Determination of molecular masses:

It is rated as one of the most significant applications of size exclusion chromatography. A determination of molecular masses of proteins is particularly cited as an important example. The underlined principle is based on the basic mechanism of the technique. It has been observed that for most proteins a good close correlation between molecular mass and elution behaviour is observed; very few proteins show anomalous behaviour. Relationship between elution behaviour and molecular masses has been obtained for carbohydrates and peptides. The Sephadex G-100 and Sephadex G-200 have been used most often. Agar and agrose gels have also been used and so is Bio-Gel P. The eluant in use with proteins is usually buffer or saline. Long beds should be used for better accuracy and precision of the measurement.

Although with some gels, calibration measurements are available but it is recommended to make calibration curve with each bed. Adsorption phenomena limit the use of gel filtration chromatography for large molecules and particles. This technique has lower accuracy for molecular mass measurement than some other available methods. Instead, this method of determination has the advantage of rapid measurement, is easy to perform and requires fairly inexpensive equipment.