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Determination of melatonin in human plasma with solid-phase extraction, high-performance liquid chromatography and fluorescence detection
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry.
2003 (English)In: Scandinavian Journal of Clinical and Laboratory Investigation, ISSN 0036-5513, E-ISSN 1502-7686, Vol. 63, no 1, 81-88 p.Article in journal (Refereed) Published
Abstract [en]

A new bioanalytical method for the determination of melatonin in plasma with high-performance liquid chromatography (HPLC) and fluorescence detection preceded by solid-phase extraction has been developed and validated. Melatonin was extracted from 3 mL plasma using a Waters Oasis HLB solid-phase extraction cartridge and the elute was evaporated to dryness and dissolved in 200 microl mobile phase; acetonitrile-phosphate buffer, 0.01 M pH 7.2 (25:75, v/v). 125 microL was injected into the HPLC system and separation was carried out on a Waters SymmetryShield RP18 column 5 microm (250 x 4.6 mm). Excitation and emission wavelengths were set to 285 nm and 345 nm, respectively. The HPLC system was able to separate melatonin and internal standard (5-fluorotryptamine) from other endogenous indole compounds such as serotonin and tryptophan. Determination down to 0.10 nmol/L was possible, with an intra-assay precision of about 13%. Melatonin was stable in plasma for at least 30 days at about 23 degrees C.

Place, publisher, year, edition, pages
2003. Vol. 63, no 1, 81-88 p.
National Category
Analytical Chemistry
Identifiers
URN: urn:nbn:se:uu:diva-48687PubMedID: 12729073OAI: oai:DiVA.org:uu-48687DiVA: diva2:76593
Available from: 2008-10-17 Created: 2008-10-17 Last updated: 2017-12-05
In thesis
1. Development and Validation of Bioanalytical Methods: Application to Melatonin and Selected Anti-Infective Drugs
Open this publication in new window or tab >>Development and Validation of Bioanalytical Methods: Application to Melatonin and Selected Anti-Infective Drugs
2010 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

This thesis describes bioanalytical methods for measuring melatonin and some anti-infective drugs in biological fluids. Solid-phase extraction (SPE) or protein precipitation was used for enrichment and purification of the analytes and Liquid Chromatography (LC) was used to analyze the samples. Developed methods were validated according to international guidelines.

Melatonin is a hormone secreted by the pineal gland with a robust circadian rhythm. Bioanalytical methods for determination of melatonin in plasma and saliva have been developed which were used for monitoring melatonin levels in volunteers and patients suffering from sleep related diseases.

Eflornithine (DFMO) is a chiral drug used for the treatment of human African trypanosomiasis. A bioanalytical method for determination of the DFMO enantiomers in plasma, after precolumn derivatization with o-phtalaldehyde and N-acetyl-L-cystein has been developed. The method has been used to study the L- and D-DFMO pharmacokinetics, in order to investigate the possible development of an oral treatment of DFMO.

A method for simultaneous determination of three antiretroviral drugs i.e. Lamivudine (3TC), Zidovudine (AZT) and Nevirapine (NVP) in dried blood spots (DBS) was developed. The method was used for drug determination in two subjects after receiving standard antiretroviral treatment. The method seemed well suitable for the determination of 3TC and NVP and in some extent for AZT.

Lumefantrine (LF) is one of the active components in a new fixed drug combination recommended by the WHO as a replacement to older drugs that has lost their effect. A method for the determination of LF in DBS was developed. The method is suitable for monitoring of drug treatment in rural settings.

Tafenoquine is a new promising antimalarial drug under development. A method for the determination of Tafenoquine in plasma and in DBS is described. The method may be useful in future clinical studies in laboratory environment as well as in rural settings.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2010. 53 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 773
Keyword
african trypanosomiasis, melatonin, malaria, antiretroviral drugs, lumefantrine, tafenoquine, lamivudine, nevirapine, zidovudine, sampling paper, dried blood spots, capillary blood, antimalarial drugs, solid-phase extraction, liquid chromatography
National Category
Chemical Sciences
Research subject
Analytical Chemistry
Identifiers
urn:nbn:se:uu:diva-131519 (URN)978-91-554-7908-4 (ISBN)
Public defence
2010-11-16, Clas Ohlson room, Tenoren, Högskolan Dalarna, Borlänge, 13:00 (Swedish)
Opponent
Supervisors
Note
Felaktigt tryckt som Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology 703Available from: 2010-10-26 Created: 2010-10-04 Last updated: 2011-03-21Bibliographically approved

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