Biology HL Internal Assessment

Research Question: What is the effect on the use of different antioxidants (40g
dissolved in 100ml of water) in the growth rate of Raphanus sativus (3mg)?

Candidate Code: hgf427

Date of Submission: 16 of November, 2020
Group 4: Sciences - Biology HL
Table of Contents
Personal Statement………………………………………………………………………..3
1: Introduction…………………………………………………………………………….3
2: Investigation
2.1: Hypothesis………………………………………....………………………....3
2.2: Background Knowledge……………………………………………………...3
2.3: Variables……………………………………………………………………...4
3: Procedure
3.1: Apparatus……………………………………………………………………..6
3.2: Photography of set-up………………………………………………………...6
3.3: Methodology………………………………………………………………….7
3.4: Justification…………………………………………………………………...8
3.5: Risk Assessment……………………………………………………………...8
4: Raw Data………………………………………………………………………………..9
5: Processed Data………………………………………………………………………….9
5.1: Statistical Test……………………………………………………………….10
5.2: Notes and qualitative observations………………………………………….11
6: Evaluation
6.1: Conclusion…………………………………………………………………..12
6.2: Weaknesses and Improvements……………………………………………..13
6.3: Extensions…………………………………………………………………...14
Appendices
Appendix 1A……………………………………………………………………..14
Appendix 1B……………………………………………………………………..14
Appendix 1C……………………………………………………………………..15
Appendix 2A……………………………………………………………………..15
Appendix 2B……………………………………………………………………..16
Appendix 2C……………………………………………………………………..16
Bibliography (APA 6th Edition)………………………………………………………...16

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Research Question: What is the effect on the use of different antioxidants (40g dissolved in
100ml of water) in the growth rate of Raphanus sativus (3mg)?
Personal Statement
On a personal level, the connection between helping me understand possible solutions and considerations
for the growth and maintenance of crops. On a personal level my family owns lands and crops in which
the use of fertilizer is both expensive and in big quantities, for so these compounds are more corrosive to
soil and complicate near future growth of crops. On the other hand, getting to know how plants work and
its effect from an antioxidant, I get to know close the effect of a type of drug which later on can come in
hand in case I decide to study Medicine or in the case of studying Biochemistry and pharmaceutical
studies, I have the opportunity to understand or increase my basic knowledge for a better performance
later on in my career.
1: Introduction
Raphanus sativus (radish) is an annual or biennial plant with the special ability to grow at a faster rate
than other plants.(Encyclopædia Britannica, 2019). R.sativus is usually used to treat respiratory and liver
illnesses (R.Perez & R.Lule, 2004), as known for matter of photosynthesis, plants absorb nutrients from
the soil through their xylem in order to produce the necessary nutrients for their survival. Plants age and
die from several situations, some of them are that cells under oxidative stress die or last shorter, but for
these conditions antioxidants exist. Antioxidants repair DNA and maintain cells as healthy as possible to
prevent oxidative conditions and free radicals which damage DNA instructions, production of amino
acids and lead to oxidative stress (damage cells and leads to diseases). Through giving electrons to the
free radicals, antioxidants detain the seeking of free radicals for electrons and damaging the cell. (Harvard
T.H. Chan School of Public Health, n.d). Hence, the increase of antioxidants inside a cell or plant cell in
this case, leads to a longer longevity of cells and thus, increases cell growth rate, and general growth of an
organism.To increase cells longevity and its growth we could have the implementation of vitamins into
their watering in order for these seeds to store these compounds, since there is a bigger chance for cells to
last longer and plants to grow faster. The usage of antioxidants in agriculture and plants, leads to a
healthier and more biological product than fertilizers that lead to soil corrosion and future crops damage.
2: Investigation
2.1: Hypothesis
It is believed that the use of antioxidants in the growth process of radish (Raphanus sativus) will increase
both seed size therefore it will increase its speed growth. According to scientific bases and investigations,
antioxidants play an important role in plants, low molecular weight antioxidants in plants between those
Vitamin C or ascorbic acid and vitamin E, function as a redox (reduction and oxidation) buffer which
influences its oxidation, ribosomal malfunction through a lower process of ageing, and increases the
development of the plant, in processes such as cell elongation and mitosis. (Kasote et al, 2015). Thus,
under the presence of various antioxidants (Vitamin A, C & E), R.sativus growth will have an increase in
its growth rate compared to a normal amount of water used for its care. We might observe a major change
in growth within samples of vitamin C, followed by vitamin E and after vitamin A.
2.2: Background Knowledge
Vitamin A (C20H30O) is a micronutrient which can’t be produced and thus it must be ingested, its main
objective is to maintain all surface tissues integrity and function, finally, it supports the replacement of
dead cells for new cells. (Gilbert, 2013); vitamin A needs to be processed by carotenoids in order for the
production of the vitamin. Vitamin C or ascorbic acid (C6H8O6) is another micronutrient which performs
the repariment and growth of tissue in an organism, promoting both a faster growth and development of

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the plant. As many other vitamins they work as a free radical scavenger, decreasing oxidative stress,
hence preventing cells and the organism to oxidise and die faster. (Abdullah et al, 2020). Vitamin E
(C20H30O) as mentioned before, works to prevent oxidation in cell membranes and lipoproteins by free
radicals, and prevents the oxidation of lipids preventing further chain reactions from a free radical.
(Böhm, 2018).

ROS or reactive oxygen species are those that cause during germination, aging and desiccation, oxidative
stress (oxidation) in higher concentrations leading to a major cellular damage and the complete
deterioration of seeds and organisms. Although plant cells have internal antioxidant systems (enzymes)
which scavenge ROS, limiting the oxidative conditions and supporting a longer life for cells, antioxidants
provide these features in major quantities. (Pehlivan, 2017). Through germination time the antioxidant
properties of edible seeds such as R.sativus increase slightly (times 5.14) although free radicals and
oxidative stress persists its existence ((Tarasevičienė et al, 2019). Hence, the use of antioxidants leads to
plants store these compounds and reduce the oxidative condition, leading to a longer lasting of cells and
tissue.

