高效液相色谱蒸发光散射检测法测定聚乙二醇干扰素α-2a注射液中游离聚乙二醇的含量

Quantitation of Free Polyethylene Glycol in PEGylated Interferon Alpha-2a Injection by HPLC-ELSD

建立聚乙二醇干扰素α-2a注射液中游离聚乙二醇含量的HPLC测定方法。以XBridge BEH 300 C4（4.6 mm×15.0 cm,3.5μm）色谱柱为固定相,以水-乙腈为流动相,梯度洗脱,流速1.0 mL/min,蒸发光散射检测器检测。游离聚乙二醇与聚乙二醇干扰素α-2a及其它辅料成分得到很好分离,聚乙二醇在20~100μg/mL范围内,进样量的对数值与峰面积的对数值之间有良好的线性关系（r=0.999）,检测限和定量限分别为0.3μg和0.5μg,平均回收率为102.2%（RSD=2.1%）。结果表明,所建立的含量测定方法简便、快速、准确,可用于聚乙二醇干扰素α-2a注射液中游离聚乙二醇的含量测定。

A liquid chromatographic method was developed to quantitatively determine the free polyethylene glycol （PEG） in PEGy- lated interferon alpha-2a injection. This method separated and quantified free PEG upon an XBridge BEH 300 CA column （4.6 wm×15.0 cm,3.5 mm） using gradient elution of water-acetonitrile at the flow rate of 1.0 mL/min with an evaporative light scattering detector （ELSD）. Drift tube temperature of the ELSD system was set to 90℃ and flow rate of nebulizing gas was 2.0 L/min. Free PEG was baseline separated from PEGylated interferon alpha-2a and other excipient ingredients. The logarithm of peak area from ELSD was linearly correlated to the logarithm of the injected quantity of PEG at the range of 0.4 - 2μg （ r =0. 999）. The limit of detection （LOD） and limit of quantitation （LOQ） of the method were 0.3 μg and 0.5 i.Lg,respectively. The average recovery of PEG was 102.2% with RSD of 2.1% （ n = 6）. The results show that the established method is simple, sensitive and accurate, and suggest applicability in the determination of free PEG in PEGylated interferon alpha-2a injection.