Described are an interface module for two-dimensional chromatography and a method of performing a chromatographic separation that may use the interface module. The interface module includes a valve module, a collection needle, a modifier module and a sample storage element. The valve module has a first port configured to receive an eluent from a first chromatography system, a second port configured to provide a fraction obtained from the eluent, a third port and a fourth port. The collection needle and the modifier module are in fluidic communication with the valve module at the third and fourth ports, respectively. The modifier module includes a source of a modifier solvent. The sample storage element is in fluidic communication with the valve module and is configured to receive a volume of the fraction for injection as a sample into a second chromatography system.

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 chromatography system includes a first chromatography column for receiving and separating a flow stream, a plurality of traps configured to trap a plurality of distinct flow segments exiting the first chromatography column during separation of the flow stream, and a second chromatography column operatively associated with the plurality of traps for receiving and separating the distinct flow segments. The system can include at-column dilution at trapping and separating stages thereof. A chromatography method for operating the chromatographic system includes measuring a plurality of time segments corresponding to a plurality of peaks of a fluid sample flowing through the first chromatographic column, and sequentially fluidly coupling the plurality of distinct flow segments with the corresponding plurality of traps during time segments corresponding to the plurality of peaks.

The present application relates to systems and methods for assaying presence of large molecule analytes, such as proteins, e.g., antibodies, antigens, receptors, and the like, using a targeted two-dimensional liquid chromatography, tandem mass spectrometry (2D-LC-MS/MS) system, optionally combined with affinity capture. In some aspects, the system is partially or fully automated. In some aspects, the system may allow detection of protein biomarkers (e.g., antibodies or antigens) from clinical or nonclinical biological tissue or fluid samples in the pg/mL to ng/mL range.

Methods for identifying viral protein constituents and quantifying the relative abundance of such viral protein constituents in a sample of viral particles are disclosed. In embodiments, the methods include first-dimension chromatography to separate intact viral capsid components of the sample, online denaturation of the viral capsid components to produce intact viral proteins, second-dimension chromatography to separate the viral proteins, and mass spectrometry to determine the masses of the viral proteins and identify the viral protein constituents of the sample.

Described are an interface module for two-dimensional chromatography and a method of performing a chromatographic separation that may use the interface module. The interface module includes a valve module, a collection needle, a modifier module and a sample storage element. The valve module has a first port configured to receive an eluent from a first chromatography system, a second port configured to provide a fraction obtained from the eluent, a third port and a fourth port. The collection needle and the modifier module are in fluidic communication with the valve module at the third and fourth ports, respectively. The modifier module includes a source of a modifier solvent. The sample storage element is in fluidic communication with the valve module and is configured to receive a volume of the fraction for injection as a sample into a second chromatography system.

The present invention relates to a novel analytical method for detecting one or more analytes in a source sample by continuous flow 2D LC-MS/MS using a single LC system.

Methods of purifying targeted oligonucleotides within a reaction mixture using hydrophilic interaction liquid chromatography (HILIC) is disclosed. One of the methods in accordance with the present disclosure includes screening the targeted oligonucleotides within the reaction mixture with HILIC to create an initial reaction mixture profile; determining an elution percentage for the targeted oligonucleotides; focusing a HILIC elution gradient around the elution percentage of the targeted oligonucleotides; and purifying the targeted oligonucleotides with HILIC using the focused elution gradient at room temperature. Some embodiments can utilize mass triggering for fraction collection of the targeted oligonucleotides. Some embodiments can utilize UV triggering when the mass falls outside of the mass range of the MS detector.

A sampling unit for handling a sample fluid includes a sample container having a length and being configured for receiving and storing the sample fluid, and a sample segment dispatching unit configured for providing a plurality of individual sample packages of the fluidic sample, each contained in a respective volume segment along the length of the sample container, and for individually dispatching each of the plurality of individual sample packages for further processing in a fluid processing unit.

A chromatography system includes a first chromatography column for receiving and separating a flow stream, a plurality of traps configured to trap a plurality of distinct flow segments exiting the first chromatography column during separation of the flow stream, and a second chromatography column operatively associated with the plurality of traps for receiving and separating the distinct flow segments. The system can include at-column dilution at trapping and separating stages thereof. A chromatography method for operating the chromatographic system includes measuring a plurality of time segments corresponding to a plurality of peaks of a fluid sample flowing through the first chromatographic column, and sequentially fluidly coupling the plurality of distinct flow segments with the corresponding plurality of traps during time segments corresponding to the plurality of peaks.