LD 50 Lab

Collaborators: Logan Lazaras, Sam Smithers, Amber Mitchel and Maggie

Introduction:
LD50 is a lethal amount or concentration that kills half of a sample population. The
purpose of this lab is to see how much salt in the water will kill 50% of the radish seeds in that
samples population of radish seeds. A dose response curve is a curve plotting the relationship
between the dose of a drug administered and its pharmacological effect. It will be used in this
lab to graph the data and show the effects of various doses of salt on the growth of radish seeds.
Serial dilution is a series of solutions, each of which are half as concentrated with salt to see its
effect on the growth of the plant. The serial dilution of this particular experiment is when the
seeds are diluted in the various water and salt solutions. Furthermore, salt is applied to
highways to see how it impacts the growth of the plants along the roadside and aquatic plants
nearby in streams. The importance of this lab is to not only discover how much absorption of
salt the plants can tolerate, but to see the amounts of plant damage high concentrations of salt
can cause.
Problem: What is the LD50 of salt for radish seeds? How does salt affect the initial of radish
plants?
Hypothesis: If the solution contains more than 25% of salt, then half of the radish seed
population will not germinate.

Parts of the experiment:

1. Independent variable: Amount of salt in the solution
2. Dependent variable: How many seeds germinated and by how much they grew
3. Control variable: the water with no salt
Materials:

60 Radish seeds
Paper towels
Water
Saltwater
Plastic baggies

 Ruler
Test tubes
Markers
Methods:
1. Using 20 ML of the original concentration of salt, perform a serial dilution to create five
different salt concentrations, plus a six with plain water.
2. Count out 60 Radish seeds, 10 for each sample.
3. Put 10 seeds into a folded paper towel, Space the seeds out evenly on the paper towels so
that they do not touch each other.
4. Wet the paper towels/seeds and seal in a baggie, be sure to label the bags with the salt
concentration for that sample.
5. After 4-5 days examine the seeds, counting the number of each that germinated.
6. For the germinated seeds, measure the length of the radicle.
Data:

Data analysis:
Due to the fact that there was not enough salt added to the seeds, this experiment did not
produce an LD50 for the radishes. However, it was possible to approximate that half the seeds
in a sample population will not germinate at 288g of salt. The highest percent of germination
occurred in the bags with 6.25% solution and 1.25 grams of salt. The average length of the
seeds was as little as 30mm but the longest one was 39mm in 50% concentration of salt. Despite
the high concentration of salt at 100%, more than half of the seeds were still able to germinate.
However, they were mostly affected by this high salt concentration because 75% of them only
grew to be 5mm long. Overall, the data shows that high salt concentrations limit plant growth.

Conclusions:
In this experiment, the hypothesis stated that the 25% salt concentration will be the LD50
of the radish seeds. This means that at a 25% concentration of salt, half of the sample seed
population will die. Due to the fact that there was not an LD50 for the radish seeds, the
hypothesis was proven to be incorrect. The lack of a lethal dose was a result of not having
enough salt to kill exactly half of the radish population. Making an estimate through the
documented trends of growth shown on the data charts, the LD50 of the radish seeds would
have been at 288/g of salt. Although the experiment didnt portray an apparent lethal dose, it
showed that salt did, in fact, have an impact on the growth of the radishes. The seeds that grew
the least were in 100% concentration of salt, 18g., proving that salt particles affected the growth
of the seeds. Connecting this experiment to a real world situation, hundreds of reports have
been documented in the past three decades about the impact of road salt on the environment.
According to the Transportation Research Board, roadside trees and other vegetation are
injured by salt primarily through two mechanisms. They are increased salt concentrations in soil
and soil water, which can result in salt absorption through roots, and salt accumulation on
foliage and branches due to splash and spray. The slope of the roadside is a key factor in
determining where salt reaches vegetation, either from splash and spray or root absorption.
Injury due to salt is typified first by inhibition of general growth, followed by specific injuries
to foliage and limbs, and, in some cases, plant death. This information directly correlates with
what the LD50 lab is all about. It further the supports the fact that salt particles can damage the
root systems of plants, thus preventing plants from growing to their potential. In conclusion,
the hypothesis was not supported by the lab results because an LD50 of the radish seeds was not
found. Had there been a higher concentration of salt used for the experiment, it is most likely
that a lethal dose would have been apparent. Accordingly, the salt did affect the radish seeds
growth but the lab results were simply a rough estimate. In relation to this experiment, adverse
effects of roadside vegetation make a clear connection that show how lethal doses of salt impact
plants in the world today.

Citations:
"Definition of Dose Response Curve." Dictionary.com. Dictionary.com, n.d. Web. 13 Mar.
2015. <http://dictionary.reference.com/browse/dose-response+curve>.
Road Salt Impacts on the Environment. Web. 13 Mar. 2015. N.p.: Transportation Research
Board, n.d. PDF.< http://onlinepubs.trb.org/onlinepubs/sr/sr235/069-082.pdf>.