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

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 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.

An automated sample preparation and analysis system comprising:

• (a) an automated sample preparation station for preparing a plurality of samples in accordance with a plurality of assays that are selected from a database containing a plurality of unique assays;
• (b) a sample analysis station for automatically performing the plurality of assays;
• and
• (c) a transport mechanism for transporting prepared samples from the sample preparation station to the sample analysis station,
• wherein the sample analysis station comprises:
• (i) a source of a first chromatographic mobile phase solvent;
• (ii) a source of a second chromatographic mobile phase solvent;
• (iii) a first chromatographic column fluidically coupled to the first chromatographic mobile phase solvent and configured to receive a portion of the prepared samples from the sample transport mechanism;
• (iv) a second chromatographic column fluidically coupled to the first chromatographic column and to the source of the second chromatographic mobile phase solvent;
• and
• (v) a detector fluidically coupled to the second chromatographic column,
• wherein the first chromatographic column and the second chromatographic column are operable to trap, respectively, a first portion and a second portion of a panel of drug compounds and their metabolites during a trapping step and to elute, respectively, the first and second portions of the drug compounds and their metabolites to the detector.