Antioxidants provide protection against oxidative conditions in cells, according to the University of
Exeter (2007), “vitamin C is able to provide protection against side-effects of light during
photosynthesis”. This vitamin is able to contribute enzymes that protect cells during photosynthesis and
from radical cells, providing a faster growth for plants. In addition, vitamin E “is able to limit non
enzymatic lipid oxidation during seed storage, germination and early seedling development” (Sattler et al,
2004), which as much as vitamin C provides, coverage for any type of oxidative conditions in cells,
ensuring a both better and faster growth for plants during their seed fase until their lasting fase as a plant
that has already bore fruit or flowered. On the other hand, vitamin A which is changed during digestion
from carotenoids, provides the green colour to plants, although it still has antioxidant properties which
help for the ROS but in a smaller proportion compared to other vitamins. (Gilbert, 2013).
2.3: Variables
Independent Variable: 40g of different Antioxidants (Vitamin A, Vitamin C, Vitamin E) dissolved in
100ml. Through the use of multiple antioxidants, we will observe which one is more effective during the
growth process of a plant in this case the R.sativus. All antioxidants will be placed under the same amount
of circumstance although the type of antioxidant will vary. It will be measured through the use of an
electronic scale (± 0.01g) and a beaker of 100ml (±0.1ml) in order to reach the values established and
create the solution. The variable hasn’t been chosen as quantitative since we are measuring the effect of
different antioxidants on the growth of R.sativus rather than the effect on the change of mass of
antioxidants in plant growth.
Dependent Variable: Growth of 3mg of Raphanus sativus seeds. Under the topic of seed growth and the
effect of different biotic or abiotic factors in its rate, we search the effect on its speed of the seed to grow.
Thus, the size will be measured by the use of a ruler (±0.1cm) with a time interval of 48 hours between
every measure. Calculations needed in order to understand the effect of antioxidant on the growth of
plants is through a comparison of change between hour 0 and hour 336, helping compare growth for each
sample, values will then determine an average of growth per hour (cm/hour) under the effect of each
antioxidant.
Controlled Variables:

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Name of How will it affect the results?
Controlled
Variable
Type of If we consider the use of various types of Raphanus
Raphanus sativus, this will have a different growth rate between
sativus each other. Some might grow bigger and faster while
others might grow slower and smaller. The use of
different types of radish won’t help for valuable and
precise data in order to determine a conclusion from
the experiment/investigation.
Mass of The variation in the mass of antioxidants will lead to
antioxidant a major variation in the concentration of antioxidants
used in the solution. This variation might lead to the
possibility of certain seeds having more antioxidant
during their growth leading to certain variations and
diverse results. This type of variation won’t make our
results trustworthy.
Volume of As much as the possible effects of using a major mass
water poured of antioxidant in the solution with distilled water.
to seed sets Using more of the solution in the seeds will lead to
the possibility of more antioxidants being in reaction
with the seed affecting its growth and consequently
data being different from the other leading to a major
error and less reliable data collected.
Times of Spraying either more or less times the sets of seeds in
spraying every repetition will lead to some sets of seeds
solution reacting at different times with the antioxidant and
leading to variation in the growth speed of the
R.sativus during its development. Having different
amounts of antioxidant solutions reacting will lead to
certain sets growing bigger or smaller than others.
Time Varying between the days or intervals in which plants
Intervals for will be measured its growth, values won’t lead to
Growth prove or portray a trend in which can be calculated
measurements exactly the change of growth under the effect of
antioxidants.
Mass of The use of different masses of Raphanus sativus
Raphanus seeds will lead or cause significant contrast between
sativus seeds the results taken during the experimentation. This
(Raphanus will cause uneven and disbelief in the results. Bigger
sativus) sized seeds have both more volume for nutrient and
material storage and therefore have a better chance of
How are we going to control them?
The type of Raphanus sativus will be controlled
through using samples from the same bag or type
bought in the market. Using Raphanus sativus seeds
bought from the same place and sampled from the
same bag will reduce the uncertainty to vary in type
of Raphanus sativus leading to reliable results.
With the use of an electric scale (±0.01g) we will
measure or weigh the exact amount of antioxidant
(40g) that will be poured in the solution with 100ml
of distilled water. Being able to set the same
standards during the experiment will give a major
certainty in our results.
With the use of a beaker of 100ml we will be taking
the amount set to be poured over the seeds every 48
hours (14ml) from the solution of distilled water and
the 40g of antioxidant.
We will spray the seeds with the set amount 14ml of
the solution created from the 40g of antioxidant and
100ml of distilled water every 48 hours starting at
hour 0 and finishing at the hour 336. Using the same
intervals for spraying the water will lead to the same
amount of solution reacting with the seeds, and the
seeds having the same time to grow with an equal
volume of solution.
As well as the times of spraying solution I will be
measuring the plants growth starting at hour 0 and
going on every 48 hours. Giving the chance for all of
the plants to be measured at the same time and
having the same opportunity to grow between time
intervals.
This variable will be controlled through the use of an
electronic scale (±0.01g) which will help us get an
almost precise value of the mass of each seed of
Raphanus sativus (3mg) being used during the
experimentation, so maturation conditions are equal
for all other seeds.
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 growing or gaining more nutrients than any other.

