title: A high-throughput method for liquid chromatography-tandem mass spectrometry determination of plasma alkylresorcinols, biomarkers of whole grain wheat and rye intake.
abstract: Plasma alkylresorcinols are increasingly analyzed in cohort studies to improve estimates of whole grain intake and their relationship with disease incidence. Current methods require large volumes of solvent (>10 ml/sample) and have relatively low daily sample throughput. We tested five different supported extraction methods for extracting alkylresorcinols from plasma and improved a normal-phase liquid chromatography coupled to a tandem mass spectrometer method to reduce sample analysis time. The method was validated and compared with gas chromatography-mass spectrometry analysis. Sample preparation with HybridSPE supported extraction was most effective for alkylresorcinol extraction, with recoveries of 77-82% from 100 μl of plasma. The use of 96-well plates allowed extraction of 160 samples per day. Using a 5-cm NH2 column and heptane reduced run times to 3 min. The new method had a limit of detection and limit of quantification equivalent to 1.1-1.8 nmol/L and 3.5-6.1 nmol/L plasma, respectively, for the different alkylresorcinol homologues. Accuracy was 93-105%, and intra- and inter-batch precision values were 4-18% across different plasma concentrations. This method makes it possible to quantify plasma alkylresorcinols in 100 μl of plasma at a rate of at least 160 samples per day without the need for large volumes of organic solvents.
authors.affiliation: Division of Food and Nutrition Science, Department of Biology and Biological Engineering, Chalmers University of Technology, 412 96 Gothenburg, Sweden. Electronic address: alastair.ross@chalmers.se.
authors.forename: Alastair B
authors.initials: AB
authors.lastname: Ross
authors.affiliation: Division of Food and Nutrition Science, Department of Biology and Biological Engineering, Chalmers University of Technology, 412 96 Gothenburg, Sweden.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Svelander
authors.affiliation: Division of Food and Nutrition Science, Department of Biology and Biological Engineering, Chalmers University of Technology, 412 96 Gothenburg, Sweden.
authors.forename: Otto I
authors.initials: OI
authors.lastname: Savolainen
authors.affiliation: Division of Food and Nutrition Science, Department of Biology and Biological Engineering, Chalmers University of Technology, 412 96 Gothenburg, Sweden; Department of Nutrition, Exercise, and Sports, Faculty of Science, University of Copenhagen, 1870 Frederiksberg, Denmark.
authors.forename: Mads Vendelbo
authors.initials: MV
authors.lastname: Lind
authors.affiliation: Department of Pathobiology, Lerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USA.
authors.forename: John P
authors.initials: JP
authors.lastname: Kirwan
authors.affiliation: Nestlé Research Centre, Vers-chez-les-Blanc, 1000 Lausanne 26, Switzerland.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Breton
authors.affiliation: Nestlé Research Centre, Vers-chez-les-Blanc, 1000 Lausanne 26, Switzerland.
authors.forename: Jean-Philippe
authors.initials: JP
authors.lastname: Godin
authors.affiliation: Division of Food and Nutrition Science, Department of Biology and Biological Engineering, Chalmers University of Technology, 412 96 Gothenburg, Sweden.
authors.forename: Ann-Sofie
authors.initials: AS
authors.lastname: Sandberg
mesh_terms: D015415:Biomarkers / Q000097:blood; D002851:Chromatography, High Pressure Liquid; D004435:Eating; D057166:High-Throughput Screening Assays; D006801:Humans; D012118:Resorcinols / Q000097:blood*; D012434:Secale / Q000737:chemistry*; D053719:Tandem Mass Spectrometry; D000067348:Whole Grains / Q000737:chemistry*
chemical_list: D015415:Biomarkers; D012118:Resorcinols
keywords: Alkylresorcinols; Biomarkers; Blood plasma; Liquid chromatography–tandem mass spectrometry; Whole grain