Fluorescence enhancement of dansyl derivatives
AssigneeVarian Associates, Inc.
InventorsSeth R. Abbott, Robert Cunico
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Abstract
A method for fluorescence enhancement of dansyl derivatives of active hydrogen compounds which are easily separated in a liquid chromatography column using a polar solvent system but whose fluorescence efficiency is diminished in such polar solvents, by changing the environment of the dansyl derivative without changing the solvent.
Claims28
A method for enhancing the fluorescence detectability of dansyl derivatives of active hydrogen compounds, which are ideally separated by liquid chromatography using a non-polar column packing material and a polar solvent system but whose fluorescence efficiency is diminished by the presence of polar solvents, said method comprising the steps of
- (a) passing a polar eluant bearing at least one of said dansyl derivatives through a flow cell packed with a non-polar packing material whereby there is caused a dynamic equilibrium of at least one of said dansyl derivatives between the said polar solvent system and the said non-polar cell packing material;
- wherein said eluant is eluted from a liquid chromatography column packed with an appropriate non-polar packing material using a polar solvent system;
- (b) irradiating said flow cell with electromagnetic radiation of a frequency that causes said dansyl derivative to fluoresce in the non-polar environment;
- and
- (c) detecting the emitted fluorescence;
- whereby the environment of at least one of said dansyl derivatives is changed to a non-polar environment without changing the solvent.
A method for enhancing the fluorescence detectability of dansyl derivatives of active hydrogen compounds, which cannot be separated from impurities or contaminants by liquid chromatography using a non-polar column packing material and a polar solvent system, but the fluorescence efficiency of said dansyl derivative is diminished by the presence of polar solvents and the fluorescence efficiency of said impurity or contaminant is diminished by the presence of non-polar solvents, said method comprising the steps of:
- (a) passing a polar eluant bearing at least one of said dansyl derivatives and the said impurity or contaminant through a flow cell packed with a non-polar packing material whereby there is caused a dynamic equilibrium of at least one of said dansyl derivatives and the said impurity or contaminant between the said polar solvent system and the said non-polar cell packing material;
- wherein said eluant is eluted from a liquid chromatography column packed with an appropriate non-polar packing material using a polar solvent system;
- (b) irradiating said flow cell with electromagnetic radiation of a frequency that causes said dansyl derivative to fluoresce in the non-polar environment and causes said impurity or contaminant fluorescence to be diminished in the non-polar environment;
- and
- (c) detecting the emitted fluorescence;
- whereby the environment of at least one of said dansyl derivatives is changed to a non-polar environment without changing the solvent.
Dependencies are inferred from the claim text.