METHOD FOR BASELINE CORRECTION IN A CHROMATOGRAM

摘要

The invention discloses a method for correcting a baseline in a chromatogram obtained using a buffer with at least one UV-absorbing component which is a weak acid or a weak base. The method comprises the steps of:;a) providing a chromatography system with a chromatography column and a UV detector downstream of the chromatography column;

b) providing UV extinction coefficients at a wavelength λ for the acid form and the base form of the UV-absorbing component(s) and a dissociation constant for the component(s),

c) obtaining a chromatogram with the UV detector at wavelength λ using the buffer under a set of conditions where pH and/or conductivity is varied between these conditions,

d) for a plurality of said conditions calculating the concentrations of the acid and base forms using the dissociation constant(s) and values for pH and conductivity values, and

e) calculating UV absorption values of the acid and base forms from the concentrations and UV extinction coefficients under the conditions in question and subtracting the UV absorption values from the chromatogram.

主权项

1. A method for correcting a baseline in a chromatogram obtained using a buffer with at least one UV-absorbing component which is a weak acid or a weak base, said method comprising the steps of:
a) providing a chromatography system with a chromatography column and a UV detector downstream of said chromatography column; b) providing UV extinction coefficients at a wavelength λ for an acid form and a base form of said at least one UV-absorbing component and a dissociation constant for said component, c) obtaining a chromatogram with said UV detector at wavelength λ using said buffer under a set of conditions where pH and/or conductivity is varied between said conditions, d) for a plurality of said conditions calculating the concentrations of said acid and base forms using said dissociation constant and pH and conductivity values, and e) calculating UV absorption values of said acid and base forms from said concentrations and said UV extinction coefficients under said conditions and subtracting said UV absorption values from said chromatogram.