Automated solid-phase extraction and liquid chromatographic method for retinoid determination in biological samples.
In this study, a method for partly automated sample preparation and fully automated solid-phase extraction method for plasma, kidney and liver samples for various retinoids like all-trans-4-oxo-retinoic acid, 13-cis-4-oxo-retinoic acid, 13-cis-retinoic acid, 9-cis-retinoic acid, all-trans-retinoic acid, retinol and retinyl palmitate was established. Plasma, embryo-, kidney-and liver-homogenates were automatically mixed and extracted on multiple usage solid-phase (C2) extraction cartridges immediately before HPLC analysis. Automated cleaning, preconditioning and incorporation of the loaded cartridge to fully automated HPLC separation and quantification of the various retinoids in a single HPLC run was established. The recovery of the retinoids was generally between 80 and 90%. Intra-day repeatability was < 11.7%. As little as 1.2 ng/ml could be quantified in lipid-mixture standard samples. This method allows a highly automated sample preparation and a fully automated solid-phase extraction with good selectivity for the study of endogenous retinoids and retinoids after nutritional supplementations and pharmacological applications in several biological samples.
Rühl
Ralph
R
Institute of Nutritional Science, University of Potsdam, Arthur Scheunert-Allee 114-116, D-14558 Potsdam-Rehbrücke, Germany. ralphruehl@web.de
Schweigert
Florian J
FJ
D000818:Animals; D001331:Automation; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D051379:Mice; D015203:Reproducibility of Results; D012176:Retinoids / Q000032:analysis*; D012680:Sensitivity and Specificity
D012176:Retinoids