Biology Internal Assessment

Research question: Does the presence of organic (3, 6, 9%) or synthetic fertilizer (3, 6, 9%)
provide more support to Fenugreek plants in terms of leaf surface area, plant height, and iron
content.
Topic: Plant biology.
Independent Variable: Organic multipurpose fertilizer, synthetic multipurpose fertilizer.
Dependent Variable: Iron content, height, and leaf surface area.
Controlled variables How were they controlled Why were they controlled
Amount of water. The amount of water given The amount of water
to the plants was controlled provided was controlled
by using a measuring because all the plants were
cylinder. required to receive water in
the same quantities.
The concentration of the The concentration of the The concentrations of the
fertilizers. fertilizers was controlled by fertilizers were controlled to
measuring the amount of gain data on how different
water mixed with different concentrations affect the
quantities of fertilizer. plants.
Location of the plants. The location of the plant The location of the plants
was controlled by placing was controlled because they
the plant in a location where require indirect sunlight.
it would be safe with enough Direct sunlight or no
sunlight and shade. sunlight would both kill the
plants or hinder their
growth.
Frequency of watering the The number of times the The number of times the
plants. plant was watered was plant was watered was
controlled by following a set controlled because too little
schedule on when to water or too much water would
the plant. hinder the plants' growth or
kill them.

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Hypothesis:
 The plant will grow taller in height for the synthetic fertilizer variable.
 The plant will have a larger surface area with the synthetic fertilizer variable.
 The plant will have more iron content with the synthetic fertilizer variable.

Introduction
Iron is extremely important for us. In the body, red blood cells carry oxygen around. Iron aids in the
production of these blood cells. Iron deficiency anaemia is caused by a lack of iron.
Over the pandemic, my mother picked up the hobby of growing plants and homegrown vegetables. I
often noticed her searching the web for the best and fastest way to grow her plants. Normally she
would use a homemade fertilizer which consists of leftovers, kitchen scraps, coffee grounds,
eggshells, etc. She was happy with how her homemade fertilizer worked but they used to take far too
long to gather so she decided to just buy a natural fertilizer instead. I heard it was going well. This got
me curious to find and prove which is the most effective way to nurture and grow plants. Is it a natural
store-bought fertilizer or a synthetic fertilizer? Natural store-bought fertilizer is a professionally
created fertilizer made from natural resources, homemade fertilizer which is the mix of leftovers,
kitchen scraps, and veg-based supplies that can be found at home. Synthetic fertilizer would be a
fertilizer made mainly using chemicals that unnaturally boost growth and the rate of growth.
Background information
Natural fertilizer contains carbon and is produced naturally. It is a substance that supplies nutrients
and sustains plant life by being added to soil or plants. The fertilizer I used for my experiment is from
the brand Criyagen Agri & Biotech Pvt. Ltd. The fertilzier contains Azospirillum sp.−5 X 107 CFU
per gram, Phosphate solubilising bacteria (PSB)−5 X 107 CFU per gram, and potassium mobilizing
bacteria (KMB)−5 X 107 CFU per gram. Azospirillum is used for its nitrogen-fixing abilities,
phosphate solubilising bacteria are used to solubilize inorganic phosphorus from insoluble
compounds.
Inorganic fertilizers are manufactured from inorganic compounds derived primarily from petroleum
products. Nutrients are added to the soil by chemical fertilizers, but they add no other components. To
live, plants need many other elements as well as nutrients. Living organisms and organic matter are
also essential. Organic matter in the soil does not support microbiology.
The reason Fenugreek or Methi was used is that this plant grows rapidly and are competitive enough
against most weeds. As winter progresses, the small-leafed variety will continue to grow slowly while
the larger variety will typically die off.
The reason I have used Fenugreek for my experiment is that it is cheap, easily available and grows
quickly. I had to use a plant that grew quickly to test it within the limited time provided. Leaf surface
area is important to measure how much sunlight is acquired and how much water is lost to carry out

