Convenient experimental determination of adsorption isotherms in a Hydrophobic Interaction Chromatography system |
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Authors: | R J Klimchak S Wang |
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Institution: | (1) Department of Chemical and Biochemical Engineering, Rutgers University, 08855-1360 New Brunswick, New Jersey, USA |
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Abstract: | Summary HPLC was combined with a packable microbore guard column to obtain the adsorption isotherm of lysozyme in a Hydrophobic Interaction Chromatography system. The equipment configuration enabled isotherm determination of the protein on a relatively low pressure chromatographic media (TosoHaas 650M Phenyl).Notation Cm,i
is the mobile phase concentration of protein. (M/L3
(liquid))
- Cm,0
=0
- Cs,i
is the stationary phase concentration of protein. It is the concentration of protein on the chromatographic media. (M/L3
(solid))
- Cs,0
=0
- M,L
is the dimensions mass and length
- Vr,i
is the retention volume of the peak front that corresponds to a mobile phase protein on the concentration Cm,i. (L3
(liquid))
- i
i is a counter that is used to keep track of Cm, Cs, and Vr.For example, i=1 in the term Cm,i denotes the first, and lowest, mobile phase protein concentrations are described by higher values of i.
- Vd
is the system dead volume. It consists of all of the system volume that the mobile phase "sees" or contacts, includingchromatographic media interparticle and pore volume. (L3
liquid)
- Vs
the stationary phase volume. Vs is the nonporous bead volume. For porous beads, Vs is the bead volume - the porevolume. (L3
(solid))
- Ve
is the empty column volume. (L3
liquid)
- Vm
is the packed column mobile phase volume and consists of the pore volume and the excluded volume. (L3
(liquid))
- Ve system
is the empty column system volume. (L3
(liquid))
- Vfrit
the volume of mobile phase that fills the column frits. (L3
(liquid))
- Vwoc
the system volume without the column connected. (L3
(liquid)) |
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Keywords: | |
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