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Development of a rapid UHPLC method for the in-process determination of coupling reagents used in peptide synthesis

Barry, Deborah (2013) Development of a rapid UHPLC method for the in-process determination of coupling reagents used in peptide synthesis. PhD thesis, Dublin City University.

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Abstract

The following thesis details the extensive development of a rapid liquid chromatography method for the in-process determination of peptide coupling reagents used in peptide synthesis. The determination of peptide coupling reagents, additives and associated by-products is important during peptide synthesis to ensure the concentration of these products are below the threshold of toxicological concern in the final peptide. A number of column technologies and mobile phases were evaluated for the separation of the analytes using an ultra-high performance liquid chromatography system and, a 15 minute method for the simultaneous determination of fourteen peptide coupling reagents, additives and by-products was established. This method was determined to be selective and capable of accurately quantitating the amount of TMU, HOBt, HCTU, HBTU, 6-ChloroHOBt, TBTU, Oxyma Pure, COMU, DIU, DIC, PyBOP, and TCTU in the presence of two peptides from Ipsen Manufacturing Ireland LTD. Analytical method validation was performed as per ICH guidelines for specificity, accuracy, linearity, precision, detection limit, quantitation limit and robustness. This rapid UHPLC method is directly transferable onto LC-MS and the intended application of this method is for evaluation of development peptides in Ipsen Manufacturing Ireland LTD.

Item Type:Thesis (PhD)
Date of Award:November 2013
Refereed:No
Supervisor(s):Heise, Andreas
Uncontrolled Keywords:rapid liquid chromatography method; peptide coupling reagents; peptide synthesis
Subjects:Physical Sciences > Analytical chemistry
DCU Faculties and Centres:DCU Faculties and Schools > Faculty of Science and Health > School of Chemical Sciences
Research Initiatives and Centres > Irish Separation Science Cluster (ISSC)
Use License:This item is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 3.0 License. View License
Funders:Ipsen Manufacturing Ireland Ldt
ID Code:19367
Deposited On:25 Nov 2013 14:56 by Andreas Heise. Last Modified 16 Sep 2017 01:02

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