Determination of Retinol in Serum by High-Performance Liquid Chromatography with Fluorometric Detection Using a Column-Switching System

Abstract

We developed a high-performance liquid chromatographic method with fluorometric detection using a column-switching system for the determination of retinol levels in human serum. The lower limit of detection of retinol in serum was 30 ng/ml. Retinol in sample solutions was stable at room temperature in room lighting for at least 30 h. The recovery of retinol in serum was 99% over the range 19.6 to 196 ng/0.2ml serum. Withinand between-assay reproducibilities (CV) for samples containing three concentrations of retinol (low, medium and high) were 0.8, 1.4 and 1.1% (n=10 each), and 5.0, 2.3 and 1.8% (n=5 each), respectively. A high degree of correlation was present between serum levels of retinol as determined using the present method and those determined using the conventional method (r=0.95, Syx=61, n=35).