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photosynthesis. The larger the leaf surface area, the more sunlight collected and less water lost. The
height of the plant is important because it allows the plant to gain access and compete with other
plants for more sunlight. Iron is an important component in plants because plants require iron to
synthesise chlorophyll. They also need to maintain the structure and function of chloroplast to
survive. The reason I am doing my exploration on this topic is that I would like people to figure out
how, when and what fertilizer to use when they want to grow a plant. If they use the right fertilizer for
their needs, cases of eutrophication in nearby water bodies will decrease significantly and it saves a
plethora of animals and fishes along with the food chain in that area and the ecosystem. Also to help
my mother with her interests.
Materials
Quantity Materials required Uncertainty in instruments
used for this study
Approx. 100 Fenugreek seeds
5 (100ml) Beakers
2 (100ml) Measuring cylinder (10cm3) ± 0.2 c m3
(100cm3) ± 0.5 c m 3
1 Scientific weighing scale
5kgs Potting soil
50 Pots
500g NPK organic fertilizer multipurpose
500g NPK synthetic fertilizer multipurpose
1 pair Rubber gloves
For measuring Leaf surface area
5 pages Graph paper
For checking iron content
1 Electronic weighing balance Digital balance ±0.01g
5 Crucible
5 Bunsen burner
1 Tongs
2 Stirring rod
500ml Dilute Hydrochloric acid
1 Stopwatch
1L Distilled water
10 (100ml) Conical flask
2 Dropper

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 1 Marker
5ml Ferrous chloride solution
50g Ammonium Thiocyanate
1 Colorimeter (Vernier logger pro)
5 Cuvette

Procedure
Germination of Fenugreek seeds
1. Germinate the Fenugreek seeds for around 3-5 days.
2. To germinate the seeds, simply put some potting soil into a pot and put a few seeds into the
pot.
3. Then water the seeds with 5ml every day till you see the seeds germinating.
4. Be careful not to waterlog the plant and only lightly dampen the soil while in the germination
phase.
Preparation of fertilizer concentration
Concentration (%) Organic fertilizer (g) Synthetic fertilizer (g)
3 0.03 0.03
6 0.06 0.06
9 0.09 0.09
5. For this particular experiment, I have picked 3 concentrations, 3, 6, and 9%
6. To create the concentrations, measure 100ml of distilled water with a measuring
cylinder.
7. Using an electronic measuring balance, measure the quantity of fertilizer required to
make the concentrations.
8. Mix it with water.
9. Mix it thoroughly by shaking it.
Experimental setup to grow the plants
1. To grow the plants, collect garden soil and put specific quantities (half a cup) of it
into 100 c m3 paper cups according to the number of trials but for this experiment, 30
cups were used.
2. Then put approximately 10 Fenugreek seeds in the cups.
3. Water the plants with 5 ml of the fertilizers every three days and avoid water logging
or under-watering. If noticed, act accordingly.
4. Fenugreek normally takes two to three weeks to fully grow.

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Procedure to measure iron content in the plant
 Creating Standard Iron solution
1. The procedure to measure iron content starts with finding the standard iron test.
2. To do this, we need to mix different quantities of ferric chloride solution with 10ml of
distilled water.
3. The concentrations I used were 0.4, 0.8, 1.2, 1.6, and 2.0% These concentrations
required me to use 0.04, 0.08, 0.12, 0.16, and 0.20 grams of ferrous chloride solution.
Concentration (percentage%) Quantity of ferrous chloride (c m 3)
0.4 0.04
0.8 0.08
1.2 0.12
1.6 0.16
2.0 0.20

Measuring iron content

1. To measure the iron content in the Fenugreek leaves, pluck leaves from each trial and
concentration.
2. The number of leaves required from each trial should amount to 2 grams.
3. Clean the leaves plucked from the plants with distilled water and filter paper to
ensure that there is no iron contamination on the leaf.
4. Put leaves from each trial in separate crucibles on a Bunsen burner.
5. Burn the leaves till it has turned into powdery ash.
6. Use a stirring rod to crush the leaves into a finer powder.
7. Leave to cool for 5 minutes.
8. Then thoroughly mix the ash with 10ml of 1 mol L−1hydrochloric acid.
9. Add 5ml of distilled water to the mixture and mix thoroughly again.
10. Filter to solution into a conical flask through a filter paper.
11. The filtered solution is used for the analysis.
12. The analysis starts with measuring and collecting data for the standard iron test.
13. Put the cuvettes with the standard solution in the colorimeter while calibrating the
colorimeter each time with a blank cuvette filled with distilled water.
14. After collecting data for the standard solution, we measure the iron content in the
cuvettes with the experimental solution.

