title: A fast and sensitive method for quantifying lumefantrine and desbutyl-lumefantrine using LC-MS/MS.
abstract: A sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed for quantification of lumefantrine (LUM) and its metabolite desbutyl-lumefantrine (DBL) in human plasma. Sample preparation was done by protein precipitation using acetonitrile containing deuterated lumefantrine (LUM-d18) and desbutyl-lumefantrine (DBL-d9) as internal standards. Total chromatography time was 2.2min using an Hypersil Gold C18 column (20×2.1mm, 1.9μm), with a gradient using 0.5% formic acid in water (mobile phase A) and 0.5% formic acid in methanol (mobile phase B) at a flow rate of 0.5mL/min. The mass spectrometric quantification was performed in positive electro spray ionization (ESI+) mode using selected reaction monitoring (SRM). Measuring range was 21-529ng/mL for LUM and 1.9-47ng/mL for DBL in plasma. Inter- and intra-assay precision was within 10% coefficient of variation (CV) for all levels of both LUM and DBL. Accuracy was within ±10% for all levels of both LUM and DBL. This method requires 100μL plasma volume and its short retention times allow a high throughput. Samples were stable for a week at +5°C, and up to six months -20°C. The method was successfully applied for plasma LUM and DBL determination in children under 5 years of age with uncomplicated malaria, up to 28 days after a standard 3-day treatment with artemether-lumefantrine.
authors.affiliation: Division of Clinical Pharmacology, Department of Laboratory Medicine, Karolinska Institutet Huddinge, Sweden.
authors.forename: A V
authors.initials: AV
authors.lastname: Silva
authors.affiliation: Division of Clinical Pharmacology, Department of Laboratory Medicine, Karolinska Institutet Huddinge, Sweden; Department of Pharmacology and Therapeutics, Makerere University, 7062 Kampala, Uganda.
authors.forename: N
authors.initials: N
authors.lastname: Mwebaza
authors.affiliation: Department of Pharmacology and Therapeutics, Makerere University, 7062 Kampala, Uganda.
authors.forename: M
authors.initials: M
authors.lastname: Ntale
authors.affiliation: Division of Clinical Pharmacology, Department of Laboratory Medicine, Karolinska Institutet Huddinge, Sweden.
authors.forename: L L
authors.initials: LL
authors.lastname: Gustafsson
authors.affiliation: Division of Clinical Pharmacology, Department of Laboratory Medicine, Karolinska Institutet Huddinge, Sweden. Electronic address: anton.pohanka@karolinska.se.
authors.forename: A
authors.initials: A
authors.lastname: Pohanka
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods*; D004983:Ethanolamines / Q000097:blood*; D005449:Fluorenes / Q000097:blood*; D006801:Humans; D057230:Limit of Detection; D000078102:Lumefantrine; D012015:Reference Standards; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D004983:Ethanolamines; D005449:Fluorenes; C535063:desbutyllumefantrine; D000078102:Lumefantrine
keywords: Desbutyl-lumefantrine; Liquid chromatography; Lumefantrine; Malaria; Plasma; Plasmodium falciparum; Sensivity; Tandem mass spectrometry