High Performance Size Exclusion Chromatography HPLC-SEC

7 High Performance Size Exclusion Chromatography (HPLC-SEC) 

Size exclusion chromatography (SEC) enables the determination of the molecular size of organic molecules. Samples were filtered through a 0.45 pm filter (Gelman Sciences Acrodiscs) prior to analysis. The membrane filter material was Super (Polyether-sulphone). 

SEC was performed according to the method of Chin et al. (1994). A Shodex KW802.5 SEC column (Waters Corp., Milford, MA., USA) was used and a Waters liquid chromatography system consisting of the following components was used for the analysis Waters 501 high pressure pump. Waters 717 autosampler, InterAction column temperature control oven, Waters 484 UV/VIS detector, and Waters Millenium 2.0 computer software package. 

The mobile phase consisted of 200 mM phosphate at pH 6.8, adjusted to an ionic strength of 0.1 M with high purity NaCl. The eluent was filtered through a preconditioned 0.22 pm membrane filter to prevent interference from particulates. The system was operated at 1.0 mL/min and 30 C, with 200 pL 

The log of the molecular weight versus peak retention time for the PSS standards were plotted and consistently yielded a straight line. By using the calibration equation  

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See also in sourсe #XX -- [ ]

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