Supercritical Fluid Extraction/Chromatography - Applications Handbook

Significance of Online Supercritical Fluid Extraction/Chromatography

Analytical methods for complex samples often demand time-consuming and solvent-intensive pretreatment steps. The integration of supercritical fluid extraction (SFE) with supercritical fluid chromatography (SFC) leverages the unique properties of supercritical carbon dioxide—low viscosity, high diffusivity, tunable polarity—to streamline workflows. This approach enables rapid, high-resolution separations with reduced organic solvent consumption, mild extraction conditions for labile compounds, and full automation from sample to data.

Objectives and Overview of Studies

These whitepapers and application notes describe:

• Automated optimization of chiral separations by switching among multiple columns and modifiers.
• Application of online SFE-SFC for cleaning validation and trace pesticide analysis.
• Offline SFE pretreatment for vitamins, pharmaceuticals, and environmental contaminants.
• Direct analysis of biological fluids and unstable target compounds using online SFE-SFC.
• Development of UHPLC/SFC switching systems for laboratories.

Methodology and Instrumentation

The core platform is the Shimadzu Nexera UC series:

• SFC modules (LC-30ADSF CO₂ pumps, SFC-30A back-pressure regulators).
• SFE modules (SFE-30A extraction units) with dedicated extraction vessels (0.2–5 mL) and rack changers for up to 48 samples.
• Switching valves for dynamic split control to optimize column loading.
• SFC-UV or SFC-MS (8050/8060 or 8040) detectors with make-up solvent delivery for ionization support.
• Method Scouting Solution software for automated chiral screen on up to 12 columns and multi-modifier gradients.

Main Results and Discussion

1. Chiral Separations: Automatic scouting of 12 Daicel CHIRALPAK/CHIRALCEL columns and up to four modifiers yielded optimized enantiomeric separations of model compounds (omeprazole) in minutes.
2. Online SFE-SFC: Single-pass extraction and analysis of surfactants on swabs and vitamins in supplements demonstrated complete extraction, high recovery (≥ 95%), and low RSD (< 6%).
3. Trace Pesticide Analysis: Combined SFE-SFC-MS/MS achieved multi-pesticide screening (327 compounds) in agricultural products with LOQs of 0.01–1 ng/g and ≤ 30% RSD.
4. Environmental Samples: Offline SFE-GC/MS of over 300 pesticides in soil yielded automated extraction in 30 min, recoveries of 70–120%, and ≤ 10% RSD.
5. Biological Matrices and Labile Compounds: Direct SFC-MS/MS of cerebrospinal fluid enabled analysis of choline/acetylcholine; online SFE-SFC preserved the reduced coenzyme Q10 form without oxidation.

Benefits and Practical Applications

• Workflow Integration: From extraction to separation to detection within one system.
• Solvent Savings: Up to 95% reduction in organic solvent use compared to LC.
• Speed: Analysis times cut to one-third by high-flow SFC.
• Automation: Continuous processing of up to 48 samples reduces manual labor and errors.
• Compatibility: Easy upgrade of existing UHPLC to UHPLC/SFC switching platforms.
• Versatility: Analysis of chiral drugs, vitamins, surfactants, pesticides, metabolites, and unstable compounds.

Future Trends and Possibilities

• Expanded online coupling of other pretreatment techniques (e.g., SFE-GC, SFE-LC).
• Further miniaturization for lower sample volumes (dried blood spots, microdialysis).
• Advanced software-driven scouting for multimodal separations.
• Wider application in metabolomics, lipidomics, and polymer analysis.
• Integration of greener modifiers and CO₂ recycling.

Conclusion

Online SFE-SFC platforms represent a paradigm shift in sample preparation and analysis by unifying extraction and chromatography under supercritical conditions. The Shimadzu Nexera UC systems have demonstrated enhanced speed, sensitivity, reproducibility, and environmental benefits across a range of applications from pharmaceutical chiral screening to food safety and environmental monitoring.

References

1. Shimadzu Application News LAAN-A-LC-E271, LAAN-A-LC-E275, LAAN-A-LC-E277, LAAN-A-LC-E279, LAAN-A-LC-E291.
2. QuPPe—Quick Polar Pesticides (EURL-SRM, Version 9.1, 2016).
3. Pittcon Posters PO-CON1618E, PO-CON1652E, PO-CON1624E, PO-CON1667E.
4. Shimadzu Technical Reports C10G-E059, C190-E209, C190-E200, C190-E205.