Size-exclusion chromatography temperature effects

The effect of temperature on column efficiency, however, is frequently exploited, particularly in size exclusion chromatography (SEC). As has already been discussed, high efficiencies are essential in SEC due to the limited peak capacity of the column and consequently, the very small separation ratios. However the effect of temperature on column efficiency is not well understood by many analysts and consequently, will be discussed in some detail. It was shown on page... 

The high salt concentrations used can present a challenge for the HPLC equipment. Pumps with seal-wash are prrferred. If chloride buffers are used, HPLC systems with nonmetallic fluid paths are recommended. It is also advisable to flush the salt solution out of the system when it is not in use. Temperature and pH can affect the separation and should therdbre be controlled. But both the temperature effect and pH effect are smaller than in other forms of chromatography (with the exception of size-exclusion chromatography). 

Mori, S. Suzuki, T. Effect of column temperatures on molecular weight determination by high performance size exclusion chromatography. Anal. Chem. 1980, 52, 1625. 

Mori, S. Suzuki, M. Hydronic volume fluctuation of polystyrene by column temperature and its effect to retention volume in size exclusion chromatography. J. Liquid Chromatogr. Relat. Technol. 1984, 7 (9), 1841. 

Figure 24 Size exclusion chromatography patterns of high-temperature (solid Une) and high-salt (dotted line) fractions desorbed form damaged (a) and virgin hair (b) after application of a collagen hydrolysate. A higher amount of large peptides was adsorbed on bleached/waved hair than on virgin hair. Large peptides were found to be less effectively bound than smaller peptides, as they were selectively removed by hot water (32).

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