title: Simultaneous determination of moxifloxacin and ofloxacin in physiological fluids using high performance liquid chromatography with ultraviolet detection.
abstract: A novel, sensitive and validated RP-HPLC-UV method was developed for simultaneous determination of moxifloxacin and ofloxacin using timolol maleate as internal standard in physiological fluids. Different experimental parameters were optimized and validated according to international guidelines. Complete separation of the analytes was achieved with Kromasil 100-5C18 analytical column (250mm×4.6mm×5μm), methanol and 0.05% trifloroacetic acid (TFA) (38:62v/v) were used as mobile phase, pumped at flow rate of 1.1ml/min in isocratic phase, column oven temperature maintained at 45°C and detection wavelength of 290nm. Protein precipitation method was applied to extract the drugs from human plasma and bovine aqueous humor samples using methanol as precipitating solvent. This method is linear in concentration range of 0.018-100μg/ml for moxifloxacin and 0.014-20μg/ml for ofloxacin. The recoveries of the method were 97.52 and 97.39% in human plasma for MX and OFN respectively, while in aqueous humor 94.48% for MX. The LOD values in plasma were found to be 10.0 and 8.00ng/ml for MX and OFN respectively, while their respective LOQ values were 18.0 and 14ng/ml. In aqueous humor the LOD and LOQ for MX were 16.0 and 24ng/ml respectively. In future, this method will be used to study the pharmacokinetic profile of moxifloxacin and ofloxacin in biological fluids and pharmaceutical products.
authors.affiliation: Department of Pharmacy, University of Peshawar, Peshawar 25120, Pakistan.
authors.forename: Fahim Ullah
authors.initials: FU
authors.lastname: Khan
authors.affiliation: Department of Pharmacy, University of Peshawar, Peshawar 25120, Pakistan. Electronic address: fazlinasir@upesh.edu.pk.
authors.forename: Fazli
authors.initials: F
authors.lastname: Nasir
authors.affiliation: Department of Pharmacy, University of Peshawar, Peshawar 25120, Pakistan.
authors.forename: Zafar
authors.initials: Z
authors.lastname: Iqbal
authors.affiliation: Department of Pharmacy, University of Peshawar, Peshawar 25120, Pakistan.
authors.forename: Ismail
authors.initials: I
authors.lastname: Khan
authors.affiliation: Department of Pharmacy, University of Peshawar, Peshawar 25120, Pakistan.
authors.forename: Naila
authors.initials: N
authors.lastname: Shahbaz
authors.affiliation: Department of Pharmacy, University of Peshawar, Peshawar 25120, Pakistan.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Hassan
authors.affiliation: Department of Pharmacy, University of Peshawar, Peshawar 25120, Pakistan.
authors.forename: Farhad
authors.initials: F
authors.lastname: Ullah
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis*; D001826:Body Fluids / Q000737:chemistry*; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D024841:Fluoroquinolones / Q000032:analysis*; D006801:Humans; D057230:Limit of Detection; D000077266:Moxifloxacin; D015242:Ofloxacin / Q000032:analysis*; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*
chemical_list: D000900:Anti-Bacterial Agents; D024841:Fluoroquinolones; D015242:Ofloxacin; D000077266:Moxifloxacin
keywords: HPLC–UV; Moxifloxacin; Ofloxacin