Method For Determination Of Levofloxacin Research Articles

Development and Validation of a RP-HPLC Method for Simultaneous Determination of Levofloxacin and Moxifloxacin in Pharmaceutical Dosage Forms

A simple, fast and economic reversed phase high performance liquid chromatographic (HPLC) method has been successfully developed and validated for simultaneous determination of fluoroquinolone analogs namely levofloxacin and moxifloxacin in both pure form (as API) and in pharmaceutical dosage forms. The method was validated according to the guidelines of ICH, FDA and USP with respect to accuracy, precision and linearity. For method development a C-18 bonded silica column (250 x 4.6 mm, 5μ, Phenomenex, Inc) was used with a mobile phase comprising of 10% aqueous solution of acetic acid and acetonitrile in a ratio of 80:20 v/v. The flow rate was 0.5 mL/min and effluents were monitored at 300 nm and the retention times were found to be at 7.0±0.1 min and 10.59±0.1 min for levofloxacin and moxifloxacin, respectively. The recovery was found to be more than 99% for each spiked samples of levofloxacin and moxifloxacin, demonstrating the accuracy of the protocol. Intra-day and inter-day precisions of the new method were less than the maximum allowable limit (RSD% £ 2.0) according to FDA. The method showed linear response with correlation coefficient value of 0.9975 in both the cases.
 Therefore, the developed method was found to be simpler, accurate, reproducible, efficient and less time consuming and can be successfully applied for the simultaneous assay of levofloxacin and moxifloxacin formulations.HPLC, method development, validation, levofloxacin, moxifloxacin.

Evaluation of intraocular penetration of levofloxacin by high performance liquid chromatography

AIM: To evaluate the levofloxacin eye drop into human eye penetration, levofloxacin eye drop concentrations in human ocular aqueous of 33 patients undergoing cataract surgery were measured by high performance liquid chromatography (HPLC). METHODS: Totally 33 volunteer patients who scheduled for phacoemulsification surgery received one drop of levofloxacin every 6h for 3d before and on the day of surgery, administration of drug was stopped 1h before surgery. Levofloxacin concentration in aqueous humor was measured by HPLC method with fluorescence detector. RESULTS: A simple, effective and sensitive HPLC method for determination of levofloxacin in human ocular aqueous was validated. Linearity was shown for levofloxacin concentration over a wide range of 1.95×10-3-1.50 µg/mL. The mean aqueous level of levofloxacin was 0.3399±0.03405 µg/mL. CONCLUSION: Results from the present study demonstrate that topical administration of levofloxacin 0.5% before cataract surgery with routine dose (one drop every 6h) unable to reach MIC90 for most common microorganism causing acute bacterial endophthalmitis.

Comparison of high‑performance liquid chromatography and ultraviolet‑visible spectrophotometry to determine the best method to assess Levofloxacin released from mesoporous silica microspheres/nano‑hydroxyapatite composite scaffolds

An assessment of Levofloxacin by high-performance liquid chromatography (HPLC) or ultraviolet-visible spectrophotometry (UV-Vis) and its pharmacokinetics in serum or plasma was made in a previous study by the present authors. Levofloxacin-loaded mesoporous silica microspheres/nano-hydroxyapatite (n-HA) composite scaffolds comprise a novel synthetic composite scaffold that may be utilized as a drug-delivery system for clinical usage. However, few studies have been published concerning a comparison of HPLC with UV-Vis, which is the preferred method for determination of Levofloxacin. In the present study, an HPLC method was first established, and subsequently a comparison of HPLC with the UV-Vis method was performed. The standard curve was established, and recovery rate from simulated body fluid was calculated. The linear concentration range for Levofloxacin was 0.05–300 µg/ml. The regression equation for HPLC was y=0.033x+0.010, with R2=0.9991, whereas that for UV-Vis was y=0.065x+0.017, with R2=0.9999. The recovery rates of low, medium and high (5, 25 and 50 µg/ml) concentrations of Levofloxacin determined by HPLC were 96.37±0.50, 110.96±0.23 and 104.79±0.06%, respectively, whereas those for low, medium and high concentrations according to UV-Vis were 96.00±2.00, 99.50±0.00 and 98.67±0.06%, respectively. Taken together, these findings demonstrated that it is not accurate to measure the concentration of drugs loaded on the biodegradable composite composites by UV-Vis. HPLC is the preferred method to evaluate sustained release characteristics of Levofloxacin released from mesoporous silica microspheres/n-HA composite scaffolds. The present study also provides guidance on which methods should be selected for investigating the sustained release properties of drugs in tissue engineering. The accurate determination of drug concentration in the drug delivery system provides guidance for the treatment of infectious diseases.

