Comprehensive RP-HPLC Analysis of Sulopenem Etzadroxil and Probenecid: Development, Validation and Stability Studies

A highly precise and validated RP-HPLC method was established for the simultaneous estimation of Sulopenem Etzadroxil and Probenecid in both pure form and pharmaceutical formulations, including stability studies.The RP-HPLC analysis was performed using a Hyperclone 5µ BDS C18 column with an isocratic mobile phase of Acetonitrile and Ammonium Formate Buffer (40:60, pH 3.0). Detection was carried out at 272 nm with a 1.0 mL/min flow rate, 10 µL injection volume, and a 5-minute runtime. The method, validated as per ICH guidelines, exhibited excellent linearity (25–150 µg/mL) with high correlation coefficients. Accuracy studies demonstrated recoveries of 100.7% for Sulopenem Etzadroxil and 99.9% for Probenecid. The robustness evaluation indicated minimal variation in retention time and peak area under slight changes in chromatographic conditions. Sulopenem Etzadroxil exhibited maximum degradation under thermal, hydrolytic, and peroxide conditions, while Probenecid showed notable degradation under peroxide, acid, and photolytic stress. Minimal degradation was observed under alkali and reduction conditions for both drugs.