Solid-phase extraction protocol for isolating retinol-d4 and retinol from plasma for parallel processing for epidemiological studies

Abstract

Solid-phase extraction permits the parallel processing of samples in large numbers. We have applied this technique to the isolation of retinol isotopomers from plasma of humans participating in a study of vitamin A stable isotope dilution. The isotopomers were analyzed by gas chromatography/mass spectrometry. The extraction involves the separation of retinol from its aqueous matrix with a C18 silica-based sorbent followed by removal of lipid contaminants with an aminopropyl silica-based sorbent. Overall recovery of retinol from plasma was 47.2% +/- 1.8%. Purity of the retinol isolated from plasma is comparable with that obtained with a single HPLC method. This method permits the preparation of 32 samples per day by one analyst. Elimination of the need for HPLC permits sample preparation in the field with a minimum of equipment and technical skill.