Methods for Analysis of Phase-I and Phase-II Metabolites and Parent Compounds without Hydrolysis

Patent metadata

Field	Value
Publication number	US-2016334375-A1
Application number	US-201615221462-A
Country	US
Kind code	A1
Filing date	Jul 27, 2016
Priority date	Dec 21, 2012
Publication date	Nov 17, 2016
Grant date	—

Abstract

A method for analyzing and quantifying a panel of drugs in a clinical sample comprises: trapping a first portion of drug parent compounds and their metabolites on a first chromatographic column; trapping a second portion of the drug parent compounds and their metabolites on a second chromatographic column; separately eluting the first and second portions of the drug parent compounds and their metabolites from the first and second chromatographic columns; detecting concentrations of each of the drug parent compounds and metabolites eluted from each of the first and second chromatographic columns with a detector; and summing the detected concentration of each drug parent compound together with the detected concentrations of all of its respective analytes so as to derive a respective total concentration of each drug in the sample.

First claim

What is claimed is:

1 . A method for analyzing and quantifying a panel of drugs in a clinical sample, the method comprising:
trapping a first portion of drug parent compounds and metabolites of the drug parent compounds on a first chromatographic column; trapping a second portion of the drug parent compounds and their metabolites on a second chromatographic column; separately eluting the first and second portions of the drug parent compounds and their metabolites from the first and second chromatographic columns; detecting concentrations of each of the drug parent compounds and metabolites eluted from each of the first and second chromatographic columns with a detector; and summing the detected concentration of each drug parent compound together with the detected concentrations of all of its respective analytes so as to derive a respective total concentration of each drug in the sample.

2 . A method as recited in claim 1 ,
wherein the trapping of the first portion of the drug parent compounds and their metabolites on the first chromatographic column comprises trapping the drug parent compounds on the first chromatographic column, and wherein the trapping of the second portion of the drug parent compounds and their metabolites on the second chromatographic column comprises trapping the metabolites on the second chromatographic column.

3 . A method as recited in claim 2 ,
wherein the trapping of the drug parent compounds on the first chromatographic column comprises flowing a solution of the sample in a first mobile phase comprising a first mobile phase composition into the first chromatographic column, and wherein the trapping of the metabolites on the second chromatographic column comprises flowing an effluent from the first chromatographic column into the second chromatographic column.

4 . A method as recited in claim 3 , wherein the first chromatographic column comprises a stationary phase that traps the drug parent compounds thereon upon receiving the flow of the sample in solution in the first mobile phase.

5 . A method as recited in claim 4 , wherein the first mobile phase comprises a Mobile Phase A, wherein the Mobile Phase A comprises a solution of ammonium formate and formic acid in water.

6 . A method as recited in claim 2 ,
wherein the trapping of the drug parent compounds on the first chromatographic column comprises flowing a solution of the sample in a first mobile phase comprising a first mobile phase composition into the first chromatographic column, and wherein the trapping of the metabolites on the second chromatographic column comprises flowing, into the second chromatographic column, a mixture between an effluent from the first chromatographic column and a second mobile phase comprising a second mobile phase composition, wherein the second mobile phase composition increases preferential partitioning of the drug metabolite compounds onto a stationary phase of the second chromatographic column.

7 . A method as recited in claim 6 , wherein the first chromatographic column is a reversed-phase chromatographic column comprising a C18 stationary phase and the second chromatographic column is a reversed-phase chromatographic column that comprises a stationary phase comprising a polar endcapped, ether-linked phenyl phase.

8 . A method as recited in claim 5 , wherein the first chromatographic column is a reversed-phase chromatographic column comprising a C18 stationary phase and the second chromatographic column is a reversed-phase chromatographic column that comprises a stationary phase comprising a polar endcapped, ether-linked phenyl phase.

9 . A method as recited in claim 8 , wherein the eluting of the second portion of the drug parent compounds and their metabolites from the second chromatographic column comprises:
breaking a fluidic coupling between an outlet of the first chromatographic column and an inlet of the second chromatographic column; and flowing, into the inlet of the second chromatographic column and through the second chromatographic column, a second mobile phase that comprises a Mobile Phase B, wherein the Mobile Phase B comprises a solution of ammonium formate and formic acid in methanol.

10 . A method as recited in claim 8 , wherein the eluting of the first portion of the drug parent compounds and their metabolites from the first chromatographic column comprises:
breaking a fluidic coupling between an outlet of the first chromatographic column and an inlet of the second chromatographic column; and flowing, into an inlet of the first chromatographic column and through the first chromatographic column, a second mobile phase that comprises a Mobile Phase B, wherein the Mobile Phase B comprises a solution of ammonium formate and formic acid in methanol.

11 . A method as recited in claim 1 ,
wherein the trapping of the first portion of the drug parent compounds and their metabolites on the first chromatographic column comprises trapping the metabolites on the first chromatographic column by flowing a solution of the sample in a first mobile phase comprising a first mobile phase composition into the first chromatographic column, and wherein the trapping of the second portion of the drug parent compounds and their metabolites on the second chromatographic column comprises trapping the drug parent compounds on the second chromatographic column by flowing an effluent from the first chromatographic column into the second chromatographic column.

12 . A method as recited in claim 11 , wherein the first chromatographic column is a hydrophilic interaction liquid chromatography column and the second chromatographic column is a reversed-phase chromatographic column comprising a C18 stationary phase.

13 . A method as recited in claim 12 , wherein the first mobile phase comprises Mobile Phase A, wherein the Mobile Phase A comprises a solution of ammonium formate and formic acid in methanol.

14 . A method as recited in claim 13 , wherein the eluting of the first portion of the drug parent compounds and their metabolites from the first chromatographic column comprises:
breaking a fluidic coupling between an outlet of the first chromatographic column and an inlet of the second chromatographic column; and flowing, into an inlet of the first chromatographic column and through the first chromatographic column, a second mobile phase that comprises a Mobile Phase B, wherein the Mobile Phase B comprises a solution of ammonium formate and formic acid in water.

15 . A method as recited in claim 1 ,
wherein the trapping of the first portion of the drug parent compounds and their metabolites on the first chromatographic column comprises flowing a solution of the sample in a first mobile phase comprising a first mobile phase composition into the first chromatographic column, wherein the trapping of the second portion of the drug parent compounds and their metabolites on the second chromatographic column comprises flowing an effluent from the first chromatographic column into the second chromatographic column, wherein the eluting of the second portion of the drug parent compounds and their metabolites from the second chromatographic column comprises:
breaking a fluidic coupling between an outlet of the first chromatographic column and an inlet of the second chromatographic column; and
flowing a second mobile phase having a second mobile phase composition into the inlet of the second chromatographic column and through the second chromatographic column, and
wherein the eluting of the first portion of the drug parent compounds and their metabolites from the first c

Assignees

• Thermo Finnigan Llc

Inventors

• Herman Joseph
• Fair Sarah J
• Argoti Dayana

Classifications

• G01N30/02

  Column chromatography · CPC title

• G01N30/462 Primary

  with different eluents or with eluents in different states (G01N30/463 takes precedence) · CPC title

• G01N2030/027

  Liquid chromatography · CPC title

• G01N2030/143

  selective absorption · CPC title

• G01N30/468

  involving switching between different column configurations · CPC title