Simultaneous determination of newly developed antiviral agents in pharmaceutical formulations by HPLC-DAD

TLDR

Viekira Pak® is a newly approved oral combination of ombitasvir, paritaprevir, ritonavir, and dasabuvir for treating hepatitis C genotype 1 infection. The study aimed to develop and validate a rapid, selective, and economical HPLC‑DAD method for simultaneous determination of these four antivirals in bulk and pharmaceutical preparations. Using an RPC18 column with a 10 mM phosphate buffer (pH 7)/acetonitrile (35:65) mobile phase at 1 mL min⁻¹ and 254 nm detection, sorafenib as internal standard, the method was validated for specificity, linearity, LOD, LOQ, accuracy, precision, robustness, and stability. Calibration curves were linear (2.5–60 µg mL⁻¹ for PAR, 1.25–30 µg mL⁻¹ for DAS, 1.7–40 µg mL⁻¹ for RIT, 0.42–10 µg mL⁻¹ for OMB) with R² > 0.999, and the method accurately quantified the drugs in tablets without excipient interference, making it suitable for routine quality control.

Abstract

Ombitasvir/paritaprevir/ritonavir/dasabuvir (Viekira Pak®) are the newest medicines approved for use in the treatment of hepatitis C virus (HCV) and are available in tablet form as an oral combination. Specifically, these agents are indicated in the treatment of HCV in patients with genotype 1 infection. Due to the therapeutic importance and increased use of Viekira Pak, proper methods for its determination in bulk and pharmaceutical formulations must be developed. The present study describes the development and validation of a simple, rapid, selective and economical reverse phase high performance liquid chromatography-diode array detection (HPLC-DAD) method for the simultaneous determination of paritaprevir (PAR), ombitasvir (OMB), dasabuvir(DAS) and ritonavir (RIT) in bulk and pharmaceutical preparations. The proposed method was carried out using an RPC18 column (150 × 4.5 mm, 3.5 μ), with a mobile phase consisting of 10 mM phosphate buffer (pH 7)and acetonitrile (35:65, v/v) at a flow rate of 1 ml/min and a detection wavelength of 254 nm. Sorafenib (SOR) was selected as the internal standard to ensure that the quantitative performance was high. The method was validated based on its specificity, linearity, limit of detection, limit of quantitation, accuracy, precision, robustness and stability. The calibration curves for PAR, DAS, RIT and OMB were linear at 2.5–60, 1.25–30, 1.7–40 and 0.42–10 μg/ml, respectively, and all of the correlation coefficients were >0.999. The proposed method was successfully applied for the determination of ombitasvir/paritaprevir/ritonavir/dasabuvirin tablets, without interference from the excipient peaks. Hence, the method can be applied for the routine quality control analysis of the studied drugs, either in bulk or dosed forms.