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Improved High‐Performance Liquid Chromatography Method with Diode Array Detection for Simultaneous Analysis of Retinoic Acid Isomers, Retinol, Retinyl esters, Vitamin E, and Selected Carotenoids in Guinea Pig Tissues
Author(s) -
Schäffer Michael W,
Roy Somdutta Sinha,
Mukherjee Shyamali,
Das Salil K
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.621.3
Subject(s) - lutein , chemistry , retinol , retinyl palmitate , chromatography , lycopene , retinyl acetate , high performance liquid chromatography , retinoic acid , detection limit , chromatography detector , carotenoid , vitamin , biochemistry , gene
We report the validation of a versatile reversed‐phase gradient HPLC method allowing simultaneous detection of retinoic acid (RA) isomers (13‐ cis , 9‐ cis , all‐ trans ‐RA), retinol, retinyl esters (retinyl palmitate (RP), stearate and acetate as internal standard), tocopherols (T) (α,γ,δ) and selected carotenoids (lutein, lycopene, β‐carotene) in Hartley guinea pig (GP) tissues, using a diode array detector. Under the tested conditions, the method yielded low intra‐ and inter‐assay variability, with good recovery and sufficient sensitivity. Limits of detection, tested linear range and coefficient of correlation (ng injected; μg/mL; r 2 ‐values, respectively) were: 13‐ cis RA (0.451; 0.023–22.562; 0.9981), 9‐ cis RA (0.5044; 0.022–21.617; 0.9974), all‐ trans ‐RA (0.4930; 0.018–17.609; 0.9964), retinol (0.8615; 1.436–14.359; 0.9916), RP (0.3587; 0.028–28.181; 0.9981), α–T (6.6214; 0.368–367‐857; 0.9996), γ–T (5.9224; 0.279–279.669; 0.9982), δ–T (4.2148;), lutein (0.1159; 0.009–9.273; 0.9711), lycopene (0.6975; 0.074–7.363; 0.9992), and β‐carotene (1.1847; 0.01–10.202; 0.9758). This method was used to quantify the cited analytes in several GP tissues. It should be noted that lutein, a dietary component, was detected in GP lung. The significance of the presence of lutein in lung remains to be elucidated. (Supported by US Army Grant W81XWH‐06‐2‐0044).