Polymer HPLC

In this chapter, high-performance liquid chromatography of oligomers and (high) polymers (polymer HPLC) will be briefly presented. As mentioned in Section 16.1, there exist several monographs, chapter in books, and review papers on this subject, for example [1-33], Most of them contain numerous examples of the HPLC separation and molecular characterization of particular macromolecular substances. Therefore, this chapter discusses almost exclusively the general principles of polymer HPLC and only few selected examples of practical applications will be mentioned for illustration. 

The dissolution of macromolecules is a prerequisite for the application of liquid chromatography for their separation and characterization. Compared to HPLC of small molecules, concentration of the polymer solutions injected into the analytical HPLC columns is higher and usually assumes lmg mL and more. This is mainly due to detection problems the detectors used in polymer HPLC are much less sensitive (Section 16.9.1) than detectors for small molecules, which often carry the UV-absorbing chromophores. This means that samples subject to polymer HPLC must exhibit rather high solubility. 

RETENTION MECHANISMS IN POLYMER HPLC 16.3.1 General Considerations... 

The most straightforward way for understanding the retention phenomena in polymer HPLC is afforded by thermodynamics. It is widely accepted that the retention of any kind of substance in... 

Depending on the particular method of polymer HPLC, is defined in different ways. It is the total volume of pores, Vp, in a porous packing but it can be also related to the total surface of packing (mostly to the surface situated within the pores) or to the effective volume of bonded phase. The volume of pores is relatively well defined in the case of many packings applied in polymer HPLC and plays an especially important role in the exclusion-based separations (Sections 16.3.3, 16.3.4, and 16.4.1). The exclusion processes, however, play an important role in the coupled techniques of polymer HPLC (Section 16.5). In the latter cases, the surface of packings and the effective volume of bonded phase are to be taken into account. In some theoretical approaches also, surface exclusion is considered. 

Numerous theories attempted to define the quantitative relation between the distribution constant K and the molecular characteristic of polymer samples so far such attempts have not been successful. The uncertainty of is one of the reasons why methods of polymer HPLC are considered nonabsolute. 

Selected Properties of Some Solvents Used in Polymer HPLC... 

The thermodynamic quality of mobile phase may be so poor that macromolecules precipitate within column or at least exhibit a tendency to phase separation. This is a specific feature of certain procedures of polymer HPLC (Sections 16.3.7,16.5.3, and 16.5.6). 

In conclusion, two different parameters of mobile phase are to be distinguished in polymer HPLC namely its strength toward column packing and its quality toward column packing and especially toward separated macromolecules. 

FIGURE 16.3 Dependences of the polymer retention volume on the logarithm of its molar mass M or hydrodynamic volume log M [T ] (Section 16.2.2). (a) Idealized dependence with a long linear part in absence of enthalpic interactions. Vq is the interstitial volume in the column packed with porous particles, is the total volume of liquid in the column and is the excluded molar mass, (b) log M vs. dependences for the polymer HPLC systems, in which the enthalpic interaction between macromolecules and column packing exceed entropic (exclusion) effects (1-3). Fully retained polymer molar masses are marked with an empty circle. For comparison, the ideal SEC dependence (Figure 16.3a) is shown (4). (c) log M vs. dependences for the polymer HPLC systems, in which the enthalpic interactions are present but the exclusion effects dominate (1), or in which the full (2) or partial (3,4) compensation of enthalpy and entropy appears. For comparison, the ideal SEC dependence (Figure 16.3a) is shown (5). (d) log M vs. dependences for the polymer HPLC systems, in which the enthalpic interactions affect the exclusion based courses. This leads to the enthalpy assisted SEC behavior especially in the vicinity of For comparison, the ideal SEC dependence (Eigure 16.3a) is shown (4). 

In order to explain the dependences of log M vs. Vr depicted in Figure 16.3b through d and to nnderstand particnlar processes in the polymer HPLC columns, the qualitative thermodynamic consideration, Eqnation 16.6 can be nsed. 

It can be concluded that adsorption is an important tool for controlling retention volumes of samples in polymer HPLC. Eluent composition and temperature are the most feasible variables to affect adsorption in the given polymer-column packing systems. The thermodynamic quality of eluent plays less important role. 

In conclusion, the enthalpic partition processes in the columns for polymer HPLC substantially differ from the adsorption processes. Enthalpic partition can be employed for the separation of polymers of the low-to-medium polarity in combination with the alkyl bonded phases on silica gels. The extent of the enthalpic partition and consequently also of the polymer retention is controlled primarily by the thermodynamic quality of eluent toward separated species and by the extent of the bonded phase solvation. 

Phase separation (precipitation) of a polymer strongly depends on all its molecular characteristics. On the one hand, this allows very efficient separations in polymer HPLC utilizing phase separation and re-dissolution processes [20]. On the other hand, due to complexity of phase separation phenomena, the resulting retention volumes of complex polymers may simultaneously depend on several molecular characteristics of separated macromolecules. This may complicate interpretation of the separation results. Both precipitation and redissolution of most polymers is a slow process. It may be affected by the presence of otherwise inactive surface of the column packing. Therefore, the applicability and quantitative control of the phase separation phenomena may be limited to some specific systems of polymer HPLC. 

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