HPLC determination of Besifloxacin in Ophthalmic Suspension using Chromolith^® SpeedROD RP-18e Column

Scope

Here we demonstrate an application for Besifloxacin in ophthalmic suspension on RP-HPLC using Chromolith^® SpeedROD RP-18e column for system suitability, linearity, repeatability, percentage recovery, LOD and LOQ.

Introduction

Besifloxacin (Figure 1) is a bactericidal fluoroquinolone-type antibiotic, used for the treatment of bacterial conjunctivitis caused by susceptible bacteria. It was approved by US FDA in May 2009. Here, a solution is demonstrated for the determination of Besifloxacin in ophthalmic suspension dosage using HPLC with a monolith column and UV Detection at 289 nm. At present, there are no pharmacopeial monographs available for the assay of Besifloxacin in ophthalmic suspension dosage forms.

Instrumentation

• Waters Alliance™ e2695 with Empower 3 Software
• Milli-Q^® Integral 3 Water purification system
• Ultrasonic bath from PCI analytics
• Millex^® HV Durapore membrane filter (PVDF) 0.45 µm

Chromatographic Data of Besifloxacin Analysis

Results and Discussion

An isocratic RP-HPLC method for the determination of Besifloxacin in ophthalmic suspension was developed using Chromolith^® SpeedROD RP-18 endcapped column with UV detection at 289 nm. The experimental data indicated an excellent linearity with an r² value of 0.999 (Figure 4) for the selected concentration range (0.005 – 75 µg/mL, Table 4). The LOD and LOQ values were estimated as 2.3 µg/mL and 7 µg/mL, respectively. The method precision for assay was below 1% RSD and the percentage recovery found to be ranging from 102.1 -104 % (Table 5).

In conclusion, the data for linearity, system suitability, repeatability of the method suggests its aptness for the assay of Besifloxacin in opthalmic suspension using Chromolith^® SpeedROD RP-18 endcapped column.