the pH of the sucrose solutions used in this investigation could be adjusted to
a required value.

varying the (concentration of) calcium ion solution (1)

(ii) Comment on the conclusion that both percentage germination and mean pollen tube
growth have the same optimum sucrose concentration.

● both (appear to) have optimum at 0.4 mol dm−3 (1)

● (but for either) it could be anywhere between above 0.2 mol dm−3 and below 0.8 mol
dm−3 (1)
● values for optimum could be different (to each other) / they might not have the same
optima / one optimum might not be 0.4 mol dm−3 (1)
● more concentrations of sucrose (between 0.2 and 0.8 mol dm−3) should be investigated
(1)
 ENZYMES ,PROTEINS AND PH
QUESTIONS/ANSWERS

In the production of glucose, cellulose from water hyacinths is mixed with the enzyme
cellulase. Cellulase breaks down the cellulose to produce glucose.

In an investigation, different masses of cellulose were added to beakers and the volume
made up to 100 cm3, using distilled water. Cellulase was also added to each beaker.

After 15 minutes, the concentration of glucose in each beaker was determined. (a) (i)
State the independent variable in this investigation.

the (mass / weight / concentration) of cellulose ;

In this investigation, the optimum temperature for cellulase was used. Explain how the
optimum temperature was determined.

1. find the rate of production of glucose / find the {concentration / eq} of

glucose after set time ;
2. at a (suitable) range of temperatures/ quoted range ;
3. credit further detail of method ;

4. idea that optimum temperature is the temperature at which the reaction is

{quickest / eq} ;

(iii) Name one variable, other than temperature, that should be controlled in
this investigation.

Describe how this variable could be controlled.

Variable

How the variable could be controlled

1. pH ;
2. use of buffer ;
Or
3. {enzyme / cellulase} {concentration/ mass} ;
 4. add a known {mass / volume} of cellulase to a known volume of water / use
stock solution ;
Or
5. volume of enzyme solution added ;
6. use of {graduated pipette / burette / eq} ;

The enzyme urease catalyses the following reaction:

urea → ammonia + carbon dioxide

The effect of pH on this reaction was investigated using the following

method.

(a) (i)

One test tube containing 5 cm3 of urease solution was placed in a water bath
at 40 °C and left for 10 minutes.

Another test tube containing 5 cm3 of urea solution in a buffer at pH 3.0 was
placed in the same water bath and left for 10 minutes.

After 10 minutes, the contents of both tubes were mixed together in one test
tube. This test tube was replaced in the water bath.

The concentration of ammonia was measured after 15 minutes and again

after 60 minutes.

The procedure was repeated for pH values of 4.0, 6.5, 6.8, 7.3, 8.0 and 9.0.
Explain why the urease solution and the urea solution were kept in the water
bath at 40 °C before and after being mixed.

​ for equilibration (1)

​ because mixing the solutions at different temperatures would make
the temperature ≠ 40 °C (1)
​ (40 °C could be) optimum for urease
(1)
​ (so if the mixture) {cooled down / warmed up} rate would change
during experiment
(1)
​ ensures only pH is varied
(1)
(ii) Explain how you could determine suitable concentrations of urea and
urease solutions to use in this investigation.

​ run the experiment with a range of different concentrations (1)

​ choose a combination in which the rate of change is not too {high /
fast} / too {low / slow} / time taken is not too {long / short} (1)

The enzyme invertase breaks down sucrose into its two component
monosaccharides. (a) (i) Describe the structure of sucrose.

glucose and fructose (1)

joined by glycosidic bond (1)

The effect of pH on the initial rate of the reaction catalysed by invertase was
investigated at 25 °C.

State the independent variable in this investigation.

PH

(iii) Suggest why the temperature was kept at 25 °C.

​ temperature affects {action / reaction rate / number of (S and E) collisions} of

enzyme (1)
​ (25°C) {may be / is} optimum (for invertase) / gives rate which is not too fast or
slow / (1)

iv) Name one non‐biological (abiotic) variable, other than temperature, that
should be kept constant in this investigation.

invertase / sucrose concentration (1)

(v) Describe how you would keep the variable you have named in (a)(iv) at
a constant value.

​ dissolve a stated mass of {invertase / sucrose} in a stated volume of water / use a

stock solution of {sucrose / invertase} (1)
​ use the same volume of this solution for each pH (1)
(vi) Explain why the initial rate of this reaction was determined.

the rate slows down as {sucrose is used up / sucrose becomes limiting} (1)

so (rates / results} cannot be compared (1)

(vii) Suggest how the initial rate of this reaction could be determined.

​ measure the mass of {sucrose broken down / products formed} at intervals over
time (1)
​ plot a graph of mass (y) against time (x) (1)
​ find the gradient of the graph at its start (1)

An investigation compared the protein content of some foods.

Each food was crushed in distilled water to produce a suspension. This was
filtered

and the liquid filtrate was tested for protein.

(a) (i) Describe how you could test for protein in this filtrate.

addition of {biuret reagent / base and copper sulfate} (1)

purple colour (1)

(ii) Describe how the filtrates should have been prepared to allow a valid
comparison of the protein content of these foods.

​ same mass of (food) sample (1)

​ same volume of water (for extraction) (1)
​ control of another aspect of extraction method (1)
A- A semi-quantitative test gives {an estimate / a range} of the
concentration of a substance (1)

​ A quantitative test determines the exact concentration of a substance

(1)

B- (the diagram shows that) a darker colour = more protein (1)

​ the {solution / result} of a ({biuret / food}) test is compared with

(the tubes in) the diagram (1)
​ therefore the concentration is estimated by deciding which of the
(known) protein solutions
in the diagram is closest to the (food test)
result (1)