Ion pair-based mobile phase additives to improve the separation of alkaloids in supercritical fluid chromatography.

In this study, a supercritical fluid chromatography (SFC) method based on ion pair reagents was used to separate alkaloids. The chromatographic parameters, including the stationary phase, additive type, additive concentration, outlet pressure, temperature and flow rate, were optimized. Baseline separation was completed in 20 min on an Agilent Pursuit 5 PFP column (4.6 × 150 mm) using carbon dioxide as the mobile phase and 7.5 mM sodium 1-pentanesulfonate as an additive with gradient elution at 140 bar, 60 °C, and a flow rate of 1.5 mL/min. The retention rate and resolution of the analytes were satisfactory. The limits of detection were 27.04-298.03 ng/mL, and the limits of quantification were 90.15-993.42 ng/mL. The recoveries of low and high concentrations were 77.46-111.86% and 83.84-111.00%, respectively. This ion pair additive greatly improved the separation efficiency of alkaloids. Consequently, this SFC method was successfully applied to the separation of alkaloids from Rhizoma corydalis.