HPLC METHOD DEVELOPMENT AND VALIDATION FOR THE ESTIMATION OF LOTEPREDNOL ETABONATE AND LEVOFLOXACIN IN COMBINED DOSAGE FORM
Vijay Kumar Singh, Prem Prasad, Dr. Sanjay Kumar Kushwaha
ABSTRACT
A simple, precise, accurate, and robust reverse-phase high-performance liquid chromatography (RP-HPLC) method was developed and validated for the simultaneous estimation of Loteprednol Etabonate (LPE) and Levofloxacin (LFC) in combined ophthalmic dosage form. The chromatographic separation was achieved using a C18 column with a mobile phase composed of acetonitrile and phosphate buffer (pH 3.5) in a suitable ratio, at a flow rate of 1.0 mL/min and detection wavelength set at 254 nm. The retention times for Loteprednol Etabonate and Levofloxacin were found to be approximately 3.452 min and 5.614 min, respectively. The method was linear in the concentration range of 2–10 µg/mL for both drugs, with correlation coefficients (r²) greater than 0.999. Accuracy was confirmed by recovery studies, with mean recoveries ranging from 97.56% to 99.14%. The method demonstrated excellent precision with %RSD values less than 1% for intra- and inter-day studies. LOD and LOQ were found to be 0.20 µg/mL and 0.45 µg/mL for LPE, and 0.25 µg/mL and 0.75 µg/mL for LFC, respectively. The method was also found to be robust and specific, and successfully applied to assay analysis of marketed formulations. These results suggest that the developed method is suitable for routine quality control of LPE and LFC in combined dosage forms.
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