Centrifugal Partition Chromatographic (CPC) T. VINAY KUMAR (08171S0408) M- PHARMACY (MPAQA), Bharat institute of technology, (pharmacy)

 History of CPC  Introduction to counter current chromatography (CCC)  Principle  Instrumentation  Selection of solvent  The advantages of CPC technology  Applications of CPC technology Topics Covered in Presentation

History of CPC Centrifugal partition chromatography is pioneered by Ito in This is basically out growth of counter current chromatography as developed by “Craig and post”. Sanki engineering LTD. Develops the CPC(1982) Dr. Yoichiro develops HSCCC

Counter Current Chromatography (CCC) A separation technique that involves two liquid phases flowing in opposite direction. The stationary phase and the mobile phase result in a relative countercurrent motion. Both stationary and mobile phases are liquids. Separation is based on the partition co- efficient of the components in mixture.

Principle The principle of separation involves the partition of a solute between two immiscible solvents (mobile phase and stationary phases). The relative proportion of solute passing into each of the two solvents determined by partition coefficient.

Instrumentation  Two types of instruments I.CPC manufacture by Sanki engineering ltd II.Multicoil countercurrent system by Ito ( mfg by Pharma Tech Research Corp)

Sanki engineering's CPC instrument

CPC Rotor Elements Partition Disc Pack Partition Channels Partition Disc

The Inner Workings of the CPC

Operation Initially pump the stationary phase solvent into the channel while the rotor is spinning at low speed Followed by mobile phase solvent at the rotation speed required for separation. As the mobile phase flows steadily sample was applied Speed of the rotor

Multi coil counter current chromatography This instrument consists of two or three identical multilayer coils arranged symmetrically around the rotor frame of the centrifuge Each coil column under goes planetary motion.

Selection of solvent systems Critical points in selection are Sample solubility Partition coefficient Chloroform based system (or) Ternary phase diagram is used for the selecting the solvent system

Foucault suggested three criteria to follow ternary phase diagrams - Select the best solvent in which sample can be completely dissolved, - Select two solvents (one is less polar another is more polar), best solvent will partition into the two other solvents, - The less polar fraction of the sample will preferentially go into the less polar phase and the more polar fraction will preferentially go into the more polar phase so that average partition coefficient stay around 1.

Advantages No need of solid support as the stationary phase, 100% recovery of the compounds, Many partition system can be prepared, Low solvent consumption, Same solvent pair can be used for the reveres phase chromatography,

We can used for the large scale operations, Low price of stationary phase solvents compared to that of column packing's. Faster and less expensive product development.

Applications PharmaceuticalBio-Medical BiotechnologyFats and Oils Fermentation Chemicals

Pharmaceutical Fractional purification of natural Tocopherols, fatty acids and phospholipids, Hydrophobicity parameters of drugs, Separation of metal ions, Purification and separation of vitamins A,D and E.

Chemicals Separation of Chiral compound, Synthesis of lipid A, Separation of Saturated and Unsaturated fatty acids, Separation of Alkaloids, Fatty acid esters, Herbmedicine, Monosaccharide, Quinones, etc.

Biotechnology Purification of antibiotics from fermentation broth, Enzymes from Yeast extract, Fungous toxin Nivalenol, Separation of salmon sperm DNA, Separation of serum Proteins.

References CPC article by Udaya wnasundara and Paul fedec. Pos piolot plant crop. chromatography