Automated semi-preparative gradient recycling liquid chromatography
AssigneeWaters Technologies Corporation
InventorFabrice Gritti
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Abstract
The disclosure relates to a gradient twin column recycling chromatography method that is used to separate a mixture containing closely eluting compounds. In one embodiment, a sample includes a primary organic compound and one or more impurities that closely elute with the primary organic compound. A gradient mobile phase is initially used to remove unwanted early eluting and late eluting impurities from the sample. After the gradient removal of some of the impurities is complete, the remaining mixture of the primary organic compound and the closely eluting impurities are separated using recycle chromatography methodology with an isocratic mobile phase.
Claims10
A method of performing recycling chromatography comprising:
- injecting a sample into a mobile phase flow stream of a chromatography system to create a combined flow stream, the sample comprising:
- an organic compound and a plurality of impurities, the chromatography system comprising:
- a first chromatographic column and a second chromatographic column coupled to each other in series;
- and
- a fraction collector coupled to the first chromatographic column and the second chromatographic column;
- passing the sample through the first chromatographic column using a gradient mobile phase stream, wherein during passage of the sample through the first chromatographic column low retention time impurities are transferred to the fraction collector, wherein the low retention time impurities are impurities that have a lower retention time than the organic compound when using the gradient mobile phase and;
- wherein the organic compound and a subset of the plurality of impurities having a similar elution to the organic compound when using the gradient mobile phase are transferred to the second chromatographic column as an analyte band using the gradient mobile phase;
- and
- wherein high retention time impurities remain in the first chromatographic column, wherein the high retention time impurities are impurities that have a higher retention time than the organic compound when using the gradient mobile phase;
- passing the analyte band through the second chromatographic column using an isocratic mobile phase stream;
- recycling the analyte band from the second chromatographic column to the first chromatographic column and from the first chromatographic column to the second chromatographic column using the isocratic mobile phase stream until a baseline resolution of the organic compound and the impurities, having a similar elution to the organic compound, present in the analyte band is achieved;
- passing the organic compound to the fraction collector.
Dependencies are inferred from the claim text.