反相高效液相色谱法检测重组类病毒颗粒疫苗原液中二硫苏糖醇的残留量

发布时间：2018-06-23 22:13

本文选题：反相高效液相色谱法 + 重组类病毒颗粒疫苗　；参考：《中国生物制品学杂志》2015年12期

【摘要】：目的反相高效液相色谱法(reverse phase-high performance liquid chromatography,RP-HPLC)检测重组类病毒颗粒(virus-like particles,VLPs)疫苗原液中二硫苏糖醇(DL-dithiothreitol,DTT)的残留量,并对方法进行验证及初步应用。方法按设定的色谱条件对检测重组VLPs疫苗原液中DTT含量的RP-HPLC法进行专属性、精密性、准确性验证,确定方法的最佳线性范围及最低检测限,用该方法检测3批重组HPV单价疫苗原液中的DTT残留量。结果该方法检测DTT时,不受溶液中溶剂和蛋白的干扰;线性范围为1.5~15.0μg/ml(R~2=0.997),最低检测限为1.5μg/ml;该方法检测不同浓度DTT的平均相对标准偏差(relative standard deviation,RSD)均5%;重组HPV疫苗原液分别加入3、5、10μg/ml的DTT对照品溶液后,经该方法检测的回收率在80%~120%之间。3批次重组HPV单价疫苗原液中的DTT含量均小于1.5μg/ml。结论本实验的RP-HPLC法专属性、精密性良好,且准确可靠,可用于重组HPV疫苗原液中DTT残留量的检测。
[Abstract]:Objective to determine the residue of DL-dithiothreitol (DL-dithiothreitol) in the original solution of recombinant viral particles (virus-like particlesus) vaccine by reverse phase-high performance liquid chromatography-RP-HPLC, and to verify the method and its preliminary application. Methods the RP-HPLC method for detecting DTT content in recombinant VLPs vaccine was determined by RP-HPLC according to the set chromatographic conditions. The specificity, accuracy and accuracy of the method were verified. The optimum linear range and the lowest detection limit of the method were determined. DTT residues in three batches of recombinant HPV monovalent vaccine were detected by this method. Results when DTT was detected by this method, it was not interfered by solvent and protein in solution. The linear range was 1.5 ~ 15.0 渭 g/ml (R ~ (2 +) 0.97) and the lowest detection limit was 1.5 渭 g 路ml ~ (-1). The recoveries of this method were between 80% and 120%. The DTT content in the original solution of recombinant HPV vaccine was less than 1.5 渭 g / ml. Conclusion the RP-HPLC method is specific, accurate and reliable, and can be used for the determination of DTT residues in recombinant HPV vaccine.
【作者单位】：北京生物制品研究所有限责任公司;
【基金】：国家科技部重大新药创制(2013ZX09402302)
【分类号】：O657.72;R392-33

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