Distilled When doing certain experiments with water solutions, We will manipulate or control this variable through
Water the use of normal water won’t be successful enough the revision of a laboratory scientist that will provide
to react accordingly to planned due to the fact that tap most of our materials to reduce any error
water drags salts and other minerals present in (systematic). The distilled water will be used in all
pipelines which can have an effect both in growth and of the antioxidant solutions but it will be in its pure
in the reaction of water with the antioxidant, affecting state in order to calculate a normal growth of a
antioxidant properties and qualities. (Burg & Oshrat, Raphanus sativus seed without any external
2015) compounds reacting with it.
Uncontrolled Variables:
Light Light will be another factor which can affect the Although measuring the light being received or directed
photosynthesis of the seeds and its germination into the plant can be complex to measure. I will be
due to the fact that light can make the process creating the experiment in indoor conditions. Thus, light
faster or slower and thus, the growth of the plant will be equally distributed or recieved between all of the
can vary significantly leading to the disbelief and seeds without any alteration of light during the day if so
annulment of our results seeds were planted with direct sunlight.

Temperature Temperature variations will make the germination Temperature variables will be minimized by the
process and simultaneously the photosynthesis experiment being placed indoors, the temperature will
process in order for the plant to grow, either range between 24.0°C - 28.0°C. Temperature will vary
slower or faster leading to different values and between the range depending on the atmospheric
certain big variation between repetition. conditions.
3: Procedure
3.1: Apparatus
❏ 100 seeds of radish (Raphanus sativus) of 3mg
❏ 1 electronic scale (± 0.01g)
❏ 2 liter of distilled water
❏ 1 Chronometer (± 1 h)
❏ 1 Lab Coat
❏ 1 Thermometer (± 0.1°C)
❏ 4 plastic plates
❏ 1 Pair of safety goggles
❏ 20 Paper Towels
❏ 4 beakers of 100ml
❏ 1 syringe (±0.1ml)
❏ 1 Notebook for Data Collection
❏ 200 g of pulverized Vitamin A, Vitamin C and Vitamin E
❏ 1 Magnetic Stirrer
❏ 1 Ruler (±0.1cm)
❏ 1 Marker
3.2: Photograph of set-up
A picture taken by myself using an iphone XR, on 03/10/2020, it displays how the experiment should look like with
the Raphanus sativus sample seeds from all 5 repetitions for each solution before starting

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3.3: Methodology
1. Collect all of the necessary materials and place the materials in a safe and spacious space
2. Make sure that the instruments of measurement such as rulers and chronometer are in order and
well prepared. Measure constantly the room temperature to ensure it is between 24°C and 28°C.
3. Put on lab coat and goggles
4. With the use of the electronic scale (± 0.01g) weigh 40g from the 200g sample of Vitamin C
5. Take a beaker of 100ml and fill it with 100ml of distilled water. Simultaneously, with the use of a
marker write on the beaker which repetition the beaker represents and which antioxidant it has
(Vitamin A, C or E).
6. Pour the 40g of Vitamin C in the beaker of 100ml of distilled water
7. Take the beaker and place it in the magnetic stirrer. Turn it on and wait until all of the vitamin has
dissolved in the water, once it is dissolved turn it off and place the solution aside.
8. Take 5 seeds, from the 100 seeds of R.sativus, and with an electronic scale weigh 3mg each.
9. Take a paper towel and place it over one of the plastic plates. After, place the 5 seeds with a
certain separation between each other.
10. With the use of a syringe (±0.1ml) take 14ml from the solution of antioxidant and water from the
step #8, and pour it evenly over the seeds. Place the seeds in a location with enough light for all
of them to simultaneously have the same amount of light.
11. Once poured the seed with the solution measure its length if any and start the chronometer
12. While time passes by collect all type of qualitative data during the experiment
13. When the chronometer has set 48 hours have passed since poured, measure the seeds growth with
a ruler (±0.1cm) and record it in the Table #1.
14. Take the syringe and extract another 14ml from the solution and pour it evenly in the seeds. Once
poured start the chronometer again until it reaches 48 hours again.
15. Repeat step #11 to step #14 until 228 hours have passed from the first time the seeds where
poured with the solution
16. Start the chronometer for 48 more hours, and collect the last data of how much each seed grew.
Data will be based on 336 hours after the seeds where poured with the solution for the first time
17. Repeat step #5 to step #16 for the other antioxidants (Vitamin A and E) being conscious that the
data being recorded is well organized and understandable for further analysis. Record the data
with similar tables as Raw Data Table #1. (raw Data Tables #2 and #3).
18. For water only skip the solution making process and skip the weighing of antioxidants, although
measure 100ml with the use of a beaker of 100ml.
19. Repeat the whole experiment from step #1 to step #18, 4 more times in order to make the results
more precise and reliable. Record the results in the correct raw data table. (It is recommended to
do all of the 5 repetitions at the same time for time saving and to ensure a more equal

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 circumstance for uncontrollable variables.). In case that all seeds don’t grow, repeat the
experiment until reaching the complete 25 seeds for each solution (5 seeds for 5 repetitions).
20. Once finished, organize and clean materials, get rid of any waste from liquids or other materials.
21. Make a second table for each raw data table, where you state the average growth of the 5 seeds
from each repetition. Add 2 more columns where you state another average of the 5 repetitions
and standard deviations. (Look at Table #5,6,7 & 8).
22. Make an ANOVA test based on the column 7 of the results from Tables #5,6,7 and 8, in order to
determine any statistical significance between the data.
23. Make a scatter plot graph were graph error bars represent standard deviations and a trendline
24.
25. Check the graph for any inconsistent data and check why such data isn’t consistent, refer to
qualitative data recorded for such data taken during the experimentation to make a conclusion of
possible effect from external environmental variables
26. Consider the graph slope to conclude on the increase in the speed growth of R.sativus with the use
of the various antioxidants. Take into account, the values used are averages which might
influence the ending result of the experiment.
27. Evaluate the experiment and make conclusions taking into account both results and method.
Consider possible changes for future investigations that came out from the experiment.
3.4: Justification
The independent variable previously presented were used for the main specific reasons which are Vitamin
A, Vitamin C and Vitamin E. the most known and currently easily accessible antioxidants in the market
which allow a more lifelike comparison of antioxidants in seed development. The absence of antioxidants
in the experiment represents the conventional watering process during the growth of a seed which will
lead to the comparison of the growth effect of antioxidants vs. a normal watering process.

I’ve decided to realize the experiment with 5 seeds per antioxidant in order to have many data and
repeated the experiment 5 times, reaching a total of 25 seeds per antioxidant in order to reach reliable and
accurate data from the experimentation. Distilled water was used in order to prevent any type of metal,
chemicals or external compounds present in colombian water that might affect both water and its reaction
with antioxidants during its solution creation.(Instituto Nacional de Vigilancia de Medicamentos y
Alimentos- INVIMA, 2015).

The growth rate or the dependent variable was established with these certain measurements because these
measurements will lead to the comparison of distance or growth by a measurement of time, in other words
a speed like measurement (cm/hours). Results graph will lead to the analysis of the data for the speed rate
for each antioxidant solution (40g in 100ml of distilled water) in R.sativus seeds of 3mg.
3.5: Risk Assessment
Safety Issues: Although all materials used weren’t corrosive or dangerous to our health, the use of
goggles and lab coats was used for more protection and to prevent any type of harm.
Ethical Issues: There were no ethical issues needed to be taken into account due to the fact that
antioxidants were used rather than antibiotics that might affect bacteria resistance.
Environmental Issues: Using antioxidants won’t affect a plant by leading it to die faster. Since it won’t
be grown in soil, meaning that the use of vitamins won’t be near soil that can get damaged from such.

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 4: Raw Data
Raw Data Table #1: A table showing the growth (± 0.1cm) of Raphanus sativus seeds of 3mg (±0.01g)
during 336 hours while pouring 14ml of water every 48 hours 5 seeds in each of the 5 repetitions for
water only.
Water Only
Growth - G
Time - T δG = ± 0.1cm
δT = ± 1 hour Repetition #1 Repetition #2 Repetition #3 Repetition #4 Repetition #5

1 2 3 4 5 1 2 3 4 5 1 2 3 4 5 1 2 3 4 5 1 2 3 4 5
0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0
48 0,7 0,6 0,6 0,8 0,9 0,6 0,7 0,7 0,9 0,5 0,8 0,8 0,6 0,5 0,7 0,7 0,8 0,7 0,7 0,9 0,6 0,7 0,6 0,7 0,9
96 1,5 1,5 1,4 1,7 1,7 1,5 1,6 1,4 1,8 1,4 1,8 1,6 1,4 1,6 1,7 1,7 1,7 1,5 1,6 1,9 1,5 1,8 1,6 1,7 1,8
144 2,4 2,6 2,3 2,7 2,6 2,5 2,5 2,2 2,5 2,3 2,7 2,6 2,4 2,6 2,5 2,8 2,7 2,4 2,5 3,0 2,6 2,7 2,4 2,5 2,8
192 3,4 3,6 3,3 3,5 3,6 3,6 3,4 3,2 3,6 3,3 3,7 3,5 3,2 3,7 3,4 3,8 3,7 3,3 3,5 3,8 3,5 3,7 3,3 3,6 3,7
240 4,6 4,5 4,4 4,5 4,3 4,6 4,4 4,1 4,5 4,4 4,8 4,4 4,1 4,6 4,5 4,7 4,7 4,3 4,5 4,7 4,6 4,6 4,4 4,6 4,8
288 5,6 5,5 5,5 5,6 5,3 5,5 5,6 5,2 5,4 5,3 5,6 5,3 5,0 5,5 5,6 5,7 5,5 5,2 5,4 5,6 5,7 5,3 5,4 5,8 5,7
336 6,5 6,6 6,4 6,8 6,3 6,5 6,4 6,1 6,3 6,1 6,6 6,3 6,0 6,5 6,5 6,6 6,4 6,1 6,3 6,5 6,6 6,1 6,2 6,7 6,6
Total Increase 6,5 6,6 6,4 6,8 6,3 6,5 6,4 6,1 6,3 6,1 6,6 6,3 6,0 6,5 6,5 6,6 6,4 6,1 6,3 6,5 6,6 6,1 6,2 6,7 6,6
Note: Move on to appendices section 1 (appendix 1A, 1B and 1C) to observe the rest of raw data tables
for the reaction of Raphanus sativus seeds under the reaction of vitamins A, C and E.
5: Processed Data
Processed Data Table #1: A table showing an average of the samples for Raw Data Table #1 (water
only) Raphanus sativus seeds for all 5 repetitions across 336 hours
Water Only Standard
Average
Time - T Deviation
Average Growth of the 5 seeds of Raphanus sativus from each repetition - G Growth - G
δT = ± 1 Growth - G
δG = ± 0.1cm δG = ±
hour δG = ±
0.1cm
Repetition #1 Repetition #2 Repetition #3 Repetition #4 Repetition #5 0.01cm
0 0,0 0,0 0,0 0,0 0,0 0,0 0,00
48 0,7 0,7 0,7 0,8 0,7 0,7 0,03
96 1,6 1,5 1,6 1,7 1,7 1,6 0,06
144 2,5 2,4 2,6 2,7 2,6 2,6 0,09
192 3,5 3,4 3,5 3,6 3,6 3,5 0,07
240 4,5 4,4 4,5 4,6 4,6 4,5 0,08
288 5,5 5,4 5,4 5,5 5,6 5,5 0,07
336 6,5 6,3 6,4 6,4 6,4 6,4 0,08
Note: Move on to appendices section 2 (appendix 2A, 2B & 2C) to observe the rest of processed data
tables for the reaction of Raphanus sativus seeds under the reaction of vitamins A, C and E.

