determination of related substances in ex4c by uplc 超高效液相 .pdf

Advances in Analytical Chemistry 分析化学进展, 2017, 7(2), 92-101 Published Online May 2017 in Hans. /journal/aac /10.12677/aac.2017.72013 Determination of Related Substances in Ex4c by UPLC 1 1,2* 1 1 1 Xinru Yang , Guimin Zhang , Li Ma , Siguang Liu , Liangliang Zhao 1 Lunan Pharmaceutical Group Co., Ltd, Linyi Shandong 2 National Engineering and Technology Research Center of Chirality Pharmaceutical, Linyi Shandong th th th Received: Apr. 25 , 2017; accepted: May 14 , 2017; published: May 17 , 2017 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 µm) 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 µl 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 ma- terial. Conclusion: The validation results showed that the established metho