HPLC METHOD FOR THE SIMULTANEOUS
DETERMINATION OF THE COMPONENTS OF
AN AQUEOUS ANTIDOTE SOLUTION
LAVINIA G. HINESCU1, CRISTINA M. RANETTI1, MIHAELA
IONESCU1, ELENA IONICA1, CONSTANTIN DRAGHICI1,
CONSTANTIN MIRCIOIU1,2, CRISTIANA COSMESCU1, VICTOR A.
VOICU1,3
1Army Center for Medical Research, 37 C.A. Rosetti St., Bucharest, Romania
2University of Medicine and Pharmacy “Carol Davila”, Faculty of
Pharmacy, 6 Traian Vuia Street, Bucharest, Romania
3University of Medicine and Pharmacy “Carol Davila”, Faculty of
Medicine, Department of Toxicology, Bucharest, Romania
*corresponding author: lhinescu@yahoo.com
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Abstract:

The objectives of this study were to develop and validate a method for the
simultaneous HPLC-UV determination of obidoxime (OBI), scopolamine (SCA), Atropine (ATR) and imipramine (I) in an aqueous antidote solution. An ion-pair mechanism with DAD (diode array detector) detection was used to quantitatively determine the four drugs.
The proposed HPLC method uses a Kromasil 100-5C8 (250x4.6mm) column, gradient elution, at 25°C column temperature. The mobile phase consisted of 25% acetonitrile and 75% aqueous phase (5mM sodium 1-heptanesulfonate monohydrate, pH=3.5). The analyte detection was an UV detection at λ=230±4 nm, with reference λ=360± 8 nm. The calibration curve was linear between 1.3-152.5 μg/mL (r=0.9959, n=6) for OBI, between 0.13–13.75 μg/mL (r=0.9951, n=6) for SCA, between 0.25–25.5 μg/mL (r=0.9999, n=6) for ATR and between 0.58–50.80 μg/mL (r=0.9996, n=6) for I. In conclusion, this chromatographic assay fulfilled all the requirements for being a reliable and feasible method, including linearity accuracy, precision and stability.

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