Improvement of total analytical workflow by using online SFE-SFC

Total analysis time is composed of sample preparation, a chromatographic separation of the target compounds and column equilibration. Online SFE-SFC can decrease the total analysis time, because after simply placing a sample in a special extraction vessel, it automatically performs the entire process from extraction of target components to acquisition of data. Furthermore, SFC can be expected to separate isomers in a shorter time than HPLC. Online SFE-SFC is particularly time-saving in the pharmaceutical industry. More than half of low molecular-weight drugs have stereoisomers, and pharmacological activities of each enantiomer are different. Therefore, it is important that the efficacy and safety of compounds are accurately evaluated as enantiomers, especially in pharmaceutical formulations and its related industries. Chiral separation using SFC and HPLC is one of the typical methods for purifying enantiomers from racemic mixtures. A suitable column and mobile phase for targeted chiral separation have to be evaluated before starting the analysis. To determine the optimized analytical conditions, a large number of candidate conditions have to be examined and this process requires extensive method development. A more prompt and simplistic system for determining the optimized analytical conditions has been needed. A HPLC/SFC switching system meets and exceeds those needs. We demonstrated high-resolution separations using a HPLC/SFC switching system, which automatically switches analytical modes between HPLC and SFC in a single sequence. Here, we report the process of a high-efficiency method development workflow for chiral compounds by using SFC and HPLC in a single sequence. In addition, an online SFE-SFC method was developed and a plasma sample was analyzed under the optimized conditions.