SAIntS Biology Department 2013/14

AS Core Practicals Handbook

Edexcel Specification
Edexcel AS Biology – Core Practical Handbook 1
 AS Biology – Core Practical Handbook

Contents Page:

AS Core Practical Titles Page: 2

Practical 1.1 - The effect of caffeine on heart rate. Page: 3

Practical 1.2 - The vitamin C content of fruit juice. Page: 6

HSW activity - Lifestyle factors & heart disease Page: 8

Practical 2.1 - The effect of temperature on membranes. Page: 9

Practical 2.2 - Enzyme concentration and rate of reaction. Page: 12

Practical 3.1 - Observing mitosis. Page: 14

Practical 3.2 - Totipotency and plant tissue culture. Page: 19

Practical 4.1 - The strength of plant fibres. Page: 22

Practical 4.2 - Investigating plant mineral deficiencies. Page: 25

Practical 4.3 - The antimicrobial properties of plants. Page: 26

HSW Criteria / Learning Outcomes Page: 30

AS Core Practicals - Key Expressions (Paper 3B) Page: 33

AS Core Practicals - Summary Sheet Page: 35

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Alternative practical set-
up for catalase & H2O2
concentration

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Key Word /
Definition / Notes
Aspect
The difference between the actual values and the measured values. If the difference
is high, then accuracy is low and vice versa. Accuracy can be improved by using
Accuracy
appropriate apparatus and methods for making measurements. There is little
difference between your results and the recorded “true” results
Anomalous
A result that appears „out of place‟, often as a result of human error.
Result

When drug companies / scientists (who have a big stake in the outcome of an
Bias experiment) promote certain hypothesis / website / source / article; without
reference or cross checking.

Calculations Show the Working out – it may get you marks!

Choosing the Data is a shown as a percentage that adds up to 100% - Pie Chart
correct graph Both variables are quantitative (i.e. numerical) or one is discrete – Bar Graph
/ chart Both Variables are continuous – Line Graph
Control A factor that is kept constant so that its effects on the DV are consistent throughout
Variable all experiments
Dependent The factor that is measure to give the results & is affected by the IV. DV depends
Variable (DV) upon the IV.
Describe the trend in the graph without making theoretical assumptions. Make
conclusions based on the experimental data. Refer to the hypothesis being
Describing
investigated, while making conclusions. Limitations are genuine sources of error. Look
Trends
out for variables like temperature, pH, etc. not being controlled. Also check if the
experiment has been replicated.
Economic
An issue that is money / business related – e.g. loss of revenue / jobs
Implications
Environmental
An issue that is effects the environment (animals and plants) – e.g. deforestation.
Implications

Whether an issue is right or wrong (moral or immoral). E.g. keeping animals in zoos;
Ethical
testing medicines / drugs on animals; hunting animals (wrong of for ornaments /
Implications
trophies / dubious medications). Could lead to extinction.

The working hypothesis that the IV does have an effect on the DV. By disproving the
Null Hypothesis you can accept your Experimental Hypothesis. NB. It‟s virtually
Experimental
impossible to prove something correct, yet very simple to prove something incorrect.
Hypothesis
Therefore, scientists aim to disprove their Null Hypothesis, which then allows them to
accept their Experimental Hypothesis.

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Key Word /
Definition / Notes
Aspect
Axes: IV on X-axis. DV on Y-axis. Label both appropriately with units.
Scale: The curve should cover more than 50 % of the graph paper.
Plot all points accurately. Mark the plotted point with a dot and a circle or with a
Graph
cross.
Line: Either draw a best fit line / curve to show trends or join each point with a neat
straight line, passing exactly through the point. Do not extrapolate.
Graph / table
Must refer to the IV & the DV
titles
Independent
The factor that affects the DV. The factor you change.
Variable (IV)

Limitations Experimental failings or restrictions

Null Opposite of the working hypothesis: i.e. that the IV has no effect on the DV. Aim to
Hypothesis disprove this hypothesis experimentally

Work that is carried out prior to actually starting to collect results in an

Preliminary investigation. Allows consideration of amounts, equipment suitability & range of
Work results to take. Adjustments would then be considered before starting the actual
procedure.

A mistake in the method or malfunction in the equipment which leads to the production
Random Error of a single anomalous result, inconsistent with the trend. Once spotted, a random error
should be either repeated or ignored.