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 Σ 𝐺𝑟𝑜𝑤𝑡ℎ 𝑜𝑓 𝑠𝑒𝑒𝑑𝑠 𝑓𝑟𝑜𝑚 1 𝑟𝑒𝑝𝑒𝑡𝑖𝑡𝑖𝑜𝑛 𝑎𝑡 𝑎 𝑐𝑒𝑟𝑡𝑎𝑖𝑛 𝑡𝑖𝑚𝑒
Calculation of average growth per repetition at certain time: 𝑁𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑠𝑒𝑒𝑑𝑠
Example calculation for average growth at certain time:
0,7 + 0,6 + 0,6 + 0,8 + 0,8
❏ Average growth at 48 hours for repetition #1 with Water Only: 5
= 0, 7
(Rounded to one decimal place for all values, because the uncertainty of the growth is ±0.1cm).
❏ The same calculation process was carried out for raw data tables #2, #3 and #4 as seen in
appendix section 2 (appendix 2A, 2B and 2C).
Calculation of standard deviation growth (STDEV.P) for all repetitions at a certain time:
2
Σ(𝑥 1− µ)
σ= 𝑁
❏ Standard deviation growth at 48 hours for repetition #1 with Water Only:
2
Σ(𝑥 1− 0,7)
❏ Average is known thus it can be simplified in the fórmula : σ = 𝑁
❏ Subtract from each value of the set the mean, and square the result:
❏ (0,7 - 0,7)^2 = 0 // (0,7 - 0,7)^2 = 0 // (0,7 - 0,7)^2 = 0 // (0,8 - 0,7)^2 = 0,01 //
(0,7 - 0,7)^2 = 0
0+0+0+0,1+0
❏ Add all number and divide by n-1 or 5-1 in this case: 5−1
= 0, 025
❏ Square root the previous answer 0, 025 = 0, 03
Standard deviation example was calculated with rounded samples, although the real calculations in the
table were done with the complete not rounded result with help of a statistical software (Microsoft Excel).
Calculation of average growth for all repetitions at a certain time (Column 7 of Processed Data Tables):
Σ 𝐴𝑣𝑒𝑟𝑎𝑔𝑒 𝑔𝑟𝑜𝑤𝑡ℎ 𝑓𝑟𝑜𝑚 𝑒𝑎𝑐ℎ 𝑟𝑒𝑝𝑒𝑡𝑖𝑡𝑖𝑜𝑛 𝑎𝑡 𝑎 𝑐𝑒𝑟𝑡𝑎𝑖𝑛 𝑡𝑖𝑚𝑒
𝑁𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑟𝑒𝑝𝑒𝑎𝑡𝑠
Example calculation for average growth for all repetitions at a certain time:
0,7 + 0,7 + 0,7 + 0,8 + 0,7
❏ Average growth for all repetitions at 48 hours with Water Only:: 5
= 0, 7
(Rounded to one decimal place for all values, because the uncertainty of the growth is ±0.1cm).
❏ The same calculation process was carried out for raw data tables #2, #3 and #4 as seen in
appendix section 2 (appendix 2A, 2B and 2C).
5.1: Statistical Test
In order to be able to denote a statistically significant difference between the various groups and values
recollected from the variation of antioxidants and its effect in Raphanus sativus, a one-way ANOVA test
was performed based on the 7th column of the processed data tables #1, #2, #3 and #4, through the use of
Microsoft Excel. The test provides a statistical correlation between the use of different antioxidants and
its effect in Raphanus sativus growth rate, plus the use of only one independent variable defines the use of
a One-way ANOVA rather than a Two-way ANOVA.
Null Hypothesis (H0): It is believed that there is no notable relation between the seed growth rate of
Raphanus sativus, and the use of different antioxidants.
Alternative Hypothesis (H1): It is believed that there is a remarkable relation between the use of
antioxidants and seed growth rate of Raphanus sativus, and the use of different antioxidants.

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 Processed Data #5 (Screenshot from Excel): A table showing the ANOVA test results

Based on values of F crit being higher than F value provided by the ANOVA test (One Way), the null
hypothesis will be rejected and thus, the alternative hypothesis will be accepted. For instance, there is a
significant statistical difference between the use of different antioxidants and the growth of Raphanus
sativus over the course of 336 hours. For this reason, we could consider that there is a relationship
between the established hypothesis and the hypothesis accepted according to the statistical test.
Graph #1: A graph showing the effect of antioxidants and water only on the growth of Raphanus
sativus (3mg) during a period of 336 hours

The error bars of the graph represent the standard deviation on each data point calculated in the processed
data tables, whilst the R2 value determines fitting of the line of best fit. The small values of both error
bars and R2 value represent a major accuracy and certainty of the data collected through the experiment.
5.2: Notes and qualitative observations
1. Vitamin A & E solutes didn’t dissolve completely in the solution

