Determination of carotenoids in microalgae using supercritical fluid extraction and chromatography

Abstract

A method was developed based on supercritical fluid chromatography for quantitative determination of carotenoids in extracts of Scenedesmus sp. By utilizing the low backpressure in supercritical fluid chromatography, a C18 column and a 2-ethyl pyridine column were coupled in series. It was concluded that even minor changes in temperature had a substantial effect on selectivity. A standard mixture of 8 carotenoids and microalgae extracts obtained through supercritical fluid extraction with and without 10% ethanol as a co-solvent were successfully separated. All of the carotenoids were separated within 10 minutes, while the total analysis time was 20 minutes. The method was validated and the carotenoids of microalgae extracts were quantified. Furthermore, the method should be seen as a more rapid and environmentally sustainable alternative to traditional high-performance liquid chromatography methods utilizing organic solvents.

Impact of injection solvents on supercritical fluid chromatography

“…obtained by SFE as described in a previous paper [10]. overlaid with chromatograms with 1.5 mL added dead volume to the SFC system (dashed line).…”

“…Ethanol ( ethanol as a co-solvent, at a pressure of 300 bar, a temperature of 60 °C, a flow of 2 mL min -1 , and extraction time was 60 minutes, as have previously been described [10]. The extract was diluted 10 times in each of the solvents except in acetone, dichloromethane, chloroform and dimethyl sulfoxide.…”

“…The chromatographic analysis of the SFE extract of microalgae containing carotenoids was performed as has been previously described, but with the modification that injection volume was 10 µL instead of 50 µL [10]. The carotenoids were separated on the C18 column coupled in front of the 2-EP column.…”

Purification and Formulation of Xanthophyll for Pharmaceutical Use: Current Strategies and Future Trends

“…SC follows the principle of solid stationary phase chromatography with super critical fluids as mobile phase. Stationary phases comprising C18 columns, 2-ethyl pyridine columns and monolithic silica columns have been employed, where a tandem of columns has proven to increase resolution while maintaining short running times [57]. Monolithic silica columns have facilitated analysis since higher flow rates can be achieved [58].…”

Fast Separation Method Development for Supercritical Fluid Chromatography Using an Autoblending Protocol

“…In recent years, supercritical fluid chromatography (SFC) has been widely used for the analysis of a wide range of analytes, especially hydrophobic molecules [1][2][3][4]. Due to the relatively low toxicity, high diffusivity, low viscosity and high solvating power of the mobile phase used (supercritical CO 2 , SCCO 2 ), SFC is regarded as a greener separation tool complementing or replacing gas chromatography (gC) and high-performance liquid chromatography (HPlC) [5].…”