A reliable chromatographic method for the simultaneous determination of ciprofloxacin and moxifloxacin in human serum
2020
Purpose: To develop and validate a simple chromatographic method for the analysis of ciprofloxacin and moxifloxacin in human serum.Methods: After protein precipitation had been performed, high performance liquid chromatography (HPLC) with UV detection was utilized for the analysis of ciprofloxacin and moxifloxacin in human serum. Analytical column Zorbax SB-C18 (150 mm x 4.6 mm i.d., particle size 3.5 μm) was used as a stationary phase. Chromatographic separation was realized with the mobile phase 0.1% trifluoroacetic acid in water for chromatography - methanol (66:34, v/v), at the flow rate of 1 mL/min, temperature of 35 oC and detection at 280 nm. The method validation was performed according to the guidelines of the European Medicines Agency (EMA).Results: The chromatographic run time was about 12 minutes and no interference was observed. For ciprofloxacin, the method was linear over a concentration range of 0.5-50 μg/mL, with a correlation coefficient of 0.9874. For moxifloxacin, the method was linear over a concentration range of 0.5-50 μg/mL, with a correlation coefficient of 0.9946. Since relative standard deviation (RSD) and relative recovery values were within acceptable limits according to EMA guidelines, good intra-day precision, inter-day precision, as well as the accuracy of the method, were observed.Conclusion: A simple and reliable HPLC-UV method has been developed and validated for the simultaneous determination of ciprofloxacin and moxifloxacin in human serum. The method can be applied for therapeutic drug monitoring but also and pharmacokinetic studies of ciprofloxacin and moxifloxacin.
Keywords: Human serum, Ciprofloxacin, Moxifloxacin, Protein precipitation, HPLC, UV detection, Method validation
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