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Determination of eflornithine enantiomers in plasma, by solid-phase extraction and liquid chromatography with evaporative light-scattering detection
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Physical and Analytical Chemistry.
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Physical and Analytical Chemistry.
2007 (English)In: Journal of chromatography. B, ISSN 1570-0232, E-ISSN 1873-376X, Vol. 846, no 1-2, 98-104 p.Article in journal (Refereed) Published
Abstract [en]

A bioanalytical method for determination of eflornithine (DFMO) in 1000 μL human plasma has been developed and validated. DFMO and the internal standard (IS) were analysed by liquid chromatography with evaporative light-scattering detection (ELSD). Separation was performed on a Chirobiotic TAG (250 mm × 4.6 mm) column with ethanol (99.5%):0.01 mol/L acetic acid-triethylamine buffer at the rate of 25:75% (v/v) with flow rate of 1.0 mL/min. For d-DFMO in plasma the inter-assay precision was 6.5% at 75 μmol/L, 6.6% at 375 μmol/L and 5.8% at 750 μmol/L. For l-DFMO in plasma the inter-assay precision was 10.4% at 75 μmol/L, 6.5% at 375 μmol/L and 5.0% at 750 μmol/L. The lower limit of quantification (LLOQ) was determined to 25 μmol/L where the precision was 4.3% and 5.7%, respectively.

Place, publisher, year, edition, pages
2007. Vol. 846, no 1-2, 98-104 p.
Keyword [en]
Eflornithine, DFMO, 2-Fluoromethyl-dl-ornithine, Chiral chromatography, Chirobiotic TAG, Human African Trypanosomiasis, Evaporative light-scattering detection, ELSD, HPLC, Chirality
National Category
Chemical Sciences
Identifiers
URN: urn:nbn:se:uu:diva-96863DOI: 10.1016/j.jchromb.2006.08.030ISI: 000244281100013OAI: oai:DiVA.org:uu-96863DiVA: diva2:171587
Available from: 2008-03-19 Created: 2008-03-19 Last updated: 2011-02-15Bibliographically approved
In thesis
1. Drug Analysis: Bioanalytical Method Development and Validation
Open this publication in new window or tab >>Drug Analysis: Bioanalytical Method Development and Validation
2008 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

This thesis describes bioanalytical methods for drug determination in biological matrixes, with drugs in focus used against diseases largely affecting low-income countries.

Solid-phase extraction is used for sample cleanup, and processed samples are analyzed by liquid chromatography. Developed bioanalytical methods are validated according to international guidelines.

Eflornithine (DFMO) is a chiral drug, used for treating human African trypanosomiasis. A bioanalytical method for determination of DFMO enantiomers in plasma is presented. The enantiomers are detected by evaporative light-scattering detection. The method has been applied to determination of D-DFMO and L-DFMO in rats, after intravenous and oral administration of racemic DFMO. It is concluded that DFMO exhibits enantioselective absorption, with the more potent enantiomer L-DFMO being less favored.

Sulfadoxine (SD) and sulfamethoxazole (SM) are sulfa-drugs used for malaria and pneumonia respectively. Two methods are described for simultaneous determination of SD and SM in capillary blood sampled on filter paper. The former method allows direct injection of extracts from dried blood spots (DBS), while for the latter method solid-phase extraction is added. Pre-analytical factors contributing to measurement uncertainty is also discussed, and it is concluded that it is of high importance that homogeneity in type of sampling paper and sampling volume is assured.

Piperaquine (PQ) is an antimalarial, increasingly used in artemisinin combination therapy. A method for determination of piperaquine in DBS is presented. By using a monolithic LC column, a very short LC analysis of two minutes per sample is achieved.

A method for simultaneous determination of three antiretroviral drugs i.e. lamivudine (3TC), zidovudine (AZT) and nevirapine (NVP), in DBS samples is described. The method is applied to drug determination in two subjects after receiving standard antiretroviral treatment. Conclusion is that the method is suitable for determination of 3TC and NVP, and to some extent for AZT.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2008. 59 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 406
Keyword
Analytical chemistry, African trypanosomiasis, calibration model, chiral chromatography, dried blood spots, eflornithine, evaporative light-scattering detection, HIV/AIDS, lamivudine, liquid chromatography, malaria, nevirapine, piperaquine, pneumonia, solid-phase extraction, sulfadoxine, sulfamethoxazole, validation, zidovudine, Analytisk kemi
Identifiers
urn:nbn:se:uu:diva-8547 (URN)978-91-554-7125-5 (ISBN)
Public defence
2008-04-18, Clas Ohlson room, Tenoren, Högskolan Dalarna, Borlänge, 13:00 (English)
Opponent
Supervisors
Available from: 2008-03-19 Created: 2008-03-19 Last updated: 2009-03-24Bibliographically approved

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