Raw Data Data that has not yet been organized, analysed or evaluated

Always need specific dates / details of website; authors; reference to additional of

References /
suitable non-web resources; ICUN red list reference to book; title of article /
Bibliographies
newspaper / journal

When the variability in replicated results is very high, the reliability is relatively low.
Due to certain variables not being controlled or faulty experimental procedures. Range
bars / error bars on graphs give a fair indication of the reliability. If there is
Reliability considerable overlap between error bars, then variability is very high and reliability is
low. Students should be able to identify factors that may decrease the reliability of
the results. The same results are recorded if the experiment is repeated. Standard
deviation / standard error are excellent measures of reliability.
1) Identify risk (e.g. powdered enzyme could affect respiratory system / get into eyes
Risk / skin cuts
Assessments 2) Minimise risk - e.g. wear mask / goggles / gloves / cover any cuts with plasters
3) Action if risk occurs – e.g. wash with copious water
Social
An issue that affects people – e.g. loss of homes; increase in disease / famine
Implications

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Key Word /
Definition / Notes
Aspect
Spearman Rank Statistical test which allows students to find out whether 2 variables are correlated
Correlation (i.e does increasing one cause the other to increase or decrease?)
Usually down to an uncontrolled factor, a systematic error affects the entire
Systematic experiment, usually shifting the results by a consistent amount each experiment.
Error Systematic errors always produce inaccurate results, but in some cases the data
produced may still be reliable & as a trend may still be observable, valid to a degree.
The IV comes in the 1st column. Arrange values in ascending order.
Label all columns and rows appropriately and accurately.
Tables Include SI units (International Standard units – i.e. Metric units) in the headings of
the columns and rows.
Be consistent with significant figures / decimal places.
Used to compare 2 means (mean = normal "average"). Involves choosing between 2
hypotheses. For the t-test, the null hypothesis (H0) is always that the 2 means are
equal – e.g. "mean length of leaves at the top of the tree = mean length of leaves at the
t-test
bottom of the tree". It can never be that the means are different, or that one is
bigger/smaller than the other. You always start out assuming that the null hypothesis
is true, and only change your mind if the evidence is good enough.
A combination of accuracy and reliability. Valid results are representative and can be
Validity
used to make accurate predictions.

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A level Biology - How Science Works

HSW Criteria Learning Outcome Practical

a) Explain how the development of scientific theories involves
1) Use theories, models and ideas to develop
hypothesising, collecting & interpreting data & using creative thinking.
& modify scientific explanations
b) Explain the importance of modelling as way of developing scientific

The Core Practicals cover the HSW criteria. Complete the last column, noting which practical fulfil each criteria.

