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Title: Optimization of sensitivity and separation in capillary zone electrophoresis with indirect fluorescence detection

Abstract

Laser-induced indirect fluorescence detection can be used a general detector in capillary zone electrophoresis. Indirect fluorescence detection, where a fluorophore is used as the principal component of the electrophoretic buffer, allows the visualization of nonfluorescing ions through charge displacement of the fluorophore. Stabilization of laser power improves the dynamic reserve to 10/sup 3/ without the complication of double-beam correction techniques. A modification of the CZE power supply allows the use of dilute solutions and narrow bore capillary tubing (< 15 ..mu..m i.d.). The stability of the background fluorescence is improved by silylation of the fused silica capillary. These modifications improve the detection limits for many anions, nucleotides, and proteins to the range of 50-100 amol of sample injected. Additionally, this improvement in sensitivity results in a concomitant increase in separation efficiency. The electrophoretic separation can be optimized by calculating effective mobilities of ions based on their dissociation as a function of pH. More than 300,000 theoretical plates are observed in a separation of the nucleotide 5'-monophosphates optimized in this manner.

Authors:
;
Publication Date:
Research Org.:
Iowa State Univ., Ames (USA)
OSTI Identifier:
6357575
DOE Contract Number:  
W-7405-ENG-82
Resource Type:
Journal Article
Journal Name:
Anal. Chem.; (United States)
Additional Journal Information:
Journal Volume: 60:23
Country of Publication:
United States
Language:
English
Subject:
37 INORGANIC, ORGANIC, PHYSICAL AND ANALYTICAL CHEMISTRY; NUCLEOTIDES; ELECTROPHORESIS; CAPILLARIES; EXPERIMENTAL DATA; FLUORESCENCE SPECTROSCOPY; OPTIMIZATION; PH VALUE; QUANTITATIVE CHEMICAL ANALYSIS; BLOOD VESSELS; BODY; CARDIOVASCULAR SYSTEM; CHEMICAL ANALYSIS; DATA; EMISSION SPECTROSCOPY; INFORMATION; NUMERICAL DATA; ORGANIC COMPOUNDS; ORGANS; SPECTROSCOPY; 400105* - Separation Procedures; 400102 - Chemical & Spectral Procedures

Citation Formats

Kuhr, W G, and Yeung, E S. Optimization of sensitivity and separation in capillary zone electrophoresis with indirect fluorescence detection. United States: N. p., 1988. Web. doi:10.1021/ac00174a021.
Kuhr, W G, & Yeung, E S. Optimization of sensitivity and separation in capillary zone electrophoresis with indirect fluorescence detection. United States. https://doi.org/10.1021/ac00174a021
Kuhr, W G, and Yeung, E S. Thu . "Optimization of sensitivity and separation in capillary zone electrophoresis with indirect fluorescence detection". United States. https://doi.org/10.1021/ac00174a021.
@article{osti_6357575,
title = {Optimization of sensitivity and separation in capillary zone electrophoresis with indirect fluorescence detection},
author = {Kuhr, W G and Yeung, E S},
abstractNote = {Laser-induced indirect fluorescence detection can be used a general detector in capillary zone electrophoresis. Indirect fluorescence detection, where a fluorophore is used as the principal component of the electrophoretic buffer, allows the visualization of nonfluorescing ions through charge displacement of the fluorophore. Stabilization of laser power improves the dynamic reserve to 10/sup 3/ without the complication of double-beam correction techniques. A modification of the CZE power supply allows the use of dilute solutions and narrow bore capillary tubing (< 15 ..mu..m i.d.). The stability of the background fluorescence is improved by silylation of the fused silica capillary. These modifications improve the detection limits for many anions, nucleotides, and proteins to the range of 50-100 amol of sample injected. Additionally, this improvement in sensitivity results in a concomitant increase in separation efficiency. The electrophoretic separation can be optimized by calculating effective mobilities of ions based on their dissociation as a function of pH. More than 300,000 theoretical plates are observed in a separation of the nucleotide 5'-monophosphates optimized in this manner.},
doi = {10.1021/ac00174a021},
url = {https://www.osti.gov/biblio/6357575}, journal = {Anal. Chem.; (United States)},
number = ,
volume = 60:23,
place = {United States},
year = {1988},
month = {12}
}