A Stability Indicating UPLC Method for the Determination of Levofloxacin Hemihydrate in Pharmaceutical Dosage Form: Application to Pharmaceutical Analysis
A reliable and sensitive isocratic stability indicating RP-UPLC method has been developed and validated for quantitative analysis and content uniformity study of levofloxacin hemihydrate in tablets. An isocratic method for analysis of levofloxacin hemihydrate was archived on ACQUITY UPLC BEH C18 (100*2.1)？mm particle size 1.7？ columns within shorter runtime of 4 min with a flow rate of 0.400？mL/min and using a photodiode array detector to monitor the eluate at 294？nm. The mobile phase consisted of acetonitrile-buffer (23？:？77 v/v), (buffer: 20？mM K2HPO4 + 1？mL triethylamine in 1？L water, by orthophosphoric acid). Response was a liner function of drug concentration in the range of 0.5–80？ g/mL ( ) with a limit of detection and quantification of 0.1 and 0.5？ g/mL, respectively. Accuracy (recovery) was between 99.77% and 101.55%. The drug was subjected to oxidation, hydrolysis, photolysis, and thermal degradation. Degradation products resulting from the stress studies did not interfere with the detection of levofloxacin hemihydrate, and the assay is stability indicating. 1. Introduction Levofloxacin hemihydrate (Figure 1) is a synthetic chemotherapeutic antibiotic of the fluoroquinolone drug class and is used to treat severe life-threatening bacterial infection or bacterial infection that have failed to respond to other antibiotic classes. Figure 1: Levofloxacin hemihydrate. IUPAC name is (S)-9-fluoro-2,3-dihydro-3-methyl-10-(4-methylpiperazin-1-yl)-7-oxo-7H-pyrido[1,2,3-de]-1,4-benzoxazine-6-carboxylic acid. Levofloxacin hemihydrate is highly water soluble in nature. It is also soluble in organic solvents, soluble in glacial acetic acid and chloroform, sparingly soluble in methanol, slightly soluble in ethanol, and practically insoluble in ether. Levofloxacin hemihydrate is odourless drug. Methods for quantitative analysis of Levofloxacin by HPLC [1–7], by UV [8–10] spectroscopy in single as well as in combination, are available in the literature. There are no methods available for quantitative analysis by UPLC. The method was developed and validated as per ICH [11–13] and USP [14] guideline. 2. Experimental 2.1. Chemicals and Reagents Levofloxacin hemihydrates reference standard (claim 99.48%) was provided by Cipla pharmaceuticals. Tablets (500？mg) of Levofloxacin were produced from a pharmacy. HPLC grade methanol and ortho phosphoric acid were purchased from Merck India Limited, Mumbai, India. Analytical grade hydrochloric acid, sodium hydroxide pellets, and hydrogen peroxide solution 30% (v/v) were obtained from Ranbaxy Fine Chemicals, New Delhi,
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