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 15. The same method was followed for the experimental solution where you put the
blank before every reading and wipe the cuvette with filter paper to avoid iron
contamination.
Measuring leaf surface area and plant height
1. This step is done at the end of the growth process of the plant when the plant is fully
grown.
2. Puck one leaf from the plants from each concentration and trial.
3. Use a pencil to outline the leaf on 0.5 or 0.25 cm graph paper.
4. Then check to see how many boxes have a line passing through them. Count the
number of half boxes and complete boxes.
5. I used a 0.25cm graph paper which means that I will have to multiply the number of
complete boxes by 0.25 and a half boxes by 0.125. Then add both the values together
and that will give you the surface area of the leaf.
6. To measure the height of the plant, simply pluck the whole plant.
7. Using a scale, measure the whole length of the plant from the tip of its roots to the tip
of the stem.
8. If the plant is bent or curved, straighten it and measure the height accurately.

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Processed data
Leaf surface area Organic fertilizer (3% concentration) Synthetic fertilizer (3% concentration)
Mean 13.5 14.95
Standard deviation 66.75 74.75
Plant height Organic fertilizer (3% concentration) Synthetic fertilizer (3% concentration)

Mean 23.98 24.92

Standard deviation 119.9 124.6
Iron content Standard iron Organic fertilizer (3% concentration) Synthetic fertilizer (3%
concentration)
Mean 0.390 0.1604 0.2106
Iron concentration The absorbance of organic sample The concentration of the unknown sample
Mean 0.1604 0.308%

Standard Deviation 0.802 1.54

The absorbance of synthetic sample The concentration of the unknown sample

Mean 0.2106 0.504%

Standard Deviation 1.053 2.52

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 Calibration Curve of iron content
0.6

f(x) = 0.22025 x + 0.1259

R² = 0.853348036607194 1.65, 0.534
0.5
Synthetic, Organic,
(1.2, [Y VALUE]) (2,[Y VALUE])

0.4
Absorbance (Au)

0.3 1.2, 0.385

0.2

0.1

0
0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2

Concentration of iron (%)

N/A: The data could not be retrieved due to the death of the plants in the respective
concentrations.

T-Test for statistical calculation for standard deviation

x̄1 − x̄2

√
2 2
 Formula to calculate unequal variances : s1 s 2
+
n 1 n2

 Level of significance= 0.05

 1=synthetic fertilizer
 2=organic fertilizer
 x-bar1 and x-bar2 is the mean of the first and second samples respectively.
 s1 and s2 are the standard variations of the first and second samples respectively.
 n1 and n2 are the numbers of data in each sample.
 The H o of this test is that the organic fertilizer synthetic fertilizer is better for the iron
content and growth of the Fenugreek plant. The H 1 is that synthetic fertilizer is better.

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 If the value of the test comes up above 0.05, we accept the H 1 but if it's lower then we
accept the H o .
Leaf surface area t-test:
14.95−13.5

√ 66.752 74.752 ¿ 0.032

5
+
5

Comparative chart for leaf surface area

15.5
15
14.5
Mean

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13.5
13
12.5
1

Organic fertilizer (3% concentration)

Synthetic fertilizer (3% concentration)

The leaf surface area t-test lands below 0.05 by a difference of 0.0176 which means we
assume the null hypothesis. The difference is not significant but still has a margin to the
alternative hypothesis. This calculation tells us that synthetic fertilizer is better than organic
fertilizer when compared to organic fertilizer when it comes to the size of the leaf.
Plant height t-test:
24.92−23.98

√ 124.6 119.9 ¿ 0.012

2 2
+
5 5

Comparative chart for plant height

25
24.8
24.6
24.4
Mean

24.2
24
23.8
23.6
23.4
1

Organic fertilizer (3% concentration) Synthetic fertilizer (3% concentration)

