药学学报, 2021, 56(8): 2276-2281
引用本文:
俞小娟#, 于传飞#, 张荣建, 武刚, 崔永霏, 郭璐韵, 王兰*. HPLC-FMA法测定单抗药物中聚山梨酯80含量的方法学联合验证[J]. 药学学报, 2021, 56(8): 2276-2281.
YU Xiao-juan#, YU Chuan-fei#, ZHANG Rong-jian, WU Gang, CUI Yong-fei, GUO Lu-yun, WANG Lan*. Interlaboratory method validation of HPLC-FMA for determination of polysorbate 80 in monoclonal antibodies[J]. Acta Pharmaceutica Sinica, 2021, 56(8): 2276-2281.

HPLC-FMA法测定单抗药物中聚山梨酯80含量的方法学联合验证
俞小娟#, 于传飞#, 张荣建, 武刚, 崔永霏, 郭璐韵, 王兰*
中国食品药品检定研究院, 北京 102629
摘要:
对HPLC-FMA (high performance liquid chromatography-fluorescence micelle assay)法测定单抗药物中聚山梨酯80含量的方法开展实验室间的联合验证,以研究该方法在多个实验室间的可应用性和可转移性以及纳入现行版药典的可行性。本文使用两个厂家(J.T.Baker公司和南京威尔)来源的聚山梨酯80在7个实验室开展验证。结果显示,在蛋白浓度≤ 20 mg·mL-1,聚山梨酯80含量在0.05~0.5 mg·mL-1内时,该方法专属性良好;加标样品的准确性分别为92.20%~117.70%(J.T.Baker公司),93.90%~117.20%(南京威尔);组内精密度分别为RSD < 4.30%(J.T.Baker公司),RSD < 2.60%(南京威尔);总体精密度分别为RSD < 5.45%(J.T.Baker公司),RSD < 6.70%(南京威尔);线性相关系数分别为r > 0.98(J.T.Baker公司),r > 0.99(南京威尔)。联合验证结果证明,HPLC-FMA法准确性、精密度、线性和专属性均良好,可应用于不同实验室单抗药物中聚山梨酯80含量的放行检测及稳定性分析。
关键词:    聚山梨酯80      HPLC-FMA      方法学验证      单克隆抗体     
Interlaboratory method validation of HPLC-FMA for determination of polysorbate 80 in monoclonal antibodies
YU Xiao-juan#, YU Chuan-fei#, ZHANG Rong-jian, WU Gang, CUI Yong-fei, GUO Lu-yun, WANG Lan*
National Institutes for Food and Drug Control, Beijing 102629, China
Abstract:
The high performance liquid chromatography-fluorescence micelle assay (HPLC-FMA) method for the content determination of polysorbate 80 in monoclonal antibody drugs was validated to study its applicability and transferability between various laboratories, and the feasibility to be included in the Chinese Pharmacopoeia. Both J.T. Baker and Nanjing Well-sourced polysorbate 80 was used in the collaborative validation of polysorbate 80 content analysis in seven different laboratories. The results show that when the protein concentration was no more than 20 mg·mL-1 and the concentration of polysorbate 80 ranged from 0.05 to 0.5 mg·mL-1, the method had good specificity. The recovery rates of the spiked samples ranged from 92.20% to 117.70% for J.T.Baker and from 93.90% to 117.20% for Nanjing Well. The intra-laboratory precision (%RSD) was less than 4.30% for J.T. Baker, and less than 2.60% for Nanjing Well, while the overall precision was less than 5.45% for J.T. Baker, and less than 6.70% for Nanjing Well. The linear correlation coefficient was more than 0.98 for J.T. Baker and more than 0.99 for Nanjing Well. The results of the collaborative validation prove that the HPLC-FMA method has good accuracy, precision, linearity, and specificity, and could be used for release control analysis of polysorbate 80 content in monoclonal antibodies across different laboratories.
Key words:    polysorbate 80    HPLC-FMA    method validation    monoclonal antibody   
收稿日期: 2021-01-28
DOI: 10.16438/j.0513-4870.2021-0151
基金项目: 国家药典委员会药品标准制修订研究课题(2018S003);国家重点研发计划课题(2020YFC0860700).
