Purification methods high-pressure liquid chromatography

Measurements of the common physical constants such as boiling point or refractive index are not sufficiently sensitive to determine the trace amounts of impurities in question. Besides the common spectroscopic methods, techniques like gas chromatography (GC), high-pressure liquid chromatography (HPLC), or thin-layer chromatography (TLC) are useful. The surest criterion for the absence of interfering foreign compounds lies in the polymerization itself the purification is repeated until test polymerizations on the course of the reaction under standard conditions are reproducible (conversion-time curve, viscosity number of the polymers). 

Solid-phase parallel synthesis mimics the previously described solution phase strategy. This approach easily lends itself to both semi- and full automation. In contrast to the solution phase method, purification is easily achieved by simply washing the resin beads, and the reactions can be driven to completion by excess reagents, multiple cycles, and microwave techniques. The initial building block or scaffold is attached to the resin bead by a detachable linker. At the end of the synthesis, the final construct is released under the appropriate cleavage conditions for automated purification, usually by high-pressure liquid chromatography (HPLC). This allows bioanalysis of the final product in aqueous solution under standard assay conditions. 

From 1980 s, several research groups studied the components of EFE, independently [4,5,6]. Different purification methods were employed to isolate the enzymes, including sulfuric ammonia precipitation, gel filtration, ion exchanging, high-pressure liquid chromatography (HPLC)... 

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