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2) Use knowledge & understanding to pose
scientific questions, define scientific problems,
present scientific arguments and scientific
ideas
3) Use appropriate methodology, including
ICT, to answer scientific questions & solve
scientific problems
4) Carry out experimental and investigative
activities, including appropriate risk
management, in a range of contexts
5) Analyse & interpret data to provide
evidence, recognising correlations and causal
relationships
Edexcel AS Biology – Core Practical Handbook
understanding.
a) Distinguish between questions that science can address, and those
which science cannot address.
b) Identify scientific questions or problems within a given context.
c) Apply scientific theories to answer scientific questions or address
scientific problems.
Justify methods, techniques and processes used during scientific
investigations, including use of ICT, to collect valid & reliable data and
produce scientific theories for a chosen question or problem.
Produce a risk assessment before carrying out a range of practical work.
a) Analyse data including use of: descriptive statistics (mean, mode &
median, error bars, standard deviation identification of outliers and range);
graphic representation to identify patterns & relationships (eg correlation
and cause) with appropriate statistical tests (A2 only).
b) Interpret data with reference to methods of analysis used.
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 As you complete the core practicals - make a note in the table of which criteria the practical meets.
HSW Criteria
6) Evaluate methodology, evidence & data &
resolve conflicting evidence
7) Appreciate the tentative nature of scientific
knowledge
8) Communicate information & ideas in
appropriate ways using appropriate terminology
9) Consider applications & implications of
science & appreciate their associated benefits
and risks
10) Consider ethical issues in the treatment of
humans, other organisms & the environment
11) Appreciate the role of the scientific
community in validating new knowledge &
ensuring integrity
12) Appreciate the ways in which society uses
science to inform decision-making
Edexcel AS Biology – Core Practical Handbook
Learning Outcome Practical
Evaluate the validity of inferences made from data in terms of the methods,
techniques and processes used to collect & analyse the data, recognising
any systematic or random errors present or conflicting evidence.
Explain how scientific theories are developed, refined, supported or refuted
as new data or new interpretations of data become available.
Present scientific information using text, graphics and other media as
appropriate using scientific terminology with reference to data and credible
sources.
a) Evaluate activities in terms of their associated benefits and risks to
humans, other organisms and the environment.
b) Discuss the risk associated with an activity in terms of the actual level of
the risk and its potential consequences, associated uncertainties, and the
factors affecting people’s perception of the risk.
a) Identify ethical issues arising from the application of science as it impacts
on humans, other organisms and the environment.
b) Discuss scientific solutions from a range of ethical viewpoints.
a) Discuss the importance of critical evaluation of new data or new
interpretations of data which challenge established scientific theories or
propose new theories.
b) Describe how the process of communication through journals,
conferences & peer review contribute to validation of new scientific theories
by the scientific community.
Discuss how science influences decisions on an individual, local, national or
international level.
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 Name of practical; Other variables to be
Other equipment Method and outcome Possible evaluation issues
IV / DV controlled
Effect of caffeine  Temperature Microscope, Method: Remove 1 Daphnia and place in cavity slide. Remove pond water and replace  Ensuring Daphnia were same size
e
on Daphnia heart  Volume of solutions counter, cavity with distilled water. Leave for 5mins to acclimatise then observe & count heart rate  Left too long under microscope, temp 
rate  Stress of Daphnia slide, dropping under microscope for 30s, multiply number by 2 to calculate beats/min. Repeat with 2 (due to lamp) = increased heart rate
n n
IV: caffeine conc  Size of Daphnia pipettes, stop clock, more Daphnia. Repeat again, this time with small conc of caffeine solution in place of  Ensuring enough data is collected
n n n
DV: heart rate of  Time of distilled water, test distilled water. Carry out for 5 conc of caffeine = 3 repeats at 3 conc .  Too high conc of caffeine kills Daphnia
n
Daphnia acclimatisation tubes, stop clock Outcome: as caffeine conc increased, heart rate increased  Counting of heart beat can be inaccurate
