目的 優(yōu)化有關物質(zhì)分析方法，有效分離羅紅霉素分散片中13種雜質(zhì)，建立加校正因子的主成分自身對照法測定有關物質(zhì)，并與原研制劑進行有關物質(zhì)一致性評價研究。方法 使用Waters XBridge^® C₁₈柱（150 mm×4.6 mm，3.5 μm） ，以0.52 mol·L^-1磷酸二氫銨水溶液（用5 mol·L^-1氫氧化鈉調(diào)節(jié)pH值至4.3）為流動相A，以乙腈-水（70∶30）為流動相B，進行梯度洗脫；體積流量0.9 mL·min^-1，柱溫20℃，檢測波長205 nm，進樣20 μL。對13種雜質(zhì)成分進行線性回歸，以線性斜率計算各雜質(zhì)相對于羅紅霉素的校正因子，用加校正因子的主成分自身對照法計算雜質(zhì)含量并進行方法驗證。羅紅霉素分散片經(jīng)堿破壞、酸破壞、氧化破壞、高溫破壞、光照破壞后檢測色譜圖變化；取參比制劑及自研制劑，置于溫度40℃、相對濕度75%條件下放置6個月，分別于第1、2、3、6個月取樣測定其有關物質(zhì)含量。結(jié)果 在已建立的色譜條件下，羅紅霉素與相鄰雜質(zhì)、雜質(zhì)與雜質(zhì)之間均能有效分離，分離度均≥1.5。羅紅霉素在0.968 9～100.090 0 μg·mL^-1相關系數(shù)為0.999 4，其他已知雜質(zhì)在相應濃度范圍內(nèi)相關系數(shù)也均在0.999 0～1.000 0，線性關系均良好。各已知雜質(zhì)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對照法測定3批樣品中雜質(zhì)含量，結(jié)果與雜質(zhì)對照品外標法一致。自研制劑雜質(zhì)與原研制劑具有有關物質(zhì)一致性。自研制劑在光照條件下穩(wěn)定；在氧化、酸、堿、高溫破壞條件下均有不同程度的降解，在酸條件下主峰降解最為明顯，雜質(zhì)B、D顯著增加；氧化條件下降解產(chǎn)生氮氧化物；原研制劑與自研制劑在加速條件下均未檢出雜質(zhì)B、K及氮氧化物，在加速6個月過程中，二者雜質(zhì)D均有增長趨勢，其他雜質(zhì)無明顯變化。結(jié)論 建立的方法具有良好的專屬性、準確性和重復性，可采用加校正因子的主成分自身對照法測定羅紅霉素分散片中有關物質(zhì)。

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

國家自然科學基金面上項目(81973443)