As Edexcel unit 3 core practicals

Unit 3
plan an
investigation
for all core
practicals.
Plan an investigation

1. Core practical one for serial dilution

Carry out the experiment using 5 different concentrations of sucrose( g

dm-3)

Use serial dilution method

By using the stock solution of (. ) gm dm-3
And dilution factor of 0.1 ( 1: 10)
Where we use 9ml of water and 1 ml of solution to have total volume of each
solution in each test tube 10 ml .

Controlled variables
Same temperature using a thermostatic water bath / air conditioned room…
( abiotic)
Same type and age of pollen grains …( biotic)
Same volume of solutions of different sucrose concentrations ..( abiotic)

Time interval
Leave the pollen grains in each sucrose concentration for 2 hours

Dependent :
Use a microscope ( eyepiece graticule and stage micrometer ) to measure
the length of pollen tubes grown
 2. Core practical of vitamin C

Procedure:
1. Extraction of fruit juice ....crushing and add distilled water to extract vitamin C
2.independent ; different extracts of fruit juice with different vitamin C content
......or use a stock solution and carry serial dilution method

3. Controlled variables;
Fruits of same age . Same mass same volume of extract , same storage
conditions , same concentration of DCPIP

4. Methodology
Titration of the fruit juices by adding DCPIP to juices drop by drop and mix
gently upon adding each drop where the color change from blue to colourless

5. Dependent ; measure the volume / number of drops of DCPIP added for the
colourless to change blue / for blue color to remain

6. Standardization : compare to standard solution of known vitamin c

concentration
Repeat using 1% of vitamin C solution so compare the volume of DCPIP
decolorised by fruit juice with the volume decolorised by standard vitamin c
solution
 End point in case titrant is DCPIP

Is the blue color of DCPIP persist

To get the concentration of the unknown sample …….cross multiplication

Vitamin C sample is the titrant ;

End point …remain colorless

To get the concentration of vitamin C use the formula

Two ways of calculations

1. Vitamin C sample ( standardized ) is the titrant
End point …remain colorless
To get the concentration of vitamin C using a formula
. Concentration of
Volume of standardised solution x
standard solution
=
Volume of fruit juice

÷ 1. Known concentration
10mg ……..10 ml
2. Unknown
x mg ……. 20 ml

Reaction between DCPIP and vitamins

DCPIP gain electrons from ascorbic acid / vitamin C
&¥
Where vitamin C is described as an anti oxidant use
As vitamin C losses electrons and become oxidised
Preventing chemicals / components inside the cell from becoming oxidised
 '
We should not shake vigorously while titrating ?

To avoid splitting Vit C

Shaking too vigorously introduce oxygen to DCPIP, so will be reoxidised
So blue color will return
So difficult to judge the end point
DCPIP

DCPIP …oxidised…blue
DCPIP ….reduced ….colorless

End point
Vitamin C reduce DCPIP …colorless

Core practical 3

Investigating effect of temperature and alcohol on the cell membrane

Safety :
1. Ethanol is highly flammable ..keep it away from the flame and keep the stoppers on the
bottles .
2. Wear eye googles

A) effect of temperature :
1. Carry out the experiment at 5 different temperatures ( 10 C to 50 C) using a
thermostatic water bath
2. Get 5 test tubes of same size and add equal volume of water
3. Cut 5 pieces of beet roots ( containing Anthocyanin pigments ) using a cork borer all
of same size and from beetroots of same species and age
4. Rinse and dry to remove pigments from surface due to cutting
5. Allow equilibration of the 5 test tubes of water to reach to experimental temperature
by leaving them in the water bath at required temperature for 5 mins.
6. Add the beet root pieces to the water
7. Use the colorimeter to measure the degree of absorbance
Zero the colorimeter using a blank cuvette filled with distilled water .
Explanation as unit 1 exactly

Absorbance

Sudden steep increase in rate of diffusion when protein molecules in the cell membrane
denatures so begining to lose their tertiary structure .

A at low temperature
Cell membrane / tonoplast still intact
Pigments are large molecules
So too large to pas through membrane

As temperature increase at B :
1. Increase in kinetic energy of molecules
Increase movement and diffusion of pigment molecules
2. Phospholipid membrane becomes more fluid ….by decreasing interaction between fatty acid tails ..xo become
more loosely packed ..
SO < INCREASE IN PERMEABILITY of the membrane so more pigment molecules can diffuse outside so % of
light absorbance increase , light transmission decrease .

At ver high temperature :

The protein molecules in the cell membrane becomes totally denatured by disrupting the bonds holding
the tertiary structure in place , also the phospholipid melt .
Gaps are formed i the membrane though which pigments flood out

At C ….the level of transmission / absorbance level off /out As the concentration of pigments inside
is equal to that outside the cell ( equilibrium ) .
B) effect of alcohol :

1.Carry out the experiment at 5 different alcohol concentrations .

