目的 建立阿奇霉素顆粒中基因毒性雜質(zhì)對甲苯磺酸乙酯的高效液相色譜（HPLC）測定方法。方法 采用Waters XBridge C₁₈（250 mm×4.6 mm，5 μm）色譜柱，以磷酸鹽緩沖液（2.2 g·L^-1無水磷酸氫二鉀溶液，用稀磷酸調(diào)節(jié)pH值至8.9）為流動相A；甲醇-乙腈（25∶75）為流動相B，進(jìn)行梯度洗脫，體積流量為1.0 mL·min^-1，檢測波長為225 nm，柱溫為60℃，進(jìn)樣量為50 μL。進(jìn)行專屬性、線性關(guān)系、定量限和檢測限、精密度、重復(fù)性、回收率、穩(wěn)定性、耐用性試驗。結(jié)果 空白溶劑、空白輔料、混合溶液中鄰近對甲苯磺酸乙酯的雜質(zhì)L均不干擾對甲苯磺酸乙酯檢測；對甲苯磺酸乙酯質(zhì)量濃度在0.05～0.20 μg·mL^-1（R²＝0.999 9）與峰面積呈良好線性關(guān)系；檢測限為0.016 7 μg·mL^-1，定量限為0.050 0 μg·mL^-1；平均回收率為95.76%～96.48%，RSD為0.42%～1.53%；精密度、重復(fù)性、穩(wěn)定性、耐用性均符合要求。3批樣品中均未檢出對甲苯磺酸乙酯。結(jié)論 建立的方法專屬性強(qiáng)、靈敏度高、特異性強(qiáng)、準(zhǔn)確度高，可用于阿奇霉素顆粒中基因毒性雜質(zhì)對甲苯磺酸乙酯的限度控制與檢測。

Objective To establish a high performance liquid chromatography (HPLC) method for the determination of genotoxic impurity ethyl p-toluenesulfonate in azithromycin granules. Methods The separation was performed on Waters XBridge C₁₈ column (250 mm×4.6 mm, 5 μm) at a flow rate of 1.0 mL·min^-1. The mobile phase A was phosphate buffer (2.2 g·L^-1 anhydrous dipotassium hydrogen phosphate solution, adjusted to pH 8.9 with dilute phosphoric acid), and mobile phase B consisted of methanol-acetonitrile (25:75) and gradient elution was performed. The detection wavelength was set at 225 nm, the column temperature was 60℃, and the injection volume was 50 μL. Specificity, linearity, limit of quantitation and detection, precision, repeatability, recovery, stability and durability tests are carried out. Results Blank solvent, blank excipient and impurity L nearest to ethyl p-toluenesulfonic acid in mixed solution do not interfere with ethyl p-toluenesulfonic acid detection. Good linear relationships between the concentration and the peak area of ethyl p-toluenesulfonate in the range of 0.05-0.20 μg·mL^-1(R²=0.999 9) was achieved. The limit of detection was 0.016 7 μg·mL^-1, and the limit of quantitation was 0.050 0 μg·mL^-1. The average recoveries were 95.76%-96.48%, RSD were 0.42%-1.53%. Precision, repeatability, stability and durability all meet the requirements. No genotoxic impurities ethyl p-toluenesulfonate was detected in three batches of samples. Conclusion This method has strong specificity, high sensitivity, high specificity and high accuracy, and can be used for the limit control and detection of ethyl p-toluenesulfonate, a genotoxic impurity in azithromycin granules.

R917