Development of Simple High Performance Liquid Chromatographic Method for the Determination of Astaxanthin in Human Plasma

Abstract

A fast and simple high-performance liquid chromatographic UV-Vis method for the determination of astaxanthin in human plasma was developed and validated. The method involved direct injection of plasma sample after deproteinization using a 1:9 mixture of ethanol-tetrahydrofuran. The mobile phase comprised methanol:water:ethyl acetate (82:8:10% (v/v)) with a total run time of 5 min. Analysis was run at a flow rate of 1.5 mL/min with the detector set at 474nm. This method is specific and sensitive, with a quantification limit of approximately 0.25 µg/mL. The calibration curve was linear over a concentration range of 0.39 − 50.0 µg/mL. The method had a mean absolute recovery of 99%, while the within-day and between day coefficients of variation and percentage errors were all less than 3%. The speed, specificity, sensitivity, reproducibility and stability of this method make it suitable for the determination of human plasma astaxanthin in routine analysis.