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Title: A new post-column reactor-laser induced fluorescence detector for capillary electrophoresis

Abstract

Capillary zone electrophoresis (CZE), a powerful separation method based on the differential migration of charged species under the influence of an electric field, has been widely used for separations covering from small ions to big biomolecules. Chapter 1 describes the method, then discusses detection of the separated analytes by laser induced fluorescence and by chemical derivatization, and the use of O-phthaldialdehyde (OPA) as a post-column reagent. Chapter 2 describes a post-column reactor which uses two narrow bore capillaries connected coaxially. This reactor differs from other coaxial reactors in terms of capillary dimensions, reagent flow control, ease of construction and most importantly, better limits of detection. The derivatization reagent is electroosmotically driven into the reaction capillary and the reagent flow rate is independently controlled by a high voltage power supply. Amino acids, amines and proteins, derivatized by OPA/2-mercaptoethanol using this post-column reactor coupled with LIF detection, show low attomole mass limits of detection, and for the first time, the authors demonstrate single cell capability with a post-column derivatization scheme. The single cell capability shows that this reactor could find applications in assaying non-fluorescent or electrochemically inactive components in individual biological cells in the future.

Authors:
 [1]
  1. Iowa State Univ., Ames, IA (United States)
Publication Date:
Research Org.:
Ames Lab., IA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
206510
Report Number(s):
IS-T-1770
ON: DE96006809; TRN: AHC29607%%27
DOE Contract Number:  
W-7405-ENG-82
Resource Type:
Thesis/Dissertation
Resource Relation:
Other Information: TH: Thesis (M.S.); PBD: 2 Jan 1996
Country of Publication:
United States
Language:
English
Subject:
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; 59 BASIC BIOLOGICAL SCIENCES; AMINES; ELECTROPHORESIS; AMINO ACIDS; PROTEINS; PEPTIDES; FLUORESCENCE; LASER RADIATION; CHEMICAL REACTORS; DERIVATIZATION; ACCURACY; LIMITING VALUES

Citation Formats

Liling, Zhang. A new post-column reactor-laser induced fluorescence detector for capillary electrophoresis. United States: N. p., 1996. Web. doi:10.2172/206510.
Liling, Zhang. A new post-column reactor-laser induced fluorescence detector for capillary electrophoresis. United States. doi:10.2172/206510.
Liling, Zhang. Tue . "A new post-column reactor-laser induced fluorescence detector for capillary electrophoresis". United States. doi:10.2172/206510. https://www.osti.gov/servlets/purl/206510.
@article{osti_206510,
title = {A new post-column reactor-laser induced fluorescence detector for capillary electrophoresis},
author = {Liling, Zhang},
abstractNote = {Capillary zone electrophoresis (CZE), a powerful separation method based on the differential migration of charged species under the influence of an electric field, has been widely used for separations covering from small ions to big biomolecules. Chapter 1 describes the method, then discusses detection of the separated analytes by laser induced fluorescence and by chemical derivatization, and the use of O-phthaldialdehyde (OPA) as a post-column reagent. Chapter 2 describes a post-column reactor which uses two narrow bore capillaries connected coaxially. This reactor differs from other coaxial reactors in terms of capillary dimensions, reagent flow control, ease of construction and most importantly, better limits of detection. The derivatization reagent is electroosmotically driven into the reaction capillary and the reagent flow rate is independently controlled by a high voltage power supply. Amino acids, amines and proteins, derivatized by OPA/2-mercaptoethanol using this post-column reactor coupled with LIF detection, show low attomole mass limits of detection, and for the first time, the authors demonstrate single cell capability with a post-column derivatization scheme. The single cell capability shows that this reactor could find applications in assaying non-fluorescent or electrochemically inactive components in individual biological cells in the future.},
doi = {10.2172/206510},
journal = {},
number = ,
volume = ,
place = {United States},
year = {1996},
month = {1}
}

Thesis/Dissertation:
Other availability
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