HPLC法测定巴特芬搽剂的有关物质

Determination of related substances in Batrafen Nail Lacquer by HPLC

目的:建立测定巴特芬搽剂有关物质的高效液相色谱法。方法:采用Purospher C18柱,以流动相ψ[A(乙腈):流动相B(含0.025 mol/L枸橼酸、0.0025 mol/L乙二胺四乙酸二钠,pH6.5)]=35∶65的混合液作为流动相,流速为1.0 mL/min,检测波长为303 nm,进样量为20μL,柱温为30℃。结果:有关物质Ⅰ及有关物质Ⅱ的浓度在0.05～3.0μg/mL范围内均与峰面积呈良好的线性关系,r分别为0.9994、0.9992。平均回收率分别为100.2％、100.4％,RSD分别为0.54％、0.74％(n=9)。最低检测限量有关物质Ⅰ为0.1 ng,有关物质Ⅱ为0.3 ng。结论:本方法简便,快速,结果准确,重现性好,可用于该药有关物质的检查。

Objetive: To establish a HPLC method for the determination of related substances in Batrafen Nail Lacquer. Methods: The separation was performed on a Purospher C18 column (5 μm, 4.0 mm × 125 mm) with acetonitrile and a solution of 0.025 mol/L citric acid monohydrate and 0. 0025 mol/L Disodium ethylenediamine tetracetate in 1000 mL pure water (adjusted to pH6. 5 with 10% NaOH ) ( 35 : 65) as the mobile phase . The flow rate was 1. 0 mL/min, the detection wavelength was 303 nm, the injection volume was 20μL and the column temperature was maintained at 30℃. Results: The calibration curves were linear in the concentration range of 0. 05 -3.0 μg/mL for related substance Ⅰ and related substance Ⅱ , r= 0.9994 and 0.9992 respectively . The average recovery rates were 100.2% and 100.4% , RSDs were 0.54% and 0. 74% (n = 9) respectively. The detection limits were 0. 1 ng for related substance Ⅰ and 0. 3 ng for related substance Ⅱ. Conclusions: The method is simple, rapid, accurate and well reproducible. It may be adopted for the examination of the impurities of the drug.