US 12,078,621 B2
Automated semi-preparative gradient recycling liquid chromatography
Fabrice Gritti, Franklin, MA (US)
Assigned to Waters Technologies Corporation, Milford, MA (US)
Filed by Waters Technologies Corporation, Milford, MA (US)
Filed on Mar. 21, 2022, as Appl. No. 17/699,775.
Claims priority of provisional application 63/165,914, filed on Mar. 25, 2021.
Prior Publication US 2022/0308020 A1, Sep. 29, 2022
Int. Cl. G01N 30/46 (2006.01); G01N 30/02 (2006.01); G01N 30/32 (2006.01); G01N 30/82 (2006.01); G01N 30/30 (2006.01)
CPC G01N 30/461 (2013.01) [G01N 30/82 (2013.01); G01N 2030/027 (2013.01); G01N 2030/3007 (2013.01); G01N 2030/326 (2013.01); G01N 2030/328 (2013.01)] 10 Claims
OG exemplary drawing
 
1. 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.