Chapter 5 Size-exclusion chromatography

摘要: Publisher Summary Size-exclusion chromatography (SEC), also called “gel-permeation chromatography,” “molecular-sieve or “gel filtration,” separates molecules according to their sizes. Smaller are retarded on a column, whereas larger ones eluted more rapidly. As the retention time can be directly correlated with sizes of molecules, this method is particularly useful for determination molecular weight. SEC generally applicable separation in range 0.5–1000 kDa, but proteins other giant separated. physically separate folded macromolecules from unfolded ones, case slow equilibria. The major applications include weight and molecular-weight distribution natural synthetic polymers. Separation strongly depends many factors such as column packing, dimensions, flow rate, sample volume, mobile-phase composition. Capillary provides strategy microscale purification. columns best suited direct coupling electrospray ionization–mass spectrometry (ESI–MS) because they have comparable rates that delivered ESI source without splitting.