Simultaneous measurement of retinol, α-tocopherol and six carotenoids in human plasma by using an isocratic reversed-phase HPLC method

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 867; no. 2; pp. 226 - 232
• Main Authors: Karppi, Jouni, Nurmi, Tarja, Olmedilla-Alonso, Begoña, Granado-Lorencio, Fernando, Nyyssönen, Kristiina
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 15.05.2008; Elsevier Science
• Subjects: alpha-Tocopherol - blood; Analysis; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Carotenoids; Carotenoids - blood; Chromatography, High Pressure Liquid - methods; Freezing; Fundamental and applied biological sciences. Psychology; General pharmacology; HPLC; Humans; Medical sciences; Pharmacology. Drug treatments; Quality Control; Reference Standards; Reproducibility of Results; Retinol; Sample preparation; Stability of carotenoids; Vitamin A - blood; α-Tocopherol; Carotenoids; Sample preparation; α-Tocopherol; Freezing; HPLC; Stability of carotenoids; Retinol; Human; Biological fluid; Stability; Vitamin; HPLC chromatography; Antioxidant; Isocratic condition; Blood plasma; Reversed phase chromatography; Simultaneous measurement; Carotenoid; Physicochemical properties
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2008.04.007

A simple and sensitive isocratic reversed-phase high-performance liquid chromatography (HPLC) method for simultaneous determination of retinol, α-tocopherol and six carotenoids in human plasma was described. Sample preparation of the earlier published method was further developed by addition of ultrapure water, which enabled aqueous layer to freeze facilitating phase separation without pipetting thus also improving precision of the method. Developed method appeared to be less laborious and time consuming compared to the traditional extraction methods, which require removal of organic layer by pipetting. The recoveries (absolute and relative) were between 80% and 103%. The intra-assay CVs were 1.1–4.0% (normal level) and 3.3–9.0% (low level). Inter-assay CVs were 5.3–8.8%. Reference method for all these analytes was not available, but a comparison with another published method was carried out. The results of the comparison matched satisfactorily. The method is used routinely in our laboratory in a large population-based study.