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Miniaturization of ElectrochemicalDetection System for Biuret-Based Peptide Detection Following Capillary HPLC

Xu, Xiaomi (2009) Miniaturization of ElectrochemicalDetection System for Biuret-Based Peptide Detection Following Capillary HPLC. Master's Thesis, University of Pittsburgh. (Unpublished)

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Miniaturization of liquid chromatography, capillary HPLC, has made possible the separation of samples with volumes in the low microliter to nanoliter range. Downsizing separation column requires more sensitive detection method. Biuret-based electrochemistry has proven to be effective in peptide detection, but work needs to be done to make it feasible in capillary separation system.This thesis presents modifications of a standard electrochemical detector with dual working electrodes, which decreases bandspreading into the nL range suitable for capillary HPLC. This miniaturized detector has detection limit of 3 nM (6 fmol) for peptides on the upstream anode. The downstream cathode has high selectivity by applying reductive potential. The common carbon working electrode can be replaced with gold electrodes with a self-assembled naphthalenethiol monolayer, which suppresses the background current from Au while giving detectable current from Cu-peptide redox reaction. When the modified Au electrode is assembled to a flow cell for peptide detection, it gives stable current response and the sensitivity is comparable to a glassy carbon electrode.These advances will permit non-electrochemists to determine peptides by capillary HPLC quantitatively, and open the way for chip-based detectors.


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Item Type: University of Pittsburgh ETD
Status: Unpublished
CreatorsEmailPitt UsernameORCID
Xu, Xiaomixix11@pitt.eduXIX11
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee ChairWeber, Stephen Gsweber@pitt.eduSWEBER
Committee MemberMichael, Adrianamichael@pitt.eduAMICHAEL
Committee MemberAmemiya, Shigeruamemiya@pitt.eduAMEMIYA
Date: 22 January 2009
Date Type: Completion
Defense Date: 19 November 2006
Approval Date: 22 January 2009
Submission Date: 11 July 2008
Access Restriction: 5 year -- Restrict access to University of Pittsburgh for a period of 5 years.
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; electrochemical detection; miniaturization
Other ID:, etd-07112008-143941
Date Deposited: 10 Nov 2011 19:50
Last Modified: 15 Nov 2016 13:45


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