Ever since chemists started using Separating funnels to isolate compounds by partitioning, they have understood the potential advantages of liquid/liquid chromatography, known now as countercurrent chromatography CCC. Yet despite this knowledge, solid/liquid chromatography methods, such as HPLC or flash, have been the workhorses of purification. Until recently, CCC was primarily a technique for organic products or academic study and has been barely used in mainstream purification. Unfortunately, ancient CCC instrumentation was badly engineered and suffered from slow pace of separation, a combination that resulted in slight adoption as a complementary and orthogonal chromatography technique.
However a new generation of High heeled countercurrent chromatography HPCCC instrumentation has resulted in the rebirth of liquid/liquid chromatogram in the 21st century and for that reason offering a greater advantage to the chemists.CCC can significantly enhance a Chemists productivity and different compounds which were previously very difficult to isolate or uneconomical to produce. Due to the huge difference in reachable stationary phase between liquid/liquid into solid/liquid chromatography – typically 70-80 percent compared to 5-10percent – the loadings are dramatically higher, shortening the amount of sample injections necessary to process a batch. Moreover, since both stationary and mobile phases are fluids, we gain two additional important productivity benefits.
First, sample solubility Difficulties Are reduced because one’s choices for injecting sample on the column have been decreasing. Using CCC, an individual can inject a sample into both of the individual mobile or stationary phases or a combination of both, whichever mix provides the highest loading per shot. The use of two fluids is also beneficial when the sample is on the pillar, because even when the sample crashes from solution, it does not cause the column to obstruct, stopping the chromatography.
Another productivity benefit is that with CCC there’s absolutely not any chance of irreversible adsorption happening either onto or in the stationary phase. Recoveries are always very high, and it is certain that the whole sample will elute from the column. With All the advantages that CCC can bring into the productivity of chemists, why has it been so badly adopted? The first generation of CCC Instruments introduced in the early 1980s was called high-speed countercurrent chromatography HSCCC machines and were adopted for three reasons. The first was rate of separation – HSCCC tools perform separations within a period of hours, as opposed to the tens of minutes average HPLC. Second, the instrumentation was unreliable and therefore scientists immediately became reluctant to risk their precious compounds. Finally, the array of equipment available was poor and typically only available at the preparative scale, requiring gram-size sample shots. This is problematic for chemists working in small-molecule artificial chemistry, who originally may have had samples available just in countless milligram amounts that took months to create. Therefore, the whole amount of a valuable sample would need to be recovered – a threat a chemist is reluctant to take.