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Tosoh: The Chemistry of Innovation
Technical Support
Chromatography Calculations
Cleaning Methods
Principles of Chromatography
Size Exclusion
Hydrophobic Interaction
Reversed Phase
Ion Exchange
Affinity
Frequently Asked Questions


The physical characteristics of a molecule are exploited in liquid chromatography to affect a separation. The most basic separation is based on the size (hydrodynamic diameter) of the molecule. When separating based on size the chromatographic mode is size exclusion chromatography. If the molecule has a positive or negative charge associated with it, as is the case with all proteins, peptides, nucleic acids and some carbohydrates, cation exchange or anion exchange chromatography are employed. Hydrophobic regions on the molecule can be exploited by using normal phase, reversed phase, or hydrophobic interaction chromatography. The three dimensional structure typically defines the activity of the molecule. Using this conformation of the molecule is very powerful separation technique and it is commonly used in chiral (for smaller molecules) or affinity (for larger molecules) chromatography. The following chart defines the chromatographic modes based on the physical characteristic used.

Physical Characteristic

Mode
 Size (Hydrodynamic diameter)
 Molecular Charge
 Hydrophobic Regions
 Conformational Recognition
 Affinity
 Chiral

 


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Bioscience Division

Shiba, Minato-ku, Tokyo 105-8623, Japan
TEL:+81-3-5427-5118
FAX:+81-3-5427-5198
Email: hlc@tosoh.co.jp

Tosoh Bioscience Japan, Shiba, Minato-Ku, Tokyo 105-8623, Japan TEL:+81-3-5427-5118 FAX:+81-3-5427-5198 Email:hlc@tosoh.co.jp