title: Application of dried blood spot cards to determine olive oil phenols (hydroxytyrosol metabolites) in human blood.
abstract: In this study, a fast and simple blood sampling and sample pre-treatment method based on the use of the dried blood spot (DBS) cards and ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) for the quantification of olive oil phenolic metabolites in human blood was developed and validated. After validation, the method was applied to determine hydroxytyrosol metabolites in human blood samples after the acute intake of an olive oil phenolic extract. Using the FTA DMPK-A DBS card under optimum conditions, with 20µL as the blood solution volume, 100µL of methanol/Milli-Q water (50/50, v/v) as the extraction solvent and 7 disks punched out from the card, the main hydroxytyrosol metabolites (hydroxytyrosol-3-O-sulphate and hydroxytyrosol acetate sulphate) were identified and quantified. The developed methodology allowed detecting and quantifying the generated metabolites at low μM levels. The proposed method is a significant improvement over existing methods to determine phenolic metabolites circulating in blood and plasma samples, thus making blood sampling possible with the volunteer pricking their own finger, and the subsequent storage of the blood in the DBS cards prior to chromatographic analysis.
authors.affiliation: Food Technology Department, Agrotecnio Center, Escola Tècnica Superior d'Enginyeria Agrària, University of Lleida, Avda. Alcalde Rovira Roure 191, 25198 Lleida, Spain.
authors.forename: María Carmen López
authors.initials: MCL
authors.lastname: de Las Hazas
authors.affiliation: Food Technology Department, Agrotecnio Center, Escola Tècnica Superior d'Enginyeria Agrària, University of Lleida, Avda. Alcalde Rovira Roure 191, 25198 Lleida, Spain.
authors.forename: Maria José
authors.initials: MJ
authors.lastname: Motilva
authors.affiliation: Department of Medicine, Facultat de Medicina, University of Lleida, Avda/Alcalde Rovira Roure 80, 25198 Lleida, Spain.
authors.forename: Carme
authors.initials: C
authors.lastname: Piñol
authors.affiliation: Food Technology Department, Agrotecnio Center, Escola Tècnica Superior d'Enginyeria Agrària, University of Lleida, Avda. Alcalde Rovira Roure 191, 25198 Lleida, Spain. Electronic address: albamacia@tecal.udl.cat.
authors.forename: Alba
authors.initials: A
authors.lastname: Macià
mesh_terms: D001774:Blood Chemical Analysis / Q000295:instrumentation* / Q000379:methods*; D002853:Chromatography, Liquid; D059788:Dried Blood Spot Testing; D006801:Humans; D000069463:Olive Oil / Q000737:chemistry*; D010636:Phenols / Q000097:blood*; D010626:Phenylethyl Alcohol / Q000031:analogs & derivatives / Q000097:blood / Q000378:metabolism; D053719:Tandem Mass Spectrometry
chemical_list: D000069463:Olive Oil; D010636:Phenols; C005975:3,4-dihydroxyphenylethanol; D010626:Phenylethyl Alcohol
keywords: Blood; Dried blood spot cards; Hydroxytyrosol; Olive oil extract; UPLC-MS/MS