title: Isocratic rapid liquid chromatographic method for simultaneous determination of carotenoids, retinol, and tocopherols in human serum.
abstract: An improved isocratic and rapid HPLC method was developed for the measurement of carotenoids, retinol and tocopherols in human serum. Vitamins were extracted with hexane. Mobile phase consisted of a mixture acetonitrile:methylene chloride:methanol with 20 mM ammonium acetate. This method used a small bead size (3 microm) Spherisorb ODS2 column with titane frits. Diode array and fluorescence detectors were used respectively for the detection of carotenoids and retinol/tocopherols. Chromatographic separation was complete in 13 min for beta-cryptoxanthin, cis-trans-lycopene, alpha-carotene, beta-carotene, cis-beta-carotene, retinol, delta-tocopherol, gamma-tocopherol and alpha-tocopherol. Echinenone and tocol were employed as internal standards for diode array and fluorescence detection. Accuracy was validated using standard reference material (SRM) 968C. Intra-assay and inter-assay precision were respectively 0.2-7.3% and 3.6-12.6%. Sensitivity was verified using the ICH recommendations and the limit of detection (LOD) obtained was sufficient for routine clinical application.
authors.affiliation: Department of Diagnostic Medicine, Sir Mortimer B. Davis Jewish General Hospital, Montreal, Quebec, Canada.
authors.forename: Denis
authors.initials: D
authors.lastname: Thibeault
authors.forename: Haixiang
authors.initials: H
authors.lastname: Su
authors.forename: Elizabeth
authors.initials: E
authors.lastname: MacNamara
authors.forename: Hyman M
authors.initials: HM
authors.lastname: Schipper
mesh_terms: D002138:Calibration; D002338:Carotenoids / Q000097:blood*; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D012015:Reference Standards; D013050:Spectrometry, Fluorescence; D013232:Steel / Q000737:chemistry; D024505:Tocopherols / Q000097:blood*; D014801:Vitamin A / Q000097:blood*; D014815:Vitamins / Q000097:blood*
chemical_list: D014815:Vitamins; D014801:Vitamin A; D013232:Steel; D002338:Carotenoids; D024505:Tocopherols