通讯作者: 王兰,E-mail:wanglan@nifdc.org.cn
Email: wanglan@nifdc.org.cn
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参考文献:
[1] Dwivedi M, Blech M, Presser I, et al. Polysorbate degradation in biotherapeutic formulations identification and discussion of current root causes[J]. Int J Pharm, 2018, 552:422-436.
[2] Webster GK, Chang JC, Heflin JL. Stability indicating method for polysorbate 80 in protein formulations[J]. J Chromatogr Sci, 2020, 116:1-8.
[3] Vaclaw C, Merritt K, Pringle V, et al. Impact of polysorbate 80 grade on the interfacial properties and interfacial stress induced subvisible particle formation in monoclonal antibodies[J]. J Chromatogr Sci, 2021, 110:746-759.
[4] Kishore RS, Kiese S, Fischer S, et al. The degradation of polysorbates 20 and 80 and its potential impact on the stability of biotherapeutics[J]. Pharm Res, 2011, 28:1194-1210.
[5] Glucklich N, Dwivedi M, Carle S, et al. An in-depth examination of fatty acid solubility limits in biotherapeutic protein formulations containing polysorbate 20 and polysorbate 80[J]. Int J Pharm, 2020, 591:119934.
[6] Larson NR, Wei Y, Prajapati I, et al. Comparison of polysorbate 80 hydrolysis and oxidation on the aggregation of a monoclonal antibody[J]. J Pharm Sci, 2020, 109:633-639.
[7] Kim J, Qiu J. Quantitation of low concentrations of polysorbates in high protein concentration formulations by solid phase extraction and cobalt-thiocyanate derivatization[J]. Anal Chim Acta, 2014, 806:144-151.
[8] Cheng Y, Liu Y, Wei H, et al. Quantitation of low concentrations of polysorbates 80 in protein formulations by Coomassie brilliant blue[J]. Anal Biochem, 2019, 573:67-72.
[9] Shende N, Karale A, Bhagade S, et al. Evaluation of a sensitive GC-MS method to detect polysorbate 80 in vaccine preparation[J]. J Pharm Biomed, 2020, 183:113126.
[10] Adamo M, Dick LW, Qiu D, et al. A simple reversed phase highperformance liquid chromatography method for polysorbate 80 quantitation in monoclonal antibody drug products[J]. J Chromatogr B Analyt Technol Biomed Life Sci, 2010, 878:1865-1870.
[11] Schilling K, Pawellek R, Lovejoy K, et al. Influence of charged aerosol detector instrument settings on the ultra-high-performance liquid chromatography analysis of fatty acids in polysorbate 80[J]. J Chromatogr A, 2018, 1576:58-66.
[12] Hewitt D, Alvarez M, Robinson K, et al. Mixed-mode and reversed-phase liquid chromatography-tandem mass spectrometry methodologies to study composition and base hydrolysis of polysorbate 20 and 80[J]. J Chromatogr A, 2011, 1218:2138-2145.
[13] Degterev MB, Badaukayte RA, Shukurov RR. A development of high-throughput HPLC method for the polysorbate 80 quantitation in protein therapeutic products[J]. J Chromatogr B Analyt Technol Biomed Life Sci, 2019, 1133:121847.
[14] Nair LM, Stephens NV, Vincent S, et al. Determination of polysorbate 80 in parenteral formulations by high-performance liquid chromatography and evaporative light scattering detection[J]. J Chromatogr A, 2003, 1012:81-86.
[15] Hewitt D, Zhang T, Kao YH. Quantitation of polysorbate 20 in protein solutions using mixed-mode chromatography and evaporative light scattering detection[J]. J Chromatogr A, 2008, 1215:156-160.
[16] Mondal B, Kote M, Lunagariya C, et al. Development of a simple high performance liquid chromatography (HPLC) /evaporative light scattering detector (ELSD) method to determine polysorbate 80 in a pharmaceutical formulation[J]. Saudi Pharm J, 2020, 28:325-328.
[17] Fekete S, Ganzler K, Fekete J. Fast and sensitive determination of polysorbate 80 in solutions containing proteins[J]. J Pharm Biomed, 2010, 52:672-679.
[18] ICH Q2(R1):Validation of Analytical Procedures:Text and Methodology[M/OL]. 2005-11[2021-03-24]. https://database.ich.org/sites/default/files/Q2_R1__Guideline.pdf.