RP-HPLC法测定磷酸萘酚喹的有关物质及含量

Determination of related substances and content of naphthoquine phosphate by RP-HPLC

目的:建立同时测定磷酸萘酚喹中有关物质和含量的高效液相色谱方法,为该原料药的质量控制提供了有效的分析方法。方法:Agilent SB-C8柱(150 mm×4.6 mm,5μm);流动相组成A:0.1%三氟乙酸溶液(三乙胺调节pH=3.0),B:乙腈,有关物质梯度洗脱,流速为1.0 mL.min-1,含量测定等度洗脱,流速为0.5 mL.min-1;检测波长为275 nm,柱温为35℃。结果:合成中间体Ⅰ~Ⅳ分别在2.057~51.428,2.333~58.336,1.766~44.140和2.113~52.828μg.mL-1的浓度范围内均有良好的线性关系,平均回收率均在96.1%~99.6%,RSD均在0.8%~3.0%,最低检测限均在0.5~8.8 ng;磷酸萘酚喹的线性范围为114.16~266.38μg.mL-1(r=0.999 9),低、中、高3个浓度RSD为0.24%(n=9)。结论:该方法灵敏、准确、专属性高,可用于磷酸萘酚喹的质量控制。

Objective：To establish a RP-HPLC method for simultaneous determination of related substances and content of naphthoquine phosphate for quality control.Methods： An Agilent SB-C8 column（150mm×4.6mm,5μm） was used with 0.1% trifluoroacetic acid（pH adjusted to 3.0 with triethylamine） as mobile phase A and acetonitrile as mobile phase B.Gradient elution was used for the determination of related substances and the flow rate was 1.0mL·min-1.Isocratic elution was used for the determination of content and the flow rate was 0.5mL·min-1.The detection wavelength was 275nm,and the column temperature was maintained at 35℃.Results： The calibration curves of chemically synthetic intermediates I^IV were linear in the concentration ranges of 2.057~51.428,2.333~58.336,1.766~44.140 and 2.113~52.828μg·mL-1,respectively.The average recoveries were 96.1%~99.6% with RSDs of 0.8%~3.0%,and LODs were 0.5~8.8ng.The linear range for naphthoquine phosphate was 114.16~266.38μg·mL-1（r=0.9999）,and RSD of low,mediate and high concentrations was 0.24%（n=9）.Conclusion： The method is sensitive,reliable and specific,and can be used for the quality control of naphthoquine phosphate.