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Importance of pre-analytical factors contributing to measurement uncertainty, when determining sulfadoxine and sulfamethoxazole from capillary blood dried on sampling paper
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Physical and Analytical Chemistry, Analytical Chemistry.
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Physical and Analytical Chemistry, Analytical Chemistry.
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Physical and Analytical Chemistry, Analytical Chemistry.
2008 (English)In: Journal of Chromatographic Science, ISSN 0021-9665, Vol. 16, no 10, 837-843 p.Article in journal (Refereed) Published
Abstract [en]

A bioanalytical method is developed and validated for determination of sulfadoxine (SD) and sulfamethoxazole (SM) in 100 microL capillary blood dried on sampling paper (Whatman 31ET Chr). SD and SM are extracted with 2000 microL perchloric acid and the liquid phase is loaded onto ENV+ solid-phase extraction columns. SD, SM, and the internal standard are separated on a Purospher STAR RP-18 liquid chromatography column (150 x 4.6 mm) with a mobile phase consisting of acetonitrile-sodium acetate buffer pH 5.2, I = 0.1 (33:67, v/v). Analytes are detected with UV at 256 nm. Lower limit of quantitation is 5 micromol/L, where precisions are 4.2% and 3.9% for SD and SM, respectively. Three brands of sampling papers have been compared with respect to absorption properties, extraction recoveries, and variations. Punching out dried blood spots (DBS) instead of cutting spots into strips prior to extraction has been evaluated by examining precision and accuracy of SD and SM determinations. Importance of uniformity of types of sampling paper, sampling volume and biological matrix, benefit of punching out discs from DBS, and impact on absorption properties of different brands of sampling papers are discussed. Avoiding pre-analytical errors whenever possible results in concentrations determined being more accurate and precise.

Place, publisher, year, edition, pages
2008. Vol. 16, no 10, 837-843 p.
National Category
Chemical Sciences
Identifiers
URN: urn:nbn:se:uu:diva-96866ISI: 000260912400001PubMedID: 19007488OAI: oai:DiVA.org:uu-96866DiVA: diva2:171590
Available from: 2008-03-19 Created: 2008-03-19 Last updated: 2009-07-14Bibliographically 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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