聚乙二醇化重组人干扰素β-1a注射液中游离聚乙二醇残留量的测定

Determination of Free PEG in PEG-rhIFN-β-1a Injection

关键词:  PEG-rhIFN-β-1a注射液  游离聚乙二醇  蒸发光检测器

Key words:  PEG-rhIFN-β-1a injection  free PEG  evaporation light detector

摘要:

目的 对聚乙二醇化重组人干扰素β-1a(PEG-rhIFN-β-1a)注射液中的残留聚乙二醇丁醛(PEG)进行方法学研究。方法 采用Phenomenex Jupiter C₄色谱柱，流动相：A相，0.1%三氟乙酸-水溶液；B相，0.1%三氟乙酸-乙腈溶液，梯度洗脱；流速：1.0 mL·min^?1，柱温为25 ℃，蒸发光检测器，漂移管温度为113.0 ℃，载气流速为3.1 L·min^?1。结果 PEG在0.6~2.0 μg·mL^?1内线性良好，相关系数为0.998 4，检出限为0.20 μg，定量限为0.40 μg，精密度、稳定性和重复性试验的RSD均<2.0%，平均回收率为96%~100%。结论 运用该方法对PEG-rhIFN-β-1a注射液中游离PEG残留量测定，残留量<1.0%。

Abstract:

OBJECTIVE To develop a high performance liquid chromatography method for determination of free PEG in PEG-rhIFN-β-1a injection. METHODS Phenomenex Jupiter C₄ column was used with a mobile phase of gradient elution (A phase-0.1% trifluoroacetic acid-water; B phase: 0.1% trifluoroacetic acid-acetonitrile), gradient elution. The flow rate was 1.0 mL·min^?1, and column temperature was 25 ℃. Evaporation light detector(ELSD), drift tube temperature was 113.0 ℃, and carrier gas flow rate of 3.1 L·min^?1. RESULTS PEG concentration in the range of 0.6-2.0 μg·mL^?1 showed good linearity, correlation coefficient was 0.998 4, detection limit was 0.20 μg , quantitation limit was 0.40 μg, the RSD of precision, stability and reproducibility were all <2.0%. The average recovery was from 96% to 100%. CONCLUSION Using this method, the free PEG in PEG-rhIFN-β-1a injection is determined by this method and the free PEG is <1.0%.