Microextraction by Packed Molecularly Imprinted Polymer Combined Ultra-High-Performance Liquid Chromatography for the Determination of Levofloxacin in Human Plasma

Fluoroquinolones are considered as gold standard for the prevention of bacterial infections. To improve assessment of antibacterial efficacy, a novel method for determination of levofloxacin was developed and validated. Deep eutectic solvents (DESs) as only green solvent were used as a porogen for preparation of water-compatible molecularly imprinted polymers (MIPs) with a pseudotemplate. The DESs-MIPs were characterized in detail, including scanning electron microscope, nitrogen sorption porosimetry, and Fourier transform-infrared spectra. Clearly, the maximum binding capacity of levofloxacin on DESs-MIPs in water and methanol was 0.216 and 0.077 μmol g−1, respectively. The DESs-MIPs as adsorbing materials were applied in microextraction by packed sorbent (MEPS), and the DESs-MIPs-MEPS conditions were optimized. The DESs-MIPs-MEPS coupled with ultra-high-performance liquid chromatography (UHPLC) was used to determine levofloxacin in human plasma. The method was found linear over 0.05–10 μg mL−1 with coefficient of correlation equal to 0.9988. The limit of detection and limit of quantification were 0.012 and 0.04 μg mL−1, respectively. At three spiked levels, the precision of proposed method was between 95.3% and 99.7% with intraday and interday relative standard deviations ≤8.9%. Finally, the developed method was used to examine levofloxacin from human plasma of 20 hospitalized patients after transrectal ultrasound-guided prostate biopsy, and the average concentration (±SD) of levofloxacin was 2.35 ± 0.99 μg mL−1 in plasma.

A Rapid and Simple High-Performance Liquid Chromatographic Method for Determination of Levofloxacin in Human Plasma

A simple and rapid high-performance liquid chromatography method was developed and validated for quantifying LEV in human plasma. Chromatographic separation was performed under isocratic elution on a Luna Phenomenex® C18 (150 × 4.6 mm, 5 µm) column. The mobile phase was comprised of acetonitrile, methanol, and phosphate buffer 25 mM pH 3.0 (13:7:80 v/v/v) and pumped at a flow rate of 1.5 mL/min. Detection was performed by UV detector at a wavelength of 280 nm. Samples were pre-treated with acetonitrile followed by centrifugation, evaporation, and reconstitution step. The method proved linear (r = 0.995), sensitive (LLOQ and LOD was 1.8 and 0.6 µg/mL respectively), accurate (% error above LLOQ ≤ 12% and LLOQ ≤ 20%), precise (RSD ≤ 9%), robust in the ranges of 1.8-28.8 µg/mL, rapid (separation time not more than 10 min), and simple (use no organic additive in mobile phase). The method was showed reliable for quantifying LEV in human plasma.

Development and validation of UV spectrophotometric method for determination of levofloxacin in pharmaceutical dosage forms

The objective of this research was to develop and validate an alternative analytical method for quantitative determination of levofloxacin in tablets and injection preparations. The calibration curves were linear over a concentration range from 3.0 to 8.0 μg mL-1. The relative standard deviation was below 1.0% for both formulations and average recovery was 101.42 ± 0.45% and 100.34 ± 0.85% for tablets and injection formulations, respectively. The limit of detection and limit of quantitation were 0.08 and 0.25 μg mL-1, respectively. It was concluded that the developed method is suitable for the quality control of levofloxacin in pharmaceuticals formulations.

Rapid chiral separation and impurity determination of levofloxacin by ligand-exchange chromatography

A sensitive, simple, and accurate method for determination of levofloxacin and its ( R)-enantiomer was developed to determine the chiral impurity of levofloxacin in Cravit Tablets material by ligand-exchange high performance liquid chromatography. The effects of different kinds of ligands, concentration of ligands in mobile phase, organic modifier, pH of mobile phase, and temperature on enantioseparation were investigated and evaluated. Chiral separation was performed on a conventional C 18 column, where the mobile phase consisted of a methanol–water solution (containing10 mmol L −1 l-leucine and 5 mmol L −1 copper sulfate) (88:12, v/v) and its flow-rate was set at 1.0 mL min −1. The conventional C 18 column offers baseline separation of two enantiomers with a resolution of 2.4 in less than 20 min. Thermodynamic data (ΔΔ H and ΔΔ S) obtained by Van’t Hoff plots revealed the chiral separation is an enthalpy-controlled process. The standard curves showed excellent linearity over the concentration range from 0.5 to 400 mg L −1 for levofloxacin and its ( R)-enantiomer. The linear correlation equations are: y = 1.33 × 10 5 x + 6297 ( r = 0.9991) and y = 1.34 × 10 5 x + 3565 ( r = 0.9997), respectively. The relative standard deviation (RSD) of the method was below 2.3% ( n = 3).