Determination of Related Substances in Ex4c by UPLC
Abstract: Objective: To establish a method for the determination of related substances in Ex4c (the raw ma-terial of the pegylation of glucagon-like peptide-1) by UPLC. Methods: The UPLC method was de-veloped by using a Waters ACQUITY UPLC® Peptide CSH™C18 column (130 Å, 2.1 × 150 mm, 1.7 mm) with a gradient elution system. The mobile phase A was 0.05% trifluoroacetic acid aqueous solution/tetrahydrofuran (9:1), and the mobile phase B was 0.05% trifluoroacetic acid in acetoni-trile/tetrahydrofuran (9:1). The column temperature was maintained at 45˚C while the detection wavelength setting at 214 nm. Furthermore, the sample injection was 2 ml via keeping a flow rate of 0.3 mL∙min−1. The developed method was validated. Results: The resolution between Ex4c and adjacent impurities, between Ex4C and the known impurities were greater than 1.5. All four known impurities exhibited good linear relationships with their related coefficients ≥ 0.995. And their detection limits ranged from 4.0 ng to 5.6 ng, comprising 96.3%~101.1% of recovered material. Conclusion: The validation results showed that the established method was simple, accurate and reproducible. It is capable of determinating impurities in Ex4c (the raw material of the pegylation of glucagon-like peptide-1) and can be effectively used for quality control of this drug.
文章引用: 杨欣茹 , 张贵民 , 马 丽 , 刘思光 , 赵亮亮 (2017) 超高效液相色谱法测定Ex4c的有关物质。 分析化学进展， 7， 92-101. doi: 10.12677/AAC.2017.72013
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