2. Get 5 test tubes of same size and add equal volume of alcohol
3. Cut 5 pieces of beet roots using a cork borer all of same size and from beetroots of same species
and age
4. Rinse and dry to remove pigments from surface due to cutting
5. Leave them in the different alcohol concentrations for 15 mins.
6. Use the forceps to remove the cylinders from each tube …and discard the cylinders but keep the
supernatant liquid .
7. Use the colorimeter to measure the degree of absorbance
Zero the colorimeter using a blank cuvette filled with alcohol .

Explanation same as unit 1

Explain the results

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%
1. Phospholipid are soluble in alcohol , where alcohol dissolve the fatty acids in cell membrane
Altering the hydrophobic orientation …increasing the fluidity of cell membrane
2. At very high alcohol concentration , protein molecules will get denatured, resulting in gaps or holes in
the membrane which allow more pigments to diffuse out of the cell …increasing rate of absorbance .
Decreasing light transmission .

Variables that should be kept constant

1. Age of beet root and species

2. Size of beet root cylinders
3. Volume of water / alcohol
4. Volume of solution added to cuvette
5. Time of soaking
6. Time of equilibration
 Core practical 4

A ; effect of different temperatures on rate of enzyme catalysed reaction

1. Carry out the experiment under 5 different temperatures ( 10,20 ', 30,40,50 C) and control the
temperature using thermostatic water bath .
2. Get 5 test tubes add equal volume of ( 2Cm3) of 1 % trypsin
3. Get another 5 test tubes and add equal volume of milk
4. Allow equilibration ( leaving test tubes for 5 mins at exp temp )
5. Control pH using buffer solution
6. Set the colorimeter to zero absorbance …by getting a blank cuvette filled with water and trypsin .

7. Mix the trypsin with milk …shaking gently …..then place the solution in the cuvette to be put in the
colorimeter …read the absorbance of light
Repeat with the othefr 4 Test tubes at other temperatures
Remember to use the reference cuvette toset the colorimeter to zero absorbance

B. Effect of pH
C. Effect of substrate concentration
D. Effect of enzyme concentration

Why we measure initial rate

Substrate concentration is not limiting factor

But as the reaction proceeds , substrate is being used up becoming a limiting
factor
So we cant obtain a valid comparison .
 CP 5

Preparation of a slide
Safety
Taken from human
Safety
- cotton should be placed in a disinfectant
- wash ur hands before and after
- wear gloves
- goggles
- cotton buds should be used once for one individual
In general ( preparation of a slide ) :
- avoid skin contact with acetic orcin or methylene blue
- safely use the microscope …using a light microscope with mirror ( use
daylight ) , so don’t place where sunlight might strike the mirror as this will
damage the retina

Slide preparation

1. For cheek 2. For plants

- use cotton buds - on a chopping board cut a thin transverse

- gently rub on inside part pf ur cheeks section of the plant tissue
-thin section is needed to allow light to
- rub the cotton bud on a glass slide
penetrate / pass through it .
- add few drops of methylene blue
- mount with water to hold the sample on a slide
- then cover with a cover slip - cover with a cover slip
- remove excess water using absorbent
Examine under microscope
-Place the slide on stage under the lens ( low power lens )
-switch on the light and examine the stained slide
-looking through eye piece , adjust the coarse focus using the coarse focus
knob
- then adjust using fine focus
- change to higher power lens to observe more details and magnify
 Core practical 6

Mitotic index

Procedure Mitotic index Core practical 6

1. Removal of the last 5-10 mm of root tips ( meristem).

2. Place root tip in hydrochloric acid to separate the cells/macerate tissue/ soften tissue (making
it easier to stain).
3. Addition of appropriate stain such as toludine (blue) , acetic orcein, methylene blue to high light
chromosomes.
4. Place the root tip on microscope slide, teasing cells (using needle to spread cells),
Squashing the cells underneath a cover slip to separate cells (for easier view),
and heat the slide to intensify the stain.
5. Ensure the students view the preparation under the microscope on low power first to identify the
area of dividing cells and to focus. Higher power will be required to examine the dividing cells for the
stages of mitosis
6. Counting number of cells in mitosis, then counting total number of cells , then Calculate the
mitotic index .

Cells in mitosis
Mitotic index X 100
Total number of cells For safety : wear eye goggles
Avoid contact with HCL irritant /
Measuring how actively the cells in a tissue are dividing corrosive …wear gloves

Core practical 7

Drawing

Core practical 8 ( tensile strength )

Core practical 8 ( tensile strength )

Effect of different length on tensile strength

1. Get fibres from same plant , of same age and same diameter .
2. But of different lengths …….carry the experiment under same
temperature , humidity , and thickness ( diameter) .
3.clamp one of the 20 cm string between two retort stands
4. Add Cushion underneath the string as as not to hurt your feet / to avoid
hitting the bench when string breaks
5. Gradually add increasing masses until the fibre breaks .
6. Record the mass of which the fibre breaks
7. Convert mass of which the fibre breaks
Tensile strength = breaking force / cross sectional area
8. Repeat same steps with other fibres of other lengths .

