目的:研究曲匹地尔的有关物质,并建立高效液相色谱法测定有关物质。方法:采用SinoChrom ODS-BP(200 mm×4.6 mm,5 μm) 色谱柱,以甲醇、乙腈和磷酸盐缓冲液为流动相,等度洗脱,流速1.0 mL·min-1,检测波长205 nm。采用校正因子计算已知杂质和其它杂质的含量,并对降解试验产生杂质采用LC-MS进行确认,确证降解产物结构。结果:曲匹地尔与强制破坏产生的降解产物均分离良好;曲匹地尔杂质A和杂质B定量下限分别为0.089 ng和0.13 ng,检测下限分别为0.029 ng和0.055 ng;杂质A和杂质B的质量浓度分别在0.404 8~20.24 μg·mL-1(r=1.000)和0.456 8~22.84 μg·mL-1(r=1.000)范围内与峰面积呈良好的线性关系;杂质A和杂质B低、中、高 3种浓度的平均回收率分别为100.2%和101.0%,RSD分别为1.3%和0.76%(n=9)。4批样品有关物质测定结果显示,各已知杂质和其他杂质的含量均低于0.1%,杂质总量均低于0.5%。结论:本方法可用于曲匹地尔的有关物质测定。
Objective: To establish an HPLC method for the determination of related substances in trapidil. Methods: A SinoChrom ODS-BP (200 mm×4.6 mm, 5 μm) column was used. Methanol, acetonitrile and phosphate buffer were used as mobile phases. The isocratic elution was performed at a flow rate of 1.0 mL·min-1 and the detection wavelength was 205 nm. The correction factor is used to calculate the content of known impurities and other impurities, and the impurities generated in the degradation test are confirmed by LC-MS. Results: Trapidil was well separated from known impurities and degradation products produced by forced destruction. The limits of quantification of impurity A and impurity B were 0.089 ng and 0.13 ng, and the limits of detection were 0.029 ng and 0.055 ng, respectively; Impurity A and impurity B showed good linear relationship within the range of 0.404 8-20.24 μg·mL-1 (r=1.000) and 0.456 8-22.84 μg·mL-1 (r=1.000), respectively. The average recoveries of impurity A and impurity B at low, medium, and high concentrations were 100.2% and 101.0%, and RSDs were 1.3% and 0.76% (n=9). The determination results of related substances in 4 batches of samples showed that the content of each known impurity and any other impurity was less than 0.1%, and the total impurities was less than 0.5%.Conclusion: This method can be used for the determination of related substances in trapidil.
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