High performance size-exclusion

ZORBAX POROUS SILICA MICROSPHERE COLUMNS FOR HIGH-PERFORMANCE SIZE EXCLUSION CHROMATOGRAPHY... 

A. Development of Silica Particles for High-Performance Size Exclusion Chromatography (HPSEC)... 

Modern SEC columns are packed with material other than polystyrene gels, such as porous silica particles or highly cross-linked styrene-divinylbenzene copolymers. Because of improvements in speed and resolution, the term SEC is sometimes replaced by the term high-performance size-exclusion chromatography (HPSEC). 

High-performance size exclusion chromatography is used for the characterization of copolymers, as well as for biopolymers (3). The packings for analyses of water-soluble polymers mainly consist of 5- to 10-/Am particles derived from deactivated silica or hydrophilic polymeric supports. For the investigation of organosoluble polymers, cross-linked polystyrene beads are still the column packing of choice. 

ASTM D 5296 - 92 (1994). Standard Test Method for Molecular Weight Averages and Moleeular Weight Distribution of Polystyrene by High Performance Size-Exclusion Chromatography. Annual Book of ASTM Standards, Vol.08.03, pp. 419-431. 

Analysis of the MHR by high-performance size-exclusion chromatography (HPSEC) revealed three distinct populations to be present, which were isolated on a preparative scale. The composition of the populations differed mainly in the relative proportion of rhamnose, xylose and the amount of methyl esters and acetyl groups, although the general characteristics were rather similar (Table 2). However, degradation studies with RGase showed that, in contrast... 

Molecular weight distributions were examined by high performance size-exclusion chromatography (HPSEC) as described [22] on two serial Shodex OHpak KB-803 and KB-805 columns (0.8 x 30 cm Showa Denko, Japan) with a OHpak KB-800P guard column (0.6 x 5 cm), equilibrated at 1 mL/min in 0.1 M UNO3. The system was calibrated with a pullulan calibration kit (Showa Denko). 

Fig. 3. High performance size exclusion chromatography of MHR after degradation with RGAE from A. aculeatus, RGaseA and a combination of these enzymes.

Figure 6. High-Performance Size-Exclusion Chromatography of solubilized pectin from the juice. Control is CDTA extracted pectin from parenchyma.

These high performance size exclusion separations of alpha-chymotrypsin SI and myosin light chains compare favorably with those achieved by ion exchange chromatography but require only a fraction of the time to accomplish. Furthermore, the very short retention times allow for separation of these labile proteins at room temperature, whereas operation at 0-40 C would otherwise be mandatory to avoid the loss of enzymatic activity. 

Pfannkoch, E., Lu, K. C., Regnier, F. E., and Barth, H. G., Characterization of some commercial high-performance size-exclusion chromatography columns for water-soluble polymers, /. Chromatogr. Sci., 18, 430, 1980. 

Kirkland, J. J. and Antle, P. E., High-performance size-exclusion chromatography of small molecules with columns of porous silica microspheres, J. Chromatogr. Sci., 15,137,1977. 

Russell, D. J., Calibration of high performance size exclusion chromatography for small epoxy molecules, /. Liquid Chromatogr., 11, 383, 1988. 

Czok, M., Katti, A. M., and Guiochon, G., Effect of sample viscosity in high-performance size-exclusion chromatography and its control, /. Chromatogr., 550, 705, 1991. 

Dawkins, J. V., Forrest, M. J., and Shepherd, M. J., Separation of vinyl chloride oligomers by recycle high performance size exclusion chromatography, /. Liq. Chromatogr., 13, 3001, 1990. 

Mori, S., Wada, A., Kaneuchi, F., Ikeda, A., Watanabe, M., and Mochizuki, K., Design of a highly sensitive infrared detector and application to a high-performance size exclusion chromatography for copolymer analysis, /. Chromatogr., 246, 215, 1982. 

Gysler, J., Mazereeuw, M., Helk, B., Heitzmann, M., Jaehde, U., Schunack, W., Tjaden, U.R., and van der Greef, J., Utility of isotachophoresis-capillary zone electrophoresis, mass spectrometry and high-performance size-exclusion chromatography for monitoring of interleukin-6 dimer formation, /. Chromatogr. A, 841, 63, 1999. 

Ruiz-Calero, Puignou, L., Galceran, M. T., and Diez, M, Coupling high-performance size exclusion and ion chromatography for the analysis of low-molecular-mass heparin, /. Chromatogr. A, 775, 91, 1997. 

Figure 4.21 High-performance size-exclusion chromatograms of the polymeric plasticiser Reoplex R346 using (a) RI and (b) UV detection. After Castle et al. [792]. Reproduced from L. Castle et al., Food Addit. Contain., 8, 565-576 (1991), by permission of Taylor Francis Ltd (http //www.tandf.co.uk/journals)...

T. Alfredson and J. Tschida, High Performance Size Exclusion Analysis of Coatings and Adhesive Polymer Formulations, Publ. No. LC-122, Varian Instrument Group, Walnut Creek, CA (n.d.). 

HPSEC High-performance size-exclusion ID Identification... 

Watson and Kenney [62] describe the use of high-performance size-exclusion chromatography to examine the aggregation of interferon-y and interleukin-2 after storage at elevated temperature, after mechanical agitation, and following rapid freeze-thaw. An excellent review on SEC can be found in Ref. 63. 

E. Watson and W. C. Kenney, High performance size exclusion chromotography of recombinant derived proteins and aggregated species, J. Chromotogr., 436, 289 (1988). 

Barth, H.G. and Smith D.A. "High-Performance Size-Exclusion Chromatography of Guar Gum," J Chromatog. (1981) 206, 410-415. 

Ysambertt, F., Cabrera, W., Marquez, N., Salager, J.L. (1995). Analysis of ethoxylated nonylphenol surfactants by high performance size exclusion chromatography (HPSEC). J. Liq. Chromatogr. 18(6), 1157-1171. 

Instrumentation. A Pharmacia BioPilot Column Chromatography system was used to perform large-scale size exclusion chromatography (SEC) with an 11.3 x 90 cm BioProcess column packed with Sephacryl S-200 HR gel. High performance size exclusion (HPSEC) and ion exchange chromatography (HPIEC) were conducted with Pharmacia Superose 6 and 12 (HR 10/30) and Mono-Q (HR 5/5) columns respectively, equipped with Beckman model 520 system controller and Beckman model HOB HPLC pumps. 

High Performance Size Exclusion Chromatography (HPSEC) A Varian Model 5000 liquid chromatograph equipped with a variable wavelength UV detector, two columns in series (PL gel, 300 x7.5 mm, particle size 5/im, porosity of 50 and 500A), was used, THF being the eluent. 

Figure 8. High performance size exclusion chromatography of Indulin ATR and of the THF soluble fraction of each APPL. (Reproduced with permission from Ref. 12, 1989, CNRC.)...

RG Nielsen, EC Rickard, PF Santa, DA Shakarnas, GS Sittampalam. Separation of antibody-antigen complexes by capillary zone electrophoresis, isoelectric focusing and high-performance size-exclusion chromatography. J Chromatogr 539 177-185, 1991. 

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