Validated Reversed-Phase Liquid Chromatographic Method with Gradient Elution for Simultaneous Determination of the Antiviral Agents: Sofosbuvir, Ledipasvir, Daclatasvir, and Simeprevir in Their Dosage Forms

Abstract

A simple, rapid, sensitive, and precise reversed-phase liquid chromatographic method was developed and validated for the simultaneous determination of four direct-acting antivirals, sofosbuvir (SF), ledipasvir (LD), declatasvir (DC), and simeprevir (SM), in their respective pharmaceutical formulations. Effective chromatographic separation was achieved on an Agilent Eclipse plus C8 column (250 mm × 4.6 mm, 5 µm) at 40 °C with gradient elution using a mobile phase composed of acetonitrile:phosphate buffer (pH 6.5). The quantification of SF and DC was based on peak area measurements at 260 nm, while the quantification of LD and SM was achieved at 330 nm. The linearity was acceptable from 1.0 to 20.0 μg/mL for the studied drugs, with correlation coefficients >0.999. The analytical performance of the newly proposed HPLC procedure was thoroughly validated according to ICH guidelines in terms of linearity, precision (RSD%, 0.39–1.57), accuracy (98.05–101.90%), specificity, limit of detection (LOD) (0.022–0.039 μg/mL), limit of quantification (LOQ) (0.067–0.118 μg/mL), and robustness. The validated HPLC method was successfully used to analyze the abovementioned drugs in their pure and dosage forms without interference from common excipients present in commercial formulations.

Simultaneous spectrofluorimetic determination of remdesivir and simeprevir in human plasma

“…However, it is higher than another fluorimetric method reported for SIM 25 . As a result, the key advantage of the new spectrofluorimetric technique is energy savings, as the spectrofluorometer requires the least amount of energy (less than 0.1kWh) when compared to HPLC (approximately 1.5kWh) in addition to high sensitivity of the proposed method compared to reported HPLC 12 , 13 , 32 , 33 . Despite the fact that the chromatographic technique is widely employed in pharmaceutical quality control and research laboratories, it is still considered a sophisticated technology due to the high cost of the equipment (column and detector) and the environmental damage caused by the use of hazardous organic solvents.…”

“…For the analysis of SIM, the literature review reveals several analytical methods for its assay, whether alone or in combination. These methods involve spectrophotometric 22,23 , chemometric-assisted spectrophotometric 24 , spectrofluorimetric [25][26][27] , and LC methods [28][29][30][31][32][33] .…”

Ionophore-Based Nanosphere Emulsion Incorporating Ion-Exchanger for Picogram Potentiometric Determination of HCV Drug (Daclatasvir) in Pharmaceutical Formulations and Body Fluids

“…Therefore, there is a strong need for the development of a simple, sensitive, selective, and cost-effective method for the determination of DAC in serum and urine for pharmacokinetics studies and in pharmaceutical formulations for active pharmaceutical ingredients (API) routine testing. Previously reported methods for the determination of DAC include spectroscopy [14], chromatography [15,16], and voltammetry [17][18][19]; these methods are expensive, need sample preparation, and exhibit a narrow linear concentration range. This study aimed to develop nanospheres based on ionophores for the accurate determination of DAC in pharmaceutical formulations and biofluids using complexometric potentiometric titration.…”