Provided are two-dimensional chromatography systems and methods for separating and/or analyzing complex mixtures of organic compounds. In particularly, a two-dimensional reversed-phase liquid chromatography (RPLC)supercritical fluid chromatography (SFC) system is described including a trapping column at the interface which collects the analytes eluted from the first dimension chromatography while letting the RPLC mobile phase pass through. The peaks of interest from the RPLC dimension column are effectively focused as sharp concentration pulses on the trapping column, which is subsequently injected onto the second dimension SFC column. The system can be used for simultaneous achiral and chiral analysis of pharmaceutical compounds. The first dimension RPLC separation provides the achiral purity result, and the second dimension SFC separation provides the chiral purity result (enantiomeric excess).

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.

In an LC/MS/MS analysis: injecting a sample into a passage leading to a column group provided in a liquid chromatograph, the column group including a plurality of columns serially connected to each other and packed with different kinds of packing materials; supplying an eluant to one or a plurality of columns including a column located most downstream, to separate a portion of the target components in the sample in the one or plurality of columns, and sequentially elute those components from the most downstream column to perform mass spectrometry; and supplying a different eluant to one or a plurality of columns including the most downstream column, to separate at least a portion of the target components which stayed uneluted in the one or plurality of columns in the first analysis step, and sequentially elute those components from the most downstream column to perform mass spectrometry.

In various aspects, the present disclosure pertains to materials (e.g., kits, column assemblies, liquid chromatography systems, etc.) methods for performing liquid chromatography that employ a first column (e.g., a trapping column) and a second column (e.g., an analytical column). The first column comprises a first chromatographic material having a first chromatographic surface that comprises first hydrophobic surface groups and first ionizable surface groups having a first pKa value. The second column comprises a second chromatographic material having a second chromatographic surface that comprises second hydrophobic surface groups and (a) permanently ionized surface groups or (b) second ionizable surface groups having a second pKa value. The first hydrophobic surface groups have a hydrophobicity that is less than a hydrophobicity of the second hydrophobic surface groups. Moreover, where the second chromatographic surface comprises second ionizable surface groups, the first pKa value may differ from the second pKa value by 1-12 units.

The objective of the present invention is to provide a more accurate method and system for analyzing a sample containing an optical isomer. This method is characterized in that, with reference to results of chromatography, fractions for which complete separation from one or a plurality of chromatograph peaks corresponding to one or a plurality of other components has been achieved, from the time at which the peak starts to rise from the baseline until the peak has completely returned to the baseline, are successively fractionated separately, and if complete separation cannot be achieved for a single component, the fractionated fraction is subjected to additional chromatography, and if complete separation is achieved, the fractionated fraction is subjected to chiral chromatography.

An arrangement for preparing samples and analyzing pesticides in samples contains an HILIC chromatography column with a first pump for a predominately low-water and/or non-polar solvent; and SPE enrichment arrangement; a second chromatography column with a second pump for a predominantly water-rich and/or polar solvent; a detector; and a valve arrangement for controlling the stream of sample and matrix, which valve arrangement is designed in such a way that the sample stream, in a first switching state of the valve arrangement, can be conducted from the HILIC chromatography column to the SPE enrichment arrangement and, in a second switching state, the sample enriched in the SPE enrichment arrangement can be conducted in the opposite direction from the SPE enrichment arrangement through the second chromatography column to the detector.

Methods for achieving complete sequence coverage of monoclonal antibodies by trypsin digestion and dual-column LC-MS system are provided. The disclosed method improves upon current techniques for standard peptide mapping.

The disclosed system and method improve measurement of trace volatile chemicals, such as by Gas Chromatography (GC) and Gas Chromatography/Mass Spectrometry (GCMS). A first trapping system can include a plurality of capillary columns in series and a focusing column fluidly coupled to a first detector. The first trapping system can retain and separate compounds in a sample, including C3 hydrocarbons and compounds heavier than C3 hydrocarbons (e.g., up to C12 hydrocarbons, or compounds having a boiling point around 250 C.), and can transfer the compounds from the focusing column to the first detector. A second trapping system can receive compounds that the first trapping system does not retain, and can include a packed trap, a polar column and a PLOT column fluidly coupled to one or more second detectors. The second trapping system can remove water from the sample and can separate and detect compounds including C2 hydrocarbons and Formaldehyde.

Provided are two-dimensional chromatography systems and methods for separating and/or analyzing complex mixtures of organic compounds. In particularly, a two-dimensional reversed-phase liquid chromatography (RPLC)supercritical fluid chromatography (SFC) system is described including a trapping column at the interface which collects the analytes eluted from the first dimension chromatography while letting the RPLC mobile phase pass through. The peaks of interest from the RPLC dimension column are effectively focused as sharp concentration pulses on the trapping column, which is subsequently injected onto the second dimension SFC column. The system can be used for simultaneous achiral and chiral analysis of pharmaceutical compounds. The first dimension RPLC separation provides the achiral purity result, and the second dimension SFC separation provides the chiral purity result (enantiomeric excess).

A chromatographic purification method for the isolation of a desired product fraction from a mixture using 2 chromatographic columns, comprises, within one cycle to be carried out at least once, the following steps: a first batch step, wherein said columns are disconnected and a first column is loaded with feed and its outlet is directed to waste, and from a second column desired product is recovered and subsequently the second column is regenerated; a first interconnected step, wherein the outlet of the first column is connected to the inlet of the second column, the first column is loaded beyond its dynamic breakthrough capacity with feed, and the outlet of the second column is directed to waste, a second batch step analogous to the first batch step but with exchanged columns; and a second interconnected step, analogous to the first interconnected step but with exchanged columns.