Aim: The applicability of polysaccharide-based chiral stationary phases in modern supercritical fluid chromatography (SFC)-MS/MS for chiral bioanalysis was evaluated. Materials & methods: Ten popular polysaccharide-based chiral stationary phases (CSPs) were tested using a set of 23 drugs against three cosolvents. The effect of temperature and backpressure on separation was examined. Results: The recommended order of CSPs for screening was determined. Methanol with 0.1% NH4OH is proven to be the first choice of cosolvent. Temperature of 40°C and backpressure of 10 or 15 MPa are recommended starting conditions. Phospholipid elution profiles on the polysaccharide-based CSPs were reported for the first time under SFC conditions. Conclusion: A simplified screening protocol with straightforward method optimization approaches was generated for SFC chiral assay development in a reasonable time frame with a high success rate.
Keywords: SFC–MS/MS; chiral bioanalysis; chiral stationary phase; enantiomer; enantioselective bioanalysis; supercritical fluid chromatography.