目的 优化有关物质分析方法，有效分离罗红霉素分散片中13种杂质，建立加校正因子的主成分自身对照法测定有关物质，并与原研制剂进行有关物质一致性评价研究。方法 使用Waters XBridge^® C₁₈柱（150 mm×4.6 mm，3.5 μm） ，以0.52 mol·L^-1磷酸二氢铵水溶液（用5 mol·L^-1氢氧化钠调节pH值至4.3）为流动相A，以乙腈-水（70∶30）为流动相B，进行梯度洗脱；体积流量0.9 mL·min^-1，柱温20℃，检测波长205 nm，进样20 μL。对13种杂质成分进行线性回归，以线性斜率计算各杂质相对于罗红霉素的校正因子，用加校正因子的主成分自身对照法计算杂质含量并进行方法验证。罗红霉素分散片经碱破坏、酸破坏、氧化破坏、高温破坏、光照破坏后检测色谱图变化；取参比制剂及自研制剂，置于温度40℃、相对湿度75%条件下放置6个月，分别于第1、2、3、6个月取样测定其有关物质含量。结果 在已建立的色谱条件下，罗红霉素与相邻杂质、杂质与杂质之间均能有效分离，分离度均≥1.5。罗红霉素在0.968 9～100.090 0 μg·mL^-1相关系数为0.999 4，其他已知杂质在相应浓度范围内相关系数也均在0.999 0～1.000 0，线性关系均良好。各已知杂质平均回收率均在90.0%～108.0%，9份回收率的RSD≤5.0%。采用加校正因子的主成分自身对照法测定3批样品中杂质含量，结果与杂质对照品外标法一致。自研制剂杂质与原研制剂具有有关物质一致性。自研制剂在光照条件下稳定；在氧化、酸、碱、高温破坏条件下均有不同程度的降解，在酸条件下主峰降解最为明显，杂质B、D显著增加；氧化条件下降解产生氮氧化物；原研制剂与自研制剂在加速条件下均未检出杂质B、K及氮氧化物，在加速6个月过程中，二者杂质D均有增长趋势，其他杂质无明显变化。结论 建立的方法具有良好的专属性、准确性和重复性，可采用加校正因子的主成分自身对照法测定罗红霉素分散片中有关物质。

Objective Optimize the method of related substances to effectively separate 13 known impurities in Roxithromycin Dispersible Tablets. Establish a principal component self-control method with correction factors to determine related substances, and conduct a study on the consistency of related substances with the original research agent. Methods Waters XBridge^® C₁₈ column (150 mm×4.6 mm, 3.5 μm) was used for analysis, with a gradient elution system using 0.52 mol·L^-1 ammonium dihydrogen phosphate aqueous solution (adjust pH to 4.3 with 5 mol·L^-1 sodium hydroxide) as mobile phase A and acetonitrile-water (70:30) as mobile phase B. The flow rate was 0.9 mL·min^-1, column temperature was 20 ℃, detection wavelength was 205 nm and injection volume was 20 μL. The linear regression equations of 13 impurities were drawn to calculate the correction factor of each impurity relative to roxithromycin with linear slope. The method was used to calculate impurities contents and verify. Chromatographic changes of roxithromycin dispersive tablets were detected after alkali, acid, oxidation, high temperature and light damage. The reference preparation and self-developed preparation were placed at 40 ℃ and 75% relative humidity for six months, and Samples were taken in the 1st, 2nd, 3rd and 6th month to determine the content of related substances Results The resolution between roxithromycin and neighboring impurities, and impurities was greater than 1.5 under the above chromatographic condition. The correlation coefficient of roxith-romycin in the range of 0.968 9—100.090 μg·mL^-1 was 0.999 4, and the correlation coefficients of other known impurities in the corresponding concentration range were all between 0.999 0 and 1.000 0, respectively. The linear relationship was good. The average recovery rate of each known impurity was between 90.0%—108.0%, and the RSD of nine parts recovery rate was less than or equal to 5.0%. The impurity content in the three batches of samples was determined by the principal component reference method with correction factors, and the results were consistent with the impurity reference external standard method. Impurities of self-preparation were consistent with related substances of reference preparation. The self-developed agent was stable under light condition. The impurities B and D were significantly increased under the condition of oxidation, acid, alkali and high temperature. Oxidation conditions drop solution to produce nitrogen oxides. No impurities B, K and nitrogen oxides were detected in the original and self-developed agent under the condition of acceleration. In the process of acceleration for six months, the impurity D in both of them had an increasing trend, while the other impurities had no obvious change. Conclusion This method has good specificity, accuracy and repeatability, and the principal component reference substance method with correction factor can be used to determine the related substances in roxithromycin dispersible tablets.

R917

国家自然科学基金面上项目(81973443)