Rao R Nageswara, Nagaraju V
Analytical Chemistry Division, Indian Institute of Chemical Technology, Tarnaka, Hyderabad-500007, India.
J Pharm Biomed Anal. 2004 Nov 19;36(4):729-35. doi: 10.1016/j.jpba.2004.07.040.
A simple and rapid reversed-phase high-performance liquid chromatographic method for separation and determination of process-related impurities of difloxacin (DFL) was developed. The separation was achieved on a reversed-phase C(18) column using methanol-water-acetic acid (78:21.9:0.1, v/v/v) as a mobile solvent at a flow rate of 1.0 ml/min at 28 degrees C using UV detection at 230 nm. It was linear over a range of 0.03 x 10(-6) to 1.60 x 10(-6)g for process related impurities and 0.05 x 10(-6) to 2.40 x 10(-6)g for difloxacin. The detection limits were 0.009 x 10(-6) to 0.024 x 10(-6)g for all the compounds examined. The recoveries were found to be in the range of 97.6-102.0% for impurities as well as difloxacin. The precision and robustness of the method were evaluated. It was used for not only quality assurance, but also monitoring the synthetic reactions involved in the process development work of difloxacin. The method was found to be specific, precise and reliable for the determination of unreacted levels of raw materials, intermediates in the reaction mixtures and the finished products of difloxacin.
建立了一种简单快速的反相高效液相色谱法，用于分离和测定二氟沙星（DFL）的工艺相关杂质。分离在反相C(18)柱上进行，以甲醇 - 水 - 乙酸（78:21.9:0.1，v/v/v）作为流动相，流速为1.0 ml/min，在28℃下于230 nm处进行紫外检测。工艺相关杂质在0.03×10(-6)至1.60×10(-6)g范围内呈线性，二氟沙星在0.05×10(-6)至2.40×10(-6)g范围内呈线性。所有检测化合物的检测限为0.009×10(-6)至0.024×10(-6)g。杂质和二氟沙星的回收率在97.6 - 102.0%范围内。对该方法的精密度和稳健性进行了评估。它不仅用于质量保证，还用于监测二氟沙星工艺开发工作中涉及的合成反应。该方法被发现对于测定二氟沙星的原料未反应水平、反应混合物中的中间体以及成品具有特异性、精确性和可靠性。
