RP-HPLC-ELSD法测定康柏西普眼用注射液中辅料吐温20的含量

目的： 建立反相高效液相色谱法联用蒸发光散射检测器测定康柏西普眼用注射液中辅料吐温20含量的分析方法。 方法： 采用Agilent Poroshell 120 SB-C₁₈（2.7 μm，4.6 mm×150 mm）色谱柱，以乙腈-纯水为流动相进行梯度洗脱（0~5 min，5%B；5~6 min，5%B→60%B；6~10 min，60%B→80%B；10~15 min，80%B；15~15.01 min，80%B→5%B；15.01~20 min，5%B），流速0.5 mL·min^-1，蒸发光散射检测器漂移管温度为45 ℃，雾化压力设为2.8×10⁵ Pa。 结果： 吐温20进样量在1.12~6.72 μg范围内，与色谱峰面积呈良好线性关系（R²>0.999）；高、中、低浓度样品的回收率均在100%±10%范围内。 结论： 本法经方法学验证可用于康柏西普眼用注射液中辅料吐温20含量的检测。

Objective: To develop an RP-HPLC-ELSD method for the determination of Tween 20 in Conbercept ophthalmic injection. Methods: The analysis was carried out on the Agilent Poroshell 120 SB-C₁₈(2.7 μm,4.6 mm× 150 mm)column.The mobile phase was composed of acetonitrile and water in gradient elution(0-5 min,5%B;5-6 min,5%B→60%B;6-10 min,60%B→80%B; 10-15 min,80%B;15-15.01 min,80%B→5%B;15.01- 20 min,5%B).And the flow rate was 0.5 mL·min^-1.The ELSD was maintained at 45 ℃ throughout the determination,and the nebulizer(nitrogen)gas pressure was set at 2.8×10⁵ Pa. Results: The linear ranges of Tween 20 were 1.12-6.72 ug,and the sample size had a good linear relationship with the chromatographic peak area(R²>0.999).The recoveries were all in the range of 100%±10% at high,medium and low concentrations. Conclusion: The methodology validation showed that the method can be used for determination of Tween 20 in Conbercept ophthalmic injection.

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