This paper presents the chemometrically assisted optimization and validation of the RP-HPLC method intended
for the quantitative analysis of itraconazole and its impurities in pharmaceutical dosage forms. To reach the
desired chromatographic resolution with a limited number of experiments in a minimum amount of time, Box-
-Behnken design was used to simultaneously optimize
some important chromatographic parameters, such as the
acetonitrile content in the mobile phase, pH of the aqueous
phase and the column temperature. Separation between
itraconazole and impurity F was identified as critical
and selected as a response during the optimization. The
set optimal mobile phase composition was acetonitrile/water pH 2.5 adjusted with o-phosphoric acid (50:50,
V/V). Separations were performed on a Zorbax Eclipse
XDB-C18, 4.6 ´ 150 mm, 5 m particle size column with
the flow rate 1 mL min–1, column temperature set at 30 °C
and UV detection at 256 nm. The established method was
then subjected to method validation and the required validation parameters were tested. For the robustness evaluation, fractional factorial 24–1 design was utilized and
factors that might significantly affect the system performance were defined. As other validation parameters
were also found to be suitable, the possibility to apply
the proposed method for the determination of itraconazole, its impurities B and F in any laboratory under different circumstances has been proven.