11
 2. The seeds that were being used during the experiment with the solution of vitamin C had a brighter
color more orange-red than the rest of the Raphanus sativus seeds.
3. The size of the seeds of all the samples tested increased over the course of the experiment. This
increase was more pronounced when using the solution with vitamin C.
4. Vitamin A, vitamin E and water only seed samples had similar growth speed during the first 3 days,
after such time passed vitamin samples started growing at a faster rate.
5. On the fourth day during the use of vitamin C in repetition 3 and during the seventh day of the use
of vitamin E, seeds felt more turgid to the touch during the time of the measurement. Vitamin A
samples felt a little turgid than water only samples.
6. Through time for biological reasons seeds started growing bigger until they popped and started
growing, the time until such happened was an average of 2 days until the seeds popped up.
7. Samples of water only had a significant increase in the stems thickness. Water only samples grew
wider than vitamin solution samples. Vitamin solution samples grew longer rather than wider.
8. Vitamin A samples grew at a faster rate than water only samples during the first week but started
growing at a similar almost equal rate than water only seed samples.
9. Vitamin A samples showed a greener color roots than samples with water only, vitamins E & C
6: Evaluation
6.1: Conclusion
Based on the previous experiment and its outcome results, I could conclude that the use of antioxidants
are able to prove that there is a major growth rate when used in a plant, or in the case of this experiment in
the presence of Rapahnus sativus. Hence, the null hypothesis in section 5.1 will be rejected and the
alternative hypothesis will be accepted. Accepting this hypothesis helps evidence my initial hypothesis of
a positive effect of antioxidants in the growth of R.sativus. This is shown in graph #1 where tendency
lines within samples with antioxidants are greater than the tendency line of water only seed samples. This
difference of tendency lines or growth rate, occurs due to the antioxidants capacity to work or function
against free radicals which lead to oxidative stress (ROS), that oxidizes the organism cells and leads to a
faster decaying of the plant and leads to a slower growth rate (Pehlivan, 2017). On the other hand, certain
antioxidants are able to provide other features helping plant’s metabolism such as vitamin C, protecting
plants from extreme light conditions, and leading to a faster and more adequate circumstances for
photosynthesis, preventing enzyme denaturation. (University of Exeter, 2017).

R.sativus samples are able to store antioxidants which help it to attack free radicals, and prevent or limit
oxidative stress, leading to a chain reaction of malfunction and slower growth rate. Results corroborate
my hypothesis where vitamin C & E will have a major growth while vitamin A a smaller growth. Vitamin
C samples which proved a major growth factor had an average growth rate of 8.0 cm during the 336 hours
leading to an average of 0.024 cm/h, vitamin E samples showed a major growth rate than water samples
but lower than vitamin C with a growth rate of 7.3 cm during the 336 hours = 0.022 cm/h, finally vitamin
A had an almost equal growth rate of water samples of 6.4 cm during the 336 hours or 0.019 cm/h.

Finally, the dependent variable is able to answer the research question “What is the effect on the use of
different antioxidants (40g dissolved in 100ml of water) in the growth rate of Raphanus sativus (3mg)?”.
Although the variable was a qualitative variable rather than quantitative, the use of different antioxidants
is able to prove the positive relation between the use of them and the positive relation with growth rate as
stated in the hypothesis. Qualitative results such as the turgidity present in vitamin samples, prove the

12
 ability of antioxidants to maintain healthier cells and attack free radicals and subsequently ROS.
Uncertainty apparatus were slightly higher which might lead to systematic error presence and possible
inaccuracy, but the various repetitions and the use of 25 samples per solution, gave the possibility to
increase the reliability of the results gathered.
6.2: Weaknesses and Improvements
Evaluation of experimental errors

Weakness/Source of Error Effect on data Improvement

Temperature due to it Photosynthesis leads to the growth of a plant, The use of a thermometer to record at all time
being an uncontrolled and temperature is a factor which affects this the temperature present (±0.5°C) and the use
variable, a range was used process. Data won’t be as precise as a real life of an external apparatus as an AC in order to
rather than precise values. situation. maintain the same temperature at all time.

The chronometer High uncertainty in the chronometer led to a
apparatus had a high possible effect of values being recorded at
uncertainty value. slightly different times, leading to possible
variation in the results making vitamin E and
C have more possible similar values.
The use of a more precise chronometer with a
low uncertainty (± 0.001s) will help values to
be recorded at a more equal momen than the
rest

Method to measure the Plant roots during growth period are delicate No improvement needed
plant root is too delicate enough to be broken with ease, meaning that
which might influence in while measuring them, extending them in
certain mm being order to measure to be as less invasive and
mistaken damage the root some millimeter were added
or subtracted.

The method of growing Data won’t be as precise for real life situations Conduct the experiment through both a
seeds through a since seeds will be grown directly into soil germination process to measure easily the root
germination method with more nutrients available for the plant's growth and seeds growing in soil to have a
lowered the possibilities growth. Although, the tendency of seeds with major connection with a real life situation of
to have real life values antioxidants growing more than normal growing crops.
than growing them in soil. growing techniques (water only) will be
present.

The solution of
antioxidants wasn’t spread
with a precise instrument
rather by eye measure.
Data might vary since it is probable that
certain seeds had a major amount of
antioxidant available and thus, the amount of
it to be used was major which can be seen in
the raw data tables where certain seeds grew
faster than others and other seeds grew slower.
With the use of the same syringe (± 0.1ml),
measure that each seed receives the exact
amount of solution in order to balance all
conditions for seeds to grow and gain more
reliable results.

Vitamin A and Vitamin E The reaction between the antioxidant and the
didn’t dissolve completely seed of Rapahanus sativus wasn’t complete,
in the solution. meaning that certain values might have a
different outcome, and the antioxidant
properties weren’t completely successfully
used
Using the principles of a water solution based
experiment, search for vitamins or antioxidants
that are completely water soluble or in other
occasions apply the experiment with all
antioxidants and the proper solutions to reach
its complete solubility.

The experiment was Certain processes such as photosynthesis Conduct the experiment outdoors for real life

13
 conducted indoor rather which contribute to the growth of a plant need standard conditions (direct sun-light) or rather
than outdoor light. Indoor experiments limit the amount of conduct both indoor and outdoor and compare
light received instead of a real life condition the results.
of direct light.

