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Handbook of Size Exclusion Chromatography and Related ...

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An insurmountable limitation <strong>of</strong> SEC is that molecules are fractionated<br />

according to hydrodynamic volume rather than by molar mass. This determines<br />

that (even under perfect resolution) the instantaneous MWD is not monodisperse,<br />

except perhaps for the rather special case where both repeating unit types exhibit<br />

identical specific densities <strong>and</strong> are noninteracting. The variety <strong>of</strong> molar masses in<br />

the detector cell when a “chromatographically complex” polymer is analyzed<br />

introduces some bias in the MWD (31). The bias is further magnified under<br />

imperfect resolution. Imperfect resolution results from a combination <strong>of</strong>: (a)<br />

nonexclusion (secondary or enthalpic) fractionation (32,33), <strong>and</strong> (b) instrumental<br />

broadening in the columns, fittings, <strong>and</strong> detectors (33–37). Nonexclusion effects<br />

may shift <strong>and</strong> distort the chromatograms, yielding both positive <strong>and</strong> negative<br />

molecular weight deviations. Instrumental broadening is important when the<br />

MWD is narrow or multimodal. If the instrumental broadening is not corrected for,<br />

then the polydispersity M w=M n is typically: (a) overestimated when the molecular<br />

weights are calculated from a calibration obtained with narrow st<strong>and</strong>ards, (b)<br />

underestimated when obtained from LS sensors, <strong>and</strong> (c) under- or overestimated<br />

when obtained from IVs (37,38).<br />

In liquid adsorption chromatography (LAC), copolymer molecules are<br />

fractionated according to their enthalpic interactions with the column substrate.<br />

When the repetitive unit types exhibit a difference in their adsorption–desorption<br />

behavior <strong>and</strong> such behavior is independent <strong>of</strong> the molar mass, then LAC can be<br />

used to determine the CCD (39).<br />

In many practical situations, copolymers are mixed with their corresponding<br />

homopolymers, <strong>and</strong> it may be impossible to quantitatively isolate the copolymer<br />

prior to its SEC analysis. This involves a serious complication, because SEC<br />

detectors cannot distinguish a copolymer from a homopolymer mixture with the<br />

same hydrodynamic volume <strong>and</strong> an equivalent global composition. Even in the<br />

presence <strong>of</strong> polymer mixtures, the bivariate distribution <strong>of</strong> the molecular weights<br />

<strong>and</strong> chemical composition may still be estimated if the repeating unit types exhibit<br />

a difference in their adsorption–desorption behavior. First, preparative LAC is<br />

used to isolate the copolymer from the homopolymers <strong>and</strong> to fractionate the<br />

copolymer by composition. Then, SEC is used to determine the MWD <strong>of</strong> thin<br />

slices <strong>of</strong> the LAC eluogram (40,41). With less success, previous developments<br />

have been proposed that first fractionate by hydrodynamic volume <strong>and</strong> then<br />

analyze the eluted slices by chemical composition (42).<br />

In some special cases, SEC alone is capable <strong>of</strong> determining both the MWD<br />

<strong>and</strong> the CCD (43–45). To this effect, the following (rather hard) conditions must<br />

be verified: (1) the instantaneous distributions <strong>of</strong> the molecular weights <strong>and</strong> <strong>of</strong><br />

the chemical composition are both narrow, <strong>and</strong> (2) the instantaneous average<br />

composition varies monotonically with the molecular weights. Eventually,<br />

the second condition could be relaxed if the first condition were strictly verified.<br />

Similarly, both the MWD <strong>and</strong> the DB <strong>of</strong> a branched copolymer may be determined<br />

© 2004 by Marcel Dekker, Inc.

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