Method Development and Validation for the Simultaneous Determination of Perphenazine and Amitriptyline in Pure and Marketed Pharmaceutical Dosage Form by using RP-HPLC

Abstract

An uncomplicated, precise, rapid, selective, and stable reversed-phase high-performance liquid chromatographic (RP-HPLC)
technique has been developed and validated for the concurrent quantification of Perphenazine (PPZ) and Amitriptyline
(AMT) in pure and its pharmaceutical dosage form. The method is based on Phenomenex Gemini C18 (4.6×250mm) 5μ
column. Separation is accomplished by pumping a 65:35% v/v mixture of methanol (MeOH) and TEA buffer at a rate of
1.0 mL/min, followed by UV detection at 230 nm. The study is performed with the column set at 40 °C. The total run time is
about 6 min. According to the ICH guidelines, the approach has been verified for accuracy, specificity, precision, robustness,
linearity, ruggedness, the limit of detection (LoD), the limit of quantification (LoQ), and system suitability. The technique
showed accuracy and linearity for determining PPZ and AMT between 10–50 μg/mL and 20–100 μg/mL, respectively. The
average %recovery (100.37% for PPZ and 100.34% for AMT), ruggedness (<2%), and robustness are proven to provide
good outcomes. This technique’s benefits include strong resolution with distinct peaks and adequate precision. The outcomes
show that the approach is appropriate for regular quality control analysis of commercial pharmaceutical formulations.