Countercurrent chromatography – Media Converters – SDH Multiplexer
Droplet Countercurrent Chromatography (DCCC)
Droplet CCC is the oldest form of CCC. It uses only gravity to move the mobile phase through the stationary phase. In descending mode, droplets of the denser mobile phase and sample are allowed to fall through a column of the lighter stationary phase using only gravity. If a less dense mobile phase is used it will rise through the stationary phase, this is called ascending mode. The eluent from one column is transferred to another; the more columns that are used, the more theoretical plates can be achieved. The disadvantage of DCCC is that flow rates are low, and poor mixing is achieved for most binary solvent systems, which makes this technique both time-consuming and inefficient.
Centrifugal Partition Chromatography (CPC)
Centrifugal Partition Chromatography (CPC) was invented in the eighties by the Japanese company Sanki Engineering Ltd, whose president was the late Kanichi Nunogaki. CPC has been extensively developed in France starting from the late nineties CPC use centrifugal force to speed separation and achieves higher flow rates than DCCC (which relies on gravity). The centrifugal partition chromatograph is constituted with a unique rotor (=column). This rotor rotates on its central axis (while HSCCC column rotates on its planetary axis and simultaneously rotates eccentrically about another solar axis). With less vibrations and noise, the CPC offers a wider rotation speed range (from 500 to 2000 rpm) than HSCCC. That allows a better decantation and retention for unstable biphasic system (e.g., aqueous aqueous systems or Butanol/water systems). Basics of CPC: The CPC rotor is constituted by the superposition of disks engraved with small cells connected by head / tail ducts. These cells, where the chromatographic separation takes place, can be compared to lined-up separate funnels. The rotor is filled with the stationary phase, which stays inside the rotor thanks to the rotation speed, while the mobile phase is pumped through. CPC can be operated in either descending or ascending mode, where the direction is relative to the force generated by the rotor rather than gravity. According to the fast and permanent evolution of the cells design, the efficiency and flow rate with low back pressure are improved. The CPC offers now the direct scale up from the analytical apparatuses (few milliliters) to industrial apparatuses (some liters) for fast batch production.
High-Speed Countercurrent Chromatography
The modern era of CCC began with the development by Dr. Yoichiro Ito of the planetary centrifuge and the many possible column geometries it can support. These clever devices make use of a little-known means of making non-rotating connections between the stator and the rotor of a centrifuge. (It is beyond the scope of this discussion to describe the method of accomplishing this. Any of the several books available on CCC discuss it thoroughly.)
Functionally, the high-speed CCC consists of a helical coil of inert tubing which rotates on its planetary axis and simultaneously rotates eccentrically about another solar axis. (These axes can be made to coincide, but the most common or type J CCC is discussed here.) The effect is to create zones of mixing and zones of settling which progress along the helical coil at dizzying speed. This produces a highly favorable environment for chromatography.
There are numerous potential variants upon this instrument design. The most significant of these is the toroidal CCC. This instrument does not employ planetary motion. In some respects it is very like CPC, but retains the advantage of not needing rotary seals. It also employs a capillary tube instead of the larger-diameter tubes employed in the helices of the other CCC models. This capillary passage makes the mixing of two phases very thorough, despite the lack of shaking or other mixing forces. This instrument provides rapid analytical-scale separations, which can nonetheless be scaled up to either of the larger-scale CCC instruments.
Xanthanolide purification
Modes of Operation
head to tail: the denser phase is pumped through as the mobile phase. Derived from terminology for Archimedean screw force.
tail to head: the less dense phase is used as the mobile phase.
Dual-Mode: The mobile and stationary phases are reversed part way through the run.
Gradient Mode: The concentration of one or more components in the mobile phase is varied throughout the run to achieve optimal resolution across a wider range of polarities. For example, a methanol-water gradient may be employed using pure heptane as the stationary phase. This is not possible with all binary systems, due to excessive loss of stationary phase.
Elution Extrusion Mode (EECCC): The mobile phase is extruded after a certain point by switching the phase being pumped into the system. For example, during the Elution portion of a separation using an EtOAc-water system running head to tail, the aqueous mobile phase is being pumped into the system, . In order to switch to extrusion mode, organic phase is pumped into the system. This can be accomplished either with a valve on the inlet of single pump, or ideally with an integrated system of two or three pumps, each dedicated either to a single phase of a binary mixture, or to an intermediate wash solvent. This also allows for good resolution of compounds with high mobile-phase affinities. It requires only one column volume of solvent and leaves the column full of fresh stationary phase.
pH Zone Refining: Acidic and basic solvents are used to elute analytes based on their pKa.
References
^ Ito, Y. (1970). “Countercurrent Chromatography: Liquid-Liquid Partition Chromatography without Solid Support”. Science 167: 281. doi:10.1126/science.167.3916.281. PMID 5409709.
^ Pinel B, Audo G, Mallet S, et al. (June 2007). “Multi-grams scale purification of xanthanolides from Xanthium macrocarpum. Centrifugal partition chromatography versus silica gel chromatography”. J Chromatogr A 1151 (1-2): 149. doi:10.1016/j.chroma.2007.02.115. PMID 17433347.
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