A new, simple and rapid method for the quantitative determination of the antimicrobial preservative 2-phenoxyethanol, based on reverse phase ultra-high-performance liquid chromatography has been developed. The validation was performed according the ICH Q2 guideline “Validation of Analytical Procedures”. The desired chromatographic separation was achieved on a Waters Symmetry C18 (150 × 4.6 mm, 5 μm) column using an isocratic elution, with detection at 270 nm wavelength. The mobile phase consisted of acetonitrile/water (55:45, v/v), pumped at a flow rate of 1 mL/min. The calibration curve and the analytical procedure are linear (r² = 0.999) from the concentration of 0.07 mg/mL to 1.1 mg/mL. The percent relative standard deviation for intra- and inter-day precision was <1%. The recovery of 2-phenoxyethanol in vaccines ranged between 96.5 and 100.60%. The limits of detection and quantitation were 1.3 × 10⁻⁴ and 2.7 × 10⁻⁴ mg/mL, respectively. The method was found to be robust by changing the column working temperature, the percentage of acetonitrile of the mobile phase and the flow rate. The validated method can be successfully and reliably used to quantify as well as to exclude presence of 2-phenoxyethanol preservative in marketed vaccines.

建立了一种基于反相超高效液相色谱定量测定抗菌防腐剂 2-苯氧基乙醇的新方法，该方法简单快捷。验证是根据 ICH Q2 指南“分析程序验证”进行的。在 Waters Symmetry C18 (150 × 4.6 mm, 5 μm) 色谱柱上使用等度洗脱实现了所需的色谱分离，检测波长为 270 nm。流动相由乙腈/水 (55:45, v/v) 组成，以 1 mL/min 的流速泵入。校准曲线和分析程序呈线性 ( r ² = 0.999) 从 0.07 mg/mL 的浓度到 1.1 mg/mL。日内和日间精密度的百分比相对标准偏差 <1%。疫苗中 2-苯氧乙醇的回收率介于 96.5% 和 100.60% 之间。检测限和定量限分别为1.3 × 10 ^-4和2.7 × 10 ^-4  mg/mL。通过改变色谱柱工作温度、流动相中乙腈的百分比和流速，发现该方法是可靠的。经验证的方法可以成功可靠地用于量化和排除市售疫苗中 2-苯氧乙醇防腐剂的存在。