Since 1986,
there have been an ethical and legal obligations imposed on countries that have
signed the convention of council of Europe (European Council 1986) in view to
replace, reduce and refine the use of animals in clinical research. Since then,
several research have been carried out on water flees such as daphnia. Based on the preclinical and the data
available on the different composition of cinnamon, a lethal dose test was
carried out in order to determine the toxicity of cinnamon. This LD50
test was carried out in the aim of:-
1.Determining pre clinically the
best dose to be administered to patients in the clinical trials
2.Assessing any potential risk in
the taking of cinnamomum
3.Having an approach to the 3R
theory in the aim to reduce the use of animals in pre clinical research
Equipments:-
Cinnamomum, 5
boiling tubes, 5 beakers, a micro scale, Daphnia flees, Petri dish
Methods:-
Cinnamomum was
prepared in five different concentrations of 1µg/150ml, 2µg/150ml, 4µg/150ml,
8µg/150ml, 16µg/150ml respectively. The cinnamomum was first measured very
accurately with a decimal error of +0.05µg on a micro scale. The
different weights of cinnamomum were each boiled in 10ml of water for infusion.
Each concentration was then transferred in a beaker and left to cool down for
15minutes. the daphnia sample were then poured and the volume of water was
round up to 150ml each. The sample was then tranferred in a Petri dish for
counting. The reaction was then recorded after three hours in table 2:-
Table
2: the lethal dose test determination table
Beaker
Concentration
Alive
Dead
Lethal dose %
1µg/150ml
All
None
0
2µg/150ml
All
None
0
4µg/150ml
48
46
49
8µg/150ml
31
62
67
16µg/150ml
None
All
100
The LD50 curve based on the
above table is as follows:
Figure 1 Lethal dose test (LD50)
From this experiment, it can be
deduced that the LD50 is
4µg/150ml having a percentage nearer to 50%. Therefore, from this experiment,
the dosage to be administered to patients will go on a clinical trials at this
concentration.