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Capillary HPLC Separation of Selected Neuropeptides

Yi, Ying (2012) Capillary HPLC Separation of Selected Neuropeptides. Master's Thesis, University of Pittsburgh. (Unpublished)

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Abstract

Neuropeptides play a pivotal role in brain and peripheral nervous system function. As high performance liquid chromatography (HPLC) becomes the central tool in the separation and characterization of peptide and protein samples, its selectivity optimization has attracted increasing attention. This research program aims to develop useful, quantitative analysis methods for neuropeptides and their hydrolysis fragments by capillary HPLC. Related peptide pairs are successfully separated, such as leu-enkephalin and [Des-Tyr1] leu-enkephalin, dynorphin A and dynorphin B, galanin and its fragment Gal1-16. The hydrolysis of leu-enkephalin to [Des-Tyr1] leu-enkephalin by organotypic hippocampal slice cultures (OHSCs) can be monitored by the same HPLC system. The separation of seven hippocampal neuropeptides with similar hydrophobicity, Bj-PRO-5a, [Des-Tyr1] leu-enkephalin, leu-enkephalin, pentagastrin, Antho-RW-amide I, dynorphin A 1-6 and angiotensin II, is accomplished by thermally tuned tandem capillary columns (T3C). The chromatographic selectivity is continuously, systematically and significantly optimized by individual adjustment of each column’s temperature. The T3C concept is applied for the first time with capillary columns, which is an important step towards optimization of selectivity for separations of small samples by liquid chromatography.


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Details

Item Type: University of Pittsburgh ETD
Status: Unpublished
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Yi, Yingyiy25@pitt.eduYIY25
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee ChairWeber, Stephen G.sweber@pitt.eduSWEBER
Committee MemberMichael, Adrian C. amichael@pitt.eduAMICHAEL
Committee MemberRobinson, Renã A. S. rena@pitt.eduRENA
Date: 8 June 2012
Date Type: Publication
Defense Date: 3 October 2011
Approval Date: 8 June 2012
Submission Date: 18 April 2012
Access Restriction: No restriction; Release the ETD for access worldwide immediately.
Number of Pages: 55
Institution: University of Pittsburgh
Schools and Programs: Dietrich School of Arts and Sciences > Chemistry
Degree: MS - Master of Science
Thesis Type: Master's Thesis
Refereed: Yes
Uncontrolled Keywords: Capillary HPLC, peptide separation, thermally tuned tandem capillary columns
Date Deposited: 08 Jun 2012 19:11
Last Modified: 15 Nov 2016 13:57
URI: http://d-scholarship.pitt.edu/id/eprint/11902

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