Development of a Two-Dimensional Supercritical Fluid Chromatography System in Multiple Heart-Cutting Modes

Area of Science:

• Analytical Chemistry
• Chromatography
• Supercritical Fluid Chromatography (SFC)

Background:

• Two-dimensional chromatography (2D-LC) offers enhanced separation power for complex samples.
• Supercritical fluid chromatography (SFC) is a powerful technique for separating diverse analytes, including chiral compounds.
• Developing efficient and robust 2D-SFC systems is crucial for advanced analytical challenges.

Purpose of the Study:

• To develop a simplified loop-based multi-heart-cutting SFC (mSFC-SFC) system.
• To achieve complete peak transfer between dimensions, irrespective of mobile phase composition.
• To demonstrate the system's capability in analyzing complex natural product extracts, focusing on chiral flavonoids.

Main Methods:

• Instrument design based on a single SFC system with added 2-port 6-position valves for heart-cutting.
• Utilized diode-array and mass spectrometric detection.
• Employed an achiral polar stationary phase in the first dimension and a chiral polysaccharide stationary phase in the second dimension for flavonoid separation.

Main Results:

• Achieved excellent repeatability (RSD < 2%) for retention times and peak areas.
• Demonstrated high peak capacity and shape integrity, enabling accurate diastereomer quantification.
• Reported an efficient inter-dimensional transfer rate of 83% for compressible samples.

Conclusions:

• The developed mSFC-SFC system provides a simple yet effective solution for complex separations.
• The system demonstrates robust performance and high efficiency for analyzing chiral compounds in natural products.
• This approach facilitates the quantification of diastereomers with excellent repeatability and transfer efficiency.