Xu, Xiaomi
(2009)
Miniaturization of ElectrochemicalDetection System for Biuret-Based Peptide Detection Following Capillary HPLC.
Master's Thesis, University of Pittsburgh.
(Unpublished)
Abstract
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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Details
| Item Type: |
University of Pittsburgh ETD
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| Status: |
Unpublished |
| Creators/Authors: |
|
| ETD Committee: |
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| 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: |
http://etd.library.pitt.edu/ETD/available/etd-07112008-143941/, etd-07112008-143941 |
| Date Deposited: |
10 Nov 2011 19:50 |
| Last Modified: |
15 Nov 2016 13:45 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/8340 |
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