For accuracy use smaller mass intervals

For reliability repeat 3 more times at each length and calculate the average , SD and
plot a graph .
Core practical 9 ( aseptic technique )

1. Test for different parts of the plant/ different plants

2. Grinding plant materials to make extract
3. Prepare a lawn of bacteria .
4. Allow aseptic technique : using bunsen burner at yellow flame to creat a
convection current to carry a way air borne bacteria , sterile the inoculating loop
and the neck of flask containing bacterial lawn .
5. On agar petridish . Distribute bacteria evenly using the streaking method
6. Creat 4 equal sized wells ( opening in the gel) .
7. Place the paper discs soaked with different plant extracts . ….and the fourth
paper disc soaked with water as a control experiment .
8. Seal the lid at both sides but not all way round with cello-tape, to prevent
anaerobic conditions that might kill the bacteria in culture and cause growth growth
of anaerobic bacteria
9. Incubate at temperature 20-35 ( 27 C) for 24 hours ( to 7 days) to prevent
the growth of pathogenic organisms.
10. Measure the diameter of zone of inhibition
11. Then disposal ..putting petri dish in a plastic bag , seal and sterile at 121 C for 15
mins under high pressure and temperature then throw it away .
12. Repeat and calculate the mean value

Core practical 9

Aseptic technique :

1. Keep bunsen burner on yellow flame on bench

To creat convection current to carry away airborne bacteria

2. Sterile tools such as the inoculating loop and the neck of the flask containing the bacterial sample by
heating them in bunsen burner

3. Always when opening the lid of petri dish containing bacteria ....allow slight degree opening to avoid
contamination with another microorganisms causing disease .

4. Fastening to the lid using cello-tape, but dont tape all way round to avoid anaerobic conditions
To evaluate the data

1. Comment number of readings taken/ sample size ………….reliability

2. The range of measurements taken ………….accuracy and validity
3. Check reading that are inconsistent with other reading .. anomalies
4. Controlled variables ( same ) ……..validity
5. To improve reliability …..repeat to calculate mean value
and plot SD / error bars
Spot anomalies

Validity ;
1, controlled variables
2. Repeat

Reliability :
Repeating and getting similar results
Repeating and calculating average , SD and plotting graph and error bars

Accuracy
1. Way of measurements
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Answer ALL questions.

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Write your answers in the spaces provided.

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1 An investigation compared the protein content of some foods.

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

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and the liquid filtrate was tested for protein.
(a) (i) Describe how you could test for protein in this filtrate.
(2)

Add biuret reagent

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To the sample
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And if color turns from blue to purple color then protein is present
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(ii) Describe how the filtrates should have been prepared to allow a valid
comparison of the protein content of these foods. Controlled variables
(3)

Same mass of food sample

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Same volume of water used for extraction

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Same temperature
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Same type and size of filter paper

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*P67104A0216*
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(iii) Protein solutions of concentrations from 0.0 to 10.0 mg cm−3 were tested using

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a semi-quantitative method.

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The results of the test are shown in the diagram.

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0.0 0.1 0.4 2.0 10.0

Describe the difference between a semi-quantitative and a quantitative test.

Semiquantitative is a Rough estimation to the concentration of a (2)

substance present comparing it against standard / known

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concentration
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Which involves : subjective judgement to color change

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Quantitative method : Exact determination to the concentration of

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substance present .
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Measure the intensity of color using a colorimeter and using calibration

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curve ..
(iv) Explain how the results shown in the diagram can be used to compare the
protein concentrations of food extracts.
(2)
Get the sample and add biuret reagent
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Compare the color produced against the tubes shown in the diagram
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Where the concentration is estimated by deciding which protein solution in the

diagram has the closet color to the tested food sample
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Where the darker the color the higher the concentration of protein .
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*P67104A0316* Turn over
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(b) Another method used to determine the protein content of food is to measure the

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total nitrogen content. The total nitrogen content is multiplied by a conversion

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factor to give protein content.

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The table shows data for milk and soya beans.

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Total nitrogen Conversion Protein content
Food
content / mg factor / mg

milk 505 ✗ 6.38 3222

soya beans 3 600 5.71 20 556
✗

(i) Use the information in the table to calculate the ratio of the protein content of
soya beans to that of milk.
Give your answer to one decimal place.
(2)

20556/3222= 6.4

6.4
Answer ......................... ............... . . . . . . . . . . . . . . . . . . . . . .

(ii) Some of the total nitrogen content of a food is not due to protein.
Give one other type of organic molecule that contains nitrogen.
(1)

Nucleotides / nucleic acid / DNA

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(Total for Question 1 = 12 marks)

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*P67104A0416*