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Supercritical Fluid Chromatography
The document discusses supercritical fluid chromatography (SFC), highlighting its principles, instrumentation, advantages, challenges, and applications in the biotechnology field. It details the properties of supercritical fluids, the classification of chromatographic techniques, and case studies on the purification of various compounds using SFC. The document also lists key manufacturers and specific examples of applications in the pharmaceutical industry and biotechnology.
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Supercritical Fluid Chromatography
- 1. By Guided by VaradP. Bende Dr. S. D. Saptarshi Department of Biotechnology MGM’s Jawaharlal Nehru Engineering College, Aurangabad. MS.
- 2. Contents • Supercritical Fluids •Chromatography • Supercritical Fluid Chromatography (SFC) • Instrumentation of SFC • Advantages and Challenges • Applications of SFC • Case Studies
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- 4. • Critical Pressure(PC) • Critical Temperature (TC) Supercritical
- 5. Liquid Gas Density (g/cm3) Diffusivity (cm2/s) Viscosity (Pa .s) Gas 10-5 10-1 10-4 Supercritical fluids 10-2 - 1 10-4 - 10-3 10-4 - 10-5 Liquid 1 10-5 10-2 SCF
- 6. Effect of Pressureand Temperature
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- 9. Stationary Phase Mobile Phase Chromatographic Technique CHROMATOGRAPHY Gas Liquid Gas LiquidGLC Solid Gas Solid GSC Liquid Liquid Paper Chromtography Solid TLC Liquid Solid LC Bonded Phase Size Exclusion Ion exchange AffinitySupercritical fluid Liquid Supercritical Fluid ChromatographySolid Classification of Chromatography
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- 13. Supercritical Fluids Properties of Gases High Diffusivity LowViscosity Properties of Liuquid Density Solvating Power •Dissolution •Displacement of . matrix •Transport •Penetrability
- 14. Mobile phase -CO2 •Chemically inert •Cheap •Non toxic •Non flammable •Low critical temperature and pressure
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- 18. Advantages •Cost of operation •Nontoxic •Low back pressure •High resolution •Vast range of analytes •Simple configuration
- 19. Challenges High capital cost Cannotanalyse extremely polar analytes Detector incompatibility Use of organic solvents as modifiers
- 20. APPLICATIONS •Enantioseparations •Pharma industry •Chiral compounds •Polypeptides •Lipophiliccompounds •Pigments •Biologically active compounds
- 21. Manufacturers JASCO SHIMADZU Waters Agilent Technologies
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- 23. Purification of VitaminE •Analyte of Interest - α-tocopherol from γ- and δ -tocopherol •Mobile phase - CO 2 + 3.98% EtOH •Stationary phase - Silica Chongwen Jiang , et al. (2003) •Temperature - 40 °C Pressure-18 MPa
- 24. Purification of Polypeptide •Analyteof Interest - Isomeric polypeptides (547/541 and 541/544) •Mobile phase - CO 2 + 0.2% TFA (in MetOH) •Stationary phase - Pyridine capped silica M A Patel , et al. (2011) •Temperature - 40 °C Pressure-10 MPa
- 25. Purification of Cyclosporins •Analyteof Interest - Cyclosporin analogs A,B,C,D,H •Mobile phase - CO 2 + EtOH •Stationary phase - Silica Yuefi Shao , et al. (2016) •Temperature - 80 °C Pressure-15 MPa
- 26. Purification of Carotenoids •Analyteof Interest - Carotenoids and isomers •Mobile phase - CO 2 + 1 %EtOH •Stationary phase - Phenyl Polymethylsiloxane H H Schmitz , et al. (1989) •Temperature - 45 °C
- 27. Purification of PolyunsaturatedFatty Acids •Analyte of Interest - EPA and DHA •Mobile phase - CO 2 •Stationary phase - C18 M Alkioa , et al. (2000) •Temperature - 65 °C Pressure-14.5 MPa