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The plant height t-test lands below 0.05 with a difference of 0.0378, a significant difference
that leads us to assume the null hypothesis. This calculation tells us that synthetic fertilizer is
better than organic fertilizer when compared to organic fertilizer when it comes to the height
of the plant.
Iron content t-test
0.504−0.308

√ 2.52 1.54 ¿ 0.148The iron content t-test comes out higher than the value of significance
2 2
+
5 5
with a difference of 0.0984 which is a very significant difference. The outcome indicates that
we should assume the alternative hypothesis which is that organic fertilizer provides better
care when it comes to nurturing iron content in the plant.
Statistical Analysis
The data collected and processed shows us that synthetic fertilizer is better overall when
compared to organic fertilizer in these specific conditions while competing for leaf surface
area, height, and iron content. Although organic fertilizer proved to be far superior in the area
of iron content. The surface area of the plants which grew with a synthetic fertilizer was
larger which would allow them to catch more water and sunlight. This would enable them to
grow better and gain more nutrition compared to the plants grown with organic fertilizer. The
height of the plant is important because it helps the plant compete for light against other
plants in the wild. A taller plant will get more sunlight than other plants under it, thus making
it grow healthier and faster.

Conclusion
Qualitative observation
The plant in both conditions was growing properly until the fertilizer started getting a little
too strong for the plants in the 6 and 9% concentration. Although the visibly noticed changes
started occurring a week into the experiment, I mistakenly took it as an “adulthood” of the
said conditions. But a week later I realized that I was wrong because the plants in the 3%
concentration group were growing perfectly fine. In terms of the 3% group, it was smooth
sailing to the end. In my opinion, this concentration would work the best for Fenugreek plants
in similar quantities.

Safety

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The safety considerations taken while executing the procedures were followed thoroughly.
While germinating and growing the plants, gloves and lab glasses were worn. This is because
the soil was taken from a patch of land nearby which has not been processed. It may contain
bacteria that can potentially make a person sick if come in contact with it. Therefore, the
gloves were used while potting the plants.
The process of making the concentrations of the fertilizers also required rubber gloves and a
lab coat. That is because the fertilizers may cause an allergic reaction when it makes contact
with skin. To avoid that, we are required to wear rubber gloves and a lab coat.
The process of finding and calculating the leaf surface area and plant height only required the
safety measure of wearing gloves. And even that may not be required as only handling of the
plant, graph paper, and pencil is required.
Finding the iron content involved many safety measures. To start the process of creating the
standard iron solution requires us to wear gloves and a lab coat as we handle acids and
chemicals during this segment of the experiment. After creating the standard solution, we are
required to create the solution for the experimental solution. We use diluted acids and spirit
burners during this process. First, all flammable items such as any type of cloth, paper, or
wood were removed from the vicinity of where the experiment was held. Then the leaves
were burnt to turn them into ash. Then tongs were used to take them off the flame and put
away to cool. Rubber gloves, a lab coat, and lab glasses were used in case the flame rose or
ashes flew. While using hydrochloric acid, gloves and lab coats were used. Putting the tests
into the colorimeter did not require any safety measures.
Limitations
The data collected depicts three components; growth of the plant in terms of height, leaf
surface area, and iron content in the leaves. The readings for the 6 and 9% concentration of
both synthetic and organic fertilizer could not be retrieved due to the plant not growing
effectively. The 3% test grew perfectly fine but the readings cannot be compared due to no
other test to refer to.
The reason for the other 2 concentrations not growing could be one of three reasons or all of
them together. One of the reasons might have been that the concentrations were too strong for
the particular plants to handle. Another would be that the plants were not exposed to enough
sunlight to grow to adulthood.

Improvements

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There are many ways this experiment could have been handled better. The reason for the 6
and 9% concentration of the fertiliser was too strong for the plants to handle, I could have
looked at studies done and referred to a concentration that the plants could handle to the max.
A lot of improvement could have been done in the research part of this experiment. If the
experiment was done earlier, there would have been enough time for the plants to fully grow
and even rectify any mistakes made while growing the plants.
Discussion
For this exploration, we compared how organic and synthetic fertilisers would do against
each other. The plant used for the experiment was Fenugreek (Methi). Each fertilizer had 3
concentrations with 5 trials for each concentration. The concentrations were 3, 6, and 9%
Unfortunately, only the 3% sample grew the closest to maturity. The reasons for that are
given in the limitations section. From the data collected and processed, in conclusion, what
we can assume from these tests is that synthetic fertilizers are better in certain aspects than
organic fertilizers.

