Gel-permeation

Gel permeation chromatography, exclusion chromatography. gel filtration chromatography. A technique for separating the components of a mixture according to molecular volume differences. A porous solid phase (a polymer, molecular sieve) is used which can physically entrap small molecules in the pores whilst large molecules pass down the column more rapidly. A solvent pressure up to 1000 psi may be used. 

Two classes of micron-sized stationary phases have been encountered in this section silica particles and cross-linked polymer resin beads. Both materials are porous, with pore sizes ranging from approximately 50 to 4000 A for silica particles and from 50 to 1,000,000 A for divinylbenzene cross-linked polystyrene resins. In size-exclusion chromatography, also called molecular-exclusion or gel-permeation chromatography, separation is based on the solute s ability to enter into the pores of the column packing. Smaller solutes spend proportionally more time within the pores and, consequently, take longer to elute from the column. 

At first glance, the contents of Chap. 9 read like a catchall for unrelated topics. In it we examine the intrinsic viscosity of polymer solutions, the diffusion coefficient, the sedimentation coefficient, sedimentation equilibrium, and gel permeation chromatography. While all of these techniques can be related in one way or another to the molecular weight of the polymer, the more fundamental unifying principle which connects these topics is their common dependence on the spatial extension of the molecules. The radius of gyration is the parameter of interest in this context, and the intrinsic viscosity in particular can be interpreted to give a value for this important quantity. The experimental techniques discussed in Chap. 9 have been used extensively in the study of biopolymers. 

This chapter contains one of the more diverse assortments of topics of any chapter in the volume. In it we discuss the viscosity of polymer solutions, especially the intrinsic viscosity the diffusion and sedimentation behavior of polymers, including the equilibrium between the two and the analysis of polymers by gel permeation chromatography (GPC). At first glance these seem to be rather unrelated topics, but features they all share are a dependence on the spatial extension of the molecules in solution and applicability to molecular weight determination. 

Gel permeation chromatography (GPC) is the term found most widely in the polymer literature. In this context it is used most widely as an analytical... 

Table 9,4 Data for the Analysis of the Gel Permeation Chromatogram of a Poly disperse Polymer Used in Example 9.7...

Both preparative and analytical GPC were employed to analyze a standard (NBS 706) polystyrene sample. Fractions were collected from the preparative column, the solvent was evaporated away, and the weight of each polymer fraction was obtained. The molecular weights of each fraction were obtained usmg an analytical gel permeation chromatograph calibrated in terms of both and M. The following data were obtained ... 

Distribution Coefficients. Gel-permeation stationary-phase chromatography normally exhibits symmetrical (Gaussian) peaks because the partitioning of the solute between mobile and stationary phases is linear. Criteria more sophisticated than those represented in Figure 8 are seldom used (34). 

Gel-permeation media are extremely versatile and may be used for separation of particles such as vimses (Fig. 11) as well as proteins (34). Separations of proteins and other particles having sizes equivalent to a molecular weight of 40 x 10 are possible using the agar-based Sepharose-type gel. This particular gel has a limited temperature range for operation, however. It melts upon heating to 40°C (34). 

Salt Effects. The definition of a capacity factor k in hydrophobic interaction chromatography is analogous to the distribution coefficient, in gel permeation chromatography ... 

Bentone-34 has commonly been used in packed columns (138—139). The retention indices of many benzene homologues on squalane have been determined (140). Gas chromatography of C —aromatic compounds using a Ucon B550X-coated capillary column is discussed in Reference 141. A variety of other separation media have also been used, including phthaUc acids (142), Hquid crystals (143), and Werner complexes (144). Gel permeation chromatography of alkylbenzenes and the separation of the Cg aromatics treated with zeofltes ate described in References 145—148. 

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