盐酸阿莫罗芬搽剂中杂质Ro16-8652和Ro40-1021 HPLC分析方法的建立

HPLC method development for the determination of the impurity Ro16-8652 and Ro40-1021 in amorolfine hydrochloride liniment

目的:通过对盐酸阿莫罗芬搽剂杂质Ro16-8652和Ro40-1021的研究,建立盐酸阿莫罗芬搽剂有关物质的HPLC分析方法。方法:采用Agilent ZOBAX Plus C18(100 mm×4.6 mm,3.5μm)色谱柱,以0.02 mol·L-1磷酸氢二钾溶液(A)-甲醇(B)(20∶80)为流动相等度洗脱60 min,流速1.0 mL·min-1,柱温40℃,检测波长214 nm,进样量20μL。结果:盐酸阿莫罗芬搽剂主成分与已知杂质及其强制破坏产生的降解产物分离良好;杂质Ro16-8652和Ro40-1021质量浓度分别在0.062 82~3.141μg·mL-1、0.062 46~3.123μg·mL-1范围内线性关系良好(r2>0.999 9,n=6),校正因子分别为1.21和1.09;平均回收率分别为100.1%和98.4%,RSD分别为1.6%和5.1%;检测重复性(n=6)RSD分别为1.2%和3.0%;中间精密度试验RSD(n=12)分别为1.0%和3.2%;杂质Ro16-8652、杂质Ro40-1021待测溶液在12 h内均稳定。经检测表明,稳定工艺的3批产品中,单个杂质Ro16-8652和Ro40-1021均不大于0.2%,总杂质均不大于0.6%。结论:本法可用于盐酸阿莫罗芬搽剂有关物质的检测。

Objective:To develop an HPLC method for the determination of the impurities Ro16-8652 abd Ro40-1021 in amorolfine hydrochloride liniment.Methods:The analysis was conducted on anoctadecyl bonded silica column[Agilent ZOBAX Plus C18(100 mm×4.6 mm,3.5μm)]with the isocratic elution of 0.02 mol·L-1 potassium dihydrogen phosphate buffer(A)and methanol(B)(20∶80)at the flow rate of 1.0 mL·min-1 for 60 min.The temperature was 40℃,the detection wavelength was 214 nm and the injection volume was 20μL.Results:The principle components of amorolfine hydrochloride liniment were well sparated from the konw impurities and the degrected products produced by forced degradation.The calibration curves of impurity Ro16-8652 and Ro40-1021 were linear in the self-concentration range of 0.062 82-3.141μg·m L-1 and 0.062 46-3.123μg·m L-1(R2>0.999 9,n=6),with the correction factors of 1.21 and 1.09,respectively.The average recoveries(n=9)were 100.1%and 98.4%,respectively,and the RSDs(n=9)were 1.6%and 5.1%,respectively.The RSDs of repeatability(n=6)and the intermediate precision(n=12)for impurities Ro16-8652 and Ro40-1021 were 1.2%and 3.0%,1.0%and3.2%,respectively.The test solutions with impurities Ro16-8652 and Ro40-1021 were stable within 12 h.The results showed that every impurity of Ro16-8652 and Ro40-1021 was less than identification limit of 0.2%,and the total impurities level was no more than 0.6%in the three batches of amorolfine hydrochloride liniment.Conclusion:The established method is suitable for the determination of the related substances in amorolfine hydrochloride liniment.