Reviews the SFC technique along with applications and case studies. Use of animations disturbs the view. For full ppt, drop me a mail at varadbende96@gmail.com

1. By Guided by
Varad P. 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

3. SUPERCRITICAL
FLUIDS
Supercritical
Fluids

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 Pressure and Temperature

7. CHROMATOGRAPHY

8. Adsorption Partition

9. Stationary
Phase
Mobile Phase
Chromatographic
Technique
CHROMATOGRAPHY
Gas
Liquid
Gas Liquid GLC
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

10. Column Chromatography
Stationary phase
Column

11. SUPERCRITICAL FLUID
CHROMATOGRAPHY

13. Supercritical
Fluids
Properties of
Gases
High Diffusivity
Low Viscosity
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

15. Sensitivity
Efficiency
Selectivity

16. INSTRUMENTATION

18. Advantages
•Cost of operation
•Non toxic
•Low back pressure
•High resolution
•Vast range of analytes
•Simple configuration

19. Challenges
High capital cost
Cannot analyse
extremely polar analytes
Detector incompatibility
Use of organic solvents
as modifiers

20. APPLICATIONS
•Enantioseparations
•Pharma industry
•Chiral compounds
•Polypeptides
•Lipophilic compounds
•Pigments
•Biologically active compounds

21. Manufacturers
 JASCO
 SHIMADZU
 Waters
 Agilent Technologies

22. CASE
STUDIES

23. Purification of Vitamin E
•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
•Analyte of 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
•Analyte of 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
•Analyte of 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 Polyunsaturated Fatty 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