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Toxicology Protocols

Kelsey Turner
AP Environmental Science
Period 5
To conduct a dose/response bioassay using lettuce seeds.
I hypothesize that the predicted LD-50 will be 0.01 strength.

- 1 piece of filter paper
- 2 petri dishes with plastic bags to put them in
- 10 lettuce seeds
- 5mL of 10% bleach solution
- Tape for labeling petri dishes
- 1 2mL pipette
- 2mL of each Simple Green solution: 100%, 10%, 1%, .1%, .01%, .
- 2mL of distilled water
- Scissors
- Tweezers
- Funnel

1. Soak lettuce seeds in 10% bleach solution for 5 min. Strain through
filter paper and rinse.
2. Label two petri dishes using tape for each chemical concentration
that will be in them.
3. Cut two circles of filter paper and put them in dishes
4. Put 5 seeds in each dish
5. Add 2mL of the appropriate chemical solutions to the
corresponding dishes
6. Close dishes, place in plastic bag, and seal
7. Incubate the seeds in the dark for four days at a constant temp
8. At the end of the growth period record how many seeds in each
dish have germinated


Condition Plate 1 Plate 2 Plate 3 Plate 4 Average

Control 60 100 40 40 60

.001 60 100 100 40 75

.01 100 60 60 60 70

.1 100 80 100 80 90
1 40 40 100 0 45

10 0 0 20 6.7

100 0 0 0 0


Data Analysis:
The hypothesis was unsubstantiated because the experiment data showed that the LD-50
the cleaning solution, Simple Green, was 1% and not .01% like what was predicted. The results
from period 5 were inconsistent with the other class data. There was a general pattern in the data
with some small inconsistencies, but it should have been the exact same, as all the classes were
testing the exact same seeds in the exact same solutions, under the exact same conditions. The
errors that could have affected the labs results were: groups using the wrong number of
seeds(less than 5), group members using the wrong amount of solution in their petri dishes,
members could have mislabeled their dishes leading to the wrong solutions being added to the
wrong dishes, the data collected could have been shuffled and put in the wrong columns as the
students recorded, math mistakes could have been made, and/or group members could have
simply counted the amount of sprouted seeds wrong.


In this lab, students soaked lettuce seeds in bleach and then put them in petri dishes
containing different concentrations of the cleaning chemical, Simple Green and let them sit for
four days in the dark. After the four days were up, they took them out and measured how many
seeds in each solution sprouted, to try and find the LD-50 of the Simple Green. An LD-50 is the
concentration of a chemical solution at which around half of the test subjects the chemical is
used on will die, and half will live, or in the case of this lab, half of the seeds will sprout and half
wont. The LD-50 of Simple Green in this case turned out to be the 1% concentration of the
simple green solution, because 45% of the seeds soaked in that sprouted, whereas those soaked
in other concentrations either had far more or less sprout.
This study is important it shows how much of the Simple Green solution we can use to
get the maximum amount of seeds to sprout and possibly plants to grow. This lab did not only
showcase the LD-50 of Simple Green, but it also showed that more seeds sprouted in .1% Simple
Green concentration than did in regular distilled water. This is important information as it could
help scientists and farmers create new fertilizers to help them grow the maximum amount of
plants from their seeds. Therefore, this shows that the concept and techniques from this lab can
be used for many other real world examples and experiments.
In the future, this study should be done on a far larger scale. Instead of using four plates
for each chemical concentration and then recording them to find the average, far more sets of
lettuce seeds and petri dishes should be used to get a wider range of results. If a larger amounts
of sets had been used, the averaged data would have been more accurate and small mistakes in
data collection would not matter as much and it would be far easier to identify mistakes and
remove them from the data as outliers. A wider variety of seeds types could also be used in this
experiment in the future, to figure out if 1% is the LD-50 of just lettuce seeds, or other types of
seeds as well. The seeds could also have two sets used in the dishes, one soaked in diluted
bleach solution before and one not, so that it could be seen if that had any effect on the
percentage of seeds sprouted. This information could also be useful to scientists and farmers, as
it could help a higher percentage of their seeds sprout.