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Supercritical Fluid Chromatography

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Supercritical Fluid Chromatography

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    23. 23 Practical Advantages of SFC SFC is Normal Phase complimentary to reversed phase HPLC Diffusion is much Faster higher speed/throughput-more samples/day more rapid re-equilibration-shorter cycle time Non-Linear Solvent Strength-first small addition of polar additives dramatically increases solvent strength Viscosity is Much Lower lower pressure drop, higher flows, or longer columns possible

    24. 24 What Compounds Can SFC Separate? Any solute soluble in methanol or a less polar organic solvent will elute in SFC. Strong organic acids and bases require a modifier and an additive in the mobile phase. Most salts of organic acids and bases elute. Small lipophylic peptides elute With carbon dioxide based fluids, molecular weights up to 15,000

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    27. 27 What Compounds Will NOT Elute in SFC? Any solutes requiring an aqueous environment any solutes requiring a buffered or ionic aqueous environment I.E., Biomolecules such as Proteins will NOT elute inorganic salts will not elute With carbon dioxide, few solutes over 15,000 MW elute

    28. 28 Sample Matrix? Usually solution in organic solvent methanol or less polar allows larger injections can use DMSO small (i.e., 0.5-1 µL) aqueous samples direct often reduced sample prep because SFC is compatible with both lipids and polar molecules pills dissolved with excipients, etc. present extracts from natural products extracts from fermentation

    29. 29 Why Carbon Dioxide? Miscible with much more polar solvents-wide range of solvent strength available. program composition from 0 to 100% organic modifier Low operating temperatures (i.e., 30-50°C) Compatible with most HPLC and GC Detectors Relatively safe product of human respiration can use with the FID easy disposal low purchase price Lack of a superior alternative

    30. 30 solute families amenable toSFCsolute families amenable toSFC

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    32. 32 Schematic Diagram of a Packed Column SFC

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