Bibliography

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 1. Du, Qing-Jie, et al. “Effects of Different Fertilization Rates on Growth, Yield, Quality
and Partial Factor Productivity of Tomato under Non-Pressure Gravity Irrigation.”
PLOS ONE, Public Library of Science, journals.plos.org/plosone/article?
id=10.1371%2Fjournal.pone.0247578.
2. “Fertilizer for Money Plant: Pothos Plant Care Fertilizer.” India Gardening, 7 July
2020, indiagardening.com/gardening-tips/fertilizer-for-money-plant-pothos-plant-
care/.
3. Gitz, Jonas C, et al. “A Colorimetric Method for Measuring Iron Content in Plants.”
Journal of Visualized Experiments : JoVE, MyJove Corporation, 7 Sept. 2018,
www.ncbi.nlm.nih.gov/pmc/articles/PMC6235139/.
4. Iron Colorimeter - University of Canterbury.
www.canterbury.ac.nz/media/documents/science-outreach/iron_colorimeter.pdf.
5. “Preparation of Standard Solutions.” Pharmaceutical Guidelines,
www.pharmaguideline.com/2010/09/preparation-of-standard-
solutions.html#:~:text=Iron%20Standard%20Solution%20(10%20ppm,Contains
%20iron%20in%20ferrous%20state.
6. Sajid, M., et al. “Effect of Different Levels of Nitrogen on the Growth of Money Plant
(Epipremnum Aureum) Cuttings.” AGRIS, 1 Jan. 1999,
agris.fao.org/agris-search/search.do?recordID=PK2000000705.
7. “What Is the Difference between Fertilizer Derived from Organic and Synthetic
Sources?” Milorganite, www.milorganite.com/lawn-care/organic-lawn-care/organic-
vs-synthetic#.
8. “What Is the Difference between Fertilizer Derived from Organic and Synthetic
Sources?” Milorganite, www.milorganite.com/lawn-care/organic-lawn-care/organic-
vs-synthetic.
9. "Fenugreek Seed Germination, Time, Process (Methi) | Gardening Tips". Gardening
Tips, 2022, https://gardeningtips.in/fenugreek-seed-germination-time-process-methi.

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 Appendix

Making different concentrations of fertilizer Measuring leaf surface area

Burning the leaves to measure for absorbance The plants

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 Raw data
Leaf surface area Organic fertilizer (3% Organic fertilizer (6% Organic fertilizer (9% Synthetic fertilizer (3% Synthetic fertilizer (6% Synthetic fertilizer (9%
2 2 2 2 2
concentration) (c m ) concentration) (c m ) concentration) (c m ) concentration) (c m ) concentration) (c m ) concentration) (c m 2)

T1 15.25 N/A N/A 12 N/A N/A

T2 11.25 N/A N/A 20.25 N/A N/A

T3 12.125 N/A N/A 12.5 N/A N/A

T4 14.125 N/A N/A 14.875 N/A N/A

T5 14 N/A N/A 15.125 N/A N/A

Plant height (cm) (cm) (cm) (cm) (cm) (cm)

T1 22.5 N/A N/A 29 N/A N/A

T2 23.2 N/A N/A 25.8 N/A N/A

T3 24.2 N/A N/A 22.8 N/A N/A

T4 25 N/A N/A 20 N/A N/A

T5 25 N/A N/A 27 N/A N/A

Iron content test (mcg/dL) (mcg/dL) (mcg/dL) (mcg/dL) (mcg/dL) (mcg/dL)

absorbance
Standard Iron test

0.143 T1 0.159 N/A N/A 0.275 N/A N/A

0.386 T2 0.127 N/A N/A 0.126 N/A N/A
0.403 T3 0.159 N/A N/A 0.198 N/A N/A
0.485 T4 0.155 N/A N/A 0.303 N/A N/A
0.534 T5 0.202 N/A N/A 0.151 N/A N/A

N/A=Not Available

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