Monolithic column chromatography
AssigneeQuest Diagnostics Investments Incorporated
InventorsNigel CLARKE, Mildred M. GOLDMAN
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Abstract
Provided herein are methods of liquid column chromatography in which preparative chromatography is performed in-line with analytical chromatography. In particular aspects a monolithic preparative column is used to purify an analyte of interest from a mixture of other substances by applying the mixture to the column, reversing the flow through the column to elute the analyte, which is applied to an analytical column provided in-line with the preparative column. In other aspects, a single monolithic column is used to perform both the preparative chromatography and analytical chromatography steps in succession. In another aspect, a chromatography system is provided to perform preparative and analytical chromatography using a single monolithic column.
Claims37
A chromatographic method for detecting one or more analytes in a body fluid sample containing one or more other substances, said method comprising:
- a) applying a body fluid sample to a monolithic chromatography column comprising a monolithic sorbent having macropores and mesopores;
- b) applying a first mobile phase to said monolithic chromatography column after applying said body fluid under conditions such that one or more analytes are retained on said monolithic chromatography column and one or more other substances present in said body fluid sample are removed, wherein said one or more retained analytes have a molecular weight of less than about 5,000 Daltons;
- c) eluting said one or more retained analytes by applying a second mobile phase to said monolithic chromatography column;
- and
- d) detecting said one or more analytes eluted in step c).
A method for achieving a chromatographic separation of one or more analytes from one or more other substances in a body fluid sample, said method comprising:
- a) applying a body fluid sample to a monolithic chromatography column comprising a monolithic sorbent having macropores and mesopores;
- b) applying a first mobile phase to said monolithic chromatography column after applying said body fluid sample under conditions such that said one or more analytes in the sample are retained on said monolithic chromatography column and one or more other substances are removed, wherein said one or more retained analytes have a molecular weight of less than about 5,000 daltons;
- and
- c) eluting said one or more retained analytes by applying a second mobile phase to said monolithic chromatography column.
Dependencies are inferred from the claim text.