6.3: Extensions
A possible further investigation based on this experimentation could be understanding the effect of more
antioxidants in the growth of plants, which will help answer more precisely the research question of the
effect of antioxidants in plant growth, some antioxidants to use can be vitamin A, B (and its variances)
and K. It is possible to research if the effect is the same for other types of plants like shrubs, herbs or
climbers, and learn what are the features of plants that can be able to have their growth boosted through
the use of antioxidants, since it is a topic with few information and experimentation done.
Appendices
Appendix 1A:
Raw Data Table #2: A table showing the growth (± 0.1cm) of R.sativus seeds of 3mg (±0.01g) during
336 hours while pouring 14ml of vitamin E solution every 48 hours 5 seeds in each of the 5 repetitions
for vitamin E.
Vitamin E
Growth - G
Time - T δG = ± 0.1cm
δT = ± 1 hour Repetition #1 Repetition #2 Repetition #3 Repetition #4 Repetition #5

1 2 3 4 5 1 2 3 4 5 1 2 3 4 5 1 2 3 4 5 1 2 3 4 5
0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0
48 0,9 1,1 0,8 1,1 1,0 0,8 0,9 0,9 1,1 1,2 1,1 1,0 0,8 0,9 1,1 1,3 0,9 1,2 1,0 0,8 0,9 1,1 1,3 1,2 0,9
96 1,8 2,0 1,8 2,2 2,0 2,1 2,0 2,0 2,3 2,5 2,2 1,9 1,8 2,1 2,0 2,5 2,1 2,2 2,1 1,9 2,0 2,1 2,4 2,1 1,9
144 2,7 3,0 3,0 3,1 2,9 3,1 3,1 3,2 3,4 3,5 3,2 3,0 2,9 3,0 2,9 3,5 3,2 3,1 3,1 2,8 3,0 3,0 3,5 3,2 2,9
192 3,9 4,0 4,0 4,2 4,0 4,2 4,1 4,2 4,5 4,4 4,3 4,1 4,0 4,1 4,0 4,6 4,3 4,0 4,1 3,9 4,0 4,1 4,6 4,2 3,9
240 5,0 5,2 5,1 5,3 5,1 5,4 5,1 5,2 5,5 5,4 5,4 5,0 4,9 5,1 5,0 5,7 5,5 5,0 5,2 4,9 5,0 5,3 5,2 5,4 4,9
288 6,1 6,3 6,1 6,4 6,0 6,4 6,1 6,3 6,4 6,5 6,4 6,2 5,9 6,1 6,1 6,8 6,5 6,2 6,3 6,0 6,1 6,3 6,2 6,5 6,0
336 7,3 7,4 7,0 7,3 7,1 7,5 7,1 7,4 7,3 7,6 7,3 7,2 7,0 7,2 7,1 7,8 7,5 7,3 7,3 7,2 7,2 7,4 7,2 7,4 7,0
Total Increase 7,3 7,4 7,0 7,3 7,1 7,5 7,1 7,4 7,3 7,6 7,3 7,2 7,0 7,2 7,1 7,8 7,5 7,3 7,3 7,2 7,2 7,4 7,2 7,4 7,0
Appendix 1B:
Raw Data Table #3: A table showing the growth (± 0.1cm) of R.sativus seeds of 3mg (±0.01g) during
336 hours while pouring 14ml of vitamin C solution every 48 hours 5 seeds in each of the 5 repetitions
for vitamin C.
Vitamin C
Growth - G
Time - T δG = ± 0.1cm
δT = ± 1 hour Repetition #1 Repetition #2 Repetition #3 Repetition #4 Repetition #5

1 2 3 4 5 1 2 3 4 5 1 2 3 4 5 1 2 3 4 5 1 2 3 4 5

14
 0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0
48 1,3 1,1 1,4 1,2 1,3 1,2 1,3 1,1 1,0 1,3 1,4 1,3 1,1 1,3 1,2 1,1 1,3 1,2 1,4 1,0 1,3 1,2 1,1 1,3 1,1
96 2,5 2,2 2,5 2,4 2,4 2,4 2,5 2,3 2,1 2,4 2,6 2,4 2,2 2,5 2,3 2,4 2,5 2,3 2,6 2,1 2,4 2,4 2,3 2,5 2,2
144 3,6 3,4 3,7 3,5 3,5 3,6 3,6 3,4 3,3 3,5 3,8 3,5 3,3 3,7 3,4 3,4 3,6 3,4 3,9 3,2 3,5 3,4 3,5 3,6 3,3
192 4,8 4,5 4,8 4,6 4,6 4,9 4,8 4,6 4,5 4,6 5,0 4,7 4,5 4,8 4,6 4,5 4,7 4,6 5,1 4,5 4,7 4,6 4,7 4,8 4,6
240 6,0 5,6 5,9 5,7 5,8 6,1 5,9 5,6 5,6 5,7 6,2 5,7 5,5 6,1 5,7 5,6 5,8 5,8 6,1 5,6 6,0 5,7 5,9 6,0 5,6
288 7,1 6,8 6,9 6,9 6,9 7,1 7,0 6,7 6,6 6,7 7,2 6,9 6,7 7,1 6,8 6,6 6,9 6,8 7,0 6,8 7,2 6,8 6,9 7,1 6,7
336 8,1 7,9 8,0 7,9 8,0 8,2 8,1 7,8 7,7 7,9 8,2 8,0 7,8 8,1 7,9 7,7 7,9 7,8 8,0 8,0 8,3 8,0 7,9 8,1 7,8
Total Increase 8,1 7,9 8,0 7,9 8,0 8,2 8,1 7,8 7,7 7,9 8,2 8,0 7,8 8,1 7,9 7,7 7,9 7,8 8,0 8,0 8,3 8,0 7,9 8,1 7,8
Appendix 1C:
Raw Data Table #4: A table showing the growth (± 0.1cm) of R.sativus seeds of 3mg (±0.01g) during
336 hours while pouring 14ml of vitamin A solution every 48 hours 5 seeds in each of the 5 repetitions
for vitamin A.
Vitamin A
Growth - G
Time - T δG = ± 0.1cm
δT = ± 1 hour Repetition #1 Repetition #2 Repetition #3 Repetition #4 Repetition #5