The effect of  Temperature 1% DCPIP solution, Method: pipette 1cm3 blue DCPIP into test tube. Using burette (or accurate pipette)  Difficulty in controlling temperature
temperature on  Concentration of 1% vitamin C add 1% vitamin C solution drop by drop. Shake tube gently after each drop. Continue  Amount of shaking (too much adds oxygen
cell membranes DCPIP solution (1%) solution, range of until the blue colour just disappears. Record volume of solution needed to which will slightly restore the DCPIP to
IV: temperature of  Shake each tube fruit juices, test decolourise the DCPIP. Repeat further 2 times and calculate mean result. Repeat blue)
water same no. times tubes/conical flasks, procedure with different fruit juices.  End point difficult to judge as needs to be
3
DV: % transmission  Same end point beakers, pipette Calculations: 1cm of 1% vitamin C solution contains 10mg Vitamin C, therefore mass just when blue colour disappears especially
3 3 3
of light through colour. i.e. until blue accurate to 1cm , in 1cm = 10mg x volume of 1% vitamin C to decolourise 1cm of DCPIP. in highly coloured juices
resulting solution colour of DCPIP just burette, safety Mass in sample = mass of vitamin C to decolourise 1cm3 DCPIP volume of sample  Some loss of solution when transferring
3
disappears goggles required to decolourise 1cm DCPIP
The effect of  Volume of distilled Raw beetroot, size Method: using cork borer and knife, cut pieces of beetroot into 1 cm length cylinders. Place in  Some beetroot may have skin on affecting
temperature on water 4 cork borer, white distilled water overnight to remove any dye released on preparation. Wash and blot dry. Place 8 surface area.
boiling tubes of distilled water into 8 water baths of different temperature. Once at
cell membranes  Time left in water tile, knife, ruler,  Difficulty in maintaining temperature
temperature, add a piece of beetroot to each and leave for 30 mins. Remove beetroot and shake
IV: temperature of  Size of beetroot beaker, forceps,
tubes to disperse dye. Set colorimeter to % absorbance on blue/green filter. Calibrate using  Accurate reading of the colorimeter
water piece water baths, boiling distilled water in a cuvette first then add 2cm3 of beetroot solution from the first temp to a new  Accurate size of beetroot
DV: % transmission tubes, cuvette. Place into colorimeter to read % absorbance. Repeat for all other pieces.  From the different parts of the root
of light through thermometer, Calculations & outcome: to calculate % transmission = 100 - % absorbance.  Ensuring same amount of time at the
resulting solution colorimeter & As temperature increased, % transmission slightly increased to a point at which it greatly different temperatures
cuvettes, stop clock, increased due to membrane molecules gaining more heat energy, vibrating more to a point
distilled water, where the vibrations caused large gaps in the membrane enabling the release of dye also
syringe proteins in membrane denatured leaving large pores.
The effect of  Temperature Protease e.g.1% Method: make up different concentrations of enzyme using distilled water. Ensure different  Maintaining constant temperature
changing enzyme  Volume of enzyme trypsin, casein syringes for different chemicals to prevent cross contamination. Set up water bath for  Accurately making up the different
temperature to keep constant. Place 1 test tube of 5cm3 casein solution into water bath
concentration on  Volume of substrate solution, small concentrations
alongside second tube containing 2cm3 of 0.2% trypsin. Allow to acclimatise for 3 mins so that at
rate of reaction.  Concentration of beakers,
same temperature then add trypsin to casein, start stop clock. Time how long it takes for casein  Identifying end point consistently
IV: concentration substrate thermometer, solution to turn transparent. (mark a ‘X’ on the other side of tube, as soon as seen through  Difficult to see the cross through the
of enzyme  pH distilled water, solution stop clock). Repeat a further 2 times then repeat for next concn solution
DV: time taken for syringes, stopclock, Calculations & outcome: rate = 1/time
enzyme to break large beaker As concn of enzyme increases, rate of reaction increases until a plateau point where all enzyme
down substrate has metabolised all substrate immediately.
n
Using catalase in Method: using first conc of yeast solution, acclimatise to desired temperature  Attaching syringe can be slower allowing
yeast and hydrogen alongside separate tube of hydrogen peroxide. Set up gas syringe and set to 0. Quickly loss of gas
peroxide add peroxide to yeast and attach gas syringe. Read off the volume of O2 gas produced  Inaccurate reading of gas syringe in making
n
every 10 mins until 3 readings the same. Repeat 3x for each conc of yeast solution. up dilutions
Calculations & outcome: rate = initial rate of reaction = gradient at steepest point  Reaction going too quickly to read

