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The Theory For Gradient Chromatography Revisited
The Theory For Gradient Chromatography Revisited
Objective of the presentation Discuss the background of the traditional theory for gradient chromatography. Show how a more fundamental and general theory for gradient chromatography can be obtained. Show some applications of the general theory.
Version: 05/20/07
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Zone velocity
Version: 05/20/07
Assume that a given composition of the mobile phase migrates through the column with the same velocity as the mobile phase, i.e. u0. Let the solute be injected at x=0 and t=0. The equation for the migrating zone can now be written:
Version: 05/20/07
In many cases the retention factor of a solute decreases exponentially with F, i.e.:
Brief discussion of the traditional theory (4) For a linear gradient with slope G and for a solute with retention factor ki at t=0, integration gives:
Version: 05/20/07
A fundamental starting point for an alternative gradient theory is the mass balance equation for chromatography:
c= solute concentration in the mobile phase n= solute concentration on the stationary phase F= column phase ratio D= diffusion coefficient of the solute x= axial column coordinate t= time
Version: 05/20/07
Mass balance approach(2) The stationary phase concentration is a function of the mobile phase composition, , i.e. n=n(c,(x,t)) . This means that:
For a linear adsorption isotherm F*n/ c is equal to the retention factor k((x,t)).
Version: 05/20/07
Here, the diffusive term has been omitted. The equation is the analogue of the ideal model for chromatography. The term n/ is a function of c, i.e. In the limit c0, the traditional representation of gradient chromatography theory is obtained.
Version: 05/20/07
Where c is the concentration of the solute in the mobile phase and k0 the retention factor of the solute when =0. The function /t is known and determined by the experimenter. For a linear gradient it is equal to the slope, G, of the gradient.
Version: 05/20/07
Mass balance approach(5) For this particular case the mass balance equation is:
Where ki is the initial retention factor at t=0. The solution of this equation is of the form:
Mass balance approach(6) Example: Assume that the solute is injected at x=0 as a Gaussian profile according to
Version: 05/20/07
Solution of the gradient equation for a Gaussian profile, red line. Numeric simulation of the complete mass balance equation, H=10mm, for the same input parameters. c0=10 mmol, t0=50,s ki=10, ,ti=10s
Version: 05/20/07
Solution of the gradient equation for a Gaussian profile, red line. Numeric simulation of the complete mass balance equation, H=10mm, for the same input parameters. c0=10mmol, t0=50,s ki=10, ,ti=10s
Version: 05/20/07
Solution of the gradient equation for a Gaussian profile, red line. Numeric simulation of the complete mass balance equation, H=10mm, for the same input parameters. c0=10 mmol, t0=50,s ki=10, ,ti=10s
Version: 05/20/07
Solution of the gradient equation for a Gaussian profile, red line. Numeric simulation of the complete mass balance equation, H=10mm, for the same input parameters. c0=10 mmol, t0=50,s ki=10, ,ti=10s
Version: 05/20/07
Solution of the gradient equation for a Gaussian profile, red line. Numeric simulation of the complete mass balance equation, H=10mm, for the same input parameters. c0=10 mmol , t0=50,s ki=10, ,ti=10s
Version: 05/20/07
Mass balance approach(7) Example: Assume that the solute is injected at x=0 as a profile according to
Version: 05/20/07
Assume that the solute concentration is constant and independent of time. The solution of the differential equation is:
Version: 05/20/07
Conclusions A fundamental and general theory for gradient chromatography can be obtained from the mass balance equation for chromatography. The traditional theory for gradient chromatography is a special case of a more general theory, it is valid in the limit c(solute) 0. By neglecting the dispersive term in the mass balance equation, algebraic solutions are easily found. Practical consequences: By comparing experimental data with the exact solution, the effect of dispersion can be quantified. ..
Version: 05/20/07