Comparative study of ultra-high-performance supercritical fluid chromatography and ultra-high-performance liquid chromatography to simultaneous determination of ten components in Radix hedysari

Background: In China, dried root of Hedysarum polybotrys Hand-Mazz, known as Hong Qi (Radix hedysari [RH]), is one of the important traditional Chinese medicines (TCMs). The long analysis time was a major limitation when high-throughput techniques are required for research or quality control purposes. Materials and Methods: It is necessary to develop a high?throughput method for the determination of multiple components of TCMs by investigating the separation characteristics of the target with ultra-high-performance liquid chromatography (UHPLC) and ultra-high-performance supercritical fluid chromatography (UHPSFC). Objective: This study intends to establish a high-throughput method for simultaneous determination of ten compounds in RH by UHPSFC and to develop a standardized method to comprehensively evaluate the quality of RH samples. In addition, this study was based on UHPLC and UHPSFC systems to understand and compare the effects of different separation parameters on ten target compounds in RH. A high-throughput and reliable solid-phase extraction method for simultaneous detection of ten target compounds in RH was established and validated. Results: The optimal analysis method was achieved using HSS C18SB column, gradient elution, flow rate of 1.5 mL/min, diode array detection at 270 nm, and back pressure at 12.41 Mpa. The separation method was validated sufficiently by examining the precision, recoveries, linearity, range, limit of detection, and limit of quantification and was successfully applied to quantify the 25 RH samples. In addition, the comparative study also demonstrated the complementary nature of the two separation modes. Conclusion: This is the first study that ten compounds in RH were separated and analyzed simultaneously with UHPSFC-diode array detector. In addition, under the separation conditions of UHPSFC and UHPLC, the comparison of chromatographic parameters shows that a combination of these two separation techniques efficiently separates the ten target compounds in RH. The results obtained in this study are useful for future TCM quality control and high-throughput analysis.