COUNTER CURRENT CHROMATOGRAPHY
IS COMPLIMENTARY TO PREPARATIVE HPLC.
Strengths of Counter Current Chromatography
1. CCC can cope with a wide range of radically different
2. CCC can cope with particulates and extract solid samples.
3. CCC can have orthogonal selectivity to HPLC.
4. CCC can achieve 99 + % purity of target compounds from complex samples.
5. CCC can take extremely complex matrices, such as natural product extracts
and heart cut target polarities, bio-actives etc without risk of on-column
degradation or adsorption.
6. CCC uses exactly the same type of liquid pumps, injectors, switching
valves fraction collectors etc as HPLC, flash chromatography or SMB, but
as CCC often requires only a fraction of the solvent flow or total solvent
used, CCC offers substantial cost savings in capital equipment cost by
using lower flow pumps, plus savings in solvent and process cost.
7. Scale up is linear and predictable for the modular Quattro CCC™.
8. Small laboratory 100 ml coil CCC can rapidly process 0.5 to 2 + grams
of crude extract, but only require flows of 2 to 10 ml /min and therefore
can use a standard analytical HPLC pump.
9. Single process units of 3000 ml can rapidly prepare 15 to 60 + grams
of crude extract, but only require flows of 25 to 100 ml /min. HPLC Pumps
of this capacity are available for modest cost.
10. Any number of process units can be used in series / manifolded in
parallel / or used in SMB mode for tonne per annum production.
Weaknesses of Counter Current Chromatography
1. CCC solvent selection choice is unfamiliar to many chromatographers.
2. CCC fundamental principles are unfamiliar to many chromatographers.
Strengths of Preparative HPLC
1. Choice of phase and solvent optimization familiar to most chromatographers.
2. Traditional preparative/process chromatographers understand scale up.
Weaknesses of Preparative HPLC
1. The individual costs of high-resolution 5 or 10um
particle size columns of 250mm length by 1 or 2 inch diameter are relatively
high; yet the sample loading may be only 0.5 to 2 grams. The typical eluent
flows would be 10 to 50 + ml / min and these flows would require a dedicated
Preparative HPLC pump.
2. Columns can be easily and permanently incapacitated by injection of
crude extract containing a wide mix of polarities.
3. Equipment to pack columns of larger than 2 inch bore requires a large
one-off cost, plus requires the purchase of HPLC phase as a consumable.
4. Columns capable of loadings of 15 to 60 grams would require flows of
100’s ml /min to 1000 + ml /min, and therefore require specialist
and very expensive HPLC pumps plus solvent handling equipment.
5. Scale up from 250 x 4.6 mm id analytical columns with 3 to 5um particle
to larger preparative columns often with 7, 10, 15 or 20 um particle sizes
is not familiar to many analytical chromatographers
Complimentary Nature of Counter Current Chromatography to Preparative
When suitable biphasic solvent systems are chosen a Quattro CCC™
can prepare a highly pure ( 99 + % ) target compound, directly from a
Sometimes, however, the time and or method optimization familiarity is
not available to find a successful CCC method in an allotted time scale.
Generic Counter Current Chromatography step gradient
techniques are available to heart cut target polarities from crude, complex
mixtures. This CCC technique can both radically reduce the mass needing
to be prepared by the preparative HPLC, plus substantially reduce the
likelihood of contaminating the HPLC column.
As an example…………consider
a crude natural product extract of 50 grams with 1% active. This could
not be considered directly by preparative HPLC, but would only require
a 3000 ml Quattro CCC™ running at 50 ml /min to heart cut approximately
1 grams of a narrow polarity band. A 250 mm long x 1 inch diameter preparative
HPLC column with 10um HPLC reverse phase running at 25 ml /min using standard
HPLC solvent and phase optimization techniques, could then rapidly prepare
the target, without risk of irreversible column contamination.
Now scale this down from 50 grams of sample to 1 grams
A small 100 ml Quattro CCC™ coil running at modest
flows available from standard analytical HPLC pumps, could rapidly prepare
10mg of a narrow polarity extract.
Consider the cost savings of a 10 mg Preparative HPLC
column plus ancillary equipment, compared to a 1 gram column with the
appropriate ancillary equipment……..and factor in the lack
of risk of column contamination after Counter Current Chromatography heart
Worth Thinking About ? Require More Information…
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