Going Faster, Going Greener with High Efficiency Regis' Chiral Stationary Phases in Supercritical Fluid Chiral Chromatography (SFC)
Chiral method development and chiral separations have been the main contributors to the popularity of supercritical fluid chromatography (SFC). The low viscosity and high diffusivity of the mobile phase in SFC allows for low pressure drops and fast chromatography, making it an attractive alternative to LC, especially for pharmaceutical applications. SFC is often 3–5 times faster than HPLC and often produces higher efficiency. SFC is now regarded as a standard separation support tool for chemical synthesis and development.
Chiral method development and chiral separations have been the main contributors to the popularity of supercritical fluid chromatography (SFC). The low viscosity and high diffusivity of the mobile phase in SFC allows for low pressure drops and fast chromatography, making it an attractive alternative to LC, especially for pharmaceutical applications. SFC is often 3–5 times faster than HPLC and often produces higher efficiency. SFC is now regarded as a standard separation support tool for chemical synthesis and development.
Figure 1: Sample: Atenolol, Column: RegisCell 25 cm à 4.6 mm, Mobile Phase: 50% [MeOH + 0.1% TEA], Flow Rate: 5 mL/min, Pressure: 120 bar, Temp: 30 °C
A number of factors determine the success of a SFC separation. Perhaps the most important factor is the performance of the chiral stationary phase (CSP). This study investigates the ability of a RegisCell® and RegisPack® chiral column to generate fast separations in SFC.
Figure 2: Sample: Flubiprofen, Column: RegisPack 25 cm à 4.6 mm, Mobile Phase: 30% MeOH, Flow Rate: 3 mL/min, Pressure: 120 bar, Temp: 30 °C
Instrumentation and Columns
The RegisPack® and RegisCell® polysaccharide based CSPs are coated with (tris-(3,5-dimethylphenyl) carbamoyl amylose) and (tris-(3,5-dimethylphenyl) carbamoyl cellulose), respectively. All applications were developed using 5 µm, analytical 25 cm × 4.6 mm columns. The instrumentation consisted of a Fusion™ A5 SFC conversion module (Aurora SFC Systems, Inc., Redwood City, CA) connected to a Model 1200SL HPLC (Agilent Technologies, Waldbronn, Germany).
Table I: Summary of a number of rapid separations developed on RegisCell® and RegisPack®
Conclusion:
A number of fast applications were developed on the RegisPack and RegisCell chiral columns. RegisCell and RegisPack CSPs show very good performance in SFC, allowing for very short run times and excellent efficiencies. Simple mobile phase conditions were used. Additives can greatly affect separations, but they are not always needed. Short run times may offer significant advantages in analytical work, enabling the screening of numerous samples in a short period of time, lower system volume, and reduced cost per injection. SFC chiral chromatography continues to gain interest among scientists in the pharma and R&D community as a faster, less expensive, and greener technique.
Regis Technologies, Inc.
8210 Austin Ave, Morton Grove, IL 60053
tel. (847) 583-7661, fax (847) 967-1214
Website: www.registech.com/chiral
SEC-MALS of Antibody Therapeutics—A Robust Method for In-Depth Sample Characterization
June 1st 2022Monoclonal antibodies (mAbs) are effective therapeutics for cancers, auto-immune diseases, viral infections, and other diseases. Recent developments in antibody therapeutics aim to add more specific binding regions (bi- and multi-specificity) to increase their effectiveness and/or to downsize the molecule to the specific binding regions (for example, scFv or Fab fragment) to achieve better penetration of the tissue. As the molecule gets more complex, the possible high and low molecular weight (H/LMW) impurities become more complex, too. In order to accurately analyze the various species, more advanced detection than ultraviolet (UV) is required to characterize a mAb sample.
