Rapid high-performance liquid chromatographic method for the simultaneous determination of retinol, α-tocopherol and β-carotene in human plasma and low-density lipoproteins

• Published in: Journal of chromatography. B, Biomedical sciences and applications Vol. 758; no. 2; pp. 315 - 322
• Main Authors: Gimeno, E, Castellote, A.I, Lamuela-Raventós, R.M, de la Torre-Boronat, M.C, López-Sabater, M.C
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 15.07.2001; Elsevier Science
• Subjects: Adult; Aged; alpha-Tocopherol - analysis; alpha-Tocopherol - blood; Analysis; beta Carotene - analysis; beta Carotene - blood; Biological and medical sciences; Chromatography, High Pressure Liquid - methods; General pharmacology; Humans; Lipoproteins, LDL - chemistry; Male; Medical sciences; Middle Aged; Pharmacology. Drug treatments; Reproducibility of Results; Retinol; Sensitivity and Specificity; Vitamin A - analysis; Vitamin A - blood; Vitamins; α-Tocopherol; β-Carotene; α-Tocopherol; Vitamins; Retinol; β-Carotene; Human; Biological fluid; Vitamin; HPLC chromatography; Photodiode; Deproteinization; Blood plasma; Lipoprotein LDL; Reversed phase chromatography; Diode array; Betacarotene; Quantitative analysis
• ISSN: 0378-4347; 1387-2273
• DOI: 10.1016/S0378-4347(01)00202-X

A reversed-phase HPLC method with diode-array detection was used to simultaneously determine retinol, α-tocopherol and β-carotene in human plasma and low-density lipoproteins. An aliquot of sample was de-proteinized with ethanol containing α-tocopherol acetate as internal standard, and the analytes were extracted twice with hexane. The solvent was evaporated to dryness under a stream of nitrogen and the residue was redissolved in methanol to be injected directly into the HPLC system. A multiple solvent system based on methanol, butanol and water at a flow-rate of 2 ml/min and held at 45°C provided clear separation of these compounds in only 8 min. The method showed good linearity, precision and accuracy for all compounds. Owing to its simplicity, this method may be useful in routine clinical and epidemiological work.