Qualitative and Quantitative Estimation of Ceftriaxone in Pharmaceutical Dosage Forms Using Reverse Phase High...

As the "pharmacy of the world," India plays a critical role in global pharmaceutical supply. However, increasing reports of substandard and falsified medicines raise serious concerns. With Jammu and Kashmir witnessing repeated quality failures of ceftriaxone, this study aims to verify its quality using a refined high-performance liquid chromatography (HPLC)-based analytical method.

The ceftriaxone assay was performed using Agilent 1260 Infinity HPLC System with a C18 column and EZChromS1 software. A solvent system of acetonitrile and water (20:80, v/v) was used, with flow rate of 0.5 mL/min and ultraviolet detection at 254 nm. Standard solutions (20-70 μg/mL) were prepared from a 1,000 μg/mL stock for calibration. Nineteen injectable ceftriaxone brands were procured and coded for anonymity. Sample solutions were prepared at 50 μg/mL. The method was validated as per International Council for Harmonisation (ICH) guidelines for accuracy, precision, linearity, specificity, limit of detection (LOD), limit of quantification (LOQ), and robustness. Solution stability was assessed over 7 days at 2°C-8°C.

The developed isocratic reversed-phase HPLC method showed good linearity (R² = 0.9991) over the range 20-70 μg/mL, with LOD and LOQ of 5.88 μg/mL and 17.83 μg/mL, respectively. Intra-day and inter-day precision and accuracy showed percentage relative standard deviation (%RSD) values <2% and mean recovery within acceptable limits (96.9%-102.6%). Robustness was confirmed through variations in flow rate, mobile phase composition, detection wavelength, and column temperature, all yielding %RSD <2%. Analysis of 19 marketed formulations revealed that 89.5% complied with pharmacopoeial standards (90.0%-115.0% recovery), while 10.5% (CEF A17, CEF A19) were substandard, indicating significant quality variation among brands (p < 0.05).

A simple, sensitive, and cost-effective HPLC method was developed and validated for estimation of ceftriaxone in pharmaceutical dosage forms. Among 19 tested formulations, 89.5% complied with Indian Pharmacopoeial standards, while two formulations were found substandard, indicating variability in product quality.