1 2 3 4 5 1 2 3 4 5 1 2 3 4 5 1 2 3 4 5 1 2 3 4 5
0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0 0,0
48 0,7 0,8 0,8 1,1 0,9 0,7 0,9 0,7 0,8 1,0 1,0 0,9 0,8 0,7 0,7 0,7 0,9 1,0 1,1 0,8 0,7 0,9 0,8 0,8 0,8
96 1,6 1,7 1,9 2,0 1,8 1,7 1,8 1,6 1,7 1,9 2,0 1,7 1,6 1,7 1,6 1,5 2,0 1,8 2,0 1,7 1,5 1,8 1,6 1,7 1,6
144 2,7 2,8 2,9 2,8 2,7 2,7 2,6 2,6 2,6 2,9 3,0 2,8 2,7 2,8 2,5 2,3 2,8 2,5 2,9 2,4 2,5 2,7 2,5 2,6 2,5
192 3,5 3,6 3,8 3,5 3,5 3,4 3,3 3,4 3,2 3,7 3,9 3,7 3,5 3,6 3,4 3,1 3,6 3,5 3,7 3,5 3,4 3,7 3,5 3,5 3,4
240 4,5 4,5 4,7 4,4 4,3 4,4 4,2 4,3 4,3 4,8 4,8 4,7 4,4 4,5 4,4 4,2 4,5 4,3 4,6 4,5 4,3 4,6 4,3 4,4 4,2
288 5,3 5,4 5,7 5,2 5,5 5,3 5,2 5,3 5,4 5,8 5,9 5,6 5,4 5,5 5,3 5,2 5,6 5,3 5,6 5,3 5,3 5,6 5,2 5,3 5,1
336 6,2 6,4 6,7 6,3 6,6 6,5 6,3 6,4 6,4 6,6 6,8 6,5 6,3 6,4 6,2 6,3 6,5 6,3 6,7 6,4 6,4 6,6 6,3 6,3 6,2
Total Increase 6,2 6,4 6,7 6,3 6,6 6,5 6,3 6,4 6,4 6,6 6,8 6,5 6,3 6,4 6,2 6,3 6,5 6,3 6,7 6,4 6,4 6,6 6,3 6,3 6,2
Appendix 2A:
Processed Data Table #2: A table showing an average of the samples for Raw Data Table #2 (vitamin
E) solution Raphanus sativus seeds for all 5 repetitions across 336 hours
Vitamin E
Average Growth
Std. Dev.
Time - T Average Growth of the 5 seeds of Raphanus sativus from each repetition - G -G
Growth - G
δT = ± 1 hour δG = ± 0.1cm δG = ± 0.1cm
δG = ± 0.01cm
Repetition #1 Repetition #2 Repetition #3 Repetition #4 Repetition #5
0 0,0 0,0 0,0 0,0 0,0 0,0 0,00
48 1,0 1,0 1,0 1,0 1,1 1,0 0,04
96 2,0 2,2 2,0 2,2 2,1 2,1 0,09

15
 144 2,9 3,3 3,0 3,1 3,1 3,1 0,11
192 4,0 4,3 4,1 4,2 4,2 4,1 0,09
240 5,1 5,3 5,1 5,3 5,2 5,2 0,09
288 6,2 6,3 6,1 6,4 6,2 6,2 0,09
336 7,2 7,4 7,2 7,4 7,2 7,3 0,10
Appendix 2B:
Processed Data Table #3: A table showing an average of the samples for Raw Data Table #3 (vitamin
C) solution Raphanus sativus seeds for all 5 repetitions across 336 hours
Vitamin C
Average
Time - T Std. Dev.
Average Growth of the 5 seeds of Raphanus sativus from each repetition - G Growth - G
δT = ± 1 Growth - G
δG = ± 0.1cm δG = ± 0.1cm
hour δG = ± 0.01cm
Repetition #1 Repetition #2 Repetition #3 Repetition #4 Repetition #5
0 0,0 0,0 0,0 0,0 0,0 0,0 0,00
48 1,3 1,2 1,3 1,2 1,2 1,2 0,03
96 2,4 2,3 2,4 2,4 2,4 2,4 0,02
144 3,5 3,5 3,5 3,5 3,5 3,5 0,03
192 4,7 4,7 4,7 4,7 4,7 4,7 0,02
240 5,8 5,8 5,8 5,8 5,8 5,8 0,03
288 6,9 6,8 6,9 6,8 6,9 6,9 0,06
336 8,0 7,9 8,0 7,9 8,0 8,0 0,05
Appendix 2C:
Processed Data Table #4: A table showing an average of the samples for Raw Data Table #4 (vitamin
E) solution Raphanus sativus seeds for all 5 repetitions across 336 hours
Vitamin A
Average
Time - T Std. Dev.
Average Growth of the 5 seeds of Radish from each repetition - G Growth - G
δT = ± 1 Growth - G
δG = ± 0.1cm δG = ± 0.1cm
hour δG = ± 0.01cm
Repetition #1 Repetition #2 Repetition #3 Repetition #4 Repetition #5
0 0,0 0,0 0,0 0,0 0,0 0,0 0,00
48 0,9 0,8 0,8 0,9 0,8 0,8 0,04
96 1,8 1,7 1,7 1,8 1,6 1,7 0,06
144 2,8 2,7 2,8 2,6 2,6 2,7 0,09
192 3,6 3,4 3,6 3,5 3,5 3,5 0,08
240 4,5 4,4 4,6 4,4 4,4 4,4 0,07
288 5,4 5,4 5,5 5,4 5,3 5,4 0,08
336 6,4 6,4 6,4 6,4 6,4 6,4 0,03
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