Column size exclusion chromatography

Column Si. Size-exclusion chromatography columns are generally the largest column on a process scale. Separation is based strictly on diffusion rates of the molecules inside the gel particles. No proteins or other solutes are adsorbed or otherwise retained owing to adsorption, thus, significant dilution of the sample of volume can occur, particularly for small sample volumes. The volumetric capacity of this type of chromatography is determined by the concentration of the proteins for a given volume of the feed placed on the column. 

SIZE EXCLUSION CHROMATOGRAPHY COLUMNS FROM POLYMER LABORATORIES... 

INTERACTIVE PROPERTIES OF POLYSTYRENE/DIVINYLBENZENE AND DIVINYLBENZENE-BASED COMMERCIAL SIZE EXCLUSION CHROMATOGRAPHY COLUMNS... 

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. 

Calibration of size-exclusion chromatography columns based on the finding that the retention volume of a molecular or particulate species is usually a single-valued function of an appropriate size parameter of this molecule or particle, irrespective of its chemical nature and structure. 

Note 3 Macroporous polymers are used, for example, as precursors for ion-exchange polymers, as adsorbents, as supports for catalysts or reagents, and as stationary phases in size-exclusion chromatography columns. 

D. Berek, Interactive properties of polystyrene/divinylbenzene and divinylbenzene-based commercial size exclusion chromatography columns, in Column Handbook for Size Exclusion Chromatography, C.-s. Wu, ed.. Academic Press, San Diego, CA, 1999, p. 445. 

Adamic and Bartak [6] used high pressure aqueous size exclusion chromatography with reverse pulse amperometric detection to separate copper(II) complexes of poly(amino carboxylic acids), catechol and fulvic acids. The commercially available size exclusion chromatography columns were tested. Columns were eluted with copper(II) complexes of poly(aminocarboxylic acids), citric acids, catechol and water derived fulvic acid. The eluent contained copper(II) to prevent dissociation of the labile metal complexes. Reverse pulse electrochemical measurements were made to minimise oxygen interferences at the detector. Resolution of a mixture of DTP A, EDTA and NTA copper complexes was approximately the same on one size exclusion chromatography column as on Sephadex... 

The Dextran polymers used were Pharmacia Dextran T fractions TIO (lot No. 16026), 140 (lot No. 21945), T70 (lot No. 23155) and T500 (lot No. 19073). Size-exclusion chromatography columns were calibrated with these fractions. Molecular weight distributions of these lots were determined by Pharmacia. 

Alternatively, combined antibody fractions can be desalted using size exclusion chromatography columns such as PD-10 from GE Healthcare BioSciences. 

The dried, partially protected crude peptide is dissolved in DMF-pyridine (4 1 v/v) and treated with DMF-SO3 (125-fold molar excess) in the presence of 1,2-ethanedithiol (100-fold molar excess). After about 15 h at room temperature, the solution is applied to a size exclusion chromatography column and eluted with DMF. The target compound is collected and the solvent evaporated to dryness. After lyophilization, the crude compound is treated with a 90% TFA-based reagent at 4°C. The reaction time of this step is optimized by monitoring the acidolytic treatment of a small aliquot of the O-sulfated peptide and analysis of the synthetic products by analytical HPLC and mass spectrometry. 

This method was developed by Stefan Huber (Karlsruhe, Germany) and consists of three size exclusion chromatography columns which divide the organic carbon into several fractions as a function of size, but also hydrophobic and ionogenic characteristics. A sample of up to 3 mL is injected into the instrument and filtered in-line with a 0.45 )um filter. The deposit on the filter is backwashed after 5 minutes and directly analysed with the TOC analyser to determine the particulate organic carbon content (POC). 

Also, silica-based packings are available for aqueous size-exclusion chromatography. The commercially available ones are coated with a glycidoxypro-pylsilane bonded phase. While they do not swell or shrink, they are not compatible with basic pH values and contain residual silanols that may interact with the polymer. Also, only a limited pore-size range is available compared to the aqueous size-exclusion chromatography columns based on glycidyl methacrylate. 

S. Terabe, H. Tanaka, K. Otsuka and T. Ando, Separation of Small Molecules with Size Exclusion Chromatography Columns and Micellar Solutions, J Chromatogr. Sci., 27 653 (1989). 

Herman, D.P. Field, L.R. Calibration of size exclusion chromatography columns for molecular weight determination of polyacrylonitrile and poly(vinylpyrrolidone) in N,N-dimethylformamide. J. Chromatogr. Sci. 1981, 79, 470. 

Karaca, E, Islas, C.A., Millan, M., Behrouzi, M., Morgan, T.J., Herod, A.A., Kandiyoti, R. (2004) The cahbration of size exclusion chromatography columns molecular mass distributions of heavy hydrocarbon liquids. Energy Fuels,... 

Figure 2 Typical profile obtained from high-performance size exclusion chromatography. Column SynChropak GPC 300, 300 x 7.8 mm ID Mobile phase 0.1 M KH2PO4, pH 7. Flow rate 1.0 mL/min.

---