Disclosed are methods for improving compound detection and characterization. Methods for characterizing a sample are disclosed. The methods can include providing a sample to a liquid chromatography system capable of sample separation to generate sample components; analyzing sample components by multiplexed targeted selected ion monitoring (SIM) to generate an inclusion list; and performing iterative mass spectral data-dependent acquisition (DDA) from the inclusion list, to identify individual sample components thereby characterizing the sample. In one example, multiplexed targeted SIMs and iterative MS2 DDA acquisition is used to increase robust compound identification for cell culture medium analysis.

An interface transports ions from a first pressure environment to a lower pressure analysis instrument and may include a first region pumped to a second pressure less than the first pressure, a first ion funnel disposed in the first region, a first ion carpet in the first region opposite an ion outlet end of the first ion funnel, a second region pumped to a third pressure less than the second pressure and greater than the instrument pressure, a second ion funnel disposed in the second region and a second ion carpet in the second region opposite an ion outlet end of the second ion funnel. Ions from the environment pass sequentially through the first and second ion funnels and into the analysis instrument. Each of the first and second ion funnels define a tapered axial passageway therethrough each defining a respective virtual jet disrupter therein.

A data independent acquisition method of mass spectrometry for analyzing a sample within a mass range of interest as it elutes from a chromatography system. The method comprises selecting precursor ions within a mass range of interest to be analyzed, performing at least one MS1 scan of the precursor ions using a first, high-resolution mass analyzer and performing a set of MS2 scans by segmenting the precursor ions into a plurality of precursor mass segments, each precursor mass segment having a mass range of no greater than 5 amu, and for each precursor mass segment fragmenting the precursor ions within that precursor mass segment and performing an MS2 scan of the fragmented ions using a time of flight mass analyzer.

There is disclosed a method for eliminating an added crosstalk signal from a measured data signal, which is generated by an image current. There is further disclosed a signal processing unit for carrying out the method. There is still further disclosed a mass spectrometer and a mass analyser comprising the signal processing unit for carrying out the method. There is yet still further disclosed a Fourier transform mass spectrometer configured to eliminate the added crosstalk signal from a measured data signal.

A data independent acquisition method of mass spectrometry for analysing a sample as it elutes from a chromatography system is disclosed. The method comprises selecting a precursor mass range, and performing a plurality of MS1 scans and sets of MS2 scans across the precursor mass range. Each of the MS1 scans uses a mass analyser operated at a first, relatively higher resolution, for identification and/or quantitation of the sample in the MS1 domain. The set of MS2 scans comprises performing MS2 scans of fragmented mass range segments performed with the mass analyser, operated at a second, relatively lower resolution. In the method, the MS1 scans are interleaved throughout the performing of the set of MS2 scans such that the MS1 scans provide a mass chromatogram of the sample. The ratio of the number of MS1 scans to sets of MS2 scans performed across the chromatographic peak width is at least 3:1.

A charge detection mass spectrometer may include an electrostatic linear ion trap (ELIT) or an orbitrap, an ion source to supply ions thereto, at least one amplifier operatively coupled to the ELIT or orbitrap, a processor coupled to ELIT or orbitrap and to the amplifier(s), and processor programmed to control the ELIT or orbitrap as part of a trapping event to attempt to trap therein a single ion supplied by the ion source, to record ion measurement information based on output signals produced by the amplifier(s) over a duration of the trapping event, to determine, based on the measurement information, whether the control of the ELIT or orbitrap resulted in trapping of a single ion, no ion or multiple ions, and to compute an ion mass or mass-to-charge ratio from the measurement information only if a single ion was trapped during the trapping event.

Apparatus and methods for performing charge detection mass spectrometry are disclosed. An analyte ion is injected into an electrostatic trap, which has electrodes shaped and arranged to establish a trapping field that causes the analyte ion to undergo harmonic motion along a longitudinal axis. A time-varying signal is generated by a detector representative of the harmonic motion. A data system processes the time-varying signal to derive the frequency of ion motion and the amplitude at the harmonic motion frequency, and determines the mass-to-charge ratio (m/z) of the ion based on the derived frequency and the charge from the derived amplitude. The product of the experimentally determined m/z and charge yields the mass of the analyte ion. The electrodes preferably include an elongated inner electrode surrounded by an outer electrode, forming an orbital or non-orbital electrostatic trap.

The present invention provides rapid, sensitive high-throughput methods and systems for characterizing peptides or proteins using affinity-based chromatography-coupled native mass spectrometry to improve manufacturing process of biopharmaceutical products, such as identifying impurities during antibody purification, monitoring post-translational modification variants during production, or characterizing drug-to-antibody ratio of antibody-drug conjugates. The separation profiles of the peptides or proteins are generated and compared to identify or qualify the peptides or proteins, wherein the separation profile is based on differential affinity binding.

There is disclosed a method for eliminating an added crosstalk signal from a measured data signal, which is generated by an image current. There is further disclosed a signal processing unit for carrying out the method. There is still further disclosed a mass spectrometer and a mass analyser comprising the signal processing unit for carrying out the method. There is yet still further disclosed a Fourier transform mass spectrometer configured to eliminate the added crosstalk signal from a measured data signal.

The present invention provides a mass spectrometer comprising a first ion trap, a second ion trap, a lens stack for directing ions from the first ion trap to the second ion trap and a housing. The first ion trap is arranged to form a linear or curved potential well and the second ion trap is an electrostatic ion trap, for example, an orbital ion trap, arranged to form an annular potential well. The mass spectrometer further comprises a unitary insert comprising a first cavity which holds the lens stack and a second cavity which holds the second ion trap, wherein the insert is inserted within the housing.