Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, Zhejiang University of Technology, Hangzhou 310014, China.
School of Medicine, Zhejiang University, Hangzhou 310058, China.
J Pharm Biomed Anal. 2024 Oct 15;249:116384. doi: 10.1016/j.jpba.2024.116384. Epub 2024 Jul 29.
Etimicin is a typical aminoglycoside antibiotic (AG). High performance liquid chromatography-evaporation light scattering detector (HPLC-ELSD) method is a commonly used method for determining impurities in Etimicin. However, due to the poor reproducibility, low sensitivity and narrow linear range of the ELSD, high-throughput quantitative analysis of impurities in Etimicin currently poses a challenge. In this study, a sensitive method using hydrophilic interaction liquid chromatography coupled with charged aerosol detector (HILIC-CAD) was developed for the analysis of the impurities in Etimicin. The liquid phase conditions for determination impurities in Etimicin were optimized using Box Behnken design (BBD) and response surface methodology (RSM), resulting in satisfactory separation and optimal CAD output signal. We also studied the influence of CAD parameters on the signal-to-noise ratio and linearity of Etimicin and its impurities. This method has also been proven to be effective in separating impurities from two other typical AGs, Isepamicin and Amikacin. In the method validation, the coefficient of determination (R) of Etimicin, Isepamicin and Amikacin and their impurities were all greater than 0.999, within the range of 0.5-50 μg/mL. The average recoveries of the impurities of three typical AGs were 99.03 %-101.22 %, RSDs all were less than 2.5 % for intra-day and inter-day precision, with good precision and accuracy. The developed HILIC-CAD quantification method was sensitive, accurate and highly selective for quantitative analysis of impurities in the AGs without need ion-pairing reagents, which is ensure the public medication safety. The method is first reported application of HILIC-CAD method for quantitative analysis of the impurities in AGs.
依替米星是一种典型的氨基糖苷类抗生素（AG）。高效液相色谱-蒸发光散射检测器（HPLC-ELSD）法是测定依替米星中杂质的常用方法。然而，由于 ELSD 的重现性差、灵敏度低、线性范围窄，目前高通量定量分析依替米星中的杂质仍然具有挑战性。在本研究中，开发了一种使用亲水相互作用液相色谱-带电气溶胶检测器（HILIC-CAD）的灵敏方法来分析依替米星中的杂质。使用 Box-Behnken 设计（BBD）和响应面法（RSM）优化了用于测定依替米星中杂质的液相条件，得到了令人满意的分离和最佳的 CAD 输出信号。我们还研究了 CAD 参数对依替米星及其杂质的信噪比和线性度的影响。该方法还被证明能够有效地分离另外两种典型的 AG（异帕米星和阿米卡星）中的杂质。在方法验证中，依替米星、异帕米星和阿米卡星及其杂质的决定系数（R）均大于 0.999，在 0.5-50 μg/mL 范围内。三种典型 AG 杂质的平均回收率为 99.03%-101.22%，日内和日间精密度的 RSD 均小于 2.5%，具有良好的精密度和准确性。所开发的 HILIC-CAD 定量方法灵敏、准确、选择性高，无需离子对试剂即可定量分析 AG 中的杂质，从而确保公众用药安全。该方法首次报道了 HILIC-CAD 方法用于定量分析 AG 中杂质的应用。
