Objective. To establish a method for extraction and determination of antioxidant 264 (2,6-di-tert-butyl-4-methylphenol) in the brominated butyl rubber stopper for injection and migration study in recombinant potent antitumor and antivirus protein injection (Novaferon). Methods. Dichloromethane-ethanol was adopted as the extraction solvent during the process of reflux extraction of antioxidant 264 in the brominated butyl rubber stopper. High-performance liquid chromatography (HPLC) was used for the determination of the migration of antioxidant 264 to Novaferon. The mobile phase consisted of acetonitrile-water (80 : 20, v/v). The flow rate was 1.5 mL/min. The detection wavelength was 280 nm. Results. The linearity range was from 4.003 to 200.150 μg/mL (r² = 0.99996), and the average recovery of antioxidant 264 was 97.8%. The applicability of the methodology was good, which can be used for the determination of antioxidant 264. The results indicated that antioxidant 264 was not detected in Novaferon after the accelerated test and three months of long-term test. Conclusion. The established validated method in this study can be used for the determination of antioxidant 264 in the rubber stopper, and the brominated butyl rubber stopper has good compatibility with Novaferon.

中文翻译：

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丁基橡胶塞中抗氧剂264的测定及与重组强效抗肿瘤抗病毒蛋白注射液的配伍性。

客观。建立注射用溴化丁基橡胶塞中抗氧化剂264（2,6-二叔丁基-4-甲基苯酚）的提取测定方法及重组强效抗肿瘤抗病毒蛋白注射液（Novaferon）的迁移研究。方法。溴化丁基橡胶塞中抗氧剂264的回流萃取过程采用二氯甲烷-乙醇作为萃取溶剂。高效液相色谱法 (HPLC) 用于测定抗氧化剂 264 向 Novaferon 的迁移。流动相由乙腈-水 (80: 20, v/v) 组成。流速为 1.5 mL/min。检测波长为 280 nm。结果。线性范围为 4.003 至 200.150  μg/mL（r ²  = 0.99996），抗氧剂264的平均回收率为97.8%。该方法适用性好，可用于抗氧化剂264的测定。加速试验和3个月的长期试验结果表明，诺华龙中未检出抗氧化剂264。结论。本研究建立的经过验证的方法可用于胶塞中抗氧剂264的测定，溴化丁基胶塞与诺华龙具有良好的相容性。