SEC separation

In an ideal SEC separation, the mechanism is purely sieving, with no chemical interaction between the column matrix and the sample molecules. In practice, however, a small number of weakly charged groups on the surface of all TSK-GEL PW type packings can cause changes in elution order from that of an ideal system. Fortunately, the eluent composition can be varied greatly with TSK-GEL PW columns to be compatible with a wide range of neutral, polar, anionic, and cationic samples. Table 4.8 lists appropriate eluents for GFC of all polymer types on TSK-GEL PW type columns (11). 

Sometimes elution profiles may also depend on the number of cycles. This can cause problems in the reproducibility of SEC separations. It should be mentioned that no regeneration procedure will give a completely new SEC support. 

DESIGN, PROPERTIES, AND TESTING OF POLYMER STANDARDS SERVICE SIZE EXCLUSION CHROMATOGRAPHY (SEC) COLUMNS AND OPTIMIZATION OF SEC SEPARATIONS... 

PSS columns for medium polar or mixed solvents PSS HEMA and PSS SUPREMA Basic were designed to allow SEC separations in polar media such... 

All SEC columns have to be designed and synthesized by the polymer chemist to meet the specific requirements of the separation mechanism (3). With regard to efficient SEC separations, there are a number of important aspects to consider ... 

Eor a long time there have been discussions about which type of sorbent is the best for SEC separations in various mobile phases. In principle, organic (copolymer) and inorganic packings can be used. Each type of packing has its benefits and drawbacks. Table 9.3 summarizes major sorbent properties and reveals some interesting aspects of SEC separations and its requirements on packings. 

As is known, SEC separations require interaction-free conditions. Therefore, the enthalpic contribution to the free energy term vanishes when no enthalpic interaction is postulated between analyte and sorbent ... 

Figure 9.7 shows separations under identical conditions using PSS SDV columns with 3 (Fig. 9.7a)-, 5 (Fig. 9.7b)- and 10 (Fig. 9.7c)-/i,m particle size columns. A polystyrene oligomer standard was injected and all analyses were performed in THF as the eluent. The much higher efficiencies of small particle size columns are obvious, which is important in the SEC separation of low molecular weight compounds such as additives, by-products, and resins. The reader should note that all chromatograms are area normalized and have the same Y axis to show the differences in peak width and height. 

Table 9.9 (6) gives some guidelines for proper SEC separation conditions when analyzing polymer standards with narrow molar mass distribution on a single 30-cm column. The conditions have to be adjusted when running industrial polymers (which are normally much wider in molar mass distribution). Depending on the width of the MMD, concentrations can be increased by a factor of 3 to 10 for such samples. As a general rule, it is advisable to keep the concentration of the injected solution lower than c [ j] < 0.2. 

For SEC separations of polymers, column efficiency is better characterized by specific resolution, R,p, and efficiency, T, than by theoretical plate count. Peak resolution, R is calculated according to (7) ... 

This chapter illustrates the improvements in SEC column technology and modern applications of SEC separations. The better understanding of SEC column design and separation parameters described in the theoretical sections of this chapter will help the reader fine-tune his or her own work. The same is true for column performance tests, which should be applied regularly, especially after a column purchase. In order to obtain reproducible results, it is recommended to choose column manufacturers who can assure constant quality and performance and to invest in knowledgeable, well-trained support personnel and experienced application chemists. 

For organic SEC separations the use of polystyrene/divinylbenzene (PS/ DVB) particles is almost universal throughout the industry. Polymer Laboratories PS/DVB material, PLgel, which is produced in a series of individual pore sizes, formed the basis for the original product line of SEC columns. Developments in the refinement of particle sizing introduced the benefits of smaller particle size and more efficient columns, which significantly reduced SEC analysis time through a reduction in the number of columns required for... 

The optimum flow rate for most SEC separations using conventional PLgel column dimensions (internal diameter 7.5 mm) is 1.0 ml/min. It may be of some benefit to work with lower flow rates, particularly for the analysis of higher molecular weight polymers where the reduced flow rate improves resolution through enhanced mass transfer and further reduces the risk of shear... 

Figure 13.22 shows the resolution of the surfactants Tween 80 and SPAN. The high resolution obtained will even allow the individual unreacted ethylene oxide oligomers to be monitored. Figure 13.23 details the resolution of many species in both new and aged cooking oil. Perhaps the most unique high resolution low molecular weight SEC separation we have been able to obtain is shown in Fig. 13.24. Using 1,2,4-trichlorobenzene as the mobile phase at 145°C with a six column 500-A set in series, we were able to resolve Cg, C, Cy, Cg, C9, Cio, and so on hydrocarbons, a separation by size of only a methylene group. Individual ethylene groups were at least partially resolved out to Cjg. This type of separation should be ideal for complex wax analysis.

As known, SEC separates molecules and particles according to their hydro-dynamic volume in solution. In an ideal case, the SEC separation is based solely on entropy changes and is not accompanied with any enthalpic processes. In real systems, however, enthalpic interactions among components of the chromatographic system often play a nonnegligible role and affect the corresponding retention volumes (Vr) of samples. This is clearly evident from the elution behavior of small molecules, which depends rather strongly on their chemical nature and on the properties of eluent used. This is the case even for... 

Fig. 22.11. Because the HdC separation does not involve the stagnant mohile-phase transfer, the HdC peaks are significantly narrower than the SEC peaks. The HdC separation in columns with large-pore gels will not he obvious because its separation range will overlap and mix with the SEC separation.

B. Direct SEC-[n] Calibration. Because the SEC separation process is directly related to the size of the solvated molecules, and for a homopolymer series the molecular size is directly related to MW as well as [n]/ it is not necessary to proceed through MW calculations to study polymer intrinsic viscosity. Since... 

Size exclusion chromatography (SEC) separates molecules of a polymer sample on the basis of hydrodynamic volume. When the chromatograph is equipped only with a concentration-sensitive detector, i.e. conventional SEC, a molecular weight distribution (MWD) can be obtained from the chromatogram only through use of a calibration function relating molecular weight and elution volume V (2). 

Since the parameter Rg is known to be a basic SEC size parameter for random-coil type molecules (Equation 4), and since it is also a well-defined statistical average size parameter, applicable to solutes of any shape including the sphere and rodlike molecules, Rg has been chosen in this work to serve as a common, reduced, solute-size parameter for describing the theory of SEC separation. By definition ... 

Also at variance to other modes of LC, SEC separation is determined exclusively by particle size and pore size distribution of the stationary phase, while the mobile... 

In SEC analysis of additive extracts from polymers, the effect of the extraction solvent on the mobile phase is less critical than in HPLC analysis. The extraction solvents typically employed generally do not interfere with the SEC mobile phases. Moreover, the same solvents are often used both as extraction solvent and as mobile phase. Therefore, there is no need to evaporate the extract to dryness prior to analysis and then to redissolve it in a suitable solvent. Typical extraction procedures often produce extracts that generally contain a small amount of wax. Frequently, removal of such oligomers from an extract is necessary, e.g. by means of precipitation, centrifuging, precolumn filtration or protection (use of a reversed-phase guard column). In SEC separations the presence of polyolefin wax does not usually disturb provided that the MW of the wax is higher than that of the analysed compounds. 

In the SEC mode, the separation occurs according to the molecular size of a macromolecule in solution, which is dependent on its chain length, chemical composition, solvent, and temperature. Thus, molecules of the same chain length but different composition may have different molecular sizes (hydrodynamic volumes). Since SEC separates according to hydrodynamic volume, SEC in different eluents can separate a copolymer in two diverging directions. This principle of... 

The trade-off might be that the selectivity of the gradient HPLC method for the individual cyclic oligomers is better. One way to increase resolution of the LCCC procedure is to couple this type of separation with a SEC separation in a 2D experimental setup. The higher selectivity of the 2DLC separation could help to resolve individual oligomers of different functionalities. 

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