Malaria is the world’s most important parasitic infection with 500 millions cases annually and almost 2
millions death per year. This disease is more present in Sub-Saharan Africa where 90% of the infections
are found. Artemisinin and its semi synthetic derivatives (artemether, artesunate) have actually the most
powerful activity on malaria, even in its complicated forms and resistance cases.
Various methods have been proposed for detection and quantification of artemisinin in Artemisia annua
L. by HPLC–UV, but the plant extracts used for this quantification were extracts obtained with organic
solvents (toluene, petroleum ether, hexane). To be able to use crude A. annua extracts prepared at low
cost to formulate antipaludic drugs, we chose the use of a mixture of water and ethanol as solvent of
extraction, but no adequate analytical method for this kind of extracts is published.
The main objectives of this work were first to develop an analytical method for artemisinin quantification
in hydro alcoholic extracts of A. annua. Second, this method had to be thoroughly validated by the
research and development laboratory and, third, the transfer of this method to the routine laboratory had
to be demonstrated. The final aim was to compare the estimation of measurement uncertainty obtained
during the method validation with validation standards to measurement uncertainty estimates obtained
during the method transfer study with real samples.
The method was validated following the accuracy profile methodology and was found to be accurate
in the concentration range of 10.0–54.0 g/ml with CV < 8%. Limit of detection and of quantification were
2.73 and 10.0 g/ml, respectively. The method was then successfully transferred to a laboratory in Benin
by showing that the quality of the results that it will generate during routine application of the method is
sufficient. Finally, the measurement uncertainty of the method was estimated from the validation experiments
as well as from the transfer study with authentic unspiked samples of A. annua. The comparison
of these measurement uncertainty estimations showed that they were coherent. It confirmed thus that
the estimation of measurement uncertainty from validation experiments predicts well the measurement
uncertainty of real routine samples. This analytical method was thus shown to be convenient for routine
analysis of hydro alcoholic extracts of A. annua in Benin

Method transfer, Measurement uncertainty, Method validation, Artemisia annua, Hydro alcoholic extracts