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Simultaneous quantification of the highly lipophilic atovaquone and the hydrophilic strong basic proguanil and its metabolites, using a new mixed-mode SPE approach and Steep-Gradient LC
Dalarna University, School of Technology and Business Studies, Chemical Engineering.
Dalarna University, School of Technology and Business Studies, Chemical Engineering.
2005 (English)In: Journal of Chromatographic Science, ISSN 0021-9665, E-ISSN 1945-239X, Vol. 43, no 5, p. 259-266Article in journal (Refereed) Published
Abstract [en]

A bioanalytical method is described for the simultaneous quantitative analysis of the highly lipophilic atovaquone and the strong basic proguanil with metabolites in plasma. The drugs are extracted from protein precipitated plasma samples on a novel mixed-mode solid-phase extraction (SPE) column containing carboxypropyl and octyl silica as functional groups. The analytes are further separated and quantitated using a steep-gradient liquid chromatograhic method on a Zorbax SB-CN column with UV detection at 245 nm. Two different internal standards (IS) are used in the method to compensate for both types of analytes. A structurally similar IS to atovaquone is added with acetonitrile to precipitate proteins from plasma. A structurally similar IS to proguanil and its metabolites is added with phosphate buffer before samples are loaded onto the SPE columns. A single elution step is sufficient to elute all analytes. The method is validated according to published guidelines and shows excellent performance. The within-day precisions, expressed as relative standard deviation, are lower than 5% for all analytes at three tested concentrations within the calibration range. The between-day precisions are lower than 13% for all analytes at the same tested concentrations. The limit of quantitation is 25nM for the basic substances and 50nM for atovaquone. Several considerations regarding development and optimization of a method for determination of analytes with such a difference in physiochemical properties are discussed.

Place, publisher, year, edition, pages
2005. Vol. 43, no 5, p. 259-266
National Category
Chemical Engineering
Identifiers
URN: urn:nbn:se:du-1203DOI: 10.1093/chromsci/43.5.259ISI: 000229224600007PubMedID: 15975245OAI: oai:dalea.du.se:1203DiVA, id: diva2:519533
Available from: 2005-06-01 Created: 2005-06-01 Last updated: 2017-12-07Bibliographically approved

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CiteExportLink to record
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Citation style
  • apa
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