Considering the selectivity of pore size gradient size exclusion chromatography columns.

One of the most significant performance determining variables of a size exclusion column is the pore size of its packing material. This is most definitely the case for assigning the suitability of a given column for differently sized analytes. As technologies for particle and column manufacturing continue to advance, it is worth contemplating the value of more finely controlled manipulation of this parameter. The change in a packing material's pores across the length of a size exclusion column was thus explored. A change in average pore diameter and pore size distribution was studied by means of theoretical modeling. These parameters were investigated for independent and combinatorial effects. From our predictions, versus tandem column chromatography, a gradient column apparatus does not yield sizable increases in monomer to dimer selectivity of any given critical pair. Instead, our modeling suggests it can yield more universally effective separations of multiple pairs of species at once, as is sometimes necessary when analyzing the high molecular weight components of highly aggregated drug substances.
Competing Interests: Declaration of competing interest The authors are employed by Waters Corporation, a company that manufactures and sells chromatography columns.
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