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HomeProtein PurificationSephadex® & Darcy’s Law

Sephadex® & Darcy’s Law

Sephadex® G-10, G-25, and G-50 can be assumed to behave as rigid spheres in SEC and therefore obey Darcy’s Law. This Law describes a general relationship for flow in porous media:

U = K × ΔP × L-1 Equation (1)

U = linear flow rate expressed in cm/h
ΔP = pressure drop over the packed bed expressed in cm water
L = bed height expressed in cm
K = constant of proportionality depending on the properties of the bed material and the buffer Assuming a buffer with viscosity of 1 cP: U = Ko × ΔP × L-1 Equation (2)
Ko = the “specific permeability” depending on the particle size of the medium and the water regain

Note that flow is proportional to the pressure drop over the bed and, assuming a constant pressure head, inversely proportional to the bed height. In practice this means that the pressure/flow considerations that must be made when using other SEC media do not apply to Sephadex® and that a doubling of flow rate leads to a doubling in column pressure. To a good approximation, flow rate is independent of the column diameter.

Flow at viscosities greater than 1 cP can be obtained by using the relationship: flow rate is inversely proportional to viscosity. High buffer viscosities can be compensated for by increasing the operating pressure to maintain a high flow rate.

Theoretical flow (not maximum) can be calculated from equation (2) by inserting values for ΔP and L. Specific permeabilities (K) are given in Table A2.1.

Sephadex® typePermeability K
Sephadex® G-1019
Sephadex® G-25 Superfine9
Sephadex® G-25 Fine30
Sephadex® G-25 Medium80
Sephadex® G-25 Coarse290
Sephadex® G-50 Fine36
Table A2.1.Specific permeabilities of Sephadex®
Materials
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