title: Optimized, Fast-Throughput UHPLC-DAD Based Method for Carotenoid Quantification in Spinach, Serum, Chylomicrons, and Feces.
abstract: An improved UHPLC-DAD-based method was developed and validated for quantification of major carotenoids present in spinach, serum, chylomicrons, and feces. Separation was achieved with gradient elution within 12.5 min for six dietary carotenoids and the internal standard, echinenone. The proposed method provides, for all standard components, resolution > 1.1, linearity covering the target range (R > 0.99), LOQ < 0.035 mg/L, and intraday and interday RSDs < 2 and 10%, respectively. Suitability of the method was tested on biological matrices. Method precision (RSD%) for carotenoid quantification in serum, chylomicrons, and feces was below 10% for intra- and interday analysis, except for lycopene. Method accuracy was consistent with mean recoveries ranging from 78.8 to 96.9% and from 57.2 to 96.9% for all carotenoids, except for lycopene, in serum and feces, respectively. Additionally, an interlaboratory validation study on spinach at two institutions showed no significant differences in lutein or β-carotene content, when evaluated on four occasions.
authors.affiliation: Agroscope, Competence Division Plants and Plant Products , Schloss 1, CH-8820 Wädenswil, Switzerland.|Department of Nutrition, Exercise and Sports, University of Copenhagen , Rolighedsvej 30, DK-1958 Frederiksberg C, Denmark.
authors.forename: Jane N
authors.identifier: http://orcid.org/0000-0002-3016-7480
authors.initials: JN
authors.lastname: Eriksen
authors.affiliation: Department of Nutrition, Exercise and Sports, University of Copenhagen , Rolighedsvej 30, DK-1958 Frederiksberg C, Denmark.
authors.forename: Pia L
authors.initials: PL
authors.lastname: Madsen
authors.affiliation: Department of Nutrition, Exercise and Sports, University of Copenhagen , Rolighedsvej 30, DK-1958 Frederiksberg C, Denmark.
authors.forename: Lars O
authors.initials: LO
authors.lastname: Dragsted
authors.affiliation: Agroscope, Competence Division Plants and Plant Products , Schloss 1, CH-8820 Wädenswil, Switzerland.
authors.forename: Eva
authors.initials: E
authors.lastname: Arrigoni
mesh_terms: D002338:Carotenoids / Q000032:analysis* / Q000097:blood; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002914:Chylomicrons / Q000737:chemistry*; D005243:Feces / Q000737:chemistry*; D006801:Humans; D018724:Spinacia oleracea / Q000737:chemistry*
chemical_list: D002914:Chylomicrons; D002338:Carotenoids
keywords: UHPLC-DAD; green leafy vegetables; human samples; lutein; β-carotene