RP-HPLC and chemometric assisted UV-spectrophotometric methods for simultaneous in vitro analysis of atrovastatin calcium, ezetimibe and fenofibrate in their pharmaceutical formulation

One RP-HPLC and two chemometric assisted UV spectrophotometric methods
were developed and validated for the simultaneous in vitro analysis of Atrovastatin
calcium (ATV), Ezetimibe (EZET) and Fenofibrate (FEN) in their pharmaceutical
preparation and ternary mixtures. The chromatographic separation was achieved on
a reversed-phase, Hypersil BDS C8 column (250X4.6 mm i.d, 5 particle size)
with a mobile phase consisting of methanol and 0.05M phosphate buffer (pH-6.3
adjusted with sodium hydroxide) in the ratio of (85:15)% v/v. The total run time
was 7 min. Quantitation was achieved with UV detection at 248nm based on peak
area. Linearity was observed over concentration range of 5 - 12μg mL-1 for ATV
and EZET and 80 - 192 μg mL-1 for FEN. The two chemometric methods applied
were inverse least square (ILS) and classical least square (CLS). These approaches
were successfully applied to quantify each drug in their mixture using the
information included in the UV absorption spectra of appropriate solutions in the
wavelength range 220-310nm with the intervals of 5nm (Δλ = 5nm) at 19
wavelength points. For the chemometric calibration, 18 ternary solutions were
prepared as training set and 10 ternary solutions were prepared as validation set.
The developed methods were successfully applied for laboratory prepared mixtures
as well as commercial tablet formulation for ATV, EZET and FEN concentration.
The results obtained for pharmaceutical formulation by ILS and CLS methods were
compared with isocratic HPLC method and a good agreement was found.