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DOI | 10.1016/j.chroma.2008.11.017 |
Quantitation of polysorbate 20 in protein solutions using mixed-mode chromatography and evaporative light scattering detection | |
Hewitt, Daniel; Zhang, Taylor; Kao, Yung-Hsiang | |
通讯作者 | Zhang, Taylor |
来源期刊 | JOURNAL OF CHROMATOGRAPHY A
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ISSN | 0021-9673 |
EISSN | 1873-3778 |
出版年 | 2008 |
卷号 | 1215期号:1-2页码:156-160 |
英文摘要 | An HPLC assay requiring no complex sample preparation for the measurement of polysorbate 20 in protein solutions was developed. An on-off chromatography technique was employed involving a mixed-mode stationary phase (Waters Oasis MAX, mixed-mode anion-exchange and reversed-phase sorbent) to quantify polysorbate 20 in solutions containing >100 mg/mL of protein. With 2% formic acid mobile phase, proteins are typically positive charged and are not retained because of electrostatic repulsions from the quaternary amine in the mixed-mode resin. Other formulation components elute in void volume because of their hydrophilicity. Hydrophobic polysorbate 20 is retained, eluted with a step gradient and quantified as a single peak using an evaporative light scattering detector. The performance of the assay is evaluated according to International Conference on Harmonisation (ICH) guidelines and shown to be suitable for polysorbate quantitation. Accuracy (96-108%) and repeatability (2.3% RSD) were demonstrated using protein samples spiked with polysorbate 20. This method was used to accurately measure polysorbate 20 in at least 25 different protein solutions spanning a wide range of formulations. Although the majority of the data reported here target polysorbate 20, this methodology can also be used to assay other common non-ionic surfactants such as polysorbate 80, Brij, Igepal, and Triton X-100. (C) 2008 Elsevier B.V. All rights reserved. |
英文关键词 | Polysorbate Mixed-mode chromatography Evaporative light scattering detection Protein Sample preparation |
类型 | Article |
语种 | 英语 |
国家 | USA |
收录类别 | SCI-E |
WOS记录号 | WOS:000261845500020 |
WOS关键词 | PERFORMANCE LIQUID-CHROMATOGRAPHY ; FORMULATIONS |
WOS类目 | Biochemical Research Methods ; Chemistry, Analytical |
WOS研究方向 | Biochemistry & Molecular Biology ; Chemistry |
资源类型 | 期刊论文 |
条目标识符 | http://119.78.100.177/qdio/handle/2XILL650/158062 |
作者单位 | Genentech Inc, Prot Analyt Chem, San Francisco, CA 94080 USA |
推荐引用方式 GB/T 7714 | Hewitt, Daniel,Zhang, Taylor,Kao, Yung-Hsiang. Quantitation of polysorbate 20 in protein solutions using mixed-mode chromatography and evaporative light scattering detection[J],2008,1215(1-2):156-160. |
APA | Hewitt, Daniel,Zhang, Taylor,&Kao, Yung-Hsiang.(2008).Quantitation of polysorbate 20 in protein solutions using mixed-mode chromatography and evaporative light scattering detection.JOURNAL OF CHROMATOGRAPHY A,1215(1-2),156-160. |
MLA | Hewitt, Daniel,et al."Quantitation of polysorbate 20 in protein solutions using mixed-mode chromatography and evaporative light scattering detection".JOURNAL OF CHROMATOGRAPHY A 1215.1-2(2008):156-160. |
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