Editing Column chromatography (section)

== Column adsorption equilibrium ==
For an adsorption column, the column resin (the stationary phase) is composed of microbeads. Even smaller particles such as proteins, carbohydrates, metal ions, or other chemical compounds are conjugated onto the microbeads. Each binding particle that is attached to the microbead can be assumed to bind in a 1:1 ratio with the solute sample sent through the column that needs to be purified or separated.

Binding between the target molecule to be separated and the binding molecule on the column beads can be modeled using a simple [[equilibrium reaction]] K<sub>eq</sub> = [CS]/([C][S]) where K<sub>eq</sub> is the [[equilibrium constant]], [C] and [S] are the concentrations of the target molecule and the binding molecule on the column resin, respectively. [CS] is the concentration of the complex of the target molecule bound to the column resin.<ref name=":0" />

Using this as a basis, three different isotherms can be used to describe the binding dynamics of a column chromatography: linear, Langmuir, and Freundlich.

The linear isotherm occurs when the solute concentration needed to be purified is very small relative to the binding molecule. Thus, the equilibrium can be defined as:

:[CS] = K<sub>eq</sub>[C].

For industrial scale uses, the total binding molecules on the column resin beads must be factored in because unoccupied sites must be taken into account. The [[Langmuir isotherm]] and [[Freundlich isotherm]] are useful in describing this equilibrium. The Langmuir isotherm is given by:

:[CS] = (K<sub>eq</sub>S<sub>tot</sub>[C])/(1 + K<sub>eq</sub>[C]), where S<sub>tot</sub> is the total binding molecules on the beads.

The Freundlich isotherm is given by:

:[CS] = K<sub>eq</sub>[C]<sup>1/n</sup>

The Freundlich isotherm is used when the column can bind to many different samples in the solution that needs to be purified. Because the many different samples have different binding constants to the beads, there are many different K<sub>eq</sub>s. Therefore, the Langmuir isotherm is not a good model for binding in this case.<ref name=":0" />