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 n
from graphs of volume against time for each conc . Outcome as protease.

Name of practical; Other variables

Other equipment Method and outcome Possible evaluation issues
IV / DV to be controlled
Observing Mitosis Garlic roots, sharp Method: place test tube of 2cn3 1M HCl into 60˚C waterbath. Cut off 1-2cm of root tip  Resolution of microscope
Chromosomes knife, 1M hydrochloric from garlic root. Put in watch glass containing 2cm3 of acetic alcohol for at least 12 mins.  Human error in counting numbers of cells
acid, Ethanoic alcohol,
stained blue using Remove then place into another watch glass containing 5cm3 ice cold distilled water.  Enough time in the solutions to enable
Orcein ethanoic stain,
orcein ethanoic Leave for 4-5 mins, then remove and dry. Place tips into heated HCl for 5mins then repeat successful maceration or staining.
ice-cold distilled
stain water, water bath @ process again by placing tips back into acetic alcohol etc. Tips will be very fragile at this

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 60˚C, 2 watch glasses, point. Transfer 1 tip to microscope slide, cut 4-5mm from growing tip (site of mitosis) and
test tube, 2 pipettes, keep the tip. Gently break up (macerate) with mounted needle, add 1 small drop of orcein
microscope slides, ethanoic stain and leave for 2 mins. Add coverslip and blot with filter paper. View under
forceps, mounted
microscope and identify the stages of mitosis. Calculations: percentage of cells in each
needle, filter paper,
microscope with mag stage of mitosis
x100 & x400 Mitotic index: number of cells containing visible chromosomes total number of cells in the
field of view
Totipotency & Seeds of white Method: sprinkle seeds on damp sponge and allow to germinate. Use when just starting  Unwanted pathogens growing in the gel as
Tissue Culture mustard, agar, to unfold their cotyledons (seed leaves). Make up Agar gel and pour 2cm height of gel it is a good source of water and nutrients
distilled water, damp into McCartney bottles and allow to set. With sharp scissors, cut the tops off just below  Wrong part of the plant cut and inserted
sponge, cling film,
the shoot apex (including the cotyledons). This is called an explants. Push the stem of the into gel.
McCartney bottles,
weighing scales, explant into the gel (making sure cotyledons don’t touch agar) cover with cling film and
plastic tray, 250ml place on sunny windowsill. Observe over 10 days.
beaker, glass rod, Outcome: explant grows roots and leaves continue to grow. You need to be able to
scissors, sunny explain why they are covered in cling film and why they continue to grow even when
window sill covered. Also why they shouldn’t be opened again.
The strength of  Length of fibre Stems of stinging Method: plant material should be left to soak in a bucket of water for about a week in  Maintaining length of fibres
plant fibres  Size of each nettles or celery, order for the fibres to be easily extracted (called retting). Or celery stalks should be left in  Ensuring consistency when twisting or
IV: source and type bucket, gloves, paper beaker of coloured water in order for fibres to be easily seen and pulled out. Once fibres
individual mass plaiting
towels, clamp stands,
of fibre
slotted masses and
removed, connect between 2 clamp stands and gradually add mass in the middle until the  Using fibres of the same age (as they get
DV: mass that can holders, white tile, fibre snaps. Try with individual fibres from different plants and different ways of older they become more brittle)
be held sharp knife combining fibres eg twists and plaits. Can also compare stem to individual fibres.  Extracting whole fibres that are useful
Outcome: the more fibres combined together the stronger it is.

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 Name of practical; Other variables
Other equipment Method and outcome Possible evaluation issues
IV / DV to be controlled
Investigating plant  Volume of Mexican hat plantlets Method: half fill a tube with the ‘all nutrients present’ solution. Cover the top of the tube  Ensuring accurate measurement of
mineral mineral or geranium leaves, 7 with foil or paraffin and push down on covering so that there is a well in the centre. solutions
test tubes, test tube
deficiencies solution Gently push the geranium stem/roots of Mexican hat plantlet through the hole so it is in  No air bubble caught in xylem of geranium
holder, different
IV: minerals  Species of plant mineral solutions:-
solution below. Repeat with solutions lacking in nitrogen or phosphate or potassium or  possible microorganism growth in nutrient
present  Size of each lacking 1 nutrient magnesium or calcium or lacking all. Wrap all tubes in aluminium foil and place in tube solution
DV: physical container and 1 containing all, holder on sunny window sill. Observe regularly. Outcome: the ‘all nutrients present’ plant  Insufficient time to see an effect.
c
charact of the  Amount of light aluminium foil will look healthy whereas the others will all have some abnormality. Make sure you know
plant received what nutrient deficiencies affect plants.

n
Effect of garlic and  Conc of plant Agar plate seeded Method: make plant extract by crushing 3g of plant material with 10cm3 industrial 
mint on bacterial material with bacteria, plant denatured alcohol. Shake occasionally for 10 mins. Pipette 0.1cm3 of extract onto sterile  Growth of unwanted microbes on agar
growth  Lawn of material e.g. garlic paper disc. Allow to dry on sterile petri dish. Meanwhile label agar plates with date and plates due to bad aseptic techniques
IV: presence of bacteria on & mint, pestle & split into 4 sections. 1 for each type of plant extract. Place 1 disc of each extract in each  Not shaking extract enough to ensure
garlic or mint petri dish mortar, 10cm3 quadrant of the agar plate, close and tape with hazard tape. Leave to incubate over night enough active ingredient
DV: zone of  Contamination industrial and observe zone of inhibition. Carry out controls with just distilled water on discs.  Inconsistency when adding plant extract to
inhibition around of petri dish by denatured alcohol, Outcome: the control discs completely covered with bacteria, some plant extracts will paper discs.
disc other microbes sterile pipette, create larger zones of inhibition than others, meaning they are more effective at lower  Contaminating controls
 Same volume paper discs, sterile concentrations.  Using wrong species of bacteria for lawn
of plant petri dish, sterile
material on forceps, hazard
each disc tape, marker pen.

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