Khan Fahim Ullah, Nasir Fazli, Iqbal Zafar, Khan Ismail, Shahbaz Naila, Hassan Muhammad, Ullah Farhad
Department of Pharmacy, University of Peshawar, Peshawar 25120, Pakistan.
Department of Pharmacy, University of Peshawar, Peshawar 25120, Pakistan.
J Chromatogr B Analyt Technol Biomed Life Sci. 2016 Apr 1;1017-1018:120-128. doi: 10.1016/j.jchromb.2016.03.002. Epub 2016 Mar 4.
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.
建立了一种新颖、灵敏且经过验证的反相高效液相色谱-紫外检测法，以马来酸噻吗洛尔为内标，同时测定生理体液中的莫西沙星和氧氟沙星。根据国际指南对不同实验参数进行了优化和验证。使用Kromasil 100-5C18分析柱（250mm×4.6mm×5μm）实现了分析物的完全分离，以甲醇和0.05%三氟乙酸（TFA）（38:62v/v）作为流动相，在等度洗脱阶段以1.1ml/min的流速泵送，柱温箱温度保持在45°C，检测波长为290nm。采用蛋白沉淀法，以甲醇作为沉淀溶剂，从人血浆和牛房水样本中提取药物。该方法在莫西沙星浓度范围为0.018 - 100μg/ml、氧氟沙星浓度范围为0.014 - 20μg/ml时呈线性。该方法在人血浆中对莫西沙星和氧氟沙星的回收率分别为97.52%和97.39%，而在房水中莫西沙星的回收率为94.48%。血浆中莫西沙星和氧氟沙星的检测限分别为10.0和8.00ng/ml，其各自的定量限分别为18.0和14ng/ml。在房水中，莫西沙星的检测限和定量限分别为16.0和24ng/ml。未来，该方法将用于研究莫西沙星和氧氟沙星在生物体液及药品中的药代动力学特征。
