重组人干扰素β1b中ox-IFN_β1b含量测定

目的：建立重组人干扰素β1b（IFN β1b）中氧化型干扰素（ox-IFN β1b）的测定方法。方法：RP-HPLC色谱条件：RP-C₈色谱柱（Grace Vydac，4.6 mm×250 mm，5μm），柱温40℃，0.1% TFA-10%乙腈-水溶液为流动相A，0.1% TFA-乙腈溶液为流动相B，梯度洗脱，流速为1.0 mL·min^-1，检测波长214 nm。结果：ox-IFNβ1b峰与峰3的分离度约为4.0，理论塔板数大于20 000；6次进样的RSD小于5.0%；2个厂家共6批IFN β1b原液中均检出ox-IFN β1b，用面积归一化法计算百分含量为0.47%~0.96%。结论：新建方法可有效分离和测定不同厂家IFN β1b原液中的ox-IFN β1b，对IFNβ1b质量标准的提高具有重要意义。

Objective: To develop a method for determination of ox-IFN β1b in recombinant human interferon β1b(IFN β1b). Methods: The RP-HPLC was carried out on an RP-C₈ column (Grace Vydac,4.6 mm×250 mm,5μm),at the temperature of 40. The mobile phase consisting of solution A (0.1% TFA-10% ACN-ddH₂O) and solution B (0.1% TFA-ACN as solution B) was applied with a gradient elution at a flow rate of 1.0 mL·min^-1. The detection wavelength was 214 nm. Results: The resolution of the ox-IFNβ1b peak relative to peak 3 was about 4.0 and theoretical plate number was more than 20 000. RSD of 6 injections was below 5.0%. ox-IFNβ1b was detected in 6 batches of IFN β1b bulks from two different manufacturers,and percentage contents were between 0.47% and 0.96%,which were determined by area normalization. Conclusion: The developed method could effectively separate and determine ox-IFNβ1b in IFN β1b bulks from different manufacturers,which was of great significance to the improvement of quality control of IFNβ1b.