目的: 建立HPLC法检测伊沙匹隆原料药7个有关物质(有关物质1~7)的方法,为伊沙匹隆的质量控制提供方法依据。方法: 采用Welch XB-C18色谱柱(250 mm×4.6 mm,3 μm),以乙腈-0.04 mol·L-1乙酸铵水溶液为流动相,梯度洗脱,流速1.0 mL·min-1,柱温25 ℃,检测波长249 nm,进样量10 μL。结果: 伊沙匹隆与其7个有关物质的色谱峰均能良好分离。有关物质1~7质量浓度在0.05~2 μg·mL-1范围内与相应的峰面积呈良好的线性关系(r≥0.999 5);定量限分别为0.025、0.033、0.043、0.036、0.036、0.108、0.032 μg·mL-1;平均加样回收率(n=9)分别为98.5%、98.3%、101.2%、100.3%、97.6%、101.0%和98.6%,RSD分别为2.3%、2.8%、3.3%、2.1%、2.6%、5.7%和3.6%。3批伊沙匹隆样品检测结果显示,已知杂质及其他最大单个杂质含量均＜0.083%,杂质总含量＜0.256%。结论: 本方法专属性强,准确度高,可用于伊沙匹隆原料药有关物质的测定,同时填补了国内外关于伊沙匹隆有关物质检测方法的空白。
Objective: To establish a detection method for the seven related substances (related substances 1-7) of ixabepilone API by HPLC and provide a method basis for the quality control of ixabepilone. Methods: Welch XB-C18 column(250 mm×4.6 mm,3 μm) was adopted, the mobile phase consisted of acetonitrile-0.04 mol·L-1 ammonium acetate aqueous solution with gradient elution. The flow rate was 1.0 mL·min-1, the column temperature was 25 ℃, the detection wavelength was 249 nm and the injection volume was 10 μL. Results: The chromatographic peaks of ixabepilone and its seven related substances were well separated. A good linear relationship was shown between the mass concentration of 1-7 related substances of ixabepilone and the corresponding peak area within the concentration range of 0.05-2 μg·mL-1(r≥0.999 5). The limits of quantification were 0.025, 0.033, 0.043, 0.036, 0.036, 0.108 and 0.032 μg·mL-1, respectively, and the average recoveries (n=9) were 98.5%, 98.3%, 101.2%, 100.3%, 97.6%, 101.0% and 98.6% with RSDs of 2.3%, 2.8%, 3.3%, 2.1%, 2.6%, 5.7% and 3.6%, respectively. The detection results of 3 batches of ixabepilone samples showed that the contents of known impurities and other maximum single impuritie were less than 0.083%, and the total content of impurities was less than 0.256%. Conclusion: The detection method has demonstrated strong specificity and high accuracy, it could be used for the related substances of ixabepilone API, and fill the blank of the detection method of related substances of ixabepilone at home and abroad.
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