中国药物警戒 ›› 2025, Vol. 22 ›› Issue (10): 1104-1108.
DOI: 10.19803/j.1672-8629.20250492

• 基础与临床研究 • 上一篇    下一篇

荧光胶束法测定人乳头瘤病毒疫苗中聚山梨酯80含量方法的建立

路琼1, 王飞宇1,2△, 俞小娟1, 张杰3, 梁昊宇1, 黄维金1, 聂建辉1,*   

  1. 1中国食品药品检定研究院生物制品检定所,北京 102629;
    2沈阳药科大学制药工程学院,辽宁 沈阳 110016;
    3北京云菱生物技术有限公司,北京 100176
  • 收稿日期:2025-07-24 出版日期:2025-10-15 发布日期:2025-10-20
  • 通讯作者: *聂建辉,男,博士,研究员,生物制品检定。E-mail: niejianhui@nifdc.org.cn
  • 作者简介:路琼,女,博士,助理研究员,生物制品检定。△为并列第一作者。
  • 基金资助:
    国家重点研发计划（2025YFC2311705）

Establishment of a High-Performance Liquid Chromatography-Fluorescence Micelle Assay for Determination of Polysorbate 80 in Human Papillomavirus Vaccines

LU Qiong1, WANG Feiyu1,2△, YU Xiaojuan1, ZHANG Jie3, LIANG Haoyu1, HUANG Weijin1, NIE Jianhui1,*   

  1. 1Institute for Biological Products Control, National Institutes for Food and Drug Control, Beijing 100050, China;
    2School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang, Liaoning 110016, China;
    3Beijing Yunling Biotechnology Co., Ltd, Beijing 100176, China
  • Received:2025-07-24 Online:2025-10-15 Published:2025-10-20

PDF (PC)

56

摘要： 目的 考察高效液相色谱-荧光胶束法（HPLC-FMA）测定人乳头瘤病毒（HPV）疫苗中聚山梨酯80含量的适用性。方法 采用HPLC-FMA利用反应线圈SupelcoKnitted Reactor Coil（5 m×0.50 mm ID）进行测定;流动相为三羟甲基氨基甲烷混合溶液,流速1.5 mL·min-1;检测波长激发光350 nm、发射光420 nm,采集时间2.5 min,通过荧光强度值（峰面积）计算聚山梨酯80的含量。结果 该方法专属性良好;聚山梨酯80浓度在50~250 µg·mL-1内,线性关系良好,R2=0.999 8;仪器的精密度RSD为0.223%;供试品溶液中间精密度RSD为1.269%;6份供试品溶液重复性检测平均值为91.123µg·mL-1,RSD为0.399%。不同浓度回收率在98.725%~99.866%,RSD在0.321%~1.636%。8批供试品溶液检测值为96.500、95.850、94.940、96.049、91.847、92.147、102.060、84.667µg·mL-1。结论 HPLC-FMA无需借助有毒化学试剂和色谱柱且在几分钟内完成测定,可以快速、准确、可靠测定HPV疫苗中聚山梨酯80的含量。

关键词: HPV疫苗, 人乳头瘤病毒, 聚山梨酯80含量, 荧光胶束法, 高效液相色谱, N-苯基-1-萘胺, 《中国药典》

Abstract: Objective To evaluate the applicability of a high-performance liquid chromatography-fluorescence micelle assay (HPLC-FMA) for quantitative determination of polysorbate 80 in human papillomavirus (HPV) vaccines. Methods A SupelcoKnitted Reactor Coil (5 m×0.50 mm ID) was used as the reaction coil for HPLC-FMA analysis. The mobile phase was a tris (hydroxymethyl) aminomethane mixed solution at a flow rate of 1.5 mL·min-1. The detection wavelength was set at an excitation of 350 nm and an emission of 420 nm while the acquisition time was 2.5 minutes. The contents of polysorbate 80 were calculated using the fluorescence intensity (peak area). Results The method proved to be highly specific. Polysorbate 80 showed a good linear relationship at concentrations ranging from of 50 to 250 µg·mL-1, with an R2 value of 0.999 8. The instrument precision of RSD was 0.223%, and the intermediate precision of RSD was 1.269%. The average value from six replicate measurements was 91.123 µg·mL-1 with an RSD of 0.399%. Recovery rates at different concentrations ranged from 98.725% to 99.866%, with RSD values between 0.321% to 1.636%. The contents of polysorbate 80 in eight batches were determined as 96.500, 95.850, 94.940, 96.049, 91.847, 92.147, 102.060 and 84.667µg·mL-1, respectively. Conclusion This HPLC-FMA method requires no toxic chemicals or chromatographic columns, and can complete the assay within minutes, which can facilitate rapid, accurate, and reliable quantification of polysorbate 80 in HPV vaccines.

Key words: HPV Vaccine, Human Papillomavirus(HPV), Polysorbate 80 Content, Fluorescence Micelle Assay, High Performance Liquid Chromatography, N-Phenyl-1-Naphthylamine, Pharmacopoeia

中图分类号: