title: Fast, simple, and sensitive high-performance liquid chromatography method for measuring vitamins A and E in human blood plasma.
abstract: Vitamins A and E are fat-soluble vitamins that play important roles in several physiological processes. Monitoring their concentrations is needed to detect deficiency and guide therapy. In this study, we developed a high-performance liquid chromatography method to measure the major forms of vitamin A (retinol) and vitamin E (α-tocopherol and γ-tocopherol) in human blood plasma. Vitamins A and E were extracted with hexane and separated on a reversed-phase column using methanol as the mobile phase. Retinol was detected by ultraviolet absorption, whereas tocopherols were detected by fluorescence emission. The chromatographic cycle time was 4.0 min per sample. The analytical measurement range was 0.03-5.14, 0.32-36.02, and 0.10-9.99 mg/L for retinol, α-tocopherol, and γ-tocopherol, respectively. Intr-aassay and total coefficient of variation were <6.0% for all compounds. This method was traceable to standard reference materials offered by the National Institute of Standards and Technology. Reference intervals were established using plasma samples collected from 51 healthy adult donors and were found to be 0.30-1.20, 6.0-23.0, and 0.3-3.2 mg/L for retinol, α-tocopherol, and γ-tocopherol, respectively. In conclusion, we developed and validated a fast, simple, and sensitive high-performance liquid chromatography method for measuring the major forms of vitamins A and E in human plasma.
authors.affiliation: Department of Clinical Pathology, Cleveland Clinic, Cleveland, OH, USA.
authors.forename: Chao
authors.initials: C
authors.lastname: Yuan
authors.forename: Maria
authors.initials: M
authors.lastname: Burgyan
authors.forename: Dustin R
authors.initials: DR
authors.lastname: Bunch
authors.forename: Edmunds
authors.initials: E
authors.lastname: Reineks
authors.forename: Raymond
authors.initials: R
authors.lastname: Jackson
authors.forename: Roxanne
authors.initials: R
authors.lastname: Steinle
authors.forename: Sihe
authors.initials: S
authors.lastname: Wang
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D012680:Sensitivity and Specificity; D014801:Vitamin A / Q000097:blood*; D014810:Vitamin E / Q000097:blood*
chemical_list: D014801:Vitamin A; D014810:Vitamin E
keywords: Blood plasma; Fluorescence detection; High-performance liquid chromatography; Vitamins; ultraviolet detection