Simultaneous Profiling and Identification of Carotenoids, Retinols, and Tocopherols by High Performance Liquid Chromatography Equipped with Three-Dimensional Photodiode Array Detection

Abstract

A high performance liquid chromatography (HPLC) system was developed for the simultaneous separation and identification of carotenoids, retinol, its derivatives, and tocopherols. These lipophilic and semilipophilic products were chromatographed on a C18 Vydac column and were monitored and identified using an on-line, three-dimensional photodiode array detector, which permitted profiling on the 200 to 800 nm absorption spectrum of any chromatographic peak in less than 1 sec at a high resolution of 1.2 nm. Identification of peaks and overlapping peaks is conducted by subsequent spectra analysis by the use of a powerful computer station combining Microsoft Windows™ and a chromatography manager. This methodology was successfully applied to extracts of different origins such as plants, sera, tissues and others, memorizing each spectral analysis for processing and comparing within itself or with previously stored data. Quantitative and qualitative processing of the spectral analyses allowed the build-up of a new database of the full absorption profile for each indicated product in comparison to previously published data.