Size exclusion chromatography with reversed-phase

Other detectors being developed at Sandia National Laboratory are based on miniaturizing standard laboratory separation techniques, such as capillary zone electrophoreses, size exclusion chromatography, and reverse phase and affinity electrochromatography coupled with fluorescence (Vitko and Kottenstette, 1998 Dulay et al., 1995 Ramsey et al.. 

Figure 9.10 Three-dimensional representation of the data volume of a tryptic digest of ovalbumin. Series of planar slices through the data volume produce stacks of disks in order to show peaks. Reprinted from Analytical Chemistry, 67, A. W. Moore Jr and J. W. Jorgenson, Comprehensive three-dimensional separation of peptides using size exclusion chromatogra-phy/reversed phase liquid chromatography/optically gated capillary zone electrophoresis, pp. 3456-3463, copyright 1995, with permission from the American Chemical Society.

Besides afEnity chromatography, HPLC separations of four different modes are frequently used for proteins and peptides. These are size-exclusion, ion-exchange, reversed-phase, and hydrophobic interaction chromatographies. Each mode has a different separation mechanism with its own inherent advantages... 

Another approach is to use tips of microliter pipettes as miniature columns by filling them with size exclusion chromatography (SEC) stationary phase material or standard Cig reversed phase material [125,126]. Such tips are also commercially available as ZipTips [122]. There is a risk of sample absorption on the... 

FIGURE 1.3 Three-dimensional representation of a tryptic digest of ovalbumin. The three-dimensional separation consists of size-exclusion chromatography (first dimension), reversed-phase LC (second dimension), and capillary electrophoresis (third dimension). From Moore and Jorgenson, (1995) with permission of the American Chemical Society. 

Fig. 12. Separation of styrene oligomers by reversed-phase (left) and size-exclusion chromatography (right) (Reprinted with permission from [121]. Copyright 1996 American Chemical Society). Conditions (left) column, molded poly(styrene-co-divinylbenzene) monolith, 50 mm x 8 mm i.d., mobile phase, linear gradient from 60 to 30% water in tetrahydrofuran within 20 min, flow rate 1 ml/min, injection volume 20 pi UV detection, 254 nm (right) series of four 300 mm x 7.5 mm i.d. PL Gel columns (100 A, 500 A, 105 A, and Mixed C), mobile phase tetrahydrofuran, flow rate, 1 ml/min injection volume 100 pi, toluene added as a flow marker, UV detection, 254 nm temperature 25 °C,peak numbers correspond to the number of styrene units in the oligomers...

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