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Enzyme Immunoassay in Combination with Liquid Chromatography for Sensitive and Selective Determination of Drugs in Biosamples

Lövgren, Ulf LU (1997)
Abstract
Two different sensitive microtitre-plate based enzyme immunoassays were developed and evaluated for determination of the model drug compound, budesonide, in blood plasma. In the first approach, high sensitivity was obtained by using amperometric detection in a flow injection system for determination of the enzyme product p-aminophenol. The high sensitivity in the second approach was obtained using an enzyme amplified ELISA, based on the substrate recycling technique. Both immunoassays allowed for determination of the steroid, budesonide, in blood plasma samples at low picomolar concentrations, when combined with solid phase extraction and column liquid chromatography.



Flow injection enzyme immunoassays based on three... (More)
Two different sensitive microtitre-plate based enzyme immunoassays were developed and evaluated for determination of the model drug compound, budesonide, in blood plasma. In the first approach, high sensitivity was obtained by using amperometric detection in a flow injection system for determination of the enzyme product p-aminophenol. The high sensitivity in the second approach was obtained using an enzyme amplified ELISA, based on the substrate recycling technique. Both immunoassays allowed for determination of the steroid, budesonide, in blood plasma samples at low picomolar concentrations, when combined with solid phase extraction and column liquid chromatography.



Flow injection enzyme immunoassays based on three different formats; namely competitive with post-column reaction technique, non-competitive for haptens and the displacement format, were developed and evaluated for potential bioanalytical applications with emphasis on sensitivity, precision and throughput. Alkaline phosphatase was used as enzyme label and fluorometric or amperomtric detection was used for determination of the enzyme product.



Micellar liquid chromatography was evaluated as a biocompatible sample pretreatment procedure preceding immunoassay. Blood plasma samples were injected directly into a pre-column venting system where the analyte, cortisol, was separated from cross-reacting steroids in the sample. A biocompatible mobile phase, consisting of only buffer solution and the surfactant Tween 20, was used in the chromatographic system. This allowed for direct transfer of collected fractions to the immunoassay for quantification of the analyte. (Less)
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author
opponent
  • Professor Boos, Karl-Siegfred, Institut fur Klinische Chemie, Klinikum Grosshadern, Ludwig-Maximilians-Universität Munchen, D-81366 Munchen, Germany
organization
publishing date
type
Thesis
publication status
published
subject
keywords
Analytical chemistry, amplification, alkaline phosphatase, enzyme immunoassay, liquid chromatography, flow-injection, micellar, Analytisk kemi
pages
132 pages
publisher
Dep. of Bioanalytical Chemistry, Preclinical R&D, Astra Draco AB, P.O. Box 34, S-221 00 Lund Sweden,
defense location
Kemicentrum, K:C
defense date
1997-06-07 10:15
external identifiers
  • other:ISRN: LUNKDL/(NKAK-1037)/1-132 (1997)
language
English
LU publication?
yes
id
3ed0ee2e-2bdf-481c-a7d2-311d053c78d1 (old id 29375)
date added to LUP
2007-06-13 11:15:45
date last changed
2016-09-19 08:45:10
@phdthesis{3ed0ee2e-2bdf-481c-a7d2-311d053c78d1,
  abstract     = {Two different sensitive microtitre-plate based enzyme immunoassays were developed and evaluated for determination of the model drug compound, budesonide, in blood plasma. In the first approach, high sensitivity was obtained by using amperometric detection in a flow injection system for determination of the enzyme product p-aminophenol. The high sensitivity in the second approach was obtained using an enzyme amplified ELISA, based on the substrate recycling technique. Both immunoassays allowed for determination of the steroid, budesonide, in blood plasma samples at low picomolar concentrations, when combined with solid phase extraction and column liquid chromatography.<br/><br>
<br/><br>
Flow injection enzyme immunoassays based on three different formats; namely competitive with post-column reaction technique, non-competitive for haptens and the displacement format, were developed and evaluated for potential bioanalytical applications with emphasis on sensitivity, precision and throughput. Alkaline phosphatase was used as enzyme label and fluorometric or amperomtric detection was used for determination of the enzyme product.<br/><br>
<br/><br>
Micellar liquid chromatography was evaluated as a biocompatible sample pretreatment procedure preceding immunoassay. Blood plasma samples were injected directly into a pre-column venting system where the analyte, cortisol, was separated from cross-reacting steroids in the sample. A biocompatible mobile phase, consisting of only buffer solution and the surfactant Tween 20, was used in the chromatographic system. This allowed for direct transfer of collected fractions to the immunoassay for quantification of the analyte.},
  author       = {Lövgren, Ulf},
  keyword      = {Analytical chemistry,amplification,alkaline phosphatase,enzyme immunoassay,liquid chromatography,flow-injection,micellar,Analytisk kemi},
  language     = {eng},
  pages        = {132},
  publisher    = {Dep. of Bioanalytical Chemistry, Preclinical R&D, Astra Draco AB, P.O. Box 34, S-221 00 Lund Sweden,},
  school       = {Lund University},
  title        = {Enzyme Immunoassay in Combination with Liquid Chromatography for Sensitive and Selective Determination of Drugs in Biosamples},
  year         = {1997},
}