Download Counter Current Chromatography (CCC)

Centrifugal Partition Chromatographic
(CPC)
T. VINAY KUMAR
(08171S0408)
M- PHARMACY (MPAQA),
Bharat institute of technology,
(pharmacy)
Topics Covered in Presentation
History of CPC
Introduction to counter current
chromatography (CCC)
Principle
Instrumentation
Selection of solvent
The advantages of CPC technology
Applications of CPC technology
History of CPC
• Centrifugal partition chromatography is
pioneered by Ito in 1964.
• 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 coefficient 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
Partition Channels
Partition Disc Pack
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
• Pharmaceutical
Bio-Medical
• Biotechnology
Fats 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.
• www.wikipedia.org/wiki/counter